Plant Macromolecules as Biomaterials for Wound Healing

*Felipe Domingos de Sousa, Francisco Rogênio da Silva Mendes, Jose Jovanny Bermudez-Sierra, Ayrles Fernanda Brandão da Silva, Mirele da Silveira Vasconcelos, Tamiris de Fátima Goebel de Souza, Marília de Oliveira Nunes, Antônio Eufrásio Vieira-Neto, Marcos Roberto Lourenzoni, Rosueti Diógenes de Oliveira-Filho, Adriana Rolim Campos, Renato de Azevedo Moreira and Ana Cristina de Oliveira Monteiro-Moreira*

### **Abstract**

Natural biomolecules are increasingly relevant for biomedical applications and tissue engineering for being able to produce an effect on chemical signals, organization of cells, and restitution of extracellular matrix in lesioned tissues. In this chapter, we will address the potential of plant macromolecules, in particular, carbohydrates and proteins such as hemicelluloses and lectins. While lectins are mostly carbohydrate-binding proteins, which can interact with cell surfaces to initiate anti-inflammatory pathways, as well as immunomodulatory functions, hemicelluloses are remarkably known by their ability to form viscous solutions even at low concentrations, which makes them an excellent candidate as vehicle to carry different sorts of biomolecules. Taking into account the complexity of the whole healing process, as an overlapping and coordinated cascade of events, most of the properties presented here by those materials may be of interest to the wound-care market.

**Keywords:** lectins, galactomannans, biomolecules, polysaccharides

### **1. Introduction**

Skin is the largest organ, which presents a fairly robust arrangement, working as a natural shield against physical, chemical, and bacterial damage to the body. When it suffers any disruption in this arrangement by means of acute lesions, the skin goes through a fascinating repair process finalizing in wound closure and leaving some scars. This complex mechanism is dependent on many cell types and mediators

interacting to maintain the physiological regulation of skin [1, 2]. Chronic wound results in situations such as diabetes or vascular lesions, when defective skin usually persists causing the loss of integrity, and in some cases, the overlapping of structural layers, as well as delayed skin tissue recovery [3].

Human body tissues present a wide range of physical characteristics, which include stiffness and porosity. A multidisciplinary interface between areas such as cell biology, biotechnology, mechanics, materials science, bioengineering, and clinical research has been used to contribute significantly to tissue engineering. Many wound dressings have been developed seeking to restore and improve tissue function by generating new biocompatible substitutes or by rebuilding these tissues [4, 5]. However, these sorts of materials are quite costly, which might affect the widespread adoption.

From this perspective, an ideal wound dressing would maintain a microenvironment in the injured bed and would direct the specific healing properties for each type of wound or disease that affects the patient under treatment. The dressing should also keep the moisture balance, which works as a barrier of protection against infections and would provide thermal insulation for the wound [6]. Therefore, recent technologies for biomedical applications invest in the development of scaffolds able to mimic the natural environment for skin grown and regeneration after damage [4].

Polysaccharide-based biomaterials emerge in the field of tissue engineering, as they are mainly used as hydrogels for the effective treatment of wounds and skin burns. They can be categorized as neutral (e.g., glucans, dextran, and cellulose), acids (acid hyaluronic), basic (chitosan) or sulfated polysaccharides (heparin and chondroitin). The most popular and naturally produced biomaterials of polysaccharide origin are chitosan, hyaluronic acid, and alginate. These polysaccharides may also be subdivided into homopolysaccharides such as glucans, cellulose, dextran and chitosan, and heteropolysaccharides such as alginates, agarose, carrageenan, pectin, galactomannans and xyloglucans. All exhibit peculiar physicochemical properties and a considerable biocompatibility and biodegradability, and thus have important applications in biomedical fields [4].

In this chapter, we focus our attention to plant macromolecules such as carbohydrates and proteins (in particular hemicelluloses and lectins) as biomolecules for wound healing applications.

### **2. Wound healing**

Wound healing is a critical, complex and highly coordinated process to maintain skin function. Immediately after injury, a multitude of molecular and cellular systems are activated to suspend blood loss, eliminate microorganisms and foreign materials, and recompose injured tissue [3]. These cellular and biochemical events in wound repair can be divided into the following steps: hemostasis, inflammatory response, cell proliferation and synthesis of the elements that make up the extracellular matrix (ECM), and the later period, called remodeling. These stages are not mutually exclusive but overlapping over time [7].

In physiological conditions, platelets circulate in the vicinity of the vascular walls and are activated when the continuity of the endothelial layer is ruptured and the underlying subendothelial matrix is exposed, initiating the first stage of tissue repair, characterized by hemostasis and the formation of a matrix in the wound bed. This matrix is the result of the adhesion and aggregation of circulating platelets to the components of the underlying ECM. Damaged tissue and aggregated platelets trigger extrinsic and intrinsic coagulation pathways to stabilize the fibrin platelet

**35**

*Plant Macromolecules as Biomaterials for Wound Healing*

chemotaxis of fibroblasts and smooth muscle cells [9].

their tasks are completed and are cleansed by macrophages [9, 10].

angiogenesis, fibroplasia and re-epithelialization [7, 12].

clot. This whole process forms a framework for the migration and proliferation of other cells involved in wound healing, as well as a reservoir for cytokines and

The inflammatory phase then begins as an innate immune response to promote the elimination of cellular and extracellular debris as well as pathogenic microorganisms. Both platelets and leukocytes release inflammatory cytokines providing a chemotactic gradient for additional leukocytes to be attracted and potentiate the inflammatory process. Among the inflammatory factors are interleukin IL-1α, IL-1β, IL-6, IL-8, tumor necrosis factor (TNF-α), platelet-derived growth factor (PDGF) and transforming growth factor-β. Clearly, PDGF plays an important role early in the chemotaxis of neutrophils, monocytes, smooth muscle cells and fibroblasts, while TGF-β stimulates cytokine secretion from macrophages and enhances

The initial leukocyte response is dominated by neutrophils in the first 2–5 days, with macrophages taking over from the third day. Neutrophils have three main functions. First, they generate free radicals via the myeloperoxidase pathway to kill bacteria, an important action for healing because wounds that have bacterial infection will not heal normally. They also debride the wound through the secretion of specific proteolytic enzymes that break non-viable tissue, such as serine proteases and matrix metalloproteinase (MMP-2 and -9). Hence, the neutrophils phagocyte the dead bacteria and the remained matrix. They usually undergo apoptosis when

The monocytes begin to migrate to the wound and mature into macrophages. They become the most important regulatory cell in the inflammatory reaction. In the early stages of inflammation, M1 phenotype macrophages are associated with the phagocytic activity of remaining bacteria, sequestration and production of pro-inflammatory mediators. After this period, M1 becomes the subset M2, revealing a reparative phenotype of macrophages [1]. M2 macrophages are involved in the synthesis of anti-inflammatory mediators and tissue cleansing. These cells remove obsolete neutrophils, non-functional host cells, damaged matrix, and foreign debris. M2 cells also secrete various cytokines, growth factors and other mediators, such as TGF-α, TGF-β, basic fibroblast growth factor (β-FGF), PDGF and vascular endothelial growth factor (VEGF) to amplify and resolve inflammation. At this stage of healing, macrophages regulate the proliferative stage by stimulating fibroblasts, keratinocytes and endothelial cells to differentiate, proliferate and migrate, leading to new ECM deposition, re-epithelialization and wound neovascularization [11]. The goal of the proliferative stage is to decrease the area of contracted tissue and fibroplasia, establishing a viable epithelial barrier to activate keratinocytes. TGF-β stimulated fibroblasts differentiate into myofibroblasts rich in alpha smooth muscle actin and can amplify pseudopodia, joining fibronectin and collagen in the ECM. This event provides wound contraction, which is important in the repair process, helping the edges of the wound approach. These cells are also producers of ECM substances (collagen, fibronectin, glycosaminoglycans, proteoglycans and hyaluronic acid), which interact with the cells to mediate migration, growth and differentiation. This stage is characterized by the lesion closure, which includes

The final stage of wound healing is characterized by the development of new epithelium and formation of scar tissue, a process known as remodeling, which can last for a year or stay for a longer period. The main purpose of the remodeling step is to achieve maximum tensile strength through reorganization, degradation and resynthesis of ECM. Together with intracellular matrix maturation, collagen bundles increase in diameter, whereas hyaluronic acid and fibronectin are degraded. The force of traction of the wound progressively increases with the deposition of

*DOI: http://dx.doi.org/10.5772/intechopen.89105*

growth factors [8].

*Wound Healing*

widespread adoption.

regeneration after damage [4].

applications in biomedical fields [4].

mutually exclusive but overlapping over time [7].

wound healing applications.

**2. Wound healing**

interacting to maintain the physiological regulation of skin [1, 2]. Chronic wound results in situations such as diabetes or vascular lesions, when defective skin usually persists causing the loss of integrity, and in some cases, the overlapping of struc-

Human body tissues present a wide range of physical characteristics, which include stiffness and porosity. A multidisciplinary interface between areas such as cell biology, biotechnology, mechanics, materials science, bioengineering, and clinical research has been used to contribute significantly to tissue engineering. Many wound dressings have been developed seeking to restore and improve tissue function by generating new biocompatible substitutes or by rebuilding these tissues [4, 5]. However, these sorts of materials are quite costly, which might affect the

From this perspective, an ideal wound dressing would maintain a microenvironment in the injured bed and would direct the specific healing properties for each type of wound or disease that affects the patient under treatment. The dressing should also keep the moisture balance, which works as a barrier of protection against infections and would provide thermal insulation for the wound [6]. Therefore, recent technologies for biomedical applications invest in the development of scaffolds able to mimic the natural environment for skin grown and

Polysaccharide-based biomaterials emerge in the field of tissue engineering, as they are mainly used as hydrogels for the effective treatment of wounds and skin burns. They can be categorized as neutral (e.g., glucans, dextran, and cellulose), acids (acid hyaluronic), basic (chitosan) or sulfated polysaccharides (heparin and chondroitin). The most popular and naturally produced biomaterials of polysaccharide origin are chitosan, hyaluronic acid, and alginate. These polysaccharides may also be subdivided into homopolysaccharides such as glucans, cellulose, dextran and chitosan, and heteropolysaccharides such as alginates, agarose, carrageenan, pectin, galactomannans and xyloglucans. All exhibit peculiar physicochemical properties and a considerable biocompatibility and biodegradability, and thus have important

In this chapter, we focus our attention to plant macromolecules such as carbohydrates and proteins (in particular hemicelluloses and lectins) as biomolecules for

Wound healing is a critical, complex and highly coordinated process to maintain

skin function. Immediately after injury, a multitude of molecular and cellular systems are activated to suspend blood loss, eliminate microorganisms and foreign materials, and recompose injured tissue [3]. These cellular and biochemical events in wound repair can be divided into the following steps: hemostasis, inflammatory response, cell proliferation and synthesis of the elements that make up the extracellular matrix (ECM), and the later period, called remodeling. These stages are not

In physiological conditions, platelets circulate in the vicinity of the vascular walls and are activated when the continuity of the endothelial layer is ruptured and the underlying subendothelial matrix is exposed, initiating the first stage of tissue repair, characterized by hemostasis and the formation of a matrix in the wound bed. This matrix is the result of the adhesion and aggregation of circulating platelets to the components of the underlying ECM. Damaged tissue and aggregated platelets trigger extrinsic and intrinsic coagulation pathways to stabilize the fibrin platelet

tural layers, as well as delayed skin tissue recovery [3].

**34**

clot. This whole process forms a framework for the migration and proliferation of other cells involved in wound healing, as well as a reservoir for cytokines and growth factors [8].

The inflammatory phase then begins as an innate immune response to promote the elimination of cellular and extracellular debris as well as pathogenic microorganisms. Both platelets and leukocytes release inflammatory cytokines providing a chemotactic gradient for additional leukocytes to be attracted and potentiate the inflammatory process. Among the inflammatory factors are interleukin IL-1α, IL-1β, IL-6, IL-8, tumor necrosis factor (TNF-α), platelet-derived growth factor (PDGF) and transforming growth factor-β. Clearly, PDGF plays an important role early in the chemotaxis of neutrophils, monocytes, smooth muscle cells and fibroblasts, while TGF-β stimulates cytokine secretion from macrophages and enhances chemotaxis of fibroblasts and smooth muscle cells [9].

The initial leukocyte response is dominated by neutrophils in the first 2–5 days, with macrophages taking over from the third day. Neutrophils have three main functions. First, they generate free radicals via the myeloperoxidase pathway to kill bacteria, an important action for healing because wounds that have bacterial infection will not heal normally. They also debride the wound through the secretion of specific proteolytic enzymes that break non-viable tissue, such as serine proteases and matrix metalloproteinase (MMP-2 and -9). Hence, the neutrophils phagocyte the dead bacteria and the remained matrix. They usually undergo apoptosis when their tasks are completed and are cleansed by macrophages [9, 10].

The monocytes begin to migrate to the wound and mature into macrophages. They become the most important regulatory cell in the inflammatory reaction. In the early stages of inflammation, M1 phenotype macrophages are associated with the phagocytic activity of remaining bacteria, sequestration and production of pro-inflammatory mediators. After this period, M1 becomes the subset M2, revealing a reparative phenotype of macrophages [1]. M2 macrophages are involved in the synthesis of anti-inflammatory mediators and tissue cleansing. These cells remove obsolete neutrophils, non-functional host cells, damaged matrix, and foreign debris. M2 cells also secrete various cytokines, growth factors and other mediators, such as TGF-α, TGF-β, basic fibroblast growth factor (β-FGF), PDGF and vascular endothelial growth factor (VEGF) to amplify and resolve inflammation. At this stage of healing, macrophages regulate the proliferative stage by stimulating fibroblasts, keratinocytes and endothelial cells to differentiate, proliferate and migrate, leading to new ECM deposition, re-epithelialization and wound neovascularization [11].

The goal of the proliferative stage is to decrease the area of contracted tissue and fibroplasia, establishing a viable epithelial barrier to activate keratinocytes. TGF-β stimulated fibroblasts differentiate into myofibroblasts rich in alpha smooth muscle actin and can amplify pseudopodia, joining fibronectin and collagen in the ECM. This event provides wound contraction, which is important in the repair process, helping the edges of the wound approach. These cells are also producers of ECM substances (collagen, fibronectin, glycosaminoglycans, proteoglycans and hyaluronic acid), which interact with the cells to mediate migration, growth and differentiation. This stage is characterized by the lesion closure, which includes angiogenesis, fibroplasia and re-epithelialization [7, 12].

The final stage of wound healing is characterized by the development of new epithelium and formation of scar tissue, a process known as remodeling, which can last for a year or stay for a longer period. The main purpose of the remodeling step is to achieve maximum tensile strength through reorganization, degradation and resynthesis of ECM. Together with intracellular matrix maturation, collagen bundles increase in diameter, whereas hyaluronic acid and fibronectin are degraded. The force of traction of the wound progressively increases with the deposition of

#### *Wound Healing*

collagen, and these fibers can recover approximately 80% of the force compared to normal tissue, but the force of the original tissue can never be recovered again [12].

As demonstrated, many factors can alter positively or negatively the cell interactions and the signaling mechanisms during the wound healing process. Plantderived compounds are among these factors and are able to improve the healing process through different mechanisms.

### **3. Plant-derived compounds**

Medicinal plants have been extensively used worldwide as traditional treatment for various diseases due to being a source of phytochemicals, which are nonnutritive substances present in plants enhancing tissue regeneration and acting as pro-angiogenic agents for wound healing. In addition, bioactive products extracted from plants arouse scientific and commercial interests for the development of new drugs [13]. On the other hand, plants are also source of many macromolecules such as carbohydrate and proteins extensively used as biomaterials for wound healing applications.

#### **3.1 Essential oils**

Essential oils (EOs) are the largest group of secondary metabolites biosynthesized by plant as a complex of monoterpenes (10 carbons) and sesquiterpenes (15 carbons), mainly related to plant defense mechanisms. Also known as volatile oils or aromatic plant essences, they can be found in a multitude of plant tissues such as flowers, leaves, barks, etc. Obtained by aqueous extraction or steam distillation, or cold pressing in the case of citric fruits, they have been extensively employed in cosmeceuticals and dermaceutical products [14, 15].

In the healing process, the OEs stand out for their anti-inflammatory and antimicrobial properties. The efficacy in inhibiting bacterial development, including antibiotic-resistant strains, yeasts and filamentous fungi, has boosted the study of the antimicrobial activity of essential oils. Some oils extracted from medicinal plants demonstrated therapeutic potential in combating biofilms, a mechanism of virulence produced by pathogenic microorganisms, resistant to antibiotics [16]. Carvacrol and Thymol, for example, are monoterpenes widely found in essential oils *Origanum* genus, presenting antibacterial and antifungal activities [17] in addition to analgesic effect [18]. In several studies, it has been reported that interactions between the components of EOs, even in small concentrations, may lead to antagonistic, additive or synergistic effects [15].

Although EOs are a mix of plant molecules with many applications such as antimicrobial, anti-inflammatory, besides potential healing properties, as stated above, we draw our attention here to plant polysaccharides and lectins.

#### **3.2 Carbohydrates**

Essentially, seeds play an important role in the reproductive strategies of certain species and represent a critical phase in life cycle of plants. Also, they play a vital role in diet and human health, which fosters a wide range of potential applications explored by science and technology. Substantial contributions to human well-being and health have emerged from these applications in fields such as the development of biopharmaceuticals [19].

All cells in higher plants present in their cell wall a complex network formed by numerous polymers including cellulose, non-cellulosic polysaccharides (pectin),

**37**

**Figure 1.**

*Plant Macromolecules as Biomaterials for Wound Healing*

structural glycoproteins and, on the secondary wall, lignin. A singular characteristic of plant cell walls is the presence of cellulose, consisting of glucose chains linked to β-(1→4), organized in microfibrils [20, 21]. Intertwined with these microfibrils, a series of hemicelluloses, which are polysaccharides that have similar characteris-

Cell wall polysaccharides can be divided into structural and storage polysaccharides. Primary and secondary walls contain cellulose and hemicelluloses, pectin in addition to enzymes and structural proteins, while the secondary walls contain few proteins or pectin, but usually contain lignin. Secondary cell walls appear when the cell interrupts its growth and often exhibit elaborate specializations for which the incorporation of lignin is usually the most distinctive characteristic. Therefore, the secondary walls of cotyledonary and endospermic cells in seeds of

On the other hand, in some seeds, cell wall of storage tissues (endosperm or cotyledon) is quite thick and contains deposits of polysaccharides, which are mobilized after germination. These polysaccharides are called cell wall storage polysaccharides (CWSPs) and are split into: mannans; galactomannans and glucomannans;

Storage polysaccharides are mostly water soluble and form viscous and stable dispersions, normally absorbing a large amount of this solvent. It assures water around the embryo during the imbibition and germination process, helping protect

Galactomannans are neutral cell wall polysaccharides, commonly found in endosperm of dicotyledonous seeds. They perform the storage function, being usually catabolized to provide energy and carbon skeletons to the plant during germination. They are more abundant in seeds of *Leguminosae* family in which the four major sources of commercial importance are locust bean (*Ceratonia siliqua*), guar (*Cyamopsis tetragonoloba*), tara (*Caesalpinia spinosa* Kuntze) and fenugreek

As can be seen in **Figure 1**, galactomannan are heterogeneous polysaccharides presenting a linear chain of D-mannopyranose residues linked by β-glycoside bonds (1→4) with D-galactopyranosyl attached to these via α-type glycosidic bonds (1→6). In spite of this structure, galactomannans are also called hemicelluloses,

*General structure of galactomannan, considered a hemicellulose by sharing the equatorial β-(1→4)-linked backbone with cellulose. Galactosyl residues are attached to the main backbone by α-glycosyl bonds.*

many species do not have lignin and contain low cellulose [22, 23].

*DOI: http://dx.doi.org/10.5772/intechopen.89105*

tics to cellulose, are found crosslinked to it.

*3.2.1 Plant cell wall polysaccharides*

xyloglucans and galactans [22].

(*Trigonella foenum-graecum* L.) [24].

against dehydration [22].

*3.2.1.1 Galactomannans*

structural glycoproteins and, on the secondary wall, lignin. A singular characteristic of plant cell walls is the presence of cellulose, consisting of glucose chains linked to β-(1→4), organized in microfibrils [20, 21]. Intertwined with these microfibrils, a series of hemicelluloses, which are polysaccharides that have similar characteristics to cellulose, are found crosslinked to it.

### *3.2.1 Plant cell wall polysaccharides*

*Wound Healing*

applications.

**3.1 Essential oils**

process through different mechanisms.

cosmeceuticals and dermaceutical products [14, 15].

nistic, additive or synergistic effects [15].

**3.2 Carbohydrates**

of biopharmaceuticals [19].

**3. Plant-derived compounds**

collagen, and these fibers can recover approximately 80% of the force compared to normal tissue, but the force of the original tissue can never be recovered again [12]. As demonstrated, many factors can alter positively or negatively the cell interactions and the signaling mechanisms during the wound healing process. Plantderived compounds are among these factors and are able to improve the healing

Medicinal plants have been extensively used worldwide as traditional treatment for various diseases due to being a source of phytochemicals, which are nonnutritive substances present in plants enhancing tissue regeneration and acting as pro-angiogenic agents for wound healing. In addition, bioactive products extracted from plants arouse scientific and commercial interests for the development of new drugs [13]. On the other hand, plants are also source of many macromolecules such as carbohydrate and proteins extensively used as biomaterials for wound healing

Essential oils (EOs) are the largest group of secondary metabolites biosynthesized by plant as a complex of monoterpenes (10 carbons) and sesquiterpenes (15 carbons), mainly related to plant defense mechanisms. Also known as volatile oils or aromatic plant essences, they can be found in a multitude of plant tissues such as flowers, leaves, barks, etc. Obtained by aqueous extraction or steam distillation, or cold pressing in the case of citric fruits, they have been extensively employed in

In the healing process, the OEs stand out for their anti-inflammatory and antimicrobial properties. The efficacy in inhibiting bacterial development, including antibiotic-resistant strains, yeasts and filamentous fungi, has boosted the study of the antimicrobial activity of essential oils. Some oils extracted from medicinal plants demonstrated therapeutic potential in combating biofilms, a mechanism of virulence produced by pathogenic microorganisms, resistant to antibiotics [16]. Carvacrol and Thymol, for example, are monoterpenes widely found in essential oils *Origanum* genus, presenting antibacterial and antifungal activities [17] in addition to analgesic effect [18]. In several studies, it has been reported that interactions between the components of EOs, even in small concentrations, may lead to antago-

Although EOs are a mix of plant molecules with many applications such as antimicrobial, anti-inflammatory, besides potential healing properties, as stated above,

Essentially, seeds play an important role in the reproductive strategies of certain species and represent a critical phase in life cycle of plants. Also, they play a vital role in diet and human health, which fosters a wide range of potential applications explored by science and technology. Substantial contributions to human well-being and health have emerged from these applications in fields such as the development

All cells in higher plants present in their cell wall a complex network formed by numerous polymers including cellulose, non-cellulosic polysaccharides (pectin),

we draw our attention here to plant polysaccharides and lectins.

**36**

Cell wall polysaccharides can be divided into structural and storage polysaccharides. Primary and secondary walls contain cellulose and hemicelluloses, pectin in addition to enzymes and structural proteins, while the secondary walls contain few proteins or pectin, but usually contain lignin. Secondary cell walls appear when the cell interrupts its growth and often exhibit elaborate specializations for which the incorporation of lignin is usually the most distinctive characteristic. Therefore, the secondary walls of cotyledonary and endospermic cells in seeds of many species do not have lignin and contain low cellulose [22, 23].

On the other hand, in some seeds, cell wall of storage tissues (endosperm or cotyledon) is quite thick and contains deposits of polysaccharides, which are mobilized after germination. These polysaccharides are called cell wall storage polysaccharides (CWSPs) and are split into: mannans; galactomannans and glucomannans; xyloglucans and galactans [22].

Storage polysaccharides are mostly water soluble and form viscous and stable dispersions, normally absorbing a large amount of this solvent. It assures water around the embryo during the imbibition and germination process, helping protect against dehydration [22].

#### *3.2.1.1 Galactomannans*

Galactomannans are neutral cell wall polysaccharides, commonly found in endosperm of dicotyledonous seeds. They perform the storage function, being usually catabolized to provide energy and carbon skeletons to the plant during germination. They are more abundant in seeds of *Leguminosae* family in which the four major sources of commercial importance are locust bean (*Ceratonia siliqua*), guar (*Cyamopsis tetragonoloba*), tara (*Caesalpinia spinosa* Kuntze) and fenugreek (*Trigonella foenum-graecum* L.) [24].

As can be seen in **Figure 1**, galactomannan are heterogeneous polysaccharides presenting a linear chain of D-mannopyranose residues linked by β-glycoside bonds (1→4) with D-galactopyranosyl attached to these via α-type glycosidic bonds (1→6). In spite of this structure, galactomannans are also called hemicelluloses,

#### **Figure 1.**

*General structure of galactomannan, considered a hemicellulose by sharing the equatorial β-(1→4)-linked backbone with cellulose. Galactosyl residues are attached to the main backbone by α-glycosyl bonds.*

and variations in this Gal/Man ratio cause significant changes in physicochemical parameters such as average molecular weight, intrinsic viscosity and polydispersity of this natural polymers. In addition, the solubility in water is significantly affected by this sugar ratio, which may vary depending of source and isolation procedure. As observed, the higher is the main backbone substituted by galactosyl residues, the more is soluble the galactomannan in water [25].

#### *3.2.1.2 Xyloglucans*

Xyloglucans are plant polysaccharides with both structural and storage function, found in the primary cell wall of cotyledon of various seeds. Their main chain consists of D-glucopyranose linked by β-(1→4) and branched in O-6 by α-D-xylopyranoside units, which can also be substituted in O-2 by units of β-D-galactopyranosyl [22]. This group of polysaccharides plays a fundamental role controlling cell expansion. Experimental results evidenced xyloglucans associated with microfibrils, suggesting that like other hemicelluloses, they are able to provide mechanical resistance and physical integrity to the complex arrangements in plant cell walls [26].

With reference to this association to cellulose, xyloglucans are bound via hydrogen bonds and due to their long polysaccharide chains, they assure the maintenance of network microfibrils in cell wall expansion [22]. **Figure 2** displays the common structure of storage xyloglucans, which make them able to form hydrogels and films solutions to be molded in wound dressings able to carrier potential healing molecules [27].

**Figure 2.**

*Xyloglucan structure. As a hemicellulose, they have a backbone composed of β-(1,4)-linked glucans, and conversely to galactomannan structure, xyloglucans present β-(1,2)-linked galactosyl residues. This latter can also be found branched with an α-l-fucosyl residue. However, storage xyloglucan is not fucosylated.*

#### *3.2.2 Exudate gums (Arabic, tragacanth and cashew gum)*

Exudate gums are hydrocolloids of high molecular weight and viscous appearance, derived from the exudates of branches and bark of the trunks of some plant species. In order to obtain these compounds, a process called gummosis must occur, phenomenon generated as a physiological defense response by chemical, physical and biological stimulus [28]. Chemically, they are made of a complex structure found in arabinogalactans, galacturonans, glucoronomannan or glucoronomannan of acid nature, branched and replaced by major elements (C, H, O and N), inorganic ions and secondary metabolites synthesized from the phenylpropanoid pathway (tannins, terpenoids and other phenolic compounds) [29].

Arabic gum (AG) is a polysaccharide with complex and branched structures (adhesive and cohesive properties) composed of side and main-chains with

**39**

*Plant Macromolecules as Biomaterials for Wound Healing*

β(1,3) and (1,6), respectively, and D-galactopyranosyl, α-L-arabinofuranosyl, α-L-rhamnopyranosyl and β-D-glucopyran units. In some cases, GA is covalently associated with protein fractions and a high content of hydroxyproline, leucine, serine and proline residues [30]. Among its recognized pharmacological properties, we can cite its performance as mucous and intestinal anti-inflammatory, antibacterial and antioxidant, biochemical factors that are probably influential in the wound

systems for *controlled release of drug* and guided tissue regeneration [34].

Tragacanth gum (TG) is an anionic acid branched hetero-polymer with residual units of arabinose, glucose, xylose, galactose rhamnose, fucose and galacturonic acid [32], and TG has important biological characteristics, such as biodegradable and biocompatible biomaterials, resulting in suitable materials in the design of hybrid scaffolds with pharmaceutical applications, *development* of polymeric

Similar to the polymer gums described above, cashew tree gum is extracted from *Anacardium* genus species, which are abundant Brazilian Northeast. The chemical structure is composed of a main-chain of β-galactose (1–3) and branched-bonds (1–6), containing side-residues of glucuronic acid, 4-O-methyl glucuronic arabinose, rhamnose, xylose, glucose and mannose [32]. The biochemical characteristics associated with biological activities demonstrate anti-inflammatory properties, regulators of oxidative stress and reactive oxygen species, antibacterial and gastro-

Latex is produced and stored by laticifer ducts found in some plant species. This fluid is a rich source of natural compounds such as secondary metabolites, glycosides and proteases [36]. Latex proteins have been studied by several researchers as innovative natural compounds for biomedical applications [37]. Among the most common macromolecules from protein origin are cysteine and serine peptidases, also known as latex proteases or latex peptidases. These macromolecules provide the front line of defense against natural enemies in plants acting by synergism with

Proteases are also present in animals and humans, and these enzymes have gained notoriety in medical and pharmaceutical field due to their proteolysis functions, specificity and bioactivity [38]. In human biological systems, proteases such as metalloproteinases are endogenously released by fibroblasts, macrophages, mast cells and endothelial cells after injury in extracellular matrix [39]. These enzymes initially participate in the inflammatory phase of healing by debriding the wound necrotic tissue and contributing to collagen remodeling and reduction of scar tensile strength, the later phase of cicatricial process [40]. Proteases and their inhibitors also contribute to degradation and deposition of ECM, creating a balance that is essential for the adequate and coordinated cutaneous wound healing [41]. The modulation of ECM proteases with laticifer proteins has been used as a strategy to improve the performance of healing processes in acute and chronic wounds [42]. Recent advances in plant latex biotechnology has contributed to study the pharmacological properties of proteases-rich fractions from *Calotropis procera* latex revealing its potential role in procoagulation and blood clot hydrolysis [43], modulation of inflammation [44, 45] and enhanced wound healing in animal models using biomembranes of polyvinyl alcohol as vehicle for releasing laticifer proteins [46, 47]. In addition, a phytomodulatory galactomannan-based hydrogel has been successfully used to carrier latex proteases from *C. procera* in experimental

*DOI: http://dx.doi.org/10.5772/intechopen.89105*

healing process [31–33].

protective effect [35].

*3.3.1 Latex proteases*

others latex sap proteins [36].

**3.3 Proteins**

*Plant Macromolecules as Biomaterials for Wound Healing DOI: http://dx.doi.org/10.5772/intechopen.89105*

β(1,3) and (1,6), respectively, and D-galactopyranosyl, α-L-arabinofuranosyl, α-L-rhamnopyranosyl and β-D-glucopyran units. In some cases, GA is covalently associated with protein fractions and a high content of hydroxyproline, leucine, serine and proline residues [30]. Among its recognized pharmacological properties, we can cite its performance as mucous and intestinal anti-inflammatory, antibacterial and antioxidant, biochemical factors that are probably influential in the wound healing process [31–33].

Tragacanth gum (TG) is an anionic acid branched hetero-polymer with residual units of arabinose, glucose, xylose, galactose rhamnose, fucose and galacturonic acid [32], and TG has important biological characteristics, such as biodegradable and biocompatible biomaterials, resulting in suitable materials in the design of hybrid scaffolds with pharmaceutical applications, *development* of polymeric systems for *controlled release of drug* and guided tissue regeneration [34].

Similar to the polymer gums described above, cashew tree gum is extracted from *Anacardium* genus species, which are abundant Brazilian Northeast. The chemical structure is composed of a main-chain of β-galactose (1–3) and branched-bonds (1–6), containing side-residues of glucuronic acid, 4-O-methyl glucuronic arabinose, rhamnose, xylose, glucose and mannose [32]. The biochemical characteristics associated with biological activities demonstrate anti-inflammatory properties, regulators of oxidative stress and reactive oxygen species, antibacterial and gastroprotective effect [35].

#### **3.3 Proteins**

*Wound Healing*

*3.2.1.2 Xyloglucans*

molecules [27].

**Figure 2.**

and variations in this Gal/Man ratio cause significant changes in physicochemical parameters such as average molecular weight, intrinsic viscosity and polydispersity of this natural polymers. In addition, the solubility in water is significantly affected by this sugar ratio, which may vary depending of source and isolation procedure. As observed, the higher is the main backbone substituted by galactosyl residues, the

Xyloglucans are plant polysaccharides with both structural and storage function, found in the primary cell wall of cotyledon of various seeds. Their main chain consists of D-glucopyranose linked by β-(1→4) and branched in O-6 by α-D-xylopyranoside units, which can also be substituted in O-2 by units of β-D-galactopyranosyl [22]. This group of polysaccharides plays a fundamental role controlling cell expansion. Experimental results evidenced xyloglucans associated with microfibrils, suggesting that like other hemicelluloses, they are able to provide mechanical resistance and

With reference to this association to cellulose, xyloglucans are bound via hydrogen bonds and due to their long polysaccharide chains, they assure the maintenance of network microfibrils in cell wall expansion [22]. **Figure 2** displays the common structure of storage xyloglucans, which make them able to form hydrogels and films solutions to be molded in wound dressings able to carrier potential healing

Exudate gums are hydrocolloids of high molecular weight and viscous appearance, derived from the exudates of branches and bark of the trunks of some plant species. In order to obtain these compounds, a process called gummosis must occur, phenomenon generated as a physiological defense response by chemical, physical and biological stimulus [28]. Chemically, they are made of a complex structure found in arabinogalactans, galacturonans, glucoronomannan or glucoronomannan of acid nature, branched and replaced by major elements (C, H, O and N), inorganic ions and secondary metabolites synthesized from the phenylpropanoid

*Xyloglucan structure. As a hemicellulose, they have a backbone composed of β-(1,4)-linked glucans, and conversely to galactomannan structure, xyloglucans present β-(1,2)-linked galactosyl residues. This latter can also be found branched with an α-l-fucosyl residue. However, storage xyloglucan is not fucosylated.*

Arabic gum (AG) is a polysaccharide with complex and branched structures (adhesive and cohesive properties) composed of side and main-chains with

physical integrity to the complex arrangements in plant cell walls [26].

*3.2.2 Exudate gums (Arabic, tragacanth and cashew gum)*

pathway (tannins, terpenoids and other phenolic compounds) [29].

more is soluble the galactomannan in water [25].

**38**

#### *3.3.1 Latex proteases*

Latex is produced and stored by laticifer ducts found in some plant species. This fluid is a rich source of natural compounds such as secondary metabolites, glycosides and proteases [36]. Latex proteins have been studied by several researchers as innovative natural compounds for biomedical applications [37]. Among the most common macromolecules from protein origin are cysteine and serine peptidases, also known as latex proteases or latex peptidases. These macromolecules provide the front line of defense against natural enemies in plants acting by synergism with others latex sap proteins [36].

Proteases are also present in animals and humans, and these enzymes have gained notoriety in medical and pharmaceutical field due to their proteolysis functions, specificity and bioactivity [38]. In human biological systems, proteases such as metalloproteinases are endogenously released by fibroblasts, macrophages, mast cells and endothelial cells after injury in extracellular matrix [39]. These enzymes initially participate in the inflammatory phase of healing by debriding the wound necrotic tissue and contributing to collagen remodeling and reduction of scar tensile strength, the later phase of cicatricial process [40]. Proteases and their inhibitors also contribute to degradation and deposition of ECM, creating a balance that is essential for the adequate and coordinated cutaneous wound healing [41].

The modulation of ECM proteases with laticifer proteins has been used as a strategy to improve the performance of healing processes in acute and chronic wounds [42]. Recent advances in plant latex biotechnology has contributed to study the pharmacological properties of proteases-rich fractions from *Calotropis procera* latex revealing its potential role in procoagulation and blood clot hydrolysis [43], modulation of inflammation [44, 45] and enhanced wound healing in animal models using biomembranes of polyvinyl alcohol as vehicle for releasing laticifer proteins [46, 47]. In addition, a phytomodulatory galactomannan-based hydrogel has been successfully used to carrier latex proteases from *C. procera* in experimental excisional wound models. It was observed a synergic effect between both galactomannan and proteases macromolecules, enhancing the healing [48].

### *3.3.2 Lectins*

Lectins (from Latin "*legere,*" to pick out or choose) are proteins or glycoproteins found in nonimmune nature which can specifically recognize and reversibly bind carbohydrate moieties without altering the covalent structure of their glycosyl ligands. Indeed, this attractive characteristic distinguishes lectins from other carbohydrate binding proteins and enzymes. They are also widely distributed in the plant kingdom, usually from leguminous seed origin and play crucial roles and functions in biological processes such as molecular recognition, storage proteins and plant defense mechanisms. Their interaction with glycosyl ligands occurs mainly through hydrogen bonds, van der Waals' forces, hydrophobic interactions and, less frequently, electrostatic interactions [49]. Here, we emphasize lectins from jackfruit, breadfruit and chempedak (**Figure 3**) and their biological applications.

Lectins were first discovered in plants and, although ubiquitously distributed in nature (animals, insects, viruses, fungi and bacteria), and the majority have been characterized from plant protein extracts, reflecting ease of extraction and relatively high yields usually via a simple one-step affinity chromatographic method [50]. After the discovery of jacalin, novel lectins sharing high homology to it were placed into a family of jacalin-related lectins (JRL), now divided into two different subgroups. The first comprises galactose-specific lectins (gJRL) and a few other Moraceae lectins, which exhibit specificity toward galactose and are built up of subunits consisting of a short β chain and a long α chain. The second is the mannosebinding subgroup (mJRL), which occurs in different plant families with the lectins exhibiting exclusive specificity toward glucose/mannose residues with the binding subunits contained within a single polypeptide chain [51, 52].

#### **Figure 3.**

*Artocarpus species. (A) Jackfruit. (B) Cut section of jackfruit. (C) Jackfruit seeds. (D) Breadfruit (var. seminifera). (E) Cut section of breadfruit (var. seminifera). (F) Breadfruit (var. seminifera) seeds. (G) Breadfruit (var. apyrena). (H) Cut section of breadfruit (var. apyrena). (I) Chempedak.*

**41**

*Plant Macromolecules as Biomaterials for Wound Healing*

which are readily recovered from their seed flour.

*3.3.2.1.1 Jackfruit (jacalin, ArtinM and jackin)*

indicating therapeutic applications [56].

neutral peptide NH group [62, 63].

*Artocarpus* is a genus comprising about 60 trees and shrubs of Southeast Asian and Pacific origin, belonging to the Moraceae family; all species are lactiferous with leaves, twigs, and stems producing milky sap. The name is derived from the Greek words *artos* ("bread") and *karpos* ("fruit"). Although most species of *Artocarpus* are restricted to Southeast Asia, such as *A. hypargyreus* (kwai muk), *A. lakoocha* (lakoocha), *A. kemando* (pudau), *A. hirsutus* (anjily), *A. chama* (chaplaish) and *A. odoratissimus* (marang), several species are widely distributed and cultivated throughout the tropics due to their edible fruits and timber. These include *A. heterophyllus* (jackfruit), *A. altilis* and *A. integer* (cempedak, also known as chempedak) [53–55], well-known species conspicuous as substantial sources of plant lectins

The species has an extensive record of use in folk medicine including treatment of asthma, dermatitis, anemia, diarrhea and fever; antisyphilitic and anthelmintic properties; sedative effects in convulsions; and also wound healing properties [56]. Indeed, jackfruit are in high demand in areas such as cosmeceutical, pharmaceutical and natural food handling for supplement markets, due to numerous studies on phytochemical and pharmacological properties of all parts of the plant (pulp, leaf,

Extracts and metabolites from jackfruit possess several useful bioactive compounds and may confer multiple health-promoting effects for heart and skin disorders and to treat ulcers and cancer [59]. Moreover, new studies of jackfruit properties provide additional biological findings consistent with antibacterial, antitubercular, antiviral, antifungal, antiplatelet and antiarthritic actions, thereby

The occurrence of jacalin, the D-galactose-binding lectin from *A. integrifolia* seeds was first reported in 1979 [60] and found to exceed 50% of the protein in jackfruit crude seed extracts. This is also the case for galactose-binding lectins in the *Artocarpus* genus such as frutalin and CGB (chempedak galactose-binding). Jacalin is a tetrameric two-chain lectin of 65 kDa combining a heavy α-chain of 133 amino acids and light β-chain of 20–21 amino acid to form a 3D structure as a single globular unit. By SDS-PAGE, this lectin shows two bands between 20 and 14 kDa, which correspond to glycosylated and slightly or non-glycosylated forms,

The carbohydrate-binding site (CBS) of jacalin mainly involves residues Gly1, Tyr78, Val80, Gly121, Tyr122, Trp123 and Asp125. In D-galactose-jacalin complexes, the O4 hydroxyl group of the galactose axial position forms hydrogen bonds with the side chain of Asp125 and the terminal amino group of Gly1. If O4 is at the equatorial position, as in glucose and mannose, Asp125 can still interact with O4, but not the amino group. This explains the high specificity of jacalin for galactose compared to glucose and mannose at the primary binding site. Additionally, the post-translational cleavage and removal of the "T-S-S-N" peptide linker assure a stronger hydrogen bond connecting the α- and β-chains, as non-cleavage leaves a

Further studies post-discovery of jacalin revealed that jackfruit seed extracts also possess small amounts of a D-mannose-binding lectin [64]. Previously named artocarpin, the term was provisionally replaced by the name KM+; the lectin had become confused with distinct substances from *Artocarpus* spp. which were

*DOI: http://dx.doi.org/10.5772/intechopen.89105*

*3.3.2.1 Artocarpus lectins*

root and bark) [57, 58].

respectively [61].

### *3.3.2.1 Artocarpus lectins*

*Wound Healing*

*3.3.2 Lectins*

biological applications.

excisional wound models. It was observed a synergic effect between both galacto-

Lectins (from Latin "*legere,*" to pick out or choose) are proteins or glycoproteins found in nonimmune nature which can specifically recognize and reversibly bind carbohydrate moieties without altering the covalent structure of their glycosyl ligands. Indeed, this attractive characteristic distinguishes lectins from other carbohydrate binding proteins and enzymes. They are also widely distributed in the plant kingdom, usually from leguminous seed origin and play crucial roles and functions in biological processes such as molecular recognition, storage proteins and plant defense mechanisms. Their interaction with glycosyl ligands occurs mainly through hydrogen bonds, van der Waals' forces, hydrophobic interactions and, less frequently, electrostatic interactions [49]. Here, we emphasize lectins from jackfruit, breadfruit and chempedak (**Figure 3**) and their

Lectins were first discovered in plants and, although ubiquitously distributed in nature (animals, insects, viruses, fungi and bacteria), and the majority have been characterized from plant protein extracts, reflecting ease of extraction and relatively high yields usually via a simple one-step affinity chromatographic method [50]. After the discovery of jacalin, novel lectins sharing high homology to it were placed into a family of jacalin-related lectins (JRL), now divided into two different subgroups. The first comprises galactose-specific lectins (gJRL) and a few other Moraceae lectins, which exhibit specificity toward galactose and are built up of subunits consisting of a short β chain and a long α chain. The second is the mannosebinding subgroup (mJRL), which occurs in different plant families with the lectins exhibiting exclusive specificity toward glucose/mannose residues with the binding

subunits contained within a single polypeptide chain [51, 52].

*Artocarpus species. (A) Jackfruit. (B) Cut section of jackfruit. (C) Jackfruit seeds. (D) Breadfruit (var. seminifera). (E) Cut section of breadfruit (var. seminifera). (F) Breadfruit (var. seminifera) seeds. (G) Breadfruit (var. apyrena). (H) Cut section of breadfruit (var. apyrena). (I) Chempedak.*

mannan and proteases macromolecules, enhancing the healing [48].

**40**

**Figure 3.**

*Artocarpus* is a genus comprising about 60 trees and shrubs of Southeast Asian and Pacific origin, belonging to the Moraceae family; all species are lactiferous with leaves, twigs, and stems producing milky sap. The name is derived from the Greek words *artos* ("bread") and *karpos* ("fruit"). Although most species of *Artocarpus* are restricted to Southeast Asia, such as *A. hypargyreus* (kwai muk), *A. lakoocha* (lakoocha), *A. kemando* (pudau), *A. hirsutus* (anjily), *A. chama* (chaplaish) and *A. odoratissimus* (marang), several species are widely distributed and cultivated throughout the tropics due to their edible fruits and timber. These include *A. heterophyllus* (jackfruit), *A. altilis* and *A. integer* (cempedak, also known as chempedak) [53–55], well-known species conspicuous as substantial sources of plant lectins which are readily recovered from their seed flour.

### *3.3.2.1.1 Jackfruit (jacalin, ArtinM and jackin)*

The species has an extensive record of use in folk medicine including treatment of asthma, dermatitis, anemia, diarrhea and fever; antisyphilitic and anthelmintic properties; sedative effects in convulsions; and also wound healing properties [56]. Indeed, jackfruit are in high demand in areas such as cosmeceutical, pharmaceutical and natural food handling for supplement markets, due to numerous studies on phytochemical and pharmacological properties of all parts of the plant (pulp, leaf, root and bark) [57, 58].

Extracts and metabolites from jackfruit possess several useful bioactive compounds and may confer multiple health-promoting effects for heart and skin disorders and to treat ulcers and cancer [59]. Moreover, new studies of jackfruit properties provide additional biological findings consistent with antibacterial, antitubercular, antiviral, antifungal, antiplatelet and antiarthritic actions, thereby indicating therapeutic applications [56].

The occurrence of jacalin, the D-galactose-binding lectin from *A. integrifolia* seeds was first reported in 1979 [60] and found to exceed 50% of the protein in jackfruit crude seed extracts. This is also the case for galactose-binding lectins in the *Artocarpus* genus such as frutalin and CGB (chempedak galactose-binding). Jacalin is a tetrameric two-chain lectin of 65 kDa combining a heavy α-chain of 133 amino acids and light β-chain of 20–21 amino acid to form a 3D structure as a single globular unit. By SDS-PAGE, this lectin shows two bands between 20 and 14 kDa, which correspond to glycosylated and slightly or non-glycosylated forms, respectively [61].

The carbohydrate-binding site (CBS) of jacalin mainly involves residues Gly1, Tyr78, Val80, Gly121, Tyr122, Trp123 and Asp125. In D-galactose-jacalin complexes, the O4 hydroxyl group of the galactose axial position forms hydrogen bonds with the side chain of Asp125 and the terminal amino group of Gly1. If O4 is at the equatorial position, as in glucose and mannose, Asp125 can still interact with O4, but not the amino group. This explains the high specificity of jacalin for galactose compared to glucose and mannose at the primary binding site. Additionally, the post-translational cleavage and removal of the "T-S-S-N" peptide linker assure a stronger hydrogen bond connecting the α- and β-chains, as non-cleavage leaves a neutral peptide NH group [62, 63].

Further studies post-discovery of jacalin revealed that jackfruit seed extracts also possess small amounts of a D-mannose-binding lectin [64]. Previously named artocarpin, the term was provisionally replaced by the name KM+; the lectin had become confused with distinct substances from *Artocarpus* spp. which were

similarly designated. Additionally, artocarpin was used to name the galactosebinding lectin in *Artocarpus lakoocha* seeds [65].

The origin of the name KM+ comes from the successive affinity chromatography steps in immobilized D-galactose matrices to remove jacalin (retained fraction J). The unretained fraction was called K, while M+ refers to the retained fraction on immobilized mannose matrices. Nevertheless, there remained confusion over this adopted nomenclature, which led to the proposal for a rational name change to ArtinM based on the universal code for plant proteins. This takes into account both a lectin's origin and specificity of sugar recognition [66]. Hereafter, we will apply ArtinM to include early work and mentions of KM+ (artocarpin).

ArtinM is a single polypeptide of 150 amino acids devoid of covalently attached carbohydrates with four isolectins and a pI range of 5–6.5, sharing 52% sequence identity with jacalin [67]. Unlike jacalin, there are no aromatic residues on the CBS of ArtinM, comprising Gly15, Asp138, Leu139 and Asp141. Indeed, it is believed that the galactose specificity of jacalin was achieved by a two-step process with ArtinM as its putative precursor: mutation of key aliphatic residues close to the sugar binding pocket to aromatic ones; and then cleavage of a short loop, a key step as it creates a positively charged N-terminal which interacts specifically with O4 at the axial position [68, 69].

Besides jacalin and ArtinM, jackfruit seeds are the source of a third lectin named jackin due to its affinity for chitin. Nevertheless, its low yield from natural sources still hampers further characterization, though this might soon be overcome with high-yield heterologous production in microbial systems [70].

#### *3.3.2.1.2 Breadfruit*

The Pacific Islands are the center of origin and diversity of breadfruit (*A. altilis*), sometimes referred to as *A. communis* or *A. incisa*. The species was derived from a seeded, diploid ancestor, *A. camansi,* giving two varieties, one seeded (var. seminifera) whose rind spines are as prominent as those on jackfruit, and one seedless (var. apyrena), which presents a spineless outer layer. The latter is well-appreciated by local Brazilians when cooked, because of its large starchy content of compound fruits with high levels of minerals and provitamin A carotenoids [53, 55, 71]. Breadfruit flour was approved in 2016 by the US Food and Drug Administration (FDA) as a food Generally Recognized as Safe (GRAS).

As of 1983, our group started to survey lectins in *A. incisa* seeds and found lectins with similar behavior to those in jackfruit seeds [72]. Breadfruit seeds are composed of a high-water content (up to 60%) and moderate protein content (12.25% of dry weight). Most of this protein is recovered as frutalin in chromatographic methods using crude extracts of seed flour. Therefore, frutalin is the most abundant lectin of this species, presenting multiple-binding properties in which the same CBS recognizes a range of different ligands, although it has higher affinities for α-D-galactose monosaccharides and complex carbohydrates that contain Galα1–3 glycans [73].

The CBS of frutalin in the binding of galactose is dominated by hydrogen bonding through O1, O3, O5 and O6 and backbone/side chain hydroxyl groups. Similar to the Moraceae lectins, the CBS of frutalin is located in a domain close to the N-terminus of the α chain, consisting of four key residues Gly25, Tyr146, Trp147 and Asp149. About 10 interactions occur, involving the C1 hydroxyl to residue Tyr146, hydroxyl C3 to residue Gly25, hydroxyl C4 to residues Gly25 and Asp149, and hydroxyl C6 to residues Tyr146, Trp147 and Asp149 [74]. Moreover, there is evidence to suggest that frutalin possesses stereospecificity, capable of specifically

**43**

*3.3.2.1.3 Chempedak*

*Plant Macromolecules as Biomaterials for Wound Healing*

binding α-D-galactose, since it was previously isolated on a cross-linked galacto-

FTP is a hololectin defined as a homotetramer with an identical CBS per protomer, able to bind either the same or structurally similar sugars. The CBS is formed of four residues, namely Gly16, Asp139, Leu140 and Asp142, distributed in a few loops connecting the strands β5 and β6, β7 and β8, and β11 and β12. Several hydrogen bonds (HB) occur in FTP-glucose and FTP-mannose complexes involving Gly16, Leu90, Gly138, Asp139, Leu140, Asp142 and O3, O4, O5, O6 of the carbohydrates. In MD simulations, Lys60 plays an important role forming salt bridges with Asp139 in FTP-glucose complexes, reducing the interaction between this former residue and mannose and minimizing the repulsion of the mannose hydroxyl groups with oxygen. In this case, mannose was more completely surrounded in the carbohydrate-binding site and also stabilized by indirect interaction with Asp139 through water molecules. This local structuring is more stable in the case of mannose than glucose, indicating a higher affinity of FTP for mannose residues than glucose [78]. Further studies of breadfruit seeds also revealed a lectin similar to jackin, named

frutackin. Both lectins have a 14 kDa polypeptide chain, built up of three chains linked by disulfide bonds, the partial amino acid sequences of the two lectins showing their homology to each other in terms of molecular mass, secondary structure, and primary sequence, but not to other plant chitin-binding proteins. Both jackin

Although widely known in the tropics as chempedak or chempedak, *Artocarpus integer* (Thumb.) Merr. is native to India with similar fruits to jackfruit. The *A. integer* species is rich in phenolic compounds, presenting a strong cytotoxic activity against murine leukemia P-388 [79]. Moreover, a chempedak paste of the inner bark prevents infection and helps healing when applied on wounds, as well as heated leaf extract [80]. Chempedak galactose-binding lectin (CGB) is found in high levels *A. integer* seed flour extracts. The lectin's bioactivity was first noted when extracts were tested for selective stimulation of peripheral blood mononuclear cells; at 20 μg/mL CGB stimulated proliferation of T-lymphocyte populations [81, 82]. Like frutalin and jacalin, CGB is transcribed as a propeptide and then post-translationally processed into a typical gJRL lectin, having a 13-kDa α-chain (133 amino acids) and a 2.1-kDa β-chain (21 amino acids). CGB has six different amino-acids compared to jacalin,

As with jacalin complexes, the interactions are well conserved, presenting the same CBS. In Gal-CGB complexes, the O atoms on the sugar ring are bound with the sidechain and main-chain N and O atoms on the α chain (O3 and Gly1 N; O4 and Gly1 N and Asp125 OD1; O6 and Trp123 O, Trp123 N and Tyr122 N; and O5 and Tyr122 N). Similarly, GalNac-CGB complexes present bound disaccharide in the same region though hydrogen bonds (O3 and Gly1 N; O4 and Gly1 N and Asp125 OD1; and O6

and frutackin inhibit the growth of *F. moniliforme* and *S. cerevisiae* [70].

giving 97% identity for their amino-acid sequences [83].

Frutapin (FTP) is the next most abundant lectin in breadfruit seed extracts. Preliminary, FTP investigations began in 1998, describing three different lectins with distinct carbohydrate recognition within the same species [76]. However, further studies proved difficult since native FTP was hampered by very low yields and contamination with Frutalin, a notable factor as frutalin binds diverse sugar moieties and has high abundance in plant extracts. Recombinant FTP production is now a feasible approach to circumvent this problem, allowing large-scale heterologous expression to give continuous supplies for further characterization and to

mannan column, but not on β-galactosyl-immobilized matrices [73, 75].

improve potency, particularly for biomedical applications [77].

*DOI: http://dx.doi.org/10.5772/intechopen.89105*

#### *Plant Macromolecules as Biomaterials for Wound Healing DOI: http://dx.doi.org/10.5772/intechopen.89105*

*Wound Healing*

the axial position [68, 69].

*3.3.2.1.2 Breadfruit*

Galα1–3 glycans [73].

similarly designated. Additionally, artocarpin was used to name the galactose-

ArtinM to include early work and mentions of KM+ (artocarpin).

high-yield heterologous production in microbial systems [70].

(FDA) as a food Generally Recognized as Safe (GRAS).

The origin of the name KM+ comes from the successive affinity chromatography steps in immobilized D-galactose matrices to remove jacalin (retained fraction J). The unretained fraction was called K, while M+ refers to the retained fraction on immobilized mannose matrices. Nevertheless, there remained confusion over this adopted nomenclature, which led to the proposal for a rational name change to ArtinM based on the universal code for plant proteins. This takes into account both a lectin's origin and specificity of sugar recognition [66]. Hereafter, we will apply

ArtinM is a single polypeptide of 150 amino acids devoid of covalently attached carbohydrates with four isolectins and a pI range of 5–6.5, sharing 52% sequence identity with jacalin [67]. Unlike jacalin, there are no aromatic residues on the CBS of ArtinM, comprising Gly15, Asp138, Leu139 and Asp141. Indeed, it is believed that the galactose specificity of jacalin was achieved by a two-step process with ArtinM as its putative precursor: mutation of key aliphatic residues close to the sugar binding pocket to aromatic ones; and then cleavage of a short loop, a key step as it creates a positively charged N-terminal which interacts specifically with O4 at

Besides jacalin and ArtinM, jackfruit seeds are the source of a third lectin named jackin due to its affinity for chitin. Nevertheless, its low yield from natural sources still hampers further characterization, though this might soon be overcome with

The Pacific Islands are the center of origin and diversity of breadfruit (*A. altilis*), sometimes referred to as *A. communis* or *A. incisa*. The species was derived from a seeded, diploid ancestor, *A. camansi,* giving two varieties, one seeded (var. seminifera) whose rind spines are as prominent as those on jackfruit, and one seedless (var. apyrena), which presents a spineless outer layer. The latter is well-appreciated by local Brazilians when cooked, because of its large starchy content of compound fruits with high levels of minerals and provitamin A carotenoids [53, 55, 71]. Breadfruit flour was approved in 2016 by the US Food and Drug Administration

As of 1983, our group started to survey lectins in *A. incisa* seeds and found lectins with similar behavior to those in jackfruit seeds [72]. Breadfruit seeds are composed of a high-water content (up to 60%) and moderate protein content (12.25% of dry weight). Most of this protein is recovered as frutalin in chromatographic methods using crude extracts of seed flour. Therefore, frutalin is the most abundant lectin of this species, presenting multiple-binding properties in which the same CBS recognizes a range of different ligands, although it has higher affinities for α-D-galactose monosaccharides and complex carbohydrates that contain

The CBS of frutalin in the binding of galactose is dominated by hydrogen bonding through O1, O3, O5 and O6 and backbone/side chain hydroxyl groups. Similar to the Moraceae lectins, the CBS of frutalin is located in a domain close to the N-terminus of the α chain, consisting of four key residues Gly25, Tyr146, Trp147 and Asp149. About 10 interactions occur, involving the C1 hydroxyl to residue Tyr146, hydroxyl C3 to residue Gly25, hydroxyl C4 to residues Gly25 and Asp149, and hydroxyl C6 to residues Tyr146, Trp147 and Asp149 [74]. Moreover, there is evidence to suggest that frutalin possesses stereospecificity, capable of specifically

binding lectin in *Artocarpus lakoocha* seeds [65].

**42**

binding α-D-galactose, since it was previously isolated on a cross-linked galactomannan column, but not on β-galactosyl-immobilized matrices [73, 75].

Frutapin (FTP) is the next most abundant lectin in breadfruit seed extracts. Preliminary, FTP investigations began in 1998, describing three different lectins with distinct carbohydrate recognition within the same species [76]. However, further studies proved difficult since native FTP was hampered by very low yields and contamination with Frutalin, a notable factor as frutalin binds diverse sugar moieties and has high abundance in plant extracts. Recombinant FTP production is now a feasible approach to circumvent this problem, allowing large-scale heterologous expression to give continuous supplies for further characterization and to improve potency, particularly for biomedical applications [77].

FTP is a hololectin defined as a homotetramer with an identical CBS per protomer, able to bind either the same or structurally similar sugars. The CBS is formed of four residues, namely Gly16, Asp139, Leu140 and Asp142, distributed in a few loops connecting the strands β5 and β6, β7 and β8, and β11 and β12. Several hydrogen bonds (HB) occur in FTP-glucose and FTP-mannose complexes involving Gly16, Leu90, Gly138, Asp139, Leu140, Asp142 and O3, O4, O5, O6 of the carbohydrates. In MD simulations, Lys60 plays an important role forming salt bridges with Asp139 in FTP-glucose complexes, reducing the interaction between this former residue and mannose and minimizing the repulsion of the mannose hydroxyl groups with oxygen. In this case, mannose was more completely surrounded in the carbohydrate-binding site and also stabilized by indirect interaction with Asp139 through water molecules. This local structuring is more stable in the case of mannose than glucose, indicating a higher affinity of FTP for mannose residues than glucose [78].

Further studies of breadfruit seeds also revealed a lectin similar to jackin, named frutackin. Both lectins have a 14 kDa polypeptide chain, built up of three chains linked by disulfide bonds, the partial amino acid sequences of the two lectins showing their homology to each other in terms of molecular mass, secondary structure, and primary sequence, but not to other plant chitin-binding proteins. Both jackin and frutackin inhibit the growth of *F. moniliforme* and *S. cerevisiae* [70].

#### *3.3.2.1.3 Chempedak*

Although widely known in the tropics as chempedak or chempedak, *Artocarpus integer* (Thumb.) Merr. is native to India with similar fruits to jackfruit. The *A. integer* species is rich in phenolic compounds, presenting a strong cytotoxic activity against murine leukemia P-388 [79]. Moreover, a chempedak paste of the inner bark prevents infection and helps healing when applied on wounds, as well as heated leaf extract [80].

Chempedak galactose-binding lectin (CGB) is found in high levels *A. integer* seed flour extracts. The lectin's bioactivity was first noted when extracts were tested for selective stimulation of peripheral blood mononuclear cells; at 20 μg/mL CGB stimulated proliferation of T-lymphocyte populations [81, 82]. Like frutalin and jacalin, CGB is transcribed as a propeptide and then post-translationally processed into a typical gJRL lectin, having a 13-kDa α-chain (133 amino acids) and a 2.1-kDa β-chain (21 amino acids). CGB has six different amino-acids compared to jacalin, giving 97% identity for their amino-acid sequences [83].

As with jacalin complexes, the interactions are well conserved, presenting the same CBS. In Gal-CGB complexes, the O atoms on the sugar ring are bound with the sidechain and main-chain N and O atoms on the α chain (O3 and Gly1 N; O4 and Gly1 N and Asp125 OD1; O6 and Trp123 O, Trp123 N and Tyr122 N; and O5 and Tyr122 N). Similarly, GalNac-CGB complexes present bound disaccharide in the same region though hydrogen bonds (O3 and Gly1 N; O4 and Gly1 N and Asp125 OD1; and O6

and Asp125 OD1). Also, there are a number of hydrophobic interactions contributed by Tyr78, Gly121 and Tyr122. Although closely related to jacalin's structure, CGB was unable to bind mannose as judged by isothermal calorimetry and co-crystallization studies. It is believed that this change in CGB specificity is caused by subtle differences in the environment near the sugar-binding site, including solvent molecules.

Extracts of *Artocarpus integer* seeds are a source of CMB, previously termed chempedak lectin-M [84]. The lectin is ~20-fold more abundant than ArtinM in crude extracts of *A. heterophyllus* seeds. Structural studies reveal CMB to be a 64-kDa tetramer with some of the polypeptides being disulfide-linked to give dimers. The functional activity of CMB was assessed by studying interactions with different isotypes of human immunoglobulins: strong binding to IgE and IgM was noted, unlike CGB and jacalin which strongly interact with IgA1. The lectin also shows a similar pattern of carbohydrate binding specificity to ArtinM with Man-α-1–3Man the most potent inhibitor, followed by methyl-α-D-mannopyranose and D-mannose [84].

It appears that the *Artocarpus* genus employs a plurality of lectins, though to date few of these lectins have been purified and functionally characterized. Nevertheless, a consistent pattern is that *Artocarpus* seeds contain more than one lectin with distinct carbohydrate recognition features [70, 76]. Altogether, the JRL family is complex with a vast diversity in biochemical properties and activities, which have drawn much attention because of their pivotal biomedical applications. Owing to carbohydrate-binding interactions of plant lectins with cell wall glycoproteins, they are promising targets to selectively modulate immune responses in plants. Hence, it is important to delineate the molecular details of lectin binding to CBS domains and how downstream activation of cellular immune signaling proceeds [85]. In this context, it is of particular note that *Artocarpus* lectins have a distinct repertoire of biological activities, despite their high sequence and structural homology, as can be seen in **Figure 4**.

#### **Figure 4.**

*Sequence alignment using Clustal W and structural superposition of* Artocarpus *lectins. (A) Sequence alignment of galactose-binding lectins: Jacalin (PDB 1UGW), Frutalin (PDB 5BN6), and CGB (PDB 4AKB). (B) Sequence alignment of mannose-binding lectins: ArtinM (PDB 1J4S), frutapin (PDB 5KRP) and CMB (PDB 4AKD). (C) Structural superposition of galactose-binding lectins with their carbohydrate interacting residues in complex with D-galactose: jacalin (cyan), frutalin (red) and CGB (magenta). (D) Structural superposition of mannose-binding lectins with the carbohydrate interacting residues highlighted: ArtinM (green), frutapin (yellow) and CMB (blue).*

**45**

**Table 1.**

*Plant Macromolecules as Biomaterials for Wound Healing*

with potential for wound healing applications (**Table 1**).

Hydrolysate: synthases I, alkaline protease and papain

β-glucan and at least one secondary polysaccharide group consisting of arabinogalactan, fucoidan, pectin and galactomannan

Hemicelluloses and plant lectins

**4. Recent advances using plant biomaterials for wound healing**

The worldwide increasing number of chronic-wound patients has driven an intense effort in the wound-care market in search of low cost and effective wound healing technologies [86]. Therefore, much of this knowledge is eligible for patents that play an important role in identifying technology development

In the following table, we highlight the main efforts in the last 5 years on the development of biomaterials using plant macromolecules as source of biomolecules

**Title Plant biomaterial Description IPC code Ref.**

*C. procera* The compositions may

Xyloglucan The material is

carrier for a protease mixture for removing necrotic tissue in chronic wounds.

Compound preparations that promote blood circulation and wound healing.

be used for topical administration in wound healing.

Therapeutic composition as a potent stimulator for wound healing.

used for preventing reoccurrence of adhesion after adhesion loss operation, and meanwhile has functions of preventing bacteria and promoting wound healing.

Hydrogels and biomembranes are used to treat wounds, abrasions, burns, varicose ulcers, decubitus ulcers, open superficial wounds with or without infection.

A61K38/4873 [87]

A61K 36/00 [88]

— [89]

A61K 31/716 [90]

A61L31/041 [91]

A61K 31/736 [92]

Protease mixture An aqueous gel is

*DOI: http://dx.doi.org/10.5772/intechopen.89105*

trends [86].

Necrotic tissue composition remover for wound healing

Compound preparation for wound healing and postpartum rehabilitation

Dead Sea Water and apple of Sodom extract compositions and uses thereof

Therapeutic composition for wound healing

Anti-adhesion material with antibacterial and healing properties

Plant hemicelluloses and lectins formulations with healing activity

*IPC: International Patent Classification.*

*Recent patents of plant biomaterials for wound healing.*

*Wound Healing*

homology, as can be seen in **Figure 4**.

and Asp125 OD1). Also, there are a number of hydrophobic interactions contributed by Tyr78, Gly121 and Tyr122. Although closely related to jacalin's structure, CGB was unable to bind mannose as judged by isothermal calorimetry and co-crystallization studies. It is believed that this change in CGB specificity is caused by subtle differences

in the environment near the sugar-binding site, including solvent molecules. Extracts of *Artocarpus integer* seeds are a source of CMB, previously termed chempedak lectin-M [84]. The lectin is ~20-fold more abundant than ArtinM in crude extracts of *A. heterophyllus* seeds. Structural studies reveal CMB to be a 64-kDa tetramer with some of the polypeptides being disulfide-linked to give dimers. The functional activity of CMB was assessed by studying interactions with different isotypes of human immunoglobulins: strong binding to IgE and IgM was noted, unlike CGB and jacalin which strongly interact with IgA1. The lectin also shows a similar pattern of carbohydrate binding specificity to ArtinM with Man-α-1–3Man the most potent inhibitor, followed by methyl-α-D-mannopyranose and D-mannose [84]. It appears that the *Artocarpus* genus employs a plurality of lectins, though to date few of these lectins have been purified and functionally characterized. Nevertheless, a consistent pattern is that *Artocarpus* seeds contain more than one lectin with distinct carbohydrate recognition features [70, 76]. Altogether, the JRL family is complex with a vast diversity in biochemical properties and activities, which have drawn much attention because of their pivotal biomedical applications. Owing to carbohydrate-binding interactions of plant lectins with cell wall glycoproteins, they are promising targets to selectively modulate immune responses in plants. Hence, it is important to delineate the molecular details of lectin binding to CBS domains and how downstream activation of cellular immune signaling proceeds [85]. In this context, it is of particular note that *Artocarpus* lectins have a distinct repertoire of biological activities, despite their high sequence and structural

*Sequence alignment using Clustal W and structural superposition of* Artocarpus *lectins. (A) Sequence alignment of galactose-binding lectins: Jacalin (PDB 1UGW), Frutalin (PDB 5BN6), and CGB (PDB 4AKB). (B) Sequence alignment of mannose-binding lectins: ArtinM (PDB 1J4S), frutapin (PDB 5KRP) and CMB (PDB 4AKD). (C) Structural superposition of galactose-binding lectins with their carbohydrate interacting residues in complex with D-galactose: jacalin (cyan), frutalin (red) and CGB (magenta). (D) Structural superposition of mannose-binding lectins with the carbohydrate interacting residues highlighted: ArtinM* 

**44**

**Figure 4.**

*(green), frutapin (yellow) and CMB (blue).*

## **4. Recent advances using plant biomaterials for wound healing**

The worldwide increasing number of chronic-wound patients has driven an intense effort in the wound-care market in search of low cost and effective wound healing technologies [86]. Therefore, much of this knowledge is eligible for patents that play an important role in identifying technology development trends [86].

In the following table, we highlight the main efforts in the last 5 years on the development of biomaterials using plant macromolecules as source of biomolecules with potential for wound healing applications (**Table 1**).


*IPC: International Patent Classification.*

### **Table 1.**

*Recent patents of plant biomaterials for wound healing.*

### **5. Conclusion**

As an orchestrated sequence of biochemical and cellular events for tissue repair, the healing process becomes a challenging in a way to develop a universal dressing, attending all the sorts of wound and lesions. It is here that biomaterials are notably relevant for biomedical applications once multiple properties of them can be combined in formulations of new treatments strategies of wound healing and repair. Many plants have been used traditionally in therapeutic treatments of lesions, which make them a remarkable source of biomolecules to wound-care market.

### **Author details**

Felipe Domingos de Sousa1 \*, Francisco Rogênio da Silva Mendes2 , Jose Jovanny Bermudez-Sierra2 , Ayrles Fernanda Brandão da Silva3 , Mirele da Silveira Vasconcelos4 , Tamiris de Fátima Goebel de Souza<sup>5</sup> , Marília de Oliveira Nunes5 , Antônio Eufrásio Vieira-Neto2 , Marcos Roberto Lourenzoni6 , Rosueti Diógenes de Oliveira-Filho7 , Adriana Rolim Campos2 , Renato de Azevedo Moreira<sup>2</sup> and Ana Cristina de Oliveira Monteiro-Moreira2

1 Department of Physics, Federal University of Ceará, Fortaleza, Ceará, Brazil

2 Northeast Biotechnology Network (RENORBIO), Centre of Experimental Biology (Nubex), University of Fortaleza (UNIFOR), Fortaleza, Ceará, Brazil

3 Department of Biochemistry and Molecular Biology, Federal University of Ceará (UFC), Fortaleza, Ceará, Brazil

4 Federal Institute of Education, Science and Technology of Ceará, Baturité, Ceará, Brazil

5 Department of Physiology and Pharmacology, Drug Research and Development Center (NPDM), Federal University of Ceara, Fortaleza, Ceará, Brazil

6 Fiocruz, Fundação Oswaldo Cruz—Ceará, Drugs and Biopharmaceuticals Development Group: Evolution, in silico and in vitro of Biomolecules, Fortaleza, Ceará, Brazil

7 Department of Clinical and Toxicological Analysis, Federal University of Ceará, Fortaleza, Ceará, Brazil

\*Address all correspondence to: fdsousa@yahoo.com.br

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**47**

*Plant Macromolecules as Biomaterials for Wound Healing*

[10] Martin P, Nunan R. Cellular and molecular mechanisms of repair in acute and chronic wound healing. The British Journal of Dermatology.

[11] Hesketh M, Sahin KB, West ZE, Murray RZ. Macrophage phenotypes regulate scar formation and chronic wound healing. International Journal of Molecular Sciences. 2017;**18**(7):1-10

[12] Darby IA, Laverdet B, Bonté F, Desmoulière A. Fibroblasts and myofibroblasts in wound healing. Clinical, Cosmetic and Investigational

[13] Georgescu M, Marinas O, Popa M, Stan T, Lazar V, Bertesteanu SV, et al. Natural compounds for wound healing. In: da Fonseca CJV, editor. Worldwide Wound Healing—Innovation in Natural and Conventional Methods. Portugal:

Dermatology. 2014;**7**:301-311

IntechOpen; 2016. pp. 61-89

[14] Pérez-Recalde M, Ruiz Arias IE, Hermida ÉB. Could essential oils enhance biopolymers performance for wound healing? A systematic review. Phytomedicine. 2018;**38**:57-65

[15] Bassolé IHN, Juliani HR. Essential

[16] Raut JS, Karuppayil SM. A status review on the medicinal properties of essential oils. Industrial Crops and

[17] Pesavento G, Calonico C, Bilia AR, Barnabei M, Calesini F, Addona R, et al. Antibacterial activity of oregano, Rosmarinus and thymus essential oils against *Staphylococcus aureus* and listeria monocytogenes in beef meatballs. Food Control.

oils in combination and their antimicrobial properties. Molecules.

2012;**17**(4):3989-4006

Products. 2014;**62**:250-264

2015;**54**:188-199

2015;**173**(2):370-378

*DOI: http://dx.doi.org/10.5772/intechopen.89105*

[1] Sorg H, Tilkorn DJ, Hager S, Hauser J, Mirastschijski U. Skin wound healing: An update on the current knowledge and concepts. European Surgical Research. 2017;**58**(1-2):81-94

[2] Kathju S, Gallo PH, Satish L. Scarless integumentary wound healing in the mammalian fetus: Molecular basis and therapeutic implications. Birth Defects Research. Part C, Embryo Today: Reviews. 2012;**96**(3):223-236

[3] Han G, Ceilley R. Chronic wound healing: A review of current management and treatments. Advances

in Therapy. 2017;**34**(3):599-610

[5] Nicholas MN, Jeschke MG, Amini-Nik S. Methodologies in creating skin substitutes. Cellular and Molecular Life Sciences. 2016;**73**(18):3453-3472

[6] Das S, Baker AB. Biomaterials and nanotherapeutics for enhancing skin wound healing. Frontiers in Bioengineering and Biotechnology.

[7] Gonzalez AC, Costa TF, Andrade ZD, Medrado AR. Wound healing—A literature review. Anais Brasileiros de Dermatologia. 2016;**91**(5):614-620

[8] Golebiewska EM, Poole AW. Platelet secretion: From haemostasis to wound healing and beyond. Blood Reviews.

2016;**4**(82):1-20

2015;**29**(3):153-162

[9] Zhao R, Liang H, Clarke E, Jackson C, Xue M. Inflammation in chronic wounds. International Journal of Molecular Sciences. 2016;**17**(12):1-14

[4] Chaudhari AA, Vig K, Baganizi DR, Sahu R, Dixit S, Dennis V, et al. Future prospects for scaffolding methods and biomaterials in skin tissue engineering: A review. The International Journal of Molecular Sciences. 2016;**17**(12):1-31

**References**

*Plant Macromolecules as Biomaterials for Wound Healing DOI: http://dx.doi.org/10.5772/intechopen.89105*

### **References**

*Wound Healing*

**5. Conclusion**

**Author details**

Felipe Domingos de Sousa1

Marília de Oliveira Nunes5

Adriana Rolim Campos2

Marcos Roberto Lourenzoni6

(UFC), Fortaleza, Ceará, Brazil

Fortaleza, Ceará, Brazil

Fortaleza, Ceará, Brazil

and Ana Cristina de Oliveira Monteiro-Moreira2

Jose Jovanny Bermudez-Sierra2

Mirele da Silveira Vasconcelos4

\*, Francisco Rogênio da Silva Mendes2

As an orchestrated sequence of biochemical and cellular events for tissue repair, the healing process becomes a challenging in a way to develop a universal dressing, attending all the sorts of wound and lesions. It is here that biomaterials are notably relevant for biomedical applications once multiple properties of them can be combined in formulations of new treatments strategies of wound healing and repair. Many plants have been used traditionally in therapeutic treatments of lesions, which make them a remarkable source of biomolecules to wound-care market.

, Antônio Eufrásio Vieira-Neto2

2 Northeast Biotechnology Network (RENORBIO), Centre of Experimental Biology

3 Department of Biochemistry and Molecular Biology, Federal University of Ceará

4 Federal Institute of Education, Science and Technology of Ceará, Baturité, Ceará,

5 Department of Physiology and Pharmacology, Drug Research and Development

7 Department of Clinical and Toxicological Analysis, Federal University of Ceará,

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

, Renato de Azevedo Moreira<sup>2</sup>

1 Department of Physics, Federal University of Ceará, Fortaleza, Ceará, Brazil

(Nubex), University of Fortaleza (UNIFOR), Fortaleza, Ceará, Brazil

Center (NPDM), Federal University of Ceara, Fortaleza, Ceará, Brazil

\*Address all correspondence to: fdsousa@yahoo.com.br

provided the original work is properly cited.

6 Fiocruz, Fundação Oswaldo Cruz—Ceará, Drugs and Biopharmaceuticals Development Group: Evolution, in silico and in vitro of Biomolecules,

, Ayrles Fernanda Brandão da Silva3

, Tamiris de Fátima Goebel de Souza<sup>5</sup>

, Rosueti Diógenes de Oliveira-Filho7

,

,

,

,

,

**46**

Brazil

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[2] Kathju S, Gallo PH, Satish L. Scarless integumentary wound healing in the mammalian fetus: Molecular basis and therapeutic implications. Birth Defects Research. Part C, Embryo Today: Reviews. 2012;**96**(3):223-236

[3] Han G, Ceilley R. Chronic wound healing: A review of current management and treatments. Advances in Therapy. 2017;**34**(3):599-610

[4] Chaudhari AA, Vig K, Baganizi DR, Sahu R, Dixit S, Dennis V, et al. Future prospects for scaffolding methods and biomaterials in skin tissue engineering: A review. The International Journal of Molecular Sciences. 2016;**17**(12):1-31

[5] Nicholas MN, Jeschke MG, Amini-Nik S. Methodologies in creating skin substitutes. Cellular and Molecular Life Sciences. 2016;**73**(18):3453-3472

[6] Das S, Baker AB. Biomaterials and nanotherapeutics for enhancing skin wound healing. Frontiers in Bioengineering and Biotechnology. 2016;**4**(82):1-20

[7] Gonzalez AC, Costa TF, Andrade ZD, Medrado AR. Wound healing—A literature review. Anais Brasileiros de Dermatologia. 2016;**91**(5):614-620

[8] Golebiewska EM, Poole AW. Platelet secretion: From haemostasis to wound healing and beyond. Blood Reviews. 2015;**29**(3):153-162

[9] Zhao R, Liang H, Clarke E, Jackson C, Xue M. Inflammation in chronic wounds. International Journal of Molecular Sciences. 2016;**17**(12):1-14 [10] Martin P, Nunan R. Cellular and molecular mechanisms of repair in acute and chronic wound healing. The British Journal of Dermatology. 2015;**173**(2):370-378

[11] Hesketh M, Sahin KB, West ZE, Murray RZ. Macrophage phenotypes regulate scar formation and chronic wound healing. International Journal of Molecular Sciences. 2017;**18**(7):1-10

[12] Darby IA, Laverdet B, Bonté F, Desmoulière A. Fibroblasts and myofibroblasts in wound healing. Clinical, Cosmetic and Investigational Dermatology. 2014;**7**:301-311

[13] Georgescu M, Marinas O, Popa M, Stan T, Lazar V, Bertesteanu SV, et al. Natural compounds for wound healing. In: da Fonseca CJV, editor. Worldwide Wound Healing—Innovation in Natural and Conventional Methods. Portugal: IntechOpen; 2016. pp. 61-89

[14] Pérez-Recalde M, Ruiz Arias IE, Hermida ÉB. Could essential oils enhance biopolymers performance for wound healing? A systematic review. Phytomedicine. 2018;**38**:57-65

[15] Bassolé IHN, Juliani HR. Essential oils in combination and their antimicrobial properties. Molecules. 2012;**17**(4):3989-4006

[16] Raut JS, Karuppayil SM. A status review on the medicinal properties of essential oils. Industrial Crops and Products. 2014;**62**:250-264

[17] Pesavento G, Calonico C, Bilia AR, Barnabei M, Calesini F, Addona R, et al. Antibacterial activity of oregano, Rosmarinus and thymus essential oils against *Staphylococcus aureus* and listeria monocytogenes in beef meatballs. Food Control. 2015;**54**:188-199

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bioactive film based on cashew gum polysaccharide for wound dressing applications. Carbohydrate Polymers. 2015;**122**:69-76

*Wound Healing*

Inc.; 2011. pp. 5-13

2013;**238**:627

[20] Pauly M, Gille S, Liu L,

[21] Scheller HV, Ulvskov P.

Biology. 2010;**61**(1):263-289

2010;**154**(3):1017-1023

[23] Bento JF, Mazzaro I, De

patterns of cell wall mannan/ galactomannan occurrence in seeds of the Leguminosae. Carbohydrate Polymers. 2013;**92**(1):192-199

Almeida Silva LM, De Azevedo MR, Ferreira MLC, Reicher F, et al. Diverse

[24] Prajapati VD, Jani GK, Moradiya NG, Randeria NP, Nagar BJ, Naikwadi NN, et al. Galactomannan: A versatile biodegradable seed polysaccharide. The International Journal of Biological Macromolecules. 2013;**60**:83-92

[25] Srivastava M, Kapoor VP. Seed galactomannans: An overview. Chemistry and Biodiversity.

[26] Cosgrove DJ. Expansive growth of plant cell walls. Plant Physiology and Biochemistry. 2000;**38**(1-2):109-124

[27] Picone P, Antonietta M, Ajovalasit A, Giacomazza D, Dispenza C, Di M.

[22] Buckeridge MS. Seed cell wall storage polysaccharides: Models to understand cell wall biosynthesis and degradation. Plant Physiology.

[18] Guimarães AG, Quintans JSS, Quintans-Júnior LJ. Monoterpenes with analgesic activity—A systematic review. Phytotherapy Research. 2013;**27**(1):1-15

Biocompatibility , hemocompatibility and antimicrobial properties of xyloglucan-based hydrogel fi lm for wound healing application. International Journal of Biological Macromolecules. 2019;**121**:784-795

[28] Rana V, Rai P, Tiwary AK, Singh RS, Kennedy JF, Knill CJ. Modified gums: Approaches and applications in drug delivery. Carbohydrate Polymers.

[29] Shen T, Yuan H-Q, Wan W-Z, Wang X-L. Cycloartane-type triterpenoids from the resinous exudates of

*Commiphora opobalsamum*. Journal of Natural Products. 2008;**71**(1):81-86

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[68] Pratap JV, Jeyaprakash AA, Rani PG, Sekar K, Surolia A, Vijayan M. Crystal structures of artocarpin, a Moraceae lectin with mannose specificity, and its complex with methyl-alpha-D-mannose:

Implications to the generation of carbohydrate specificity. The Journal of Molecular Biology. 2002;**317**(2):237-247

[69] Jeyaprakash AA, Srivastav A, Surolia A, Vijayan M. Structural basis for the carbohydrate specificities of artocarpin: Variation in the length of a loop as a strategy for generating ligand specificity. Journal of Molecular Biology.

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[71] Zerega N, Wiesner-Hanks T, Ragone D, Irish B, Scheffler B, Simpson S, et al. Diversity in the breadfruit complex (Artocarpus, Moraceae): Genetic characterization of critical germplasm. Tree Genetics and

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lectin artocarpin. Biochimie. 1991;**73**(5):563-571

*Wound Healing*

2015;**117**(1):51-64

1999;**7**(12):1473-1482

2005;**30**(3):603-615

Biology. 1997;**7**(5):631-636

[50] Dang L, Van Damme EJM. Toxic proteins in plants. Phytochemistry.

oil. Industrial Crops and Products.

Haldankar PM, Kalse SB. Jackfruit and its many functional components as related to human health: A review. Comprehensive Reviews in Food Science and Food Safety. 2012;**11**(6):565-576

[60] Chatterjee B, Vaith P, Chatterjee S, Karduck D, Uhlenbruck G. Comparative studies of new marker lectins for alkalilabile and alkali-stable carbohydrate

International Journal of Biochemistry.

[61] Moreira RA, Ainouz IL. Lectins from seeds of jack fuit (*Artocarpus integrifolia L*.): Isolation and

purification of two isolectins from the albumin fraction. Biologia Plantarum.

[62] Sankaranarayanan R, Sekar K, Banerjee R, Sharma V, Surolia A, Vijayan M. A novel mode of

carbohydrate recognition in jacalin, a Moraceae plant lectin with a β-prism fold. Nature Structural Biology.

[63] Abhinav KV, Sharma K, Surolia A, Vijayan M. Distortion of the ligand molecule as a strategy for modulating binding affinity: Further studies involving complexes of jacalin with β-substituted disaccharides. IUBMB

[59] Swami SB, Thakor NJ,

chains in glycoproteins. The

1979;**10**(4):321-327

1981;**23**(3):186-192

1996;**3**(7):596-603

Life. 2017;**69**(2):72-78

1991;**140**(2):197-203

[65] Chowdhury S, Ahmed H,

studies of *Artocarpus lakoocha*

Chatterjee BP. Chemical modification

[64] de Miranda-Santos IK, Mengel JO Jr, Bunn-Moreno MM, Campos-Neto A. Activation of T and B cells by a crude extract of *Artocarpus integrifolia* is mediated by a lectin distinct from jacalin. Journal of Immunological Methods.

2018;**112**:47-52

[51] Bourne Y, Zamboni V, Barre A, Peumans WJ, Van Damme EJM, Rougé P. Helianthus tuberosus lectin reveals a widespread scaffold for mannose- binding lectins. Structure.

[52] Wright CS. New folds of plant lectins. Current Opinion in Structural

[53] Zerega NJC, Ragone D, Motley TJ. Systematics and species limits of breadfruit. Systematic Botany.

[54] de Lopes MMA, de Souza KO, de Silva EO. Cempedak—*Artocarpus champeden*. In: Rodrigues S, de Silva EO, de Brito ES, editors. Exotic Fruits Reference Guide. London: Wolff, Andre

[55] Ragone D. Breadfruit—*Artocarpus* 

[56] Jagtap UB, Bapat VA. Artocarpus: A review of its traditional uses, phytochemistry and pharmacology. Journal of Ethnopharmacology.

Hatamleh AA, Munusamy MA. From

Jackfruit peel derived pectin/apatite bionanocomposites for bone healing applications. The International Journal of Biological Macromolecules.

Chu Z. Effect of a new shell material— Jackfruit seed starch on novel flavor microcapsules containing vanilla

Gerhard; 2018. pp. 121-126

pp. 53-59

2010;**129**:142-146

2018;**106**:293-301

[57] Govindaraj D, Rajan M,

waste to high-value product:

[58] Zhu H, Zhang Y, Tian J,

*altilis* (Parkinson) Fosberg. In: Rodrigues S, de Silva EO, de Brito ES, editors. Exotic Fruits Reference Guide. London: Wolff, Andre Gerhard; 2018.

**50**

[66] Pereira-da-Silva G, Roque-Barreira MC, Van Damme EJM. Artin M: A rational substitution for the names artocarpin and KM+. Immunology Letters. 2008;**119**(1-2):114-115

[67] DaSilva LLP, de Molfetta-Machado JB, Panunto-Castelo A, Denecke J, Goldman GH, Roque-Barreira MC, et al. cDNA cloning and functional expression of KM+, the mannosebinding lectin from *Artocarpus integrifolia* seeds. Biochimica et Biophysica Acta, General Subjects. 2005;**1726**(3):251-260

[68] Pratap JV, Jeyaprakash AA, Rani PG, Sekar K, Surolia A, Vijayan M. Crystal structures of artocarpin, a Moraceae lectin with mannose specificity, and its complex with methyl-alpha-D-mannose: Implications to the generation of carbohydrate specificity. The Journal of Molecular Biology. 2002;**317**(2):237-247

[69] Jeyaprakash AA, Srivastav A, Surolia A, Vijayan M. Structural basis for the carbohydrate specificities of artocarpin: Variation in the length of a loop as a strategy for generating ligand specificity. Journal of Molecular Biology. 2004;**338**(4):757-770

[70] Trindade MB, Lopes JLS, Soares-Costa A, Monteiro-Moreira AC, Moreira RA, Oliva MLV, et al. Structural characterization of novel chitin-binding lectins from the genus Artocarpus and their antifungal activity. Biochimica et Biophysica Acta, Proteins and Proteomics. 2006;**1764**(1):146-152

[71] Zerega N, Wiesner-Hanks T, Ragone D, Irish B, Scheffler B, Simpson S, et al. Diversity in the breadfruit complex (Artocarpus, Moraceae): Genetic characterization of critical germplasm. Tree Genetics and Genomes. 2015;**11**(1):1-26

[72] de Moreira RA, de Oliveira JTA. Lectins from the genus Artocarpus. Biologia Plantarum. 1983;**25**(5):343-348

[73] de Moreira RA, Castelo-Branco CC, de Monteiro ACO, Tavares RO, Beltramini LM. Isolation and partial characterization of a lectin from *Artocarpus incisa L*. seeds. Phytochemistry. 1998;**47**(7):1183-1188

[74] Vieira-neto AE. Caracterização estrutural da frutalina, uma lectina α-D-galactose ligante de sementes de artocarpus incisa e análise das suas bases moleculares de ligação à D-galactose [PhD Thesis]. Universidade Federal do Ceará; 2015

[75] De-Simone SG, Netto CC, Silva FP. Simple affinity chromatographic procedure to purify β-galactoside binding lectins. Journal of Chromatography, B: Analytical Technologies in the Biomedical and Life Sciences. 2006;**838**(2):135-138

[76] Monteiro-Moreira ACO. Caracterização estrutural de três lectinas apresentando especificidades por açúcar distintas, isoladas de sementes de fruta-

pão (*Artocarpus incisa* L.) [PhD thesis]. Federal University of Ceará—UFC; 2002

[77] Oliveira C, Teixeira JA, Domingues L. Recombinant production of plant lectins in microbial systems for biomedical application—The frutalin case study. Frontiers in Plant Science. 2014;**5**:390

[78] de Sousa FD, da Silva BB, Furtado GP, de Carneiro IS, MDP L, Guan Y, et al. Frutapin, a lectin from *Artocarpus incisa* (breadfruit): Cloning, expression and molecular insights. Bioscience Reports. 2017;**37**(4):BSR20170969

[79] Syah YM, Juliawaty LD, Achmad SA, Hakim EH, Ghisalberti EL. Cytotoxic prenylated flavones from Artocarpus

champeden. Journal of Natural Medicines. 2006;**60**(4):308-312

[80] Farooq U, Malviya R, Sharma PK. Extraction and characterization of Artocarpus integer gum as pharmaceutical excipient. Polimery w Medycynie. 2014;**44**(2):69-74

[81] Hashim O, Gende G, Jaafar M. Lectin extracts of champedak seeds demonstrate selective stimulation of T lymphocyte proliferation. Biochemistry International. 1992;**27**(1):139-143

[82] Hashim OH, Ng CL, Gendeh GS, Jaafar MIN. Iga binding lectins isolated from distinct artocarpus species demonstrate differential specificity. Molecular Immunology. 1991;**28**(4-5):393-398

[83] Gabrielsen M, Abdul-Rahman PS, Othman S, Hashim OH, Cogdell RJ. Structures and binding specificity of galactose- and mannose-binding lectins from chempedak: Differences from jackfruit lectins. Acta Crystallographica Section F: Structural Biology Communications. 2014;**70**(6):709-716

[84] Lim SB, Chua CT, Hashim OH. Isolation of a mannose-binding and IgE- and IgM-reactive lectin from the seeds of Artocarpus integer. Journal of Immunological Methods. 1997;**209**(2):177-186

[85] Van Holle S, Van Damme EJM. Signaling through plant lectins: Modulation of plant immunity and beyond. Biochemical Society Transactions. 2018;**36**(2):221-247

[86] Gwak JH, Sohn SY. Identifying the trends in wound-healing patents for successful investment strategies. PLoS One. 2017;**12**(3):1-19

[87] Askurai E, Geblin D, Clayman M FD. Necrotic tissue composition remover for wound healing. Korea; 2018. KR20180104138A

[88] Maojun Y, Wang X, Dayuan Z GS. Compound preparation capable of promoting wound healing and postpartum rehabilitation. China; 2016. CN105770849A

[89] Cohen MP, Maor Z, Ish-Shalom E CD. Dead Sea Water and apple of Sodom extract compositions and uses thereof. 2017. WO2019130301A1

[90] Phipps W. Therapeutic composition for wound healing. United States; 2016. US20160256480A1

[91] Fanglian Y, Ershuai Z, Hong S. Antiadhesion material with antibacterial and healing properties. China; 2016. CN106362221A

[92] de Sousa FD, de Moreira RA, Monteiro-Moreira ACO, Campos AR, Vasconselos PD. Plant hemicelluloses and lectins formulations with healing activity. Brazil; 2017. BR102017006983A2

**53**

**Chapter 4**

Ulcers

**Abstract**

**1. Introduction**

Modulation of Inflammatory

Wound Recovery of Diabetic

*Pawandeep Kaur and Diptiman Choudhury*

Dynamics by Insulin to Promote

About 5% of the world population is diabetic and are at a risk of slow nonrecoverable wound formation. Estimated 15–25% of diabetic patients develop foot ulcers, 6% among them needing clinical attention among which 15–20% will need an amputation. This counts for around 50% of all traumatic amputation. Wound leads to activation of dynamic inflammatory cascade responsible for the healing process. But in diabetes, a persistent rise of pro-inflammatory cytokines and low anti-inflammatory cytokines blocks the dynamic cascade. Wounding induces various pro-inflammatory cytokines such as IL-1, IL-6, IL-12, IL-18, IFN-γ, and TNFs causing accumulation of free radicals leading to inflammation which become persistent in diabetes. Inhibition of proinflammatory cytokines drives the equilibrium towards the expression of anti-inflammatory cytokines such as IL4, IL-10, IL-11, IL-13, IFN-α, and TGF-β, which is necessary for the wound recovery process. Here in this chapter, the inflammatory modulatory roles of different drugs/formulations have been discussed to unravel their significance to promote wound recovery.

**Keywords:** diabetic wound, tissue inflammation, pro-inflammatory cytokines,

Over the last 25 years, there has been found a four-fold increase in the number of diabetes mellitus cases commonly called diabetes [1]. 422 million people worldwide in 2016 have been reported to have diabetes mellitus. Diabetes in the year 2012 was a cause of 1.5 million deaths worldwide; according to WHO (World Health Organization), diabetes becomes the 8th leading cause of death [2]. Diabetes mellitus is mainly identified by the presence free glucose at high or the chronic level in the body fluids like sweat, urine, blood, etc. [3]. The major reason for diabetes mellitus among others was the hormone-mediated metabolism regulation failure. Hormones like glucagon and insulin play an important role in regulating the level of blood sugar or maintaining its balance [4]. The sugar balance in blood is important for perfect functioning of human body [5]. High/chronic level of glucose in body fluids is responsible for different pathological conditions like infection susceptibility, leading to various diseases such as arthritis, hypertension, cardiovascular problems, cataract,

anti-inflammatory cytokines, nanoformulation for wound recovery

### **Chapter 4**

*Wound Healing*

champeden. Journal of Natural Medicines. 2006;**60**(4):308-312

Artocarpus integer gum as

Medycynie. 2014;**44**(2):69-74

[80] Farooq U, Malviya R, Sharma PK. Extraction and characterization of

[88] Maojun Y, Wang X, Dayuan Z GS. Compound preparation capable of promoting wound healing and postpartum rehabilitation. China; 2016.

[89] Cohen MP, Maor Z, Ish-Shalom E CD. Dead Sea Water and apple of Sodom extract compositions and uses thereof.

[90] Phipps W. Therapeutic composition for wound healing. United States; 2016.

[91] Fanglian Y, Ershuai Z, Hong S. Antiadhesion material with antibacterial and healing properties. China; 2016.

[92] de Sousa FD, de Moreira RA, Monteiro-Moreira ACO, Campos AR, Vasconselos PD. Plant hemicelluloses

and lectins formulations with healing activity. Brazil; 2017.

CN105770849A

2017. WO2019130301A1

US20160256480A1

CN106362221A

BR102017006983A2

pharmaceutical excipient. Polimery w

[81] Hashim O, Gende G, Jaafar M. Lectin extracts of champedak seeds demonstrate selective stimulation of T lymphocyte proliferation. Biochemistry International. 1992;**27**(1):139-143

[82] Hashim OH, Ng CL, Gendeh GS, Jaafar MIN. Iga binding lectins isolated from distinct artocarpus species

specificity. Molecular Immunology.

[83] Gabrielsen M, Abdul-Rahman PS, Othman S, Hashim OH, Cogdell RJ. Structures and binding specificity of galactose- and mannose-binding lectins from chempedak: Differences from jackfruit lectins. Acta Crystallographica

Communications. 2014;**70**(6):709-716

[84] Lim SB, Chua CT, Hashim OH. Isolation of a mannose-binding and IgE- and IgM-reactive lectin from the seeds of Artocarpus integer. Journal of Immunological Methods.

[85] Van Holle S, Van Damme EJM. Signaling through plant lectins: Modulation of plant immunity and beyond. Biochemical Society Transactions. 2018;**36**(2):221-247

[86] Gwak JH, Sohn SY. Identifying the trends in wound-healing patents for successful investment strategies. PLoS

[87] Askurai E, Geblin D, Clayman M FD. Necrotic tissue composition remover for wound healing. Korea;

demonstrate differential

Section F: Structural Biology

1997;**209**(2):177-186

One. 2017;**12**(3):1-19

2018. KR20180104138A

1991;**28**(4-5):393-398

**52**
