**10.** *In-vitro* **and** *in-vivo* **study of PBNDDS of phytoconstituents**

Sustain release of phytoconstituents was observed when delivered through PBNDDS, which can be utilized for better therapeutic outcome against certain diseases. PBNDDS of phytoconstituents demonstrate better performance during *in-vitro* cancer cell line study performed on different cell lines. *In-vitro* cell line study data suggest that PBNDDS of phytoconstituents showed better activity in terms of % cell viability against AML and leukemia cell line compared to pure

**Figure 8.**

*In-vitro cell line study of plant extracts and PBNDDS of different plant extracts after [A] 48 h and [B] 72 h.*

phytoconstituents only. It was also observed that PBNDDS of phytoconstituents showed better activity against specific AML and leukemia cell lines compared to all cell lines used in the study. This is possibly due to more permeability of PBNDDS occurred through those specific cell lines, which suggest that PBNDDS may be selective for specific cancer cell lines which may be related to the phospholipid composition, type and drug delivered through PBNDDS [82]. PBNDDS of different phytoconstituents also exhibited better activity against breast cancer cell line (MCF7) study compared to phytoconstituents in isolation (**Figure 8**).

Significant improvement was observed for *in-vivo* analgesic activity for the PBNDDS of black seed oil containing thymoquinone compared to black seed oil only and control groups. This phenomenon probably related to the improve bioavailability of thymoquinone when delivered through PBNDDS. Previous research study also showed analgesic and anti-inflammatory activity for black seed oil containing thymoquinone [80, 96].

Strong anti-inflammatory activity was observed for different plant extracts and its phytoconstituents against carrageenan induced paw edema (**Table 4**). It was also observed that plant extract and phytoconstituents delivered through PBNDDS exhibit better anti-inflammatory activity, which is possibly due to enhancement of dissolution, bioavailability and stability of phytoconstituents when delivered through PBNDDS [81–83].

Neurobehavioral study of PBNDDS of phytoconstituents was performed using open field, arm maze and water maze studies (**Figure 9**). Marked improvement in locomotor activity, ambulatory performance and memory function of dementia induced mice model was observed for PBNDDS of phytoconstituents compare to disease and plant extract groups [81]. This phenomenon probably related to strong anti-inflammatory along with antioxidant activities observed for the plant extract in different research studies. CNS inflammation is one of the pathway for developing neurodegenerative disorders, therefore by reducing inflammation significantly through PBNDDS of plant phytoconstituents in dementia induced mice model might be an option to treat neurodegenerative disease. Natural phytoconstituents may contain some ingredients which also can be effective against neurodegenerative disease through another mechanism of action which need to be confirmed in future study.


#### *Phospholipid Based Nano Drug Delivery Systems of Phytoconstituents DOI: http://dx.doi.org/10.5772/intechopen.101040*

**Table 4.**

*Anti-inflammatory studies of plant extracts, and its PBNDDS (adapted from [81–83]).*

#### **Figure 9.**

*[A] Number of total errors for different mouse groups in arm maze study [B] maximum speed into central zone for different mouse groups in open field study [C] time spent on platform for different mice groups in water maze study (where \*\*\*\* means p* ≤ *0.0001, \*\*means p* ≤ *0.01 and \* mean p* ≤ *0.05) [four different groups—1. Pre disease group 2. Post disease group 3. E 200 - extract group and 4. Formulation—PBNDDS of extracts] (adapted from [81]).*

#### **11. Conclusion**

Plant extract found to have a range of major phytoconstituents which were identified and quantified by UHPLC and GC-MS. Major phytoconstituents emonstrate marked pharmacological activities which were evident by different *in-vitro* and *in-vivo* studies. Phytoconstituents delivered through PBNDDS exhibit better performance compared to phytoconstituents in isolation. It was observed that process parameters and formulation attributes showed significant impact on average size, polydispersity and entrapment or loading of phytoconstituents for PBNDDS. Processing temperature, injection rate, solvent system properties (pH, solubility level), phospholipd concentration related to drug and cholesterol are major factors affecting the quality output of PBNDDS. PBNDDS prepared using egg phospholipid was physico-chemically stable even at ambient conditions (25°C, 60% RH). This phenomenon might be a great advantage for developing PBNDDS of different phytoconstituents for improving the bioavialabilty, stability and targeted drug delivery. PBNDDS also exhibit better selective activity against cancer cell lines which is an indication for treating different types of cancer by developing PBNDDS using different formulation attributes. PBNDDS also showed better analgesic, anti-inflammatory and neurobehavioral activities compared to phytoconstituents only. Therefore PBNDDS can be a promising platform for delivering phytoconstituents with better therapeutic outcome. PBNDDS having average size of <150 nm with ≥85% entrapment or loading might exhibit desirable performance to treat chronic inflammatory disease, cancer, and CNS diseases.

*Phospholipid Based Nano Drug Delivery Systems of Phytoconstituents DOI: http://dx.doi.org/10.5772/intechopen.101040*
