*Immunological Monitoring of Osteogenesis Disorder DOI: http://dx.doi.org/10.5772/intechopen.92099*

**Laboratory**

**162**

Total number of white blood cells, 109/l

Total number of neutrophils,

Ratio stab Total number of monocytes,

Total number of CD45+CD3+-cells,

CD45+CD19+-cells,

Leukocyte migration inhibition test with

Nitro blue tetrazolium Nitro blue tetrazolium

Nitro blue tetrazolium

Myeloperoxidase

Activity cationic proteins neutrophilic

Lactoferrin,

Ig

IgM, g/l

IgG, g/l Lysozyme, mkg/ml

Total hemolytic

IL-1α, pg/ml

IL-1ra, pg/ml

TNF-α, pg/ml

complement

 activity, arbitrary units/ml

А, g/l

 ng/ml

 activity of neutrophils,

 average coefficient

granulocytes,

 average coefficient

cytochemical

 1.6–1.9 700–1500

1.38–2.50

0.92–2.10

8.5–15.8

28.6–31.0

40.0–42.0

0–50

0–500

0–50

 11.17

6.23

 25.03

6.17

 141.25

33.75\*

39.50

2.98\*

24.37

7.02

 35.24

0.22

 40.08

1.45\*

609.65

12.54\*

81.07

2.59\*

39.66

3.08

 22.06

2.04

 21.03

1.54

 625.18

12.36\*

20.14

1.98

 19.25

1.15

 50.28

2.06

 36.35

5.00\*

45.30

2.28

 48.06

2.26

 36.71

13.38

 16.23

1.17

 20.80

1.56\*

12.25

2.37

 8.02

1.88\*

14.90

2.04

 12.35

2.05

 11.73

0.58

 12.56

0.77

 12.30

0.57

 11.97

2.56

 2.27

0.03

 1.41

0.23\*

1.62

0.24\*

1.60

0.22\*

2.10

0.63

 2.89

0.23

 2.69

0.31

 2.89

0.45

 3.18

0.18

 2.03

0.29

 825.10

12.60

 501.09

12.65\*

1250.90

18.34\* 836.21

25.16 500.03

12.89\*

 1.35

0.12

 1.22

0.11

 1.27

0.02

 1.36

0.12

 1.22

0.05

cytochemical

 test

 test stimulated,

 test, stimulation

 index

 %

spontaneous,

 %

phytohemagglutinin,

 %

 %

 %

lymphocytes,

 109/l

neutrophil/segmented

 neutrophils

 109/l

 109/l

 **parameters**

**Normal values Before surgery**

4.0–9.0

2.00–5.80

0.02–0.06

0.09–0.60

1.2–3.0

46.0–56.0

4.0–8.0

0–30

10–20

50–100

>1.3

1.9–2.8

 1.91

0.25

 0.70

0.12\*

2.28

0.33

 2.10

0.08

 0.69

0.09\*

 1.00

0.20

 1.83

0.27\*

1.52

0.26

 1.73

0.14\*

2.81

0.56\*

 31.00

4.00

 35.56

5.59

 37.13

5.94

 52.20

4.46\*

49.67

4.17\*

 34.05

9.10

 23.67

4.77

 31.50

6.36

 32.40

9.05

 24.00

9.64

 8.03

0.24

 23.02

1.25\*

13.50

1.08\*

5.06

1.03

 22.09

1.11\*

 9.50

1.50

 9.67

1.17

 9.71

0.64

 9.20

1.98

 7.33

0.33

*Clinical Implementation of Bone Regeneration and Maintenance*

 60.00

2.00

 63.33

1.67

 51.29

6.36

 62.40

7.17

 62.33

0.88

 1.89

0.09

 1.56

0.27

 2.08

0.22

 2.15

0.09\*

2.10

0.09

 0.54

0.05

 0.45

0.05

 0.69

0.05\*

0.50

0.05

 0.52

0.05

 0.03

0.01

 0.02

0.01

 0.02

0.01

 0.03

0.01

 0.03

0.01

 4.34

0.26

 4.40

0.27

 4.62

0.43

 4.70

0.30

 4.88

0.35

 6.85

0.33

 6.68

0.45

 7.30

0.51

 7.35

0.55

 7.30

0.48

 **3 days**

 **10 days**

 **1 month**

 **3 months** tetrazolium test) and oxygen-independent (neutrophil activity cationic proteins) killing mechanisms, lowering the concentration of serum lysozyme.

**3.2 Features of peripheral blood response in slow consolidation of the mandible**

differences in the recorded response of leukocyte migration inhibition (**Table 2**). Serum lysozyme was reduced by 5.8 times. Interestingly, the concentration of the receptor antagonist of IL-1ra concentration was lower for IL-1α (**Table 2**) that was not observed in patients with normal bone consolidation. IL-8 levels were reduced by 1.8-fold (p < 0.05) (**Table 2**) in comparison with patients with the other group. The postoperative period was characterized by leukocytogenesis activation: leukocytosis was marked with the change in the ratio of group cells to the segmented cells. On day 3, a reduction in the number of T cells was shown (according to the

data [60]. Recovery of these cell populations occurred in 10 days (**Table 2**). Perhaps this change in the amount of T cells in the early postoperative period resulting, ultimately, in disruption of bone regeneration, because it is known that both by generating INF-γ, and through prostaglandin mechanism of these cells involved in suppressing bone destruction and the formation of osteoclasts. Decrease in the

Important changes were noted by the indicators characterizing the phagocytic activity of neutrophils. Amplification reactions were detected at 3 days after surgery when the spontaneous and stimulated superoxide radical production by neutrophils'superior control values was in the subgroup of 1.6- to 1.7-fold (p < 0.05). It can be assumed that the changes in neutrophil phagocyte system in the blood reflect the processes occurring in bone tissue. Of interest for the study was the dynamics of the content of lactoferrin at the stages of bone tissue consolidation (**Table 2**). As noted earlier, prior to the operation, its level was significantly reduced. Due to the fact that lactoferrin causes proliferation of osteoblasts and bone growth, it is not excluded that lowering the level of lactoferrin in the blood is also one of the reasons for slow regeneration of bone tissue. Later, after the operation, the lactoferrin content was always significantly lower in patients with delayed bone formation compared with patients who have bone fusion that took place at the usual time. Slow bone formation was accompanied by significant increase (72.0%) in IgM levels (**Table 2**). The revealed changes were statistically significant differences between these patients with normal bone tissue regeneration. Inflammatory response was characterized by pronounced dynamics of slow-reacting acute-phase proteins. Specifically, on day 3, after surgery, ceruloplasmin concentration was higher by 1.9-fold (p < 0.05) than in patients with normal bone regeneration. The functional activity of T cells throughout the observation period was 2.0- to 3.3-fold higher (p < 0.05) than in patients with normal consolidation, while it exceeds the normal value of 1.5 times (**Table 1**). Active cationic proteins and myeloperoxidase neutrophilic granulocytes were higher than in patients with complication of osteogenesis, respectively, by 33.6% (p < 0.05) and 3.1-fold (p < 0.05).

functional activity of CD45<sup>+</sup>

dynamics of CD45<sup>+</sup>

**165**

CD3<sup>+</sup>

*Immunological Monitoring of Osteogenesis Disorder DOI: http://dx.doi.org/10.5772/intechopen.92099*

relative number of B cells (based on dynamics of CD45<sup>+</sup>

(p < 0.05) was observed later, on day 10 (**Table 2**).

CD3<sup>+</sup>

In this group of patients, before surgery, differences in immunological parameters from data obtained on the bone consolidation without complicating osteomyelitis were revealed. The number of monocytes (**Table 2**) was lower, than in the group with the slow regeneration of bone tissue by 59.3% (p < 0.05). There was a significant attenuation of the bactericidal activity of mechanisms: the level of lactoferrin was reduced 3.5 times (p < 0.05) was only 33.6% compared with the normal values. The same can be mentioned for complement activity-30.2% of the lower limit of normal and below 4.2 times (p < 0.05) than in the group with normal bone regeneration. Increased concentration of 37.4% IgM (p < 0.05) in peripheral blood also exceeded 49.0% and the normal values (**Table 2**). There was increased



CD19<sup>+</sup>


Major changes were detected due to the inflammatory reaction induced by damage to the bone tissue in the mandible fracture, which was evaluated on the basis of a complex of acute-phase proteins. Prior to surgery, in particular, it was revealed that there was an increase in the most sensitive C-reactive protein with increasing concentration of ceruloplasmin and complement activity, indicating the presence of acute inflammation (**Table 1**).

On day 10 after surgery, the number of monocytes increased by 27.8% (p < 0.05) compared to preoperative level, and at 1 month after surgery, the number of lymphocytes increased by 13.8% (p < 0.05); the composition of population lymphocytes did not change (**Table 1**). It is known that a recognized mitogen for assessing the activity of T cells is phytohemagglutinin. According to the stress tests with phytohemagglutinin, a gain in the functional activity of the cells during the early postoperative period was identified, that is, for 3–10 days after surgery (**Table 1**). In the same period, there was a decrease in the concentration of IgM by 37.9% (p <0.05) with the subsequent restoration of its level in the peripheral blood (**Table 1**). Postoperatively, increased activity of "early" killing mechanisms was also observed: an increase in lactoferrin levels by 51.6% (p < 0.05) and the rise of total production of reactive oxygen species (nitro blue tetrazolium test) on 83.0% (p < 0.05). From the humoral immunity, a significant decrease in complement activity was revealed to be 27.7% (p < 0.05), followed by rapid recovery at 10 days. These changes appear to have been associated with the participation of complement in the elimination of foreign substance in the development of inflammatory response.

Due to the fact that not only cellular and humoral reactions played an important role in the regulation of immunological reactions, but also the effect of a number of low molecular weight peptides (cytokines) was noted, the dynamics of some of them was studied. In particular, it was found that in the postoperative period there was an increase in the concentration of IL-1α (p <0.05), an activator of the initial stages of the immune response (**Table 1**). This is likely to determine the development and course of a large number of immunological reactions in the reduction of bone tissue. Simultaneously, the increasing concentrations of receptor antagonist IL-1–IL-1ra were revealed, which limited the development of systemic inflammatory response (**Table 1**).

Concentrations of IL-1α and IL-1ra were increased to respectively 28.4 times and 17.3 times to the 10th day of observation compared to preoperative values. A similar trend was also characteristic of TNF-α (p < 0.05), similar to having IL-1 phlogogenic properties. On day 10, there was a significant increase in the level of another factor inflammation-IL-8-whose concentration in the blood was increased 9.7 times as compared to preoperative values (**Table 1**). However, during the activation process, the decreased concentrations of IL-10 were revealed, which is a factor that inhibits the synthesis of most of the cytokines. Its concentration in the serum was reduced more than 100 times (p < 0.05). The inflammatory response in normal bone regeneration had little activity and duration.

It is known that the regeneration of bone tissue composed of several sequentially successive stages: inflammation, proliferation of osteoblasts, collagenogenesis and ossification. In this case, additional studies were conducted 3 months after the operation, at the last stage when the external fixing device was dismantled, reparative regeneration is completed, the bone tissue has formed and the mandible functioned normally. By the time of full functional bone formation, immunological parameters were within normal limits.

In a number of patients with osteomyelitis (17 people) based on clinical and radiographic data, delayed regeneration of bone tissue of the lower jaw was revealed. It occurs 1.5 times later than normal consolidation (p < 0.05).

tetrazolium test) and oxygen-independent (neutrophil activity cationic proteins)

Major changes were detected due to the inflammatory reaction induced by damage to the bone tissue in the mandible fracture, which was evaluated on the basis of a complex of acute-phase proteins. Prior to surgery, in particular, it was revealed that there was an increase in the most sensitive C-reactive protein with increasing concentration of ceruloplasmin and complement activity, indicating the

On day 10 after surgery, the number of monocytes increased by 27.8% (p < 0.05)

compared to preoperative level, and at 1 month after surgery, the number of lymphocytes increased by 13.8% (p < 0.05); the composition of population lymphocytes did not change (**Table 1**). It is known that a recognized mitogen for assessing the activity of T cells is phytohemagglutinin. According to the stress tests with phytohemagglutinin, a gain in the functional activity of the cells during the early postoperative period was identified, that is, for 3–10 days after surgery (**Table 1**). In the same period, there was a decrease in the concentration of IgM by 37.9% (p <0.05) with the subsequent restoration of its level in the peripheral blood (**Table 1**). Postoperatively, increased activity of "early" killing mechanisms was also observed: an increase in lactoferrin levels by 51.6% (p < 0.05) and the rise of total production of reactive oxygen species (nitro blue tetrazolium test) on 83.0% (p < 0.05).

From the humoral immunity, a significant decrease in complement activity was revealed to be 27.7% (p < 0.05), followed by rapid recovery at 10 days. These changes appear to have been associated with the participation of complement in the elimination

Due to the fact that not only cellular and humoral reactions played an important role in the regulation of immunological reactions, but also the effect of a number of low molecular weight peptides (cytokines) was noted, the dynamics of some of them was studied. In particular, it was found that in the postoperative period there was an increase in the concentration of IL-1α (p <0.05), an activator of the initial stages of the immune response (**Table 1**). This is likely to determine the development and course of a large number of immunological reactions in the reduction of bone tissue. Simultaneously, the increasing concentrations of receptor antagonist IL-1–IL-1ra were

revealed, which limited the development of systemic inflammatory response (**Table 1**). Concentrations of IL-1α and IL-1ra were increased to respectively 28.4 times and 17.3 times to the 10th day of observation compared to preoperative values. A similar

phlogogenic properties. On day 10, there was a significant increase in the level of another factor inflammation-IL-8-whose concentration in the blood was increased 9.7 times as compared to preoperative values (**Table 1**). However, during the activation process, the decreased concentrations of IL-10 were revealed, which is a factor that inhibits the synthesis of most of the cytokines. Its concentration in the serum was reduced more than 100 times (p < 0.05). The inflammatory response in

It is known that the regeneration of bone tissue composed of several sequentially successive stages: inflammation, proliferation of osteoblasts, collagenogenesis and ossification. In this case, additional studies were conducted 3 months after the operation, at the last stage when the external fixing device was dismantled, reparative regeneration is completed, the bone tissue has formed and the mandible functioned normally. By the time of full functional bone formation, immunological

In a number of patients with osteomyelitis (17 people) based on clinical and radiographic data, delayed regeneration of bone tissue of the lower jaw was revealed. It occurs 1.5 times later than normal consolidation (p < 0.05).

trend was also characteristic of TNF-α (p < 0.05), similar to having IL-1

normal bone regeneration had little activity and duration.

parameters were within normal limits.

**164**

of foreign substance in the development of inflammatory response.

killing mechanisms, lowering the concentration of serum lysozyme.

*Clinical Implementation of Bone Regeneration and Maintenance*

presence of acute inflammation (**Table 1**).
