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

**Laboratory**

**166**

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

cytochemical

Lactoferrin,

Ig

IgM, g/l

IgG, g/l Lysozyme, mkg/ml

Total hemolytic

IL-1α, pg/ml IL-1ra, pg/ml

complement

 activity, arbitrary units/ml

А, g/l

 ng/ml

 activity of neutrophils,

 average coefficient

granulocytes,

 average coefficient

cytochemical

 test

 test stimulated,

 test, stimulation

 index

 %

spontaneous,

 %

phytohemagglutinin,

 %

 %

 %

lymphocytes,

 109/l

neutrophil/segmented

 neutrophils

 109/l

 109/l

 **parameters**

**Normal**

**Before**

**3 days**

 **10 days**

 **1 month**

 **3 months**

**surgery**

**values**

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

1.6–1.9

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

 18.36

1.17#

100.51

13.96\*,#

321.61

15.96\*,#

24.74

2.96\*,#

13.22

1.74\*,#

 30.24

3.08#

234.07

21.46\*,#

604.26

18.29\*

36.41

2.65#

18.33

2.14\*

 12.07

1.18#

44.93

2.51\*

13.46

1.49#

44.03

2.66\*

44.07

2.37\*

 2.78

0.49#

3.87

0.56#

14.21

2.33\*

3.87

0.49#

13.41

2.17\*

 15.30

2.36

 11.26

2.20\*

14.13

1.87

 6.31

0.69\*,#

14.51

1.45

 3.12

0.38#

1.05

0.19\*

2.80

0.25#

0.78

0.13\*,#

2.86

0.34

 2.48

0.41

 2.59

0.19

 2.46

0.15

 2.06

0.21#

2.68

0.31

 235.14

22.63#

451.09

17.35\*,#

956.88

16.44\*,#

566.22

23.26\*,#

355.84

14.87\*,#

 1.42

0.07

 1.63

0.08\*,#

1.36

0.12

 1.55

0.11

 1.26

0.07

 1.88

0.14

 2.19

0.18#

2.12

0.15

 1.37

0.19#

0.77

0.12\*

 0.75

0.21

 1.64

0.24\*

1.65

0.13\*

1.51

0.22\*

1.50

0.14\*,#

 31.09

3.88

 60.50

1.54\*,#

38.18

4.67

 68.34

5.46\*,#

39.96

6.13

 41.12

5.96

 37.02

3.99#

23.07

6.34\*

45.23

5.87

 25.31

5.34\*

 28.45

5.46#

45.45

5.07\*,#

44.73

6.98#

42.07

5.32#

91.09

12.34\*,#

 7.03

0.89

 9.06

2.01

 4.31

0.59\*,#

5.88

0.47#

12.71

0.36\*,#

*Clinical Implementation of Bone Regeneration and Maintenance*

 63.77

3.24

 38.50

3.51\*,#

46.47

3.54\*

63.61

2.98

 47.74

3.64\*,#

 2.09

0.20

 1.40

0.22\*

2.63

0.55

 1.70

0.10#

1.94

0.22

 0.32

0.07#

0.52

0.15

 0.30

0.11

 0.59

0.10\*

0.58

0.18

 0.04

0.01

 0.08

0.01\*,#

0.06

0.01#

0.04

0.01

 0.02

0.01

 4.37

0.38

 6.95

0.48\*,#

5.17

1.01

 3.63

0.39#

4.76

0.69

 7.08

0.50

 9.08

0.66\*,#

8.43

0.47\*

6.13

0.38

 7.50

0.51

The concentration of lysozyme in the same period was significantly reduced by 5.4 times (**Table 2**). The dynamics of cytokine concentrations also had differences: on day 3 after surgery significantly, namely more than 11-fold (p < 0.05), increased concentration of IL-1α, one of the stimulants of osteoclasts (**Table 2**). However similar changes were observed in patients with normal bone tissue regeneration. Most likely, the ratio between the level of IL-1α and IL-1ra after surgery (on day 10) played an important role in the violation of regeneration, when in patients with normal bone consolidation it was equal (**Table 1**), and at late bone consolidation, the level of IL -1ra was almost two times lower than the concentration of IL-1α (**Table 2** and **Figure 1**).

After one month, when the regeneration of bone tissue in the other group was completed, in patients with delayed consolidation, it was in the stage collagenogenesis. Features of the dynamics of immunological parameters are as follows: the number of neutrophils and lymphocytes in peripheral blood was reduced by 23.0% (p < 0.05) and 21.0% (p < 0.05) compared to those of patients with normal bone regeneration. Inhibition signs of humoral immunity were shown: IgA level was lower by 35.3% (p < 0.05), the concentration of IgM (**Table 2**), and IgG – 2.0 times (p < 0.05) was compared with the results in patients with the normal regeneration of the bone tissue. There was also a decrease in myeloperoxidase activity by 34.8% (p <0.05) and lysozyme concentration by 2 times (p < 0.05) reduced (**Table 2**). One month after surgery, the receptor level antagonist of IL-1 was higher concentration of IL-1α (**Table 2**) and TNF-α content was increased by 2.5-fold (p < 0.05), while the functional activity of the CD45<sup>+</sup> CD3<sup>+</sup> cells (**Table 2**), as already mentioned, was increased by 8.3-fold (p < 0.05).

It is interesting that in circumstances where bone regeneration has been completed, 3 months after the operation, the factors of immune responses, causing the growth of bone, are reduced. Thus, the concentration of lactoferrin was reduced by 40.5% (p < 0.05) and the number of T cells by 23.4% (p < 0.05). During this same period, the factors of immune reactions associated with the destruction of the bone tissue continue to maintain high concentrations. In particular, TNF-α level was increased by 4.2-fold (p < 0.05) (**Table 2**). The functional state of T cells in the neutrophil migration inhibition of the reaction was increased by 4.1 times (**Table 2**) metabolic functional activity of neutrophils to normal.

tissue. For each of the criteria, diagnostic sensitivity, diagnostic specificity and informative test are calculated, that is the ability to predict the possible develop-

IgM, g/l ↑ 2.7 0.9–2.6 87.7% 88.3% 89.5%

*The criteria for predicting delayed consolidation of bone tissue in the treatment of lesions of the mandible.*

To test the statistical hypotheses were recruited separate independent test samples of 49 people. It included patients with the same embodiments of bone damage and with the same kind of surgical treatment as the two major groups of the study. The criteria developed in this study showed that the coefficient of determination in this independent test sample was in the range of 84.2–97.5%. The joint use of two or more prognostic indicators allowed to increase the value of a diagnostic test for 3–4%. This fact gives more opportunities to clinicians in predicting delayed bone

The study revealed that the stages of regeneration of bone tissue (inflammation, proliferation of osteoblasts, collagenogenesis and ossification) are accompanied by changes in the immunological status. When delayed consolidation of bone tissue revealed differences of response of individual parts of the immune system, a comparative study of the immunological parameter dynamics at normal and slow osteogenesis is possible to establish criteria for delayed consolidation of bone tissue. Prognostic criteria to the operations include increasing the concentration of IgM

ment of delayed consolidation of bone tissue.

Lactoferrin, ng/ml ↓ 499.0 500.0–

formation.

**169**

**Table 3.**

**4. Conclusion**

**Prognostic criteria**

C-reactive protein,

Total number of white blood cells,

mg/l

10<sup>9</sup> /l

CD45<sup>+</sup> CD3<sup>+</sup> -cells, %

Nitro blue tetrazolium test spontaneous, %

Total hemolytic complement activity, arbitrary units/ml

**Morbidity osteogenesis**

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

> **Average for osteogenesis**

> > 1500.0

TNF-α, pg./ml ↑ 51.0 0–50.0 92.0% 90.9% 94.1% 10 days

**The diagnostic sensitivity of the test laboratory**

↓ 9.3 9.4–17.0 79.2% 81.2% 83.2%

↑ 6.9 4.0–6.8 92.3% 80.1% 93.1%

↓ 45.0 46.0–70.0 90.4% 92.3% 91.7%

↑ 33.0 10.0–32.0 91.4% 77.8% 96.3%

↓ 36.0 37.0–54.0 91.5% 93.0% 92.6%

Before surgery IgM, g/l ↑ 2.4 0.9–2.3 85.4% 94.6% 87.3%

3 days

**The diagnostic specificity of the test laboratory**

81.3% 88.4% 86.2%

**Diagnostic informative laboratory test**

Based on the study, criteria were developed by predicting delayed bone consolidation in the treatment of injuries of the lower jaw (**Table 3**).

These criteria allow for different stages of treatment (before surgery, for 3 or 10 hours after surgery) to predict the development of delayed consolidation of bone

*The ratio of the concentration of IL-1α to IL-1ra, a logarithmic scale was used to represent the data.*


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

#### **Table 3.**

The concentration of lysozyme in the same period was significantly reduced by 5.4 times (**Table 2**). The dynamics of cytokine concentrations also had differences: on day 3 after surgery significantly, namely more than 11-fold (p < 0.05), increased concentration of IL-1α, one of the stimulants of osteoclasts (**Table 2**). However similar changes were observed in patients with normal bone tissue regeneration. Most likely, the ratio between the level of IL-1α and IL-1ra after surgery (on day 10) played an important role in the violation of regeneration, when in patients with normal bone consolidation it was equal (**Table 1**), and at late bone consolidation, the level of IL -1ra was almost two times lower than the concentration of IL-1α

After one month, when the regeneration of bone tissue in the other group was

myeloperoxidase activity by 34.8% (p <0.05) and lysozyme concentration by 2 times (p < 0.05) reduced (**Table 2**). One month after surgery, the receptor level antagonist

Based on the study, criteria were developed by predicting delayed bone consol-

These criteria allow for different stages of treatment (before surgery, for 3 or 10 hours after surgery) to predict the development of delayed consolidation of bone

*The ratio of the concentration of IL-1α to IL-1ra, a logarithmic scale was used to represent the data.*

CD3<sup>+</sup> -

of IL-1 was higher concentration of IL-1α (**Table 2**) and TNF-α content was increased by 2.5-fold (p < 0.05), while the functional activity of the CD45<sup>+</sup>

cells (**Table 2**), as already mentioned, was increased by 8.3-fold (p < 0.05). It is interesting that in circumstances where bone regeneration has been completed, 3 months after the operation, the factors of immune responses, causing the growth of bone, are reduced. Thus, the concentration of lactoferrin was reduced by 40.5% (p < 0.05) and the number of T cells by 23.4% (p < 0.05). During this same period, the factors of immune reactions associated with the destruction of the bone tissue continue to maintain high concentrations. In particular, TNF-α level was increased by 4.2-fold (p < 0.05) (**Table 2**). The functional state of T cells in the neutrophil migration inhibition of the reaction was increased by 4.1 times (**Table 2**)

collagenogenesis. Features of the dynamics of immunological parameters are as follows: the number of neutrophils and lymphocytes in peripheral blood was reduced by 23.0% (p < 0.05) and 21.0% (p < 0.05) compared to those of patients with normal bone regeneration. Inhibition signs of humoral immunity were shown: IgA level was lower by 35.3% (p < 0.05), the concentration of IgM (**Table 2**), and IgG – 2.0 times (p < 0.05) was compared with the results in patients with the

completed, in patients with delayed consolidation, it was in the stage

*Clinical Implementation of Bone Regeneration and Maintenance*

normal regeneration of the bone tissue. There was also a decrease in

metabolic functional activity of neutrophils to normal.

idation in the treatment of injuries of the lower jaw (**Table 3**).

(**Table 2** and **Figure 1**).

**Figure 1.**

**168**

*The criteria for predicting delayed consolidation of bone tissue in the treatment of lesions of the mandible.*

IgM, g/l ↑ 2.7 0.9–2.6 87.7% 88.3% 89.5%

tissue. For each of the criteria, diagnostic sensitivity, diagnostic specificity and informative test are calculated, that is the ability to predict the possible development of delayed consolidation of bone tissue.

To test the statistical hypotheses were recruited separate independent test samples of 49 people. It included patients with the same embodiments of bone damage and with the same kind of surgical treatment as the two major groups of the study. The criteria developed in this study showed that the coefficient of determination in this independent test sample was in the range of 84.2–97.5%. The joint use of two or more prognostic indicators allowed to increase the value of a diagnostic test for 3–4%. This fact gives more opportunities to clinicians in predicting delayed bone formation.
