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15
TOM 17. N 5-6 2013 P.
English version: CYTOKINE PROFILE IN YOUNG PATIENTS WITH MANDIBULAR FRACTURES NOT TREATED WITH ANTIBIOTIC THERAPY*
Ruzin G., Chyryk O., Kalinichenko S.
Kharkiv National Medical University, Kharkiv
Institute of Microbiology and Immunology them. Mechnikov National Academy of Medical Sciences of Ukraine, Kharkov
Mandibular fractures are very common among the maxillofacial injuries and occur in 65-90 % cases. The aim of this research is to determine cytokine proflles of young patients with mandibular fractures to introduce more effective medication for maxillofacial Injures into clinical practice. Object and methods of research. 80 patients aged 18-35 years with mandibular fractures with no evidence of concomitant somatic diseases were examined. Patients were divided into two groups: group I (50 patients) not treated with antibiotics and group II (30 patients) treated with antimicrobials. Immunological studies included determination of serum and saiivary IL-1/S, IL-10 and IFN-y levels in patients at admission to hospital and 1 month after discharge. Results. It was determined that levels of proinflammatory cytokine IL-1/S in the patients with mandibular fractures (both in serum and in saiva) were 3.41-5.05 times higher (p<0,05) compared with those of healthy controls. IL-10 levels were 1.15 times lower (p<0.05) in serum, 2.53 times higher (p<0.05) in saiva compared with control group levels. Serum IFN-y levels in patients with mandibular fractures demonstrated no significant difference from the simHar parameters of healthy individuals, whereas salivary levels of this cytokine in the patients with fractures were 1.45 times higher (p<0.05 ). Correlation analysis of cytokine profile before treatment revealed a strong directly proportional relationship between the hospitalisation time and cytokine profile parameters that were examined (correlation coefficient ranged from 0.90 to 0.96). It has been established that one month after the treatment cytokine levels of the control group patients were significantly (p<0.01) lower compared with those of the patients from the main group. These results show that antibiotics inhibit proinflammatory and anti-inflammatory cytokines synthesis.
Key words: mandibular fractures, cytokines, immunity, antibiotics.
Introduction
Mandibular fractures are the most common among maxillofacial traumatic injuries (65-90 %) [2, 3, 10]. The incidence of post-operative complications is quite high and varies from 11 to 37 % despite constant improvement of complex treatment, development of research techniques and increasing number of drugs [4, 8].
The immune status of a patient with mandibular fractures and its post-operative infection were studied by numerous researchers, since the body nonspecific resistance state and the patient's immunity are important for effectiveness of the treatment [1].
It is well known that in spite of the basic patient's general condition and immune reactivity level, the injury itself causes unfavorable effect on activity of the most of regulatory systems of the body which manifests in complex disorders and induces inhibition of cellular and humoral immunity [1, 7, 11].
The imbalance of local and system immune reactivity is manifested first of all in changing cytokine profile [7, 11]. These changes range depending on the extent of tissue damage and reflect the processes of local inflammation and activation of immune protection mechanisms, and changes in saliva and in blood do not always coincide with each other [5].
Current routine medication treatment of patients with mandibular fractures is a drug complex which includes preventive antibiotics regardless of age of the patient, presence of systemic diseases, interval between the moment of fracture and start of treatment, etc. However, recent publications give evidence of potent immune suppressive properties of antibiotics [9, 12]. That is, against the imbalanced immune reactivity, they suppress additionally the immune system. This intentional weakening
of the body's defense, in turn, can cause infection of the fracture fissure with oral cavity pathogenic and opportunistic microorganisms resulting in the further development of inflammatory complications [6].
According to the literature, cytokines concentration level provides information on functional activity of different immunocompetent cells types, development and severity of inflammatory process, its switching to a level of the system and allows to predict the course of the disease [1, 5, 7, 11].
The aim of this research is to determine cytokine profiles in the young patients with mandibular fractures to introduce more effective medication for maxillofacial injures into clinical practice.
Object and methods of research
80 patients aged 18-35 years with mandibular fractures with no evidence of concomitant somatic diseases were examined. Patients were divided into two groups: group I (50 patients) not treated with antibiotics and group II (30 patients) treated with antimicrobials. Control group consisted of 20 healthy individuals with no chronic diseases in history and not treated with any drug therapy one month prior to the research.
Immunological studies included determination of serum and salivary IL-1p, IL-10 and IFN-y levels in patients at admission to hospital and 1 month after discharge.
Statistical analysis of data was carried out according to the rules of ordinary and alternative variation statistics.
Results and discussion
It is known that salivary glands are highly sensitive to neural and humoral factors. Development of pathological changes in oral cavity are always accompanied by a response of various parts of its defense: emergence of oral
* To cite this English version: Ruzin G., Chyryk O., Kalinichenko S. Cytokine profile in young patients with mandibular fractures not treated with antibiotic therapy // Problemy ekologii ta medytsyny. - 2013. - Vol 17, № 5-6. - P. 60 -62.
npoSAeMH eKOAoriï Ta MejHUHHH
foci of chronic infection, decreased reactivity, microbial sensitization, etc. [1, 5, 6].
Cytokines are regulatory peptides that are involved in formation and regulation of the body's defenses. Thus, interleukin 1p (IL-1p) launches an inflammatory response cascade, stimulates collagen synthesis, plays an important role in local inflammatory process, is involved in the formation of osseous tissue and mediates interactions of immune and nervous systems. Interferon gamma (IFN-y) is involved directly in the formation and development of cellular immune response (Th1) and performs regulatory functions in development of humoral response (Th2). De-
creased cellular ability to produce IFN-y prevents elimination of pathogens which can lead to chronic infection. Interleukin 10 (IL-10) has anti-inflammatory and immunosuppressive effects and causes Th1- to Th2-cellular "shift" in immune response. In addition, it can inhibit antimicrobial response at early stages of inflammation, but it protects the body from hyperinflammation and tissue damage [11].
Study of cytokine profile in patients of two main groups before treatment allowed to determine values of significant differences compared with those in relatively healthy patients (Table 1).
Table 1
Cytokine profile in patients of two main groups before treatment (M±m)
Cytokine level (pg/ml) Group of patients
Patients with mandibular fractures (n=80) Relatively healthy patients (n=20) Reference laboratory index
serum IL-1 p 9,47±0,86* 1,72±0,58 0-11,2
IL-10 4,69±0,48* 5,41±0,13 0-31
IFN-Y 10,61 ±0,75 12,76±1,89 0-10
saliva IL-1 p 7,96±0,76* 2,33±0,59 0-3,5
IL-10 3,60±0,39* 1,42±0,19 0-5
IFN-Y 9,06±0,70* 6,26±1,21 0-4
times increase in IFN-y levels (p<0.01) compared with those identified in the first two days after injury. 5-7 days after injury, cytokine levels increased even more: 6.3-6.9-fold IL-1 p level increase (p<0.01), 7.8-8.2 times in IL-10 (p<0.01), and 4.7-5.2 in IFN-y values (p<0.01). The above mentioned indicates an active immune response.
3-4 day hospital admission
|--*--H-1p •••■••• IL-10 -
Note: * - significant difference between the indicators of patients with mandibular fractures and healthy individuals (p<0,05).
According to the results, levels of proinflammatory cytokine IL-1 p, both in serum and saliva of the patients with mandibular fractures were significantly (p<0.05) higher (3.41-5.05 times the average) compared with those of healthy individuals. These elevated IL-1 p levels indicate the development of local inflammatory process, possibly by action of stress and modification of permeability of capillary walls in the area of fracture.
It was established that in patients with fractures serum IL-10 level was on average 1.15 times lower (p<0.05), whereas in saliva - vice versa, higher, on average, 2.53 times (p<0.05) compared with those in the control group. In our opinion, this is an evidence of the beginning of the local inflammatory process. Thus, changed IL-10 level in biological fluids of the patients may predict disease.
It was determined that IFN-y serum levels in the patients with mandibular fractures did not differ significantly from those of healthy individuals, whereas a significant (on average, 1.45 times) increase of this cytokine salivary level, (p<0.05), was observed in the patients with fractures compared to the controls. This elevation of IFN-y values, in our view, indicates the activation of local protection mechanisms.
Correlation analysis of cytokine profile in serum and saliva of the patients with mandibular fractures, depending on the time of their admission to hospital showed a strong proportional correlation between the day of hospitalisation and their cytokine levels before treatment (Spearman correlation coefficient ranged from 0.90 to 0.96) (Fig. 1).
It was found that patients at day 3-4 after injury develop 3.70-4.25-fold increase in IL-1 p levels (p<0.01), 3.00-3.25 times increase in IL-10 (p<0.01) and 2.7-2.9
Fig. 1. Correlation analysis of cytokine profile in patients with mandibular fractures, depending on the time of their admission to hospital.
Next research link was to determine the level of cytokines after treatment (with and with no antimicrobials in the treatment combination).
It was found that the levels of cytokines in the patients treated with antibiotics were significantly (p<0.01) lower compared with those in the patients with no antibiotic therapy in the treatment scheme. Thus, IL-1 p levels were, on average, 4.4-5.6 times lower, IL-10 values -3.0-3.5 times, and IFN-y - 6.7-9.7 times lower. It shows the inhibition in production of proinflammatory as well as anti-inflammatory cytokines in those treated with antimicrobials. These figures suggest the development of an imbalance in immune system of the patients (Table 2).
Table 2
Cytokine profile in patients studied after treatment (M±m)
The clinical group of patients Cytokine level (pg/ml)
IL-1 p IL-10 IFN-Y
saliva serum saliva serum saliva serum
1 (n=11) 3,03±0,11* 3,5±0,12* 2,82±0,11 * 3,27±0,13* 3,32±0,10* 3,83±0,10*
II (n=6) 0,63±0,11* 0,89±0,10* 0,87±0,09* 1,18±0,12* 0,27±0,06* 0,52±0,07*
Note: * - significant difference between the indicators of groups (p<0,01).
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Thus, there is a sharp decline in immune reactivity of the patients treated with antimicrobials. These data confirm the research findings concerning potent immunosup-pressive effect of antibiotics [9, 12].
In the patients not treated with antimicrobials cytokine levels decreased comparatively with the values before treatment, but more slowly than in the patients treated with antibiotics. Thus, on average, IL-1p levels were 2.22.4 times lower (p<0.05), IL-10 - 1.2-1.4 times (p<0.05), IFN-y levels- 2.5-2.6 times lower (p<0.05) compared with
Fig. 2. Cytokine profile in patients with mandibular fractures before and after treatment.
The levels of cytokines in the patients of this group were significantly higher (p<0.05) as to those treated with antimicrobial drugs. This is an evidence of protection against the development of autoimmune processes and suggests the further favorable prognosis of the disease. However, the cytokine profile values obtained in the patients of clinical group I did not reach the figures of the controls (practically healthy individuals).
Summarizing the results, we note that the patients received antimicrobials in combination therapy demonstrated significantly lower levels of all investigated cyto-kines (p<0.01) compared to the patients not treated with antibiotics. It indicates inhibition of immunity by antimicrobials, which in turn, may additionally weaken defenses. That is, the reduction in macroorganism immune reactivity may make inefficient fighting against pathogens, create the conditions for failure of local and systemic mechanisms of effective protection, which in turn may be one of the causes of secondary immune deficiency syndrome.
Conclusions
1. Study of cytokine profile in all patients with mandibular fractures before treatment found that levels of proinflammatory cytokine IL-1p were, on average, 3.415.05 times higher (p<0.05) compared with those of healthy controls.
2. IL-10 levels were in serum, on average, 1.15 times lower (p<0.05), whereas in saliva - vice versa, on average, 2.53 times higher (p<0.05) compared with those data of the control group.
3. IFN-y levels in serum did not differ significantly from those of healthy individuals, whereas in saliva of the patients with fractures a significant (1.45 times) increase of this cytokine values, on average, (p<0.05) was observed.
4. Correlation analysis of cytokine profile before treatment showed a strong direct proportional relationship between the day of hospitalisation and cytokine levels in serum and saliva (Spearman correlation coefficient ranged from 0.90 to 0.96).
5. Levels of cytokines were significantly (p<0.01) lower in the patients treated with antibiotics versus the patients treated with no antibiotics.
6. The results of studies indicate that antibiotics inhibit
the synthesis of pro- and anti-inflammatory cytokines.
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