Abstract

Heat shock protein 60 (Hsp60) is considered as one of the possible autoantigens involved in the pathogenesis of a number of chronic diseases including periodontal diseases. The application of hyaluronic acid or hyaluronan (HA) in the treatment of periodontitis has been evaluated in several clinical trials, however, the effect of hyaluronic acid on heat shock protein 60 level in periodontal soft tissues has not been studied. The aim of this work was to evaluate the effect of HA injections on levels of Hsp60 in periodontal tissue of the rats. Samples of periodontal tissue of mandibular incisors of adult male Wistar rats at 10-12 months of age were investigated. Rats were distributed into the control group and the periodontitis group. Visual manifestations of hyperemia of the gums around the incisors were the criterion for selecting animals into the periodontitis group. There were two subgroups in the control group: intact rats (I); intact rats after HA “hyaDENT BG” 1.0 MDa (BioScience GmbH, Germany) treatment (I+“G-1.0”). There were four subgroups in the periodontitis group: rats with periodontitis (P); rats with periodontitis after HA “hyaDENT BG” 1.0 MDa (BioScience GmbH, Germany) treatment (P+“G-1.0”); rats with periodontitis after HA “SERTOBEC” 2.4 MDa (S.C. Rompharm Company S.R.L., Romania) treatment (P+“S-2.4”); rats with periodontitis after HA “SERTOBEC Tendon” 2.4 МDа (S.C. Rompharm Company S.R.L., Romania) treatment (P+“ST-2.4”). There were three animals in each subgroup. Rats were injected 0.05 ml HA in the area of alveolar processus of central incisors once a week, three times. Levels of Hsp60 in total lysates of periodontal tissue were tested by Western blotting method before and after the treatment with HA (one month after the last injection). There was no significant difference between levels of Hsp60 in total lysates of periodontal tissue of intact rats and rats with periodontitis before treatment (p>0.05). Rats with periodontitis showed decreased inflammation in the periodontal tissue after treatment with HA with different molecular weight. Intact rats and rats with periodontitis which were treated with HA “hyaDENT BG” 1.0 MDa showed reduced levels of Hsp60 in total lysates of periodontal tissue comparatively with levels of Hsp60 before treatment (by 15.4 and 10.7 times respectively, p<0.001). Rats with periodontitis which were treated with HA “SERTOBEC” 2.4 MDa or HA “SERTOBEC Tendon” 2.4 MDa also showed reduced levels of Hsp60 in total lysates of periodontal tissue com­paratively with levels of Hsp60 before treatment (by 21.3 and 16.4 times respectively, p<0.001). Rats with pe­riodontitis showed the decrease in inflammation in periodontal tissue after treatment with HA with different molecular weight. Injections of HA has contributed to reduce levels of Hsp60 in periodontal tissue of intact rats and rats with periodontitis.

Highlights

  • Levels of Heat shock protein 60 (Hsp60) in periodontal tissue of rats: influence of injections of hyaluronic acid

  • There was no significant difference between levels of Hsp60 in total lysates of periodontal tissue of intact rats and rats with periodontitis before treatment (p>0.05)

  • Intact rats and rats with periodontitis which were treated with HA “hyaDENT BG” 1.0 MDa showed reduced levels of Hsp60 in total lysates of periodontal tissue comparatively with levels of Hsp60 before treatment

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Summary

MATERIALS AND METHODS

OF RESEARCH Samples of periodontal tissue of mandibular incisors of adult male Wistar rats were investigated. There were two subgroups in the control group: intact rats (I); intact rats after HA “hyaDENT BG” 1.0 MDa (BioScience GmbH, Germany) treatment (I+“G-1.0”). Levels of Hsp in total lysates of periodontal tissue were tested by Western blotting before and after the treatment with HA of different molecular weight (1 month after the last injection). After washing in PBS tween buffer (10 min x 3) the membranes were incubated with peroxidase-conjugated secondary anti-rabbit (in case of Hsp60) or anti-mouse (in case of GAPDH) antibodies for 1 h. The treatment of the membranes with secondary antibodies was followed by washing in PBS tween buffer (10 min x 3) and chemiluminescence detection according to manufacturer’s instructions by ChemiDoc. All samples of periodontal tissue were examined for levels of Hsp and GAPDH in six reproducible repetitions. Data were analyzed using Student's t-test, a p

RESULTS AND DISCUSSION
CONCLUSION
14. Hyaluronic acid
19. Psychological Stress Delays Periodontitis Healing in Rats
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