Background & Aims
Ozone therapy consists of an oxygen-ozone mixture (O2-O3; 95% O2 and 5% O3) used to treat several painful, inflammatory, and degenerative pathologies. We have recently demonstrated the analgesic and anti-inflammatory effect of the temporomandibular (TMJ) injection of oxygen-ozone mixture in the monosodium iodoacetate model of TMJ osteoarthritis (TMJOA). However, whether this effect is oxygen-mediated, at least in part, remains unclear. We know that cannabinoid CB1 receptor activation in peripheral nociceptive fibers induces analgesia and that cannabinoid CB2 receptor activation on immune cells induces the release of endogenous opioids and kappa(k)-opioid receptors mediated analgesia. In this context, we tested the hypothesis that the TMJ injection of oxygen has an analgesic effect mediated by peripheral CB1 and CB2, and ?-opioid receptors and an anti-inflammatory action
Methods
Experiments were conducted in Wistar female rats (220-280g) approved by the Ethics Committee in Animal Research of UNICAMP. TMJOA inflammatory hyperalgesia was induced by TMJ injection of monosodium iodoacetate (MIA; 2mg/30µL), and Oxygen (O2; 50µL) was TMJ injected 3 days later. TMJ hyperalgesia was assessed by measuring the mechanical nociceptive threshold with an electronic von Frey at days 0 (baseline) and 1, 3, 5, 7, 8, and 10 post-MIA injection. On day 8, animals received a TMJ injection of CB1, CB2, or ?-opioid antagonists – AM251, AM630, and nor-BNI (100µg) – or their vehicle. Mechanical hyperalgesia was assessed before and 15 min after the antagonists’ injection. AM251/AM630 were dissolved in Kolliphor®, DMSO, and saline (1:1:4; 24µL) and nor-BNI in saline (30µL). ELISA measured IL-1?, TNF-?, and IL-10 levels on TMJ periarticular tissues 7- and 11 days post-MIA injection. One- or Two-way ANOVA followed by Dunnet’s multiple comparison test was performed (p<0.05; 5-8 rats/group)
Results
MIA injection induced TMJ hyperalgesia, evidenced by a 32% decrease in the TMJ’s mechanical nociceptive threshold by day 1, and during the follow-up period (10 days) compared to saline. Oxygen injection on day 3 alleviated MIA-induced hyperalgesia as evidenced by the increase in the mechanical nociceptive threshold (from Mean±EPM: 71,37±4,90 to 103±18,05) on day 5, maintaining significantly greater than the MIA group during all the follow-up periods. Each antagonist reversed oxygen-induced analgesia, as evidenced by the decrease in the mechanical nociceptive threshold 15 min after TMJ injection on day 8 (Mean±EPM: AM251 from 88,95±12,38 to 71,79±6,97; AM630 from 86,38±5,03 to 66,10±6,35; nor-BNI from 93,95±13,40 to 68,28±11,17). The antagonists’ vehicle did not affect TMJ hyperalgesia threshold. Oxygen injection significantly increased the TMJ levels of TNF-? (2,6x higher; day 7) and IL-1? (1,7x higher; day 11) compared to the MIA group, but did not affect IL-10 levels
Conclusions
Oxygen injection into the TMJ has an analgesic effect mediated by peripheral CB1, CB2, and ?-opioid receptors despite increasing the release of the inflammatory cytokines TNF-? and IL-1?. As far as we know, this is the first study evaluating the effects of oxygen on analgesia and inflammation and its mechanisms of action on TMJ osteoarthritis.
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Presenting Author
Ana Carolina Machado
Poster Authors
Ana Carolina Machado
MSc
Universidade Estadual De Campinas (UNICAMP)
Lead Author
Tássia Machado
UNICAMP
Lead Author
César Sartori; DDs
PhD
State University of Campinas (UNICAMP)
Lead Author
Carlos A. Parada
State University of Campinas (UNICAMP)
Lead Author
Cláudia Tambeli
State University of Campinas (UNICAMP)
Lead Author
Topics
- Mechanisms: Biological-Systems (Physiology/Anatomy)