Evaluation of the anti-inflammatory and antioxidant effect of medicinal ozone on synovial joints of horses with osteochondritis dissecans

Author(s)

Cynthia do Prado Vendruscolo

• Document type: Doctoral Thesis
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Faculdade de Medicina Veterinária e Zootecnia (FMVZ/SBD)
• Defense date: 2021-06-29

Examining board members

Raquel Yvonne Arantes Baccarin; Ana Liz Garcia Alves; Roberta Carvalho Basile; Denise Tabacchi Fantoni; Maria Claudia Araripe Sucupira

• Advisor: Raquel Yvonne Arantes Baccarin

Abstract

Ozone (O3) is a highly oxidizing soluble gas. Several studies and clinical experiments have shown that ozone therapy has positive effects such as reducing pain and inflammation, improving function and benefiting bone and cartilage trophism, increasing vascularization and repair of cartilage and subchondral bone. The objective of the present study was to verify the anti-inflammatory and the oxidative stress modulation effects of O3 in the postoperative period of osteochondritis dissecans (OCD) cases that went through arthroscopic surgery. The concentrations of interleukins (1, 4, 6, 10 and TNF- α), substance P, prostaglandin E2 (PGE2), total antioxidant capacity (TAC), total oxidizing capacity (TOC), malonaldehyde (MAD), protein, urea, chondroitin sulfate (CS), hyaluronic acid (HA) and molecular weight of AH (MHHA) were measured as well as the total and differential cell count (WBC). In addition, evaluation of oxidative burst and physical, ultrasonographic, synovial membrane and lameness examinations were performed. Thirty-one joints with OCD were selected and allocated in two groups: the treated group (TG) received 5 minutes of gas lavage with 20 μg / ml of O3 after surgery and, 48h later, 5ml of O3 followed by 2ml of HA. The control (CG) was submitted to lavage with carbon dioxide and after 48h, 2 ml of HA. Synovial fluid was collected immediately before the initiation of arthroscopic procedure, and the treatment at 48 hours; the physical evaluations were performed on a daily basis until the end of the experiment and the ultrasound scans were taken before (M0h), 48 (M48h) and 96h (M96h) after the surgical procedure. Samples of synovial membranes were collected when portals were established (M0), at the end of surgical procedure (M1), and at the end of the joint lavage (M2). Results shown that at 48 hours, there was an increase, in both groups, in the concentrations of IL-1, 4, 6, 10, substance P, TAC, TOC, MAD, CS, protein, WBC and in the monocyte and neutrophil activation index, with no statistically significance only between the moments of MAD and with a difference of IL-6 between the groups (p < 0.05). It was also observed that the concentration of PGE2 decreased in the TG and increased significantly in the CG, and the concentration of TNF- α increased significantly in the TG. The MHHA and the concentrations of HA and urea decreased, but with no statistical significance between groups, only between the moments in the MHHA. The ultrasound scores decreased, being statistical significantly (p < 0.05) between M0h and M48h and 96h in both groups, with difference between groups in the scores of vascularization and synovitis. On physical examination, there was a significant difference between the groups in respiratory rate and facial pain score (p < 0.05). The treatment was able to partially control the inflammatory process, with a decrease in concentrations of inflammatory markers such as PGE2, whose concentration decreased, and in IL-6, whose concentration increased to a lesser extent in the TG. An increase in the concentrations of anti-inflammatory interleukins was also expected, however the most pronounced increase in IL-10 was not statistically significant. As for the catabolism of the extracellular matrix, the CS concentrations were lower in the TG as well as the decrease in the MHHA, although both also did not differ significantly from the CG. The facial pain score and respiratory rate were lower in the TG, demonstrating potential analgesic and anti-inflammatory effects. In the ultrasonographic evaluation, scores for vascularization and synovitis differed significantly between the groups, being smaller in the TG. The use of medicinal ozone at the end of the arthroscopic procedure in cases of osteochondritis dissecans showed anti-inflammatory and analgesic effects, decreasing the concentrations of inflammatory markers and cartilage catabolism. In addition, it provided better scores in the physical evaluation of the horses, aiding in the postoperative period. (AU)

• FAPESP's process: 17/15834-5 - Evaluation of the anti-inflammatory and antioxidant effects of medicinal ozone on synovial joints of horses with Osteochondritis Dissecans
• Grantee: Cynthia Do Prado Vendruscolo
• Support Opportunities: Scholarships in Brazil - Doctorate