Bio-Physical-Chemical evaluation of Cyclophosphamide and Methotrexate incorporated into hyaluronic acid hydrogels to regulate the inflammatory process in temporomandibular disorders.

Abstract

Temporomandibular disorder is a public health problem that affects 12% of the world population. Its clinical presentation is diverse, and may include myalgia, arthralgia, headache, malocclusion, clicking, crackles, reduced mouth opening, among other signs and symptoms. The pathophysiology of the disease is explained by immuno-inflammatory dysregulation, so treatments are based on the regulation of this process. This work aims at the development and bio-physical-chemical evaluation of hydrogels based on hyaluronic acid conjugated to bupvacaine, dexamethasone, hydrocortisone, leflunomide, aqueous extract of Matricaria recutita or aqueous extract of green propolis. For the physical-chemical analysis, scanning electron microscopy, infrared spectroscopy by Fourier transform, thermogravitometry, capture of phosphate buffer solution and rheology test in oscillatory regime will be performed. For the biological analysis, human chondrocytes (CHON-001), human skin fibroblasts (HFF-1), human macrophages (THP-1) and human osteoblasts (MG-63) will be cultured. through the alamar blue colorimetric test, apoptotic cells by terminal labeling of deoxynucleotidyl transferase mediated by deoxyuridine triphosphate, genotoxicity by micronucleus test and immunomodulatory activity by elisa. The data will be submitted to the normality test, the results that obtain normal distribution in the Gaussian curve will be evaluated by the ANOVA method complemented by the Tukey test, with a significance level of 5% (pd0.05); data that do not obtain a normal distribution will be analyzed by the Kruskal wallis test supplemented by the Dunn's test with a significance level of 5% (pd0.05).