Evaluation of treatment with TNF-alfa inhibitor (adalimumab) on the deleterial effects promoted by smoking in cartilagine and bone tissue: in vitro and in vivo studies

• Grant number: 22/07357-0
• Support Opportunities: Regular Research Grants
• Start date: December 01, 2022
• End date: November 30, 2024
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Fernanda Degobbi Tenorio Quirino dos Santos Lopes
• Grantee: Fernanda Degobbi Tenorio Quirino dos Santos Lopes
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Alexandre Povoa Barbosa ; Débora Levy ; Walcy Paganelli Rosolia Teodoro

Abstract

Smoking is one of the main risk factors for the development of cardiovascular and pulmonary diseases and different types of cancer. In addition, it is also an important factor for the exacerbation of diseases that affect the musculoskeletal system, such as osteoarthritis. In previous studies developed by our group, we have already demonstrated the deleterious effects of exposure to cigarette smoke on bone tissue, with an increase in the production of cytokines that promote an increase in osteoclastogenesis and a decrease in the activity of osteoblasts, causing changes in the tissue composition, predisposing to greater fragility. Among these cytokines that are increased by exposure to cigarette smoke is TNF-alpha. The increase in TNF-alpha is associated with increased production of reactive oxygen species (ROS) and cell death (apoptosis). In osteoarthritis, decreased production of type II collagen by chondrocytes is described as one of the main factors involved in the development and progression of this disease. In a study developed by our group, we evaluated cartilage samples taken from smokers and non-smokers who underwent total hip arthroplasty. The study is still ongoing, but we have already demonstrated an increase in chondrocyte apoptosis, as well as a decrease in the percentage of type II collagen and chemokines such as TNF-alpha, in smokers compared to non-smokers, suggesting smoking as a exacerbation factor of responses. Thus, we intend to develop two projects, one "in vivo" and another "in vitro", using a TNF alpha inhibitor already used commercially to evaluate the effects of this treatment on structural and systemic parameters, as well as cellular parameters already studied by our group in previous works. Project 1: evaluation of treatment with a TNF-alpha inhibitor (Adalimumab) in mice exposed to cigarette smoke, following an exposure protocol already extensively used by our group (daily exposures, 2X a day, 30 min in a body exposure chamber whole). Adalimumab will be administered intraperitoneally on specific days and 1 hour before the start of exposure to cigarette smoke. After the end of the protocol, the animals will be anesthetized and euthanized to remove the femurotibial joints. In this material, histomorphometric and histological analyzes will be performed regarding bone and cartilaginous tissue, as well as pro-inflammatory chemokines involved in the metabolism of these two tissues. Project 2: The treatment with Adalimumab in a culture of chondrocytes grown in a culture medium with cigarette extract (extract concentration already standardized to maintain these cells alive for 48 h) will be evaluated. We will evaluate parameters that describe cell viability, as well as expression of reactive oxygen species, cellular markers of apoptosis, TNF-alpha and collagen type II expression in cartilage homogenate. We will use a commercially used TNF-alpha inhibitor (still to standardize concentration) to treat these media exposed to cigarette extract to verify the effects of this treatment on the evaluated parameters. (AU)