Evaluation of the effect of mesenchymal stem cells from non-osteoporotic rats on the osteoblastic differentiation of mesenchymal stem cells from osteoporotic rats

• Grant number: 17/07213-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2017
• End date: June 30, 2018
• Field of knowledge: Health Sciences - Dentistry - Oral and Maxillofacial Surgery
• Principal Investigator: Márcio Mateus Beloti
• Grantee: Julia de Lima
• Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Osteoporosis can be triggered by several factors leading to bone fragility and increased susceptibility to fractures. Considering the lower frequency in men than in women, osteoporosis has been underexplored in this population. However, the osteoporosis-associated mortality and morbidity, generally related to testosterone deficiency, may not be neglected in these patients. Therefore, osteoporosis in male patients is an important global health issue and should be deeply studied. A plethora of treatments have been proposed with the aim of improving the quality of life of men with osteoporosis. Among them, a promising alternative is the use of cell therapy based on the use of mesenchymal stem cells (MSCs) and/or osteoblasts differentiated from these MSCs to treat the bone tissue damage caused by this disease. Therefore, the aim of this study will be to evaluate the influence of MSCs derived from the bone marrow of non-osteoporotic rats on osteoblastic differentiation of MCSs derived from the bone marrow of osteoporotic rats, grown under osteogenic conditions, using an indirect co-culture model. MSCs will be obtained from the bone marrow of osteoporotic rats (ORX-MSCs), in which the osteoporosis will be induced by orchiectomy, and of non-osteoporotic rats (NO-MSCs) and expanded in vitro in the growth medium. Then, the indirect co-culture will be conducted in the osteogenic medium using 24-well plates with transwells, where the NO-MSCs will be cultured inside the transwells, and ORX-MSCs will be cultured in the bottom of the wells. The controls will be non-co-cultured NO-MSCs and ORX-MSCs grown in osteogenic medium, which will be compared with ORX-MSCs grown in co-culture with NO-MSCs. We will evaluate the following cell responses: proliferation, alkaline phosphatase (ALP) activity, extracellular matrix mineralization, and gene expression of the osteoblast markers RUNX2, osterix, ALP, bone sialoprotein, and osteocalcin. The data will be submitted to the test of adherence to the normal curve to determine the appropriate statistical test. The results of this study may contribute to establishing the characteristics of candidate cells for use in cell therapy in men with osteoporosis, since, up to now this subject is underexplored by the literature.