Action of fibrin sealant derived from snake venom as a biological scaffold for mesenchymal stem cells (MSCs) in bone defects produced in the femur of rats with osteoporosis

• Grant number: 14/06001-1
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: July 01, 2014
• End date: June 30, 2017
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Clinics and Surgery
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Rui Seabra Ferreira Junior
• Grantee: Patricia Rodrigues Orsi
• Host Institution: Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP). Centro Virtual de Toxinologia. Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

The use of stem cells in tissue repair have progressively developed and this therapy tends to improve the ability or even replace the restorative capacity of bone tissue, when there is partial or complete failure in the repair mechanism. In tissue regeneration, a biomaterial can promote the necessary environment for cell differentiation and define the final form of the regenerated tissue. Studies with mesenchymal stem cells (MSCs) from patients with osteoporosis, has shown that this disease is associated with a reduction in proliferation and osteogenic differentiation of MSCs, and local injection of MSCs may improve recovery of bone structure of sites with osteoporosis. One way to avoid cell loss after application of MSCs at the site of injury is through the use of scaffolds associated with the MSC. The possibility the use a Fibrin Sealant, whose polymerizing factor is derived from snake venom, opens an important clinical-surgical option. Moreover, this sealant has advantages in relation the conventional available on the market, such as not using of human blood in its composition and the possibility to customized formulas the need for surgery, considering clotting time. Based on the above objectives of this project are: (A) Collect and grow in vitro MSCs from bone marrow of rats ; (B) characterize MSCs markers with surfaces ; (C) analyze the ability of MSCs to differentiate into osteogenic lineage ; (D) Following the cytotoxicity assay in vitro Fibrin Sealant ; (E) assess the biodegradability in vitro Fibrin Sealant ; (F) evaluate the combination of the Fibrin Sealant with MSCs and differentiated in osteogenic lineage, for the treatment of osteoporosis after a critical defect in the femur of rats ; (G) investigate serum levels of estrogen, calcium, osteocalcin and alkaline phosphatase biochemical in animal with osteoporosis after a critical defect in femur of rats; (H) Conduct analysis microcomputadorizada tomography of the femur of animals with osteoporosis after a critical defect in the femur of rats; ( I) evaluate the bone regeneration of the femur of rats with osteoporosis through histological analysis and by scanning electron microscopy; (J ) evaluate the growth and viability of MSCs and differentiated in the femur of rats by immunohistochemistry analysis. All results are expressed as mean ± standard error of the mean. Differences between means will be tested by analysis of variance (ANOVA) followed by tests of significance. Discontinuous data will be analyzed by X2 test. Statistical significance will be considered for values of P <0.05. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ORSI, PATRICIA RODRIGUES; LANDIM-ALVARENGA, FERNANDA CRUZ; JUSTULIN, JR., LUIS ANTONIO; KANENO, RAMON; GOLIM, MARJORIE DE ASSIS; DOS SANTOS, DANIELA CARVALHO; ZORZELLA CRESTE, CAMILA FERNANDA; OBA, EUNICE; MAIA, LEANDRO; BARRAVIERA, BENEDITO; et al. A unique heterologous fibrin sealant (HFS) as a candidate biological scaffold for mesenchymal stem cells in osteoporotic rats. STEM CELL RESEARCH & THERAPY, v. 8, SEP 29 2017. (14/06001-1, 09/53846-9, 12/02689-3, 12/08101-8)