Platelet-rich plasma: Evaluation of effects of the preparation method and the growth factors on differentiation of mesenchymal stem cells from human adipose tissue

• Grant number: 10/15133-8
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: March 01, 2011
• End date: November 30, 2014
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Principal Investigator: William Dias Belangero
• Grantee: Ana Amélia Rodrigues
• Host Institution: Faculdade de Ciências Médicas (FCM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Platelet-Rich Plasma (PRP) is an autologous, non-immunogenic therapy that enables the regulation and stimulation of regenerative processes, resulting in faster recovery of injured tissues. Numerous studies have shown that PRP and growth factors take part on the angiogenic process, the extracellular matrix and osseous tissue rebuilding, acting as a potent healing stimulator of these tissues. Despite the intensive use of this therapy and the numerous methods for obtaining PRP cited in the literature, scientific data regarding standardized obtainig, behavior and influence of PRP on growth and cell viability are still inconsistent. Therefore, the aim of this study is to evaluate the influence of preparation methods and growth factors on the viability and differentiation of mesenchymal cells from human adipose tissue. The study will be carried out according to the following steps: 1. Preparation of PRPs (blood collection, centrifugation, platelet activation and quantification of growth factor release) according to parameters obtained from a previous study on modulation and optimization by the proponent group; 2. Isolation of adipose-derived mesenchymal stem cells (ADMSC); 3. Maintenance and activation of cells in culture by PRP; 4. Test of cell viability by MTT method; 5. Induction of osteogenic differentiation; 6. Evaluation of osteogenic differentiation by alizarin red staining; 7. Evaluation of osteogenic differentiation by quantification of alkaline phosphatase activity. Analysis of the influence of PRP preparation and activation methods on cell proliferation and osteogenic differentiation ability will point out the most suitable method for clinical application in regenerative medicine.