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Biofunctionalization of PLDLA scaffolds immobilized with RGD peptide and bone morphogenetic protein BMP-2

Grant number: 12/22593-0
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): July 01, 2013
Effective date (End): January 31, 2016
Field of knowledge:Engineering - Biomedical Engineering
Principal Investigator:Eliana Aparecida de Rezende Duek
Grantee:Bruna Antunes Más
Home Institution: Faculdade de Engenharia Mecânica (FEM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
Associated research grant:10/17804-7 - Polymeric composites, AP.TEM

Abstract

The biofunctionalization of bioresorbable polymeric scaffolds by chemical immobilization of natural molecules with bioactive and osteoinductive properties, capable of being naturally recognized by the native bone tissue, stimulating osteogenesis process and tissue repair, from the cultivation of human mesenchymal stem cells (hCTMs ) on these materials, is a promising strategy in Bone Tissue Engineering. The RGD peptide sequence, naturally present in the adhesion proteins that compose the extracellular matrix native bone and the bone morphogenetic protein (BMP-2), with osteoinductive property, are widely recognized for promoting a significant improvement in cell-biomaterial interaction and act as signaling a cascade of biological events that stimulate proliferation, differentiation, and maintenance of osteoblastic phenotype. Thus, the present study aims to develop scaffolds of PLDLA prototyped by-layer deposition technique fibers oriented 0/90o and biofunctionalyze it by both RGD peptide and BMP-2 immobilization. The process of surface functionalization will be performed in two distinct steps and complementary where, initially, the pre-activation of the surface material is carried out by incorporating functional group carboxyl (-COOH) obtained by plasma treatment techniques, and then , the bioactive molecules are immobilized on the material surface by stable covalent bonds employing carbodiimide hydrochloride heterobifunctional crosslinking reagent. The composition of the scaffold according to the binding efficiency of the peptide RGD and BMP-2 will be analyzed by an increasing equimolar variation of biomolecules, where the concentrations of maximum binding efficiency of RGD and BMP-2 will be combined in order to obtain a PLDLA structure containing RGD (X) and BMP-2 (Y) composition. The binding efficiency of the peptide RGD and BMP-2 as well as the influence of the surface modification on the native properties of the scaffolds will be identified and evaluated physic-chemically. The potential of biological application PLDLA-RGD/BMP-2 scaffolds in the adhesion, proliferation and osteogenesis process of hCTMs will be evaluated by different molecular techniques such as flow cytometry, quantification of proteins expression involved in bone tissue by PCR real time and study of cell morphology cultured on PLDLA-RGD/BMP-2 scaffolds. (AU)

Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
MÁS, Bruna Antunes. Biofunctionalization of PLDLA scaffolds immobilized with cyclic RGDfK peptide and knuckle epitope (73-92) of BMP-2. 2016. Doctoral Thesis - Universidade Estadual de Campinas, Faculdade de Engenharia Mecânica.

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