Abstract
BACKGROUND: In this study, we report on the results from the development and early in vitro testing of a gene activated matrix encoding basic human fibroblast growth factor 2 (FGF2) in bone marrow stromal cells (BMSCs). METHODS: Polyethylenimine (PEI), a cationic polymer, was utilized as a gene delivery vector and collagen scaffolds were used as the carrier to deliver the PEI-pDNA nano-sized complexes (nanoplexes) encoding the FGF2 protein. Initially, the BMSCs were transfected in vitro with the PEI-pFGF2 nanoplexes, prepared at a N/P ratio of 10, with cells alone and naked DNA as controls. This was followed by transfection experiments using collagen scaffold containing complexes, with the scaffold alone as a control. The transfection efficacy of the nanoplexes was assessed using ELISA for the determination of FGF2 protein expressed by the transfected cells. The functionality of transfection was assessed by evaluating cellular recruitment, attachment, and proliferation of BMSCs on the scaffold using imaging techniques. RESULTS: BMSCs transfected with the PEI-pFGF2 nanoplexes (either alone or within the scaffold) led to higher expression of FGF2, compared to controls. Scanning electron microscopy and confocal imaging confirmed the recruitment and attachment of BMSCs to scaffolds containing the PEI-pFGF2 nanoplexes. Confocal microscopy showed a significantly higher number of proliferating cells within PEI-pFGF2 nanoplex-loaded scaffolds than with empty scaffolds. CONCLUSIONS: This first in vitro evaluation in BMSCs provides evidence that gene activated matrices (GAMs) encoding the FGF2 protein may have strong translational potential for clinical applications that require enhanced osseous and periodontal tissue regeneration.