Tissue engineering of a human periodontal ligament fibroblast membrane - Alveolar osteoblast scaffold double construct

Abstract

As a first step in the development of cell-scaffold constructs for periodontal regeneration, human periodontal ligament fibroblasts (hPDLF) and alveolar osteoblasts (hAO) were studied for their osteogenic potential. Both cell types produced varying levels of alkaline phosphatase and mineral-like tissue. Under culture expansion, hPDLF demonstrated a preservation of collagen-synthetic capability as seen from the synthesis of type I, III, V, XII and XIV. Subsequently, cell-substratum interaction of hPDLF on biocompatible and bioresorbable poly(e-caprolactone) (PCL) was examined. Cytocompatibility of alkali-treated PCL membranes was enhanced via a two-fold increase in cell adhesion rate, attributable to a greater accessibility of fibronectin cell-binding domain. Constructs consisting of perforated PCL membranes provided greater cell anchorage, and cell and matrix alignment than unperforated membranes, while retaining hPDLF phenotypic expression and promoting matrix maturation. hPDLF proliferated on alkali-treated, perforated PCL membranes, while hAO produced mineral-like tissue on alkali-treated PCL scaffolds at day 21. Vascularized, well-integrated hPDLF-hAO double construct was observed at day 28 of subcutaneous implantation in athymic mice, but no further osteogenesis in the earlier-mineralized matrix was seen.