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Isolation of Human Mesenchymal Stem Cells and their Cultivation on the Porous Bone Matrix
Published on: February 9, 2015
Human Amniotic Epithelial Stem Cells and Osteoblast Cells Behavior on Collagen Membranes for Bone Guided Regeneration
Antonio Pérez-Pérez1, Javier Gil2, Isabela Bueno-Bianchi3
1Departamento de Bioquímica Médica y Biología Molecular e Inmunología, Hospital Universitario Virgen Macarena, Facultad de Medicina, Universidad de Sevilla, Avenida Sánchez Pizjuán 4, 41009 Sevilla, Spain.
Abstract:
Guided bone regeneration (GBR) heavily relies on barrier membranes, with collagen being the clinical standard material. Human amniotic epithelial cells (hAECs) represent a promising, non-controversial stem cell source with substantial osteogenic and immunomodulatory potential. This study aimed to comparatively evaluate the structural characteristics of three commercial collagen membranes (Biocollagen®, Derma®, and VantyColl®) and their influence on the biological behavior, viability, and osteogenic differentiation of hAECs and hFOB 1.19 human fetal osteoblasts. The microarchitecture was assessed via scanning electron microscopy (SEM). Biological response was evaluated over 14 days, using MTT assays, calcium and phosphorus quantification, alkaline phosphatase (ALP) activity, and quantitative real-time PCR (qRT-PCR) for osteogenic markers (Runx2, Osterix, ALP, and OPN). SEM revealed a dense lamellar structure for Biocollagen®, a fibrillar and oriented architecture for Derma®, and a highly porous network for VantyColl®. Both cell types adhered to and proliferated on all membranes. Derma® provided the best long-term proliferative support for both lineages. Conversely, VantyColl® induced robust early osteoblastic differentiation, marked by exceptional upregulation of Osterix (24.93-fold) and Runx2 (2.64-fold), though it exhibited diminished long-term hAEC viability. Ultimately, collagen membrane microarchitecture dictates cell fate; dense fibrillar networks (Derma®) favor sustained growth and late matrix maturation (OPN), whereas high-porosity scaffolds (VantyColl®) amplify early osteoinductive cascades.

