Osteogenic differentiation of preosteoblasts on naturally obtained 2D biomimetic matrix

Abstract

Extracellular microenvironment is very important in cell adhesion, migration and differentiation. It is an architectural support for tissue or stem cells and provides essential components for cellular homeostasis. In this study, we prepared a naturally derived ECM obtained from preosteoblasts, naming it preosteoblast-derived matrix (PDM). The primary focus is to evaluate the effect of PDM as a biomimetic 2D substrate in upregulating osteogenic differentiation of preosteoblasts. Preosteoblasts were cultured for 1 week and then decellularized using a cocktail solution of detergents and enzymes. Both structure and composition of PDM were examined using SEM, DAPI staining, and immunofluorescent staining, respectively. The test substrates were control (TCP), gelatin-coated surface, and PDM. Once decellularized, PDM exhibited fibers in varying sizes from tens to hundreds of nanometers, many of which were interconnected in different orientations. Early cell morphology of preosteoblasts was quite different between gelatin-coated surface and PDM. Preosteoblasts were proliferating much faster on the PDM. Calcium assay also supported the beneficial effect of PDM, as compared to other substrates. The present results hint that PDM may provide a favorable surface microenvironment for preosteoblasts and this may come from some physical cues of matrix, not proven yet at this time.