Increased Osteogenic Activity of Dynamic Cultured Composite Bone Scaffolds: Characterization and In Vitro Study

Abstract

The purpose of this study was to develop and characterize beta-tricalcium phosphate (β-TCP)/polycaprolactone (PCL) scaffolds, with 2 different ratios (50/50% and 65/35%), using 3-dimensionally (3D) printed dissolvable molds, and to evaluate cellular growth and osteogenic differentiation of both groups seeded with porcine bone marrow stem cells (pBMSCs) under dynamic culture in vitro. Two different groups of scaffolds were produced: group 1 (n=40) with a ratio (wt%) of 50/50% and group 2 (n=40) with 65/35% of β-TCP/PCL. Physicochemical, morphological, and mechanical characterization of the scaffolds were performed. Scaffolds were seeded with pBMSCs and differentiated osteogenically in dynamic culture. Cell density, distribution, and viability were assessed. Osteogenic differentiation was examined through alkaline phosphatase (ALP) staining, immunofluorescence, and photospectrometry. Osteogenic differentiated constructs showed homogenous and viable cell distribution. Cell density was significantly higher (P < .05) for 65/35% scaffolds at 10 days postseeding, whereas at 6 weeks, cell number equalized for both groups. ALP activity increased over time and was significantly higher (P < .05) for 65/35% scaffolds at 14 days postseeding. The mechanical properties of the developed 65/35% scaffolds were within the range of natural trabecular bone. Moreover, the 65/35% scaffolds showed biological advantages, such as higher cell growth and higher ALP activity.