Development of a Synthetic Tissue Engineered 3D Printed Calciumalkaliphosphate-Based Bone Graft with Homogenously Distributed Osteoblasts and Mineralizing Bone Matrix In Vitro

摘要

Over the last decade there have been increasing efforts to develop adequate 3D scaffolds for bone tissue engineering from bioactive ceramics with 3D printing emerging as a promising technology. The overall objective of the present study was to generate a tissue engineered synthetic bone graft with homogenously distributed osteoblasts and mineralizing bone matrix in vitro, thereby mimicking the advantageous properties of autogenous bone grafts and facilitating usage for reconstructing segmental discontinuity defects in vivo. To this end, 3D scaffolds were developed from a silica containing calciumalkaliorthophosphate (code: GB9S14) utilizing two different fabrication processes, first a replica technique (SSM), and second 3D printing (RP). The mechanical and physical properties of the scaffolds (porosity, compressive strength, solubility) and their potential to facilitate homogenous colonization by osteogenic cells and extracellular bone matrix formation throughout the porous scaffold architecture prior to in vivo implantation were examined. To this end, murine preosteoblastic cells (MT3T3-E1) were dynamically seeded and cultured for 7 days on both scaffold types under perfusion with two different concentrations of 1.5 and 3xl06 cells per ml. The amount of cells and extracellular matrix formed and osteogenic marker expression were evaluated using hard tissue histology, immunohistochemical and histomorphometric analysis. SSM scaffolds (SSMS) displayed a significantly greater total porosity (86.9%) than RP scaffolds (RPS) (50%), while RPS exhibited significantly more open micropores, greater compressive strength and silica release. RPS seeded with a 3×10~6 cells per ml displayed greatest cell and extracellular matrix formation, mineralization and osteocalcin expression. In conclusion, RPS displayed superior mechanical and biological properties and facilitated generating a tissue engineered synthetic bone graft in vitro, which mimics the advantageous properties of autogenous bone grafts, by containing homogenously distributed terminally differentiated osteoblasts and mineralizing bone matrix and therefore is suitable for subsequent in vivo implantation for regenerating segmental discontinuity bone defects.