RESUMEN
The tendon/ligament-to-bone interface has a complex organization to enable transfer of forces through the tendon/ligament to the bone. The purpose of this study is to create a co-culture environment enabling a tissue-specific tendon region and tissue-specific bone region on a degradable scaffold, using NIH 3T3 fibroblast-deposited extracellular matrix and MC 3T3 osteoblast-deposited extracellular matrix, respectively. Before full characterization of the deposited extracellular matrix coating can be analyzed, co-culture parameters including culture medium and seeding technique should be addressed. An appropriate medium formulation was developed to reduce fibroblast to osteoblast mineralization by adjusting beta-glycerophosphate concentrations. Standard growth medium with fetal bovine serum + 3 mM beta-glycerophosphate + 25 µg/mL ascorbic acid was found to be the most suitable formulation evaluated in these study conditions. Seeding and cell migration studies of co-cultured fibroblast- and osteoblast-specific scaffolds were performed to identify whether tissue regions could be created on the scaffold. Fibroblast and osteoblast regions were successfully seeded and little to no cell migration was observed up to 42 h after seeding. Finally, a preliminary analysis of basic extracellular matrix components was measured in the fibroblast, osteoblast, and transition regions. Tissue-specific DNA, glycosaminoglycan, and collagen were found in uniform amounts on the scaffolds and were not different significantly between scaffold regions. In conclusion, initial steps to create tissue-specific fibroblast and osteoblast regions on a degradable scaffold were successful in preparation for further characterization investigations as a tendon-to-bone interface scaffold.
RESUMEN
Due to limitations of bone autografts and allografts, synthetic bone grafts using osteoconductive biomaterials have been designed. In this study, collagen-chitosan-calcium phosphate microparticle-based scaffolds fused with glycolic acid were compared to their counterparts without collagen in terms of degradation, cytocompatibility, porosity, and Young's modulus. It was found that 26-30% collagen was incorporated and that hydroxyapatite was present. Moreover, there were no differences between control and collagen scaffolds in degradation, cytocompatibility, porosity, and Young's modulus. In general, scaffolds exhibited 23% porosity, 0.6-1.2 MPa Young's modulus, 23% degradation over 4 weeks, and supported a four to seven fold increase in osteoblast cell number over 7 days in culture. Collagen can be incorporated into these bone graft substitute scaffolds, which show an improved degradation profile.