Chitosan-collagen scaffolds : characterization and evaluation on osteoblast attachment and proliferation

Background: Extracellular matrix (ECM) was used as a model for the development of three-dimensional scaffolds in bone tissue engineering since it play a major role in cellular growth and differentiation. Several ECM-like materials that combine natural or synthetic polymers with collagen have been proposed. Chitosan, natural biopolymer, is the one attractive material that combined with collagen to be fabricated into chitosan-collagen scaffolds. Although chitosan-collagen scaffolds have been widely investigated in tissue cultures, the influence of this composite scaffold on osteoblast has not been clearly evaluated. Objectives: The aim of this study was to investigate the characteristic of chitosan-collagen scaffolds and their effect on osteoblastic (MC3T3-E1) cell attachment and proliferation. Methods: The scaffolds of chitosan, collagen and chitosan-collagen composite were fabricated by freeze drying technique. Microstructures of scaffolds were examined using scanning electron microscopes. Scanning electron images were analyzed using ImagePro® Plus software program to determine the pore size and area fraction of scaffolds. After culturing MC3T3-E1 cells on the scaffolds as test and on cover slips as the control, the MTT assay was performed to assess the effect of the test scaffolds on cell attachment at 5 and 24 hours, and cell proliferation on day 1,3,5,7,10 and 14. Results: The microstructures of scaffolds had porosity with pore size ranging from 0 to 50 microns. For cell attachment, all test scaffolds showed no statistically significant difference from the control. For cell proliferation, chitosan scaffolds and chitosan-collagen composite scaffolds showed statistically significant difference from the control and collagen scaffolds (p<0.05). MC3T3-E1 cells proliferated well on the control, collagen scaffolds, chitosan-collagen composite scaffolds and chitosan scaffolds, respectively. Conclusion: Chitosan-collagen scaffolds can support osteoblast attachment and proliferation, however, the further development in microstructures and biological properties are needed.