Fabrication of hydroxyapatite-chitosan composite microspheres and their biological effect study in vitro / 安徽医科大学学报

ABSTRACT

Objective To investigate the influence of HAp-CS composite microsphere scaffold on the in vitro cell behaviors of mesenchymal cells and evaluate its potential application for bone tissue engineering.Methods Nano-hydroxyap-atite (HAp)and chitosan (CS)composites solution were assembled into microsphere scaffold through microfluidic and observed by inverted microscope.Rat bone marrow mesenchymal stem cells were co-cultured in vitro with the microspheres for calculating the adhesion rate for the first 6h.Proliferation rate was measured by cell counting in the next 1,3,6,9 d,respectively,and GraphPad Prism 6 software was used for statistical analysis.The morphology of BMSCs on the surface of HAp-CS composite microsphere was observed by scanning electron micros-copy (SEM)and confocal scanning microscopy.The cells and HAp-CS microspheres were filled into a disc mold and co-cultured for 14 ~21 d to observe the morphology.Results HAp-CS microspheres were observed to be round and with uniform size by microscope.The adhesion rate of BMSCs reached 80% after cultured for 6 h,and proliferation rate reached the highest value when cultured for 6 d.SEM observations showed that BMSCs adhered compactly to the surface of the microspheres,and the microspheres could be connected together through BMSCs.Af-ter co-culturing BMSCs with microspheres for 14 ~21 d,a complete tissue constructs could be formed.Conclusion HAp-CS microspheres are proved to be good scaffolds for promoting BMSCs adhesion and proliferation.Large a-mount of extracellular matrix can be formed to connect microspheres after co-cultured for a certain time,which paves the way for HAp-CS microspheres to be applied for bone regeneration in animal experiments.