Fabricated Hydroxyapatite/Chitosan Microsphere Miomimetic Scaffolds And The Cell Behavor In Vitro

Objective The use of microfluidic technology to prepare injectable chitosan hydroxyapatite(HAp-CS) microspheres biomimetic scaffold, and with bone marrow mesenchymal stem cells in vitro tissue engineering of bone formation, and evaluate the regenerative potential of new bone tissue engineering scaffolds.Methods Nano-hydroxyapatite(HAp) and chitosan(CS) complexes by microfluidics microspheres self-assembled into scaffolds, according to four different experimental conditions for preparing microspheres material, A1 for curing solid Na OH Prepared HAp-CS microspheres, a1 solid HAp-CS microspheres prepared for the curing of freezing; A2 HAp-CS hollow microspheres Na OH cured, a2 hollow microspheres HAp-CS freezing cured. A2, a2 hollow experimental group, A1, a1 is a solid control group. Ordinary microscope and scanning electron microscope observation of the morphology of materials to each group. Isolation and culture of rat bone marrow mesenchymal stem cells(BMSCs), a certain proportion of composite microspheres with four groups underwent dynamic culture in vitro, 6 h before calculating the adhesion rate. Train 1 d, 3 d, 6 d, 9 d, scanning electron microscopy, cytoskeletal / nucleus complex dyeing line ordinary fluorescence microscopy and confocal scanning microscope; calculated for each group of materials in the cell proliferation rate, and using Graph Pad Prism 6 software for statistical analysis. The cells were filled composite material diameter 5 mm, thickness 2 mm mold cultures 14-21 d, morphology.Results Four groups of materials prepared by inverted microscope after A2, a2 group of rules circle, the same size, transparency is slightly different, you can see the obvious hollow structure, part of the opening in the surface of the material. A1, a1 circular, similar in size, shape for solid, partial sphere rupture. Scanning electron microscopy material microstructure are four ball shape three-dimensional structure, about the size of 150-200 μm, surface micro-fiber mesh structure is highly intertwined. Four groups of materials have before 6 h have a higher adherence rate of 80%, but there was no significant difference between the groups. After each group of composite materials and BMSCs, 6d proliferation reached at the peak of the added value of a hollow set of four time points were significantly higher than a solid group, but no significant difference. Scanning electron microscopy and confocal scanning microscope hollow group were found to promote cell adhesion than a solid group of strong growth, the cells in the sphere surface is completely spread. Hollow molds in vitro experiments with cell cultures when group 18 d, to form a more complete functional tissue blocks.Conclusion HAP-CS hollow microspheres having the desired three-dimensional structure of the stent and microscopic composition, promote BMSCs adhesion, proliferation and differentiation in vitro to promote the formation of bone tissue engineering, is an ideal bone tissue engineering scaffold.