胶原多糖基纳米羟基磷灰石仿生骨支架材料的研制

目的依据仿生原理制备新型的胶原多糖基纳米羟基磷灰石（HA）复合骨支架材料，并与成骨细胞复合培养，检测其细胞相容性。方法以胶原分子与透明质酸钠的交联产物为模板，调制钙磷盐在液相中沉积其上，得到矿化胶原多糖基复合材料；采用液相分离法与少量聚乳酸复合进一步制备成为三维多孔支架，使用成骨细胞（Mc3T3-E1）接种于该支架上培养。用X—ray衍射、扫描电镜、万能材料测试机等对材料进行观察和测试分析；并用倒置相差显微镜、荧光显微镜、扫描电镜、CCK-8细胞计数试剂盒、碱性磷酸酶（ALP）活性测定等观察和分析细胞在支架材料中的生长、分化情况。结果胶原多糖基纳米HA仿生复合材料的晶粒度较低，晶体极为细小，与天然骨中羟基磷灰石的组装结构类似；该复合支架为多孔状，孔隙率约82％，孔径大小为200～650μm；抗压性能好，成骨细胞可在其上贴附、生长和繁殖，并表现出较高的成骨活性。结论所制备的胶原多糖基纳米HA仿生骨支架材料，无论从组分和结构上均与天然松质骨类似，与成骨细胞相容性好，可望成为较理想的骨组织工程支架材料。

Preparation of mineralized collagen-polyose based biomimetic scaffold material for bone tissue Engineering

Objective To prepare a novel bioactive and degradable scaffold with mineralized collagenpolyose based composite by biomimetic synthesis for bone tissue engineering and explore the compatibility of osteoblast culturing on the scaffold.Methods Using the cross-linking product of collagenⅠ and sodium hyaluronate as the template,the calcium phosphate was deposited on it to produce a mineralized composite.The 3-D porous scaffolds were prepared by liquid phase separation after the mineralized composite combining with polylactic acid (PLA) and NaCl.The materials and scaffolds were investigated by x-ray diffraction (XRD),scanning electronic microscopy (SEM) and universal testing machine.In addition,inverted microscope,fluorescence microscope,SEM,Cell Counting Kit-8 assay and alkaline phosphatase (ALP) assay were introduced to analyze the growth,function and compatibility(morphology,proliferation and differentiation ) of osteoblast-like cell on the scaffolds.Results The degree of hydroxyapatite (HA) crystals in the composite was low and the size was tiny,which were similar to that of nature bone.The SEM micrographs showed that the scaffolds possessed 82% of porosity and the pore size was about 200 μm to 650 μm.Cells on the scaffolds spread well and presented a high proliferation rate and differentiation level.Conclusion The novel scaffolds are simiar to nature cancellous (spongy) bone both on structure and in property and might be used as one of the optimal scaffolds material for bone tissue engineering.