不同交联方法制备的鹿茸多肽-胶原蛋白/壳聚糖复合材料对成骨细胞增殖的影响

目的： 检测不同交联方法制备的鹿茸多肽-胶原蛋白/壳聚糖复合材料的结构并考察其对成骨细胞增殖的影响。 方法： 将胶原蛋白溶液和壳聚糖溶液（9：1）充分混匀，分别采用真空干热交联法和0.25%戊二醛交联法制备含鹿茸多肽（1.0 mg·L^-1）的复合材料。通过扫描电镜观察复合材料的空间构象，MTT试验检测不同交联法制备的复合材料对成骨细胞hFOB1.19增殖及碱性磷酸酶（ALP）活性的影响。 结果： 干热交联复合材料的结构仍处于不均匀分布状态，但孔径明显减小；而戊二醛交联复合材料呈规则的层状皱褶结构，支架内部孔径变大，且主要以片状结构为主。交联的复合材料组能促进成骨细胞增殖，显著优于对照组和复合材料组，戊二醛交联组对成骨细胞的增殖促进作用优于干热交联组。交联后的鹿茸多肽-胶原蛋白/壳聚糖复合材料能有效地提高成骨细胞ALP活性。 结论： 戊二醛交联法能有效提高鹿茸多肽-胶原蛋白/壳聚糖复合材料的性能，促进成骨细胞增殖和ALP酶活性升高。

Effects of Polypeptides from Cervi Cornu Pantotrichum-collagen/chitosan Composite Materials Prepared by Different Crosslinking Methods on Osteoblasts Proliferation

Objective: To explore structure of polypeptides from Cervi Cornu Pantotrichum-collagen/chitosan composite materials prepared by different crosslinking methods and investigate its effects on osteoblasts proliferation. Method: Mixed collagen and chitosan solution at the ratio of 9:1,vacuum thermo-crosslinking and 0.25% glutaraldehyde cross-linking method were used to prepare composite scaffolds which contained with 1.0 mg·L^-1 of polypeptides from Cervi Cornu Pantotrichum.Structure of composite scaffolds was observed by scanning electron microscopy,MTT assay were used to test effects of different composite materials on osteoblast hFOB1.19 proliferation and activity of alkaline phosphatase(ALP). Result: Structure of thermo-crosslinking composite materials had uneven distribution but pore size significantly reduced;while glutaraldehyde cross-linking group had regular structure with layered folds,internal pore size became larger and it was mainly dominated by sheet structure.Cross-linked composite groups could facilitate osteoblast proliferation and it was significantly better than the control group and the composite materials group,anxo-action for osteoblast proliferation of glutaraldehyde crosslinking group was better than thermo-crosslinking group.Crosslinking composite materials could effectively improve ALP activity of osteoblast. Conclusion: Glutaraldehyde cross-linking method could effectively improve properties of polypeptides from Cervi Cornu Pantotrichum-collagen/chitosan composite materials,such as osteoblast proliferation promoted and ALP activity increased.