人脐带华通胶细胞外基质源性软骨组织工程支架的制备及相关研究

Preparation and its related study of the engineering scaffold from human umbilical cord Wharton's jelly extracellular matrix-derived cartilage

目的采用人脐带华通胶细胞外基质为材料制备软骨组织工程支架,并对其性能进行相关检测。方法人脐带去除外膜及动、静脉,通过机械粉碎、差速离心去细胞、冷冻干燥、紫外及化学交联等方法将华通胶组织制备成三维多孔的软骨组织工程支架,并通过组织化学和免疫组织化学方法检测其成分保留情况;将人骨髓间充质干细胞复合至支架,光镜及电镜下观察细胞在支架上的生长状况及基质分泌情况,初步了解其生物相容性;将人脐带去细胞华通胶支架植入兔背部皮下,通过组织化学及CD4^+/CD8^+T淋巴细胞免疫荧光染色,了解其炎症情况及免疫原性。结果制备得到的人脐带华通胶细胞外基质源性支架为三维多孔支架,保留了华通胶中糖胺多糖和Ⅱ型胶原等成分,糖胺多糖含量为7.05%,总胶原含量为22.5%;接种于支架上的细胞黏附于支架孔壁上,生长状态良好,并有基质分泌;在体试验未见明显免疫反应。结论人脐带华通胶细胞外基质的成分与软骨细胞外基质类似,有利于细胞黏附和生长,无明显免疫原性,是一种较有前景的软骨组织工程支架材料。

Objective To fabricate a novel scaffold of cartilage tissue engineering with human umbilical cord Wharton＇s jelly extracellular matrix and assessed it＇s function in vitro.Methods Full-term healthy human umbilical cords were collected in aseptic condition and the arteries and veins were removed. The remaining Wharton＇s jelly was pulverized, centrifugated, lyophilized and cross-linked, and thus was fabricated into a three-dimensional porous scaffold for cartilage tissue engineering. Results The 3-D porous scaffold retained glycosaminoglycan and collagen content of human umbilical cord Wharton＇s jelly extracellular matrix, as the glycosaminoglycan content accounted for 7.05% and total collagen accounted for 22.5%. The human bone marrow-derived mesenchymal stem cells adhered to the scaffold, and growed well. The human umbilical Wharton＇s jelly extracellular matrix-derived scaffold did not evoke immune rejection in the subcutaneous implantation of rabbit backs. Conclusion The human umbilical cord Wharton＇s jelly extracellular matrix has very similar chemical component with cartilage extracellular matrix. It benefits for cell adherence and proliferation, and does not evoke immune rejection in vivo, so we propose it to be an ideal material for cartilage tissue engineering.