CAJ | 학술논문

目的观察新型三维支架材料胶原复合梯度磷酸三钙在体外与软骨细胞的相容性和黏附性,评价其作为软骨组织工程支架的可行性.方法取8周龄新两兰大白兔膝关节软骨,以酶消化法获得高纯度软骨细胞,培养3代后与三维支架材料胶原复合梯度磷酸三钙在体外复合培养.用倒置相差显微镜、HE染色、免疫组织化学及扫描电镜观察软骨细胞形态、Ⅱ型胶原表达及成软骨能力,同时观察支架材料与软骨细胞的相容性.结果扫描电镜观察显示支架材料具有疏松多孔结构,孔隙结构规则,孔径100～150 μm,材料内部孔与孔之间贯通良好.支架亲水性好.软骨细胞吸附于支架表面,增殖并逐渐顺孔隙迁徙至支架内部,在孔壁贴附良好,表型维持稳定,可分泌细胞外基质.结论胶原复合梯度磷酸三钙三维支架具有良好的细胞相容性.
목적 관찰신형삼유지가재료효원복합제도린산삼개재체외여연골세포적상용성화점부성,평개기작위연골조직공정지가적가행성.방법 취8주령신량란대백토슬관절연골,이매소화법획득고순도연골세포,배양3대후여삼유지가재료효원복합제도린산삼개재체외복합배양.용도치상차현미경、HE염색、면역조직화학급소묘전경관찰연골세포형태、Ⅱ형효원표체급성연골능력,동시관찰지가재료여연골세포적상용성.결과 소묘전경관찰현시지가재료구유소송다공결구,공극결구규칙,공경100～150 μm,재료내부공여공지간관통량호.지가친수성호.연골세포흡부우지가표면,증식병축점순공극천사지지가내부,재공벽첩부량호,표형유지은정,가분비세포외기질.결론 효원복합제도린산삼개삼유지가구유량호적세포상용성.
Objective To evaluate the feasibility of collagen complex gradient tricalcium phosphate composite scaffolds for cartilage tissue engineering by observing the compatibility and adhesion of chondrocytes. Methods High purified chondrocytes from knee joints of 8-week-old New Zealand rabbits were obtained by enzyme digestion, then the chondrocytes of the third generation were cultured combined with collagen complex gradient tricalcium phosphate composite scaffolds in vitro.Chondrocyte morphology, collagen Ⅱ expression, chondrogenesis, the compatibility of chondroeyte and scaffolds were evaluated with inverted microscope, HE staining, scanning electron microscope (SEM) and immunohistochemistry.Results By SEM, the scaffold had regular, porous architecture of communicated microholes.The average pore diameter was 100-150 μm.Observation revealed good hydrophilicity of the composite scaffold.The chondrocytes adhered to the surface, proliferated and migrated into the inside of the scaffold.Chondrocytes attached to wall of microholes of the scaffold had largely maintained morphology and could secrete the extracellular matrix on the porous scaffold.Conclusions Collagen complex gradient tricalcium phosphate composite scaffold possesses good cellular compatibility.