海藻酸钠-软骨细胞培养移植修复兔关节软骨缺损的实验研究-骨科专业论文.docxVIP

山西医科大学 山西医科大学 硕士学位论文 海藻酸钠．软骨细胞培养移植修复兔 关节软骨缺损的实验研究 摘要 目的对海藻酸纳．软骨细胞复合构建的工程化软骨进行 组织形态学和超微结构观察，探讨应用工程性软骨移植修复软 骨缺损的可行性。 方法取34周龄成年新西兰兔双膝关节软骨，酶消化法 得到高纯度软骨细胞，与可注射生物材料海藻酸钠(Alginate)混 合，种植密度为4×106／ml，通过硅胶模型制成圆形柱状细胞盘 (20ul／个)，CaCl2溶液中凝胶化10min，DMEM／F12无血清培 养基加20％FBS(fatal blood solution)于24孔培养板中培养。 分别于2、4、6、8、10、12周取细胞盘，行HE、AB—PAS染 色及免疫组化分析，测定盘中蛋白多糖含量，并作电镜切片进 行投射电镜观察。同时选用成年新西兰兔28只，用电钻在兔两 侧股骨内侧髁中央造成直径3．5ram、深3mm的全层软骨缺损模 型，随机在缺损处植入体外培养4周的海藻酸钠．软骨细胞盘(实 验组)及无细胞的海藻酸钠载体(对照组)，分别于术后6、12、 24、48周取材，观察软骨缺损修复情况。 山西医科大学 山西医科大学 硕士学位论文 结果软骨细胞培养4天后开始增殖，4周时达增殖高峰， 盘中胶原及蛋白多糖的含量随培养时间延长逐渐增加；细胞移 植组软骨缺损以透明样软骨修复为主，对照组则以纤维组织填 充为主，48W时两组修复组织均有一定程度退变。 结论海藻酸钠．软骨细胞复合物体外培养可生成工程化 软骨，异体移植能以透明软骨修复软骨缺损，但在一定时间后 再生软骨容易发生退变。 【关键词1关节软骨；软骨细胞；海藻酸纳；组织工程；软 骨；移植 山西医科大学 山西医科大学 硕士学位论文 Biological characteristic study of in vitro culture of chondrocytes combined with Alginate gel from joint Articular in rabbits Abstract Objective：To observe chondrocytes histomorphology and structure in vitro culture combined injectable biocompatible matrix (Alginate)．And value reparative tissue insulted from engineered cartilage tissue in terms ofhistomorphology． Methods Articular chondrocytes from the knee joints of 34。weeks-。old new Zealand white rabbits were gain by digest cartilage using collagenase enzyme，which were mixed with alginate resulting in a cell density of 4×1 06／ml and individed into many bead，every bead contain 20vl suspensions．The beads were gelled in GaCl2 solution and cultured in 24-Multiwell．Specimens were harvested at the end of2、4、6、8、10 and 12 weeks after culture． and were stained with HE and AB—PAS，assaying proteoglycan and translating electron microscopy．Then choose 28 adult new Zealalld white rabbits，3．5×3mm Cylindrical，full-thick articular defect was produced in the femur condyle of knee jionts，engineering cartilage 1 山西医科大学 山西医科大学 硕士学位论文 constructed in vitro were implanted in one side in random，spencims was harvested at the end of 6，12，24，and 48 weeks after operation， observing repair tissue characteristic R