【摘要】：[目的]軟骨修復(fù)載體的制備是軟骨組織工程的重要課題。本實(shí)驗(yàn)以小牛皮為原料,通過脫毛-脫細(xì)胞-結(jié)構(gòu)改建的早期工藝探討,掌握最佳制備流程,為后續(xù)工藝做準(zhǔn)備。[方法]將小牛皮分別浸入0.5%dispase溶液和0.25%胰蛋白酶溶液,4℃脫毛72 h,比較脫毛效果。乙酸溶脹,刮除/切除表皮和皮下組織,獲得真皮組織。真皮組織分別浸入0.5%SDS、1%SDS與0.5%TritonX-100進(jìn)行脫細(xì)胞,充分沖洗之后凍干,HE染色比較脫細(xì)胞效果,MTT法進(jìn)行細(xì)胞毒性檢測。應(yīng)用0.25%胰蛋白酶浸泡、0.25%胰蛋白酶浸泡聯(lián)合220 V-250 W-40%功率超聲振蕩、0.5%胰蛋白酶浸泡、0.5%胰蛋白酶浸泡聯(lián)合220 V-250 W-40%功率超聲振蕩四種方法進(jìn)行結(jié)構(gòu)改建,掃描電鏡觀察、MASSON染色、孔隙率檢測比較結(jié)構(gòu)改建效果。[結(jié)果]0.5%dispase溶液和0.25%胰蛋白酶溶液均能夠良好的去除小牛皮的毛發(fā)。0.5%SDS、1%SDS脫細(xì)胞效果基本一致,TritonX-100似有將膠原纖維破壞的征兆,由于低濃度SDS更易于沖洗,后續(xù)實(shí)驗(yàn)采用0.5%SDS脫細(xì)胞。細(xì)胞毒性檢測為0級(jí)。0.5%胰蛋白酶浸泡聯(lián)合220 V-250 W-40%功率超聲是最佳的結(jié)構(gòu)改建方法,改建后的載體膠原纖維光滑、立體感明顯增強(qiáng),孔隙率顯著高于未改建的載體(P0.05)。[結(jié)論]0.25%胰蛋白酶溶液脫毛-0.5%SDS脫細(xì)胞-0.5%胰蛋白酶浸泡聯(lián)合220 V-250 W-40%功率超聲進(jìn)行結(jié)構(gòu)改建,是應(yīng)用小牛皮制備軟骨修復(fù)載體的最佳工藝。
[Abstract]:[objective] the preparation of cartilage repair carrier is an important subject in cartilage tissue engineering. In this experiment, calf skin was used as raw material, and the early process of hair removal, acellular and structural reconstruction was discussed, and the best preparation process was grasped in order to prepare for the follow-up process. [methods] the calf skin was immersed in 0.5%dispase solution and 0.25% trypsin solution respectively, and the hair removal effect was compared at 4 鈩，

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