中华创伤杂志
中華創傷雜誌
중화창상잡지
Chinese Journal of Traumatology
2014年
5期
467-470
,共4页
刘清宇%王富友%刘俊利%孙欣慰%杨柳
劉清宇%王富友%劉俊利%孫訢慰%楊柳
류청우%왕부우%류준리%손흔위%양류
组织工程%软骨%间质干细胞%脱细胞基质
組織工程%軟骨%間質榦細胞%脫細胞基質
조직공정%연골%간질간세포%탈세포기질
Tissue engineering%Cartilage%Mesenchymal stem cells%Acellular matrix
目的 构建含有天然钙化层结构的骨软骨支架,为组织工程修复骨软骨缺损提供一种理想的支架材料. 方法 选择新鲜成年猪膝关节为制备原料,切取软骨层用以制备Ⅱ型胶原水凝胶;环钻钻取直径8 mm包含天然钙化层结构的骨柱,进行脱细胞处理,组织学切片染色观察脱细胞情况;将Ⅱ型胶原水凝胶接种于含天然钙化层结构脱细胞骨柱上,冻干,10/L京尼平乙醇溶液交联,即得含天然钙化层结构的骨软骨模型,进行细胞接种及扫描电镜观察. 结果 含天然钙化层结构的脱细胞骨柱脱细胞效果良好,HE染色、甲苯胺蓝染色、固绿番红O染色及DAPI染色切片显示细胞清除彻底;构建复合支架中Ⅱ型胶原海绵支架空隙率为(91.1±3.8)％,孔径大小为(79.7±17.1)μm;脱细胞骨支架的孔隙率为(73.5±2.6)％,孔径大小为(470.2±158.8) μm;共建骨软骨模型细胞共培养扫描电镜观察可见细胞进入骨软骨模型并且生长良好. 结论 含有天然钙化层结构的骨软骨支架具有良好的生物相容性及合适的孔径及孔隙率,可能成为组织工程骨软骨缺损修复的理想材料.
目的 構建含有天然鈣化層結構的骨軟骨支架,為組織工程脩複骨軟骨缺損提供一種理想的支架材料. 方法 選擇新鮮成年豬膝關節為製備原料,切取軟骨層用以製備Ⅱ型膠原水凝膠;環鑽鑽取直徑8 mm包含天然鈣化層結構的骨柱,進行脫細胞處理,組織學切片染色觀察脫細胞情況;將Ⅱ型膠原水凝膠接種于含天然鈣化層結構脫細胞骨柱上,凍榦,10/L京尼平乙醇溶液交聯,即得含天然鈣化層結構的骨軟骨模型,進行細胞接種及掃描電鏡觀察. 結果 含天然鈣化層結構的脫細胞骨柱脫細胞效果良好,HE染色、甲苯胺藍染色、固綠番紅O染色及DAPI染色切片顯示細胞清除徹底;構建複閤支架中Ⅱ型膠原海綿支架空隙率為(91.1±3.8)％,孔徑大小為(79.7±17.1)μm;脫細胞骨支架的孔隙率為(73.5±2.6)％,孔徑大小為(470.2±158.8) μm;共建骨軟骨模型細胞共培養掃描電鏡觀察可見細胞進入骨軟骨模型併且生長良好. 結論 含有天然鈣化層結構的骨軟骨支架具有良好的生物相容性及閤適的孔徑及孔隙率,可能成為組織工程骨軟骨缺損脩複的理想材料.
목적 구건함유천연개화층결구적골연골지가,위조직공정수복골연골결손제공일충이상적지가재료. 방법 선택신선성년저슬관절위제비원료,절취연골층용이제비Ⅱ형효원수응효;배찬찬취직경8 mm포함천연개화층결구적골주,진행탈세포처리,조직학절편염색관찰탈세포정황;장Ⅱ형효원수응효접충우함천연개화층결구탈세포골주상,동간,10/L경니평을순용액교련,즉득함천연개화층결구적골연골모형,진행세포접충급소묘전경관찰. 결과 함천연개화층결구적탈세포골주탈세포효과량호,HE염색、갑분알람염색、고록번홍O염색급DAPI염색절편현시세포청제철저;구건복합지가중Ⅱ형효원해면지가공극솔위(91.1±3.8)％,공경대소위(79.7±17.1)μm;탈세포골지가적공극솔위(73.5±2.6)％,공경대소위(470.2±158.8) μm;공건골연골모형세포공배양소묘전경관찰가견세포진입골연골모형병차생장량호. 결론 함유천연개화층결구적골연골지가구유량호적생물상용성급합괄적공경급공극솔,가능성위조직공정골연골결손수복적이상재료.
Objective To establish osteochondral scaffolds with remained calcified cartilage zone for finding an ideal scaffold for tissue engineered repair of osteochondral defect.Methods Cartilage zone was harvested from fresh adult porcine knee to fabricate type Ⅱ collagen hydrogel.Bone blocks measuring 8 mm in diameter with calcified cartilage zone were prepared by trephine and acellular treatment was performed.Histological staining was used to identify complete removal of cells.To fabricate the osteochondral models containing calcified cartilage zone,type Ⅱ collagen was seeded onto the acellular bone blocks with calcified cartilage zone,lyophilized,and cross-linked with 10 g/L of genipin ethanol solution.Then cell seeding and scanning electron microscope were performed after the models were established.Results HE staining,toluidine blue staining,solid green staining,safranin O staining,and DAPI staining showed cells were completely removed from bone blocks by decellularized process.Porosity of type Ⅱ collagen sponge was (91.1 ±3.8) ％ and pore size was (79.7 ± 17.1) μm.Porosity of acellular bone blocks was (73.5 ±2.6)％ and pore size was (470.2 ± 158.8) μm.Cells seeded onto osteochondral scaffolds grew well by scanning electron microscope.Conclusion Osteochondral scaffolds with calcified cartilage zone provide good biocompatibility and suitable pore size and porosity and may be an ideal material for repairing osteochondral defect in tissue engineering.
