CAJ | 학술논문

背景：软骨细胞外基质具有众多的信号分子蛋白和因子，其成分和特性最接近天然软骨组织，因而其很可能是构建组织工程软骨的最理想原料。 　　目的：探讨海藻酸钙-软骨细胞外基质混合凝胶微球复合人脐带Wharton胶间充质干细胞在裸鼠皮下异位构建组织工程软骨的可行性。 　　方法：制备软骨细胞外基质微丝悬液，将人脐带Wharton胶间充质干细胞接种于海藻酸钙-软骨细胞外基质混合凝胶微球中体外培养后植入裸鼠背部皮下(实验组)，以人脐带 Wharton 胶间充质干细胞混合于单纯藻酸盐凝胶微球作为对照组，于4周后取材进行大体和组织学苏木精-伊红、甲苯胺蓝、番红O及Ⅱ型胶原免疫组化观察。结果与结论：体外培养时微球中干细胞呈球形软骨细胞样形态，生长、增殖情况良好；实验组术后第4周取材可见外形呈类软骨样组织块，甲苯胺蓝、番红O及Ⅱ型胶原免疫组化染色阳性，镜下观察可见大量类软骨样细胞及类软骨陷窝样结构，植入的混合凝胶微球周围组织无明显炎症反应；对照组显示微球部分降解，周围仅有少量炎症细胞及淋巴细胞。结果可见海藻酸钙-软骨细胞外基质具有良好的组织相容性，复合干细胞形成的微球植入裸鼠皮下可以构建为类软骨样组织。
배경：연골세포외기질구유음다적신호분자단백화인자，기성분화특성최접근천연연골조직，인이기흔가능시구건조직공정연골적최이상원료。 　　목적：탐토해조산개-연골세포외기질혼합응효미구복합인제대Wharton효간충질간세포재라서피하이위구건조직공정연골적가행성。 　　방법：제비연골세포외기질미사현액，장인제대Wharton효간충질간세포접충우해조산개-연골세포외기질혼합응효미구중체외배양후식입라서배부피하(실험조)，이인제대 Wharton 효간충질간세포혼합우단순조산염응효미구작위대조조，우4주후취재진행대체화조직학소목정-이홍、갑분알람、번홍O급Ⅱ형효원면역조화관찰。결과여결론：체외배양시미구중간세포정구형연골세포양형태，생장、증식정황량호；실험조술후제4주취재가견외형정류연골양조직괴，갑분알람、번홍O급Ⅱ형효원면역조화염색양성，경하관찰가견대량류연골양세포급류연골함와양결구，식입적혼합응효미구주위조직무명현염증반응；대조조현시미구부분강해，주위부유소량염증세포급림파세포。결과가견해조산개-연골세포외기질구유량호적조직상용성，복합간세포형성적미구식입라서피하가이구건위류연골양조직。
BACKGROUND:Cartilage extracelular matrix with a large number of signaling molecule proteins and factors is likely to be an ideal material for tissue engineering cartilage. OBJECTIVE: To investigate the possibility of calcium alginate and cartilage extracelular matrix combined with microencapsulated stem cels derived from human umbilical cord Wharton’s jely to construct ectopic tissue-engineered cartilage in nude mice. METHODS: Microfilament suspension of the cartilage extracelular matrix was prepared. Human stem cels derived from Wharton’s jely of the umbilical cord were inoculated in to calcium alginate and cartilage extracelular matrix gel microspheres as experimental group. Stem cels derived from human umbilical cord Wharton’s jely were incubated in simple alginate gel microspheres as control group. After in vitro culture, the microspheres wereimplanted into the dorsal subcutaneous tissue of nude mice. Samples were taken after 4 weeks, respectively, for gross and histological observation. RESULTS AND CONCLUSION:The stem cels exhibited paralel-chondrocyte morphology in microspheres, which grew and proliferated quite wel during in vitro culture. A new paralel-cartilaginous tissue was found in the subcutaneous tissue 4 weeks after surgery in the experimental group, and the tissue was positive for hematoxylin-eosin, safranine O, toluidine blue and colagen II. A large number of paralel-chondrocytes and cartilage lacuna-like structures were observed under a microscope with no obvious inflammatory reaction around the microspheres. The control group showed the partial degradation of microspheres, surrounded by only a smal number of inflammatory cels and lymphocytes. Calcium alginate and cartilage extracelular matrix microspheres have a rather good histocompatibility which can be used to construct paralel-cartilaginous tissues by implanting stem cel-microspheric compound into the subcutaneous tissue of nude mice.