目的观察皮下植入异体软骨细胞复合异种软骨微粒脱细胞基质(Cartilage microparticle acellular matrix,CMACM)和纤维蛋白胶(Fibrin glue,FG)为支架形成组织工程软骨的可能性。方法制备猪耳廓CMACM,体外培养成年兔的耳软骨细胞,将不同的混合物植人5只成年兔背部皮下。A组:异体软骨细胞复合CMACM和FG;B组:自体软骨细胞复合CMACM和FG;C组:CMACM和FG。将每只兔子背部皮肤均分为6个区,分别植入不同混合物各两个点,以备两次取材。观察并记录皮下植入体的形态变化,分别于植入后8周和12周取材,行组织学检测。结果A组和C组未能形成软骨样组织。B组8周可以形成软骨样组织,周围炎症细胞数量较多;12周时形成的软骨组织成分单一,周围没有炎症反应,类似于正常软骨,且新生的软骨组织中均长有许多小血管,新生软骨的厚度不超过1 mm。结论将同种异体软骨细胞复合CMACM和FG植入皮下,不能形成软骨样组织:而以自体软骨细胞为种子细胞则可以得到软骨样组织,但新生软骨的体积和厚度有限,并且新生的软骨组织中长有许多小血管,可能更有利于新生软骨组织的长期存活。
Objective To evaluate the possibility of formation of subcutaneously implanted tissue-engineered cartilage using allograft chondrocytes mixed with xenogenic cartilage microparticle acellular matrix(CMACM) and fibrin glue(FG)as scaffold.Methods Chondrocytes were isolated from ears of adult New Zealand white rabbits.CMACM from porcine auricle was established and combined with FG and exoteric amplified chondrocytes.The compound was transplanted into the back of rabbits subcutaneously.The experiment was divided into three groups according to compound components。Group A,allograft chondrocytes mixed with CMACM and FG;Group B,autograft chondrocytes mixed with CMACM and FG;Group C with CMACM and FG.Five adult rabbits were used,and six regions were divided on the back of every rabbit.Each rabbit was injected compound of group A,B and C in two regions.Specimens were evaluated macroscopically,then harvested and detected histologically at 8 and 12 weeks after operation.Results In group A and C compounds could not form subcutaneous cartilage tissues.While cartilage tissues formed with group B mixtures at 8 weeks;At 12 weeks,new-formed cartilage tissues seemed even more stabilized without inflammatory around it.The size of new-formed cartilage was obviously shrinked with small vessels growing into it and the thickness of new-formed cartilages was only 1 mm.Conclusion The autogenic chondrocytes combining CMACM and FG matrix is a potential material to the cartilage tissue engineering.The volume and thickness of new-formed cartilage tissue is limited and micro-vessles growing into cartilage may contribute to longer survival for new tissues.