CAJ | 학술논문

背景:设计一体化、具有过渡结构的双层支架材料,复合软骨细胞、骨髓间充质细胞,有利于新生的骨与软骨组织之间形成良好界面.目的:模仿自然骨-软骨基质构建复合支架,以软骨细胞和骨髓间充质干细胞为种子细胞,体外观察复合组织的成软骨及成骨能力.方法:制备明胶-硫酸软骨素-透明质酸及明胶-陶瓷化骨多孔复合支架,构建自然骨-软骨基质复合支架,复合兔软骨细胞与骨髓间充质干细胞,分未成骨诱导与成骨诱导两组培养,并进行 MTT、糖胺多糖含量、碱性磷酸酶活性检测,以及苏木精-伊红染色检测.结果与结论:未成骨诱导与成骨诱导两组骨髓间充质干细胞增殖及糖胺多糖含量差异无显著性意义.未成骨诱导组碱性磷酸酶活性缓慢上升,成骨诱导组诱导后碱性磷酸酶活性迅速上升,14 d 时达到稳定状态.两组苏木精-伊红染色结果无明显区别,均已形成含有双层组织的类似骨-软骨样组织,其间可见未降解支架形态,但由于基质形成不完善及支架未完全降解,此种结构不成熟,细胞分布不均匀,支架内部可见散在无细胞区域.证实采用两种细胞与双层结构的支架经体外分层复合能够形成组织工程骨软骨复合组织.
배경:설계일체화、구유과도결구적쌍층지가재료,복합연골세포、골수간충질세포,유리우신생적골여연골조직지간형성량호계면.목적:모방자연골-연골기질구건복합지가,이연골세포화골수간충질간세포위충자세포,체외관찰복합조직적성연골급성골능력.방법:제비명효-류산연골소-투명질산급명효-도자화골다공복합지가,구건자연골-연골기질복합지가,복합토연골세포여골수간충질간세포,분미성골유도여성골유도량조배양,병진행 MTT、당알다당함량、감성린산매활성검측,이급소목정-이홍염색검측.결과여결론:미성골유도여성골유도량조골수간충질간세포증식급당알다당함량차이무현저성의의.미성골유도조감성린산매활성완만상승,성골유도조유도후감성린산매활성신속상승,14 d 시체도은정상태.량조소목정-이홍염색결과무명현구별,균이형성함유쌍층조직적유사골-연골양조직,기간가견미강해지가형태,단유우기질형성불완선급지가미완전강해,차충결구불성숙,세포분포불균균,지가내부가견산재무세포구역.증실채용량충세포여쌍층결구적지가경체외분층복합능구형성조직공정골연골복합조직.
BACKGROUND: To design an integrated and biphasic scaffold that is cocultured with chondrocytes and bone marrow mesenchymal cells is beneficial to form good interface between bone and cartilage tissue. OBJECTIVE: To construct a bilayered scaffold based on imitating natural osteochondral matrix that is cocultured with chondrocyte and bone marrow mesenchymal stem cells as seed cells so as to observe the chondrogenic and osteogenic capacity of the composite tissue. METHODS: Gelatin-chondroitin sulfate-hyaluronic acid and gelatin-ceramic bone porous scaffolds were prepared to be cocultured with chondrocytes and bone marrow mesenchymal stem cells for osteogenic induction or non-osteogenic induction. MTT, glycosaminoglycan, alkaline phosphatase and hematoxylin-eosin staining were detected. RESULTS AND CONCLUSION: MTT results showed the bone marrow mesenchymal stem cel proliferation and glycosaminoglycan contents had no statistical difference between the osteogenic induction and non-osteogenic induction groups. Alkaline phosphatase activity increased gradual y in the non-osteogenic induction group, but increased rapidly in the osteogenic induction group that became stable at 14 days. No significant difference was found in the results of hematoxylin-eosin staining between the two groups. Osteochondral-like tissues were found in the two groups with non-degradable scaffolds. Cel -free regions were scattered within the scaffolds owing to the incomplete matrix formation and scaffold degradation. This study proved that these two kinds of seed cells cocultured with the bilayer scaffold in vitro can form tissue engineering osteochondral composite tissues.