Chondrogenic differentiation of human articular chondrocytes differs in biodegradable PGA/PLA scaffolds.

Joern Zwingmann, Alexander T Mehlhorn, Norbert Südkamp, Bjoern Stark, Martin Dauner, Hagen Schmal
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引用次数: 63

Abstract

Cartilage tissue engineering is applied clinically to cover and regenerate articular cartilage defects. Two bioresorbable nonwoven scaffolds, polyglycolic acid (PGA) and poly(lactic-co-glycolic acid) (PLGA) (90/10 copolymer of L-lactide and glycolide), were seeded with human chondrocytes after initial progeny in a monolayer with a serum-free medium. Two subgroups of nontreated and plasma-treated (using low-pressure plasma technique) scaffolds were investigated. The constructs were cultivated after seeding in six-well plates with serum-free medium for 7 days and implanted subcutaneously into nude mice for 6 and 12 weeks. Chondrogenic differentiations were investigated using immunhistology and reverse transcriptase-polymerase chain reaction. Cell adhesion only differed from 50% to 65% without a significant difference between the groups. During further cultivation for 7 days, the aggrecan synthesis of the seeded constructs was always higher in the PGA groups (p < 0.05). The mRNA gene expression for collagen type II was significantly higher in the PGA groups after 6 and 12 weeks (p < 0.05). A decrease in the expression of collagen type I was investigated in all groups. The expression for collagen type X and cartilage oligomeric matrix protein (COMP) increased in all groups over time. After cell proliferation in serum-free medium, the long-term chondrogenic differentiation in PGA scaffolds in vitro is cartilage specific and may be utilized in cartilage tissue engineering applications.

可生物降解PGA/PLA支架对人关节软骨细胞成软骨分化的影响。
软骨组织工程在临床上应用于关节软骨缺损的修复和再生。两种生物可吸收的无纺布支架,聚乙醇酸(PGA)和聚乳酸-共乙醇酸(PLGA) (l -丙交酯和乙醇酸酯的90/10共聚物),在初始子代后在无血清培养基中单层接种人软骨细胞。研究了未处理和等离子体处理(使用低压等离子体技术)支架的两个亚组。将构建体接种于无血清培养基的六孔板中培养7天后,分别皮下植入裸鼠6周和12周。采用免疫组织学和逆转录聚合酶链反应研究软骨分化。细胞黏附率仅为50% ~ 65%,各组间无显著差异。在继续培养7 d时,PGA组种子构建体的聚集蛋白合成始终较高(p < 0.05)。6周和12周后,PGA组ⅱ型胶原mRNA基因表达量显著升高(p < 0.05)。在所有组中都观察到I型胶原蛋白表达的减少。随着时间的推移,各组X型胶原蛋白和软骨寡聚基质蛋白(COMP)的表达均增加。细胞在无血清培养基中增殖后,PGA支架的体外长期成软骨分化是软骨特异性的,可用于软骨组织工程应用。
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来源期刊
Tissue engineering
Tissue engineering CELL & TISSUE ENGINEERING-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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