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Polyethylene terephthalate (PET) enhances chondrogenic differentiation of ovine meniscocytes in a hyaluronic acid/polycaprolactone scaffold in vitro

机译:聚对苯二甲酸乙二酯(PET)在透明质酸/聚己内酯支架中增强绵羊半月板细胞的软骨分化

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Purpose The purpose of this study was to assess the effect of polyethylene terephthalate (PET) on proliferation, differentiation, and attachment of ovine meniscocytes seeded in a hyaluronic acid/polycaprolactone biomaterial (BF-1) Methods BF-1 (30 % hyaluronic acid and 70 % polycaprolactone) cylinders with PET (CO-PET) or without PET, were seeded with 2×10 6 ovine meniscus cells. The specimens were harvested in triplets at 12 hours, seven, 14, 21 and 28 days. DNA content was measured to test proliferation, histological analysis for cell morphology, and biochemical assessment of glycosaminoglycan content and RT-PCR for type I and II collagen were used to assess differentiation, with immunohistochemistry as post-translational control. Attachment was evaluated by electronic microscopy at 28 days. Results DNA content was consistent and equal across groups, suggesting no effect of PETon cell proliferation. However, the BF-1 CO-PET showed a higher percentage of cells with spherical morphology which is typical for a chondrocytic phenotype. This biomaterial with PET also showed a higher type II collagen mRNA expression and an eightfold higher GAG-content than the material without PET. Small amounts of type I collagen mRNA expression were present on both materials at all time points. PCR results were confirmed by immunohistochemistry. Conclusion Addition of PET to a hyaluronic acid/polycaprolactone biomaterial enhances a cartilaginous phenotype, increased type II collagen mRNA expression and a higher GAG production in ovine mensicocytes.
机译:目的本研究的目的是评估聚对苯二甲酸乙二醇酯(PET)对透明质酸/聚己内酯生物材料(BF-1)中接种的绵羊半月板细胞的增殖,分化和附着的影响。方法BF-1(30%透明质酸和将装有PET(CO-PET)或不使用PET的70%聚己内酯瓶接种2×10 6个绵羊半月板细胞。在12小时,7、14、21和28天,将样品收集成三胞胎。测量DNA含量以测试增殖,细胞形态的组织学分析,糖胺聚糖含量的生化评估以及I和II型胶原的RT-PCR评估分化,并以免疫组织化学作为翻译后对照。在28天时通过电子显微镜评估附着性。结果各组的DNA含量一致且相等,表明PET对细胞增殖没有影响。但是,BF-1 CO-PET显示出较高百分比的具有球形形态的细胞,这是软骨细胞表型的典型特征。与不含PET的材料相比,这种含PET的生物材料还显示出更高的II型胶原mRNA表达和8倍的GAG含量。在所有时间点上,两种材料均存在少量的I型胶原mRNA表达。 PCR结果通过免疫组织化学证实。结论向透明质酸/聚己内酯生物材料中添加PET可增强绵羊表皮细胞的软骨表型,增加的II型胶原mRNA表达和更高的GAG产生。

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