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首页> 外文期刊>Journal of biomedical materials research. Part B, Applied biomaterials. >Biological response of chondrocytes cultured in three-dimensional nanofibrous poly(implied by-caprolactone) scaffolds
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Biological response of chondrocytes cultured in three-dimensional nanofibrous poly(implied by-caprolactone) scaffolds

机译：三维纳米纤维聚己内酯支架培养的软骨细胞的生物学反应

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Nanofibrous materials, by virtue of their morphological similarities to natural extracellular matrix, have been considered as candidate scaffolds for cell delivery in tissue-engineering applications. In this study, we have evaluated a novel, three-dimensional, nanofibrous poly(implied by-caprolactone) (PCL) scaffold composed of electrospun nanofibers for its ability to maintain chondrocytes in a mature functional state. Fetal bovine chondrocytes (FBCs), maintained in vitro between passages 2 to 6, were seeded onto three-dimensional biodegradable PCL nanofibrous scaffolds or as monolayers on standard tissue culture polystyrene (TCPS) as a control substrate. Gene expression analysis by reverse transcription-polymerase chain reaction showed that chondrocytes seeded on the nanofibrous scaffold and maintained in serum-free medium supplemented with ITS +, ascorbate, and dexamethasone continuously maintained their chondrocytic phenotype by expressing cartilage-specific extracellular matrix genes, including collagen types II and IX, aggre-can, and cartilage oligomeric matrix protein. Specifically, expression of the collagen type IIB splice variant transcript, which is indicative of the mature chondrocyte phenotype, was up-regulated. FBCs exhibited either a spindle or round shape on the nanofibrous scaffolds, in contrast to a flat, well-spread morphology seen in monolayer cultures on TCPS. Organized actin stress fibers were only observed in the cytoplasm of cells cultured on TCPS. Histologically, nanofibrous cultures maintained in the supplemented serum-free medium produced more sulfated proteoglycan-rich, cartilaginous matrix than monolayer cultures. In addition to promoting phenotypic differentiation, the nanofibrous scaffold also supported cellular proliferation as evidenced by a 21-fold increase in cell growth over 21 days when the cultures were maintained in serum-containing medium. These results indicate that the biological activities of FBCs are crucially dependent on the architecture of the extracellular scaffolds as well as the composition of the culture medium, and that nanofibrous PCL acts as a biologically preferred scaffold/substrate for proliferation and maintenance of the chondrocytic phenotype. We propose that the PCL nanofibrous structure may be a suitable candidate scaffold for cartilage tissue engineering.
机译：纳米纤维材料由于其与天然细胞外基质的形态相似性，已被认为是组织工程应用中细胞递送的候选支架。在这项研究中，我们评估了一种新颖的三维纳米纤维聚己内酯（PCL）支架，该支架由电纺纳米纤维组成，具有将软骨细胞维持在成熟功能状态的能力。将在第2到6代之间体外维持的胎牛软骨细胞（FBC）播种到三维可生物降解的PCL纳米纤维支架上，或作为单层在标准组织培养聚苯乙烯（TCPS）上作为对照基质。通过逆转录-聚合酶链反应的基因表达分析表明，软骨细胞接种在纳米纤维支架上，并维持在无血清培养基中，该培养基补充有ITS +，抗坏血酸和地塞米松，通过表达软骨特异性细胞外基质基因（包括胶原蛋白）来持续维持其软骨表型II型和IX型，聚集蛋白聚糖和软骨寡聚基质蛋白。具体而言，上调了指示成熟软骨细胞表型的IIB型胶原剪接变体转录物的表达。与在TCPS的单层培养物中观察到的平坦，分布良好的形态相反，FBC在纳米纤维支架上呈现纺锤形或圆形。仅在TCPS上培养的细胞的细胞质中观察到有组织的肌动蛋白应激纤维。组织学上，与单层培养相比，在补充的无血清培养基中维持的纳米纤维培养物产生的硫酸化程度更高的富含蛋白聚糖的软骨基质。除了促进表型分化，当将培养物保持在含血清的培养基中时，纳米纤维支架还支持细胞增殖，这可通过21天的细胞生长增加21倍来证明。这些结果表明，FBCs的生物学活性至关重要地取决于细胞外支架的结构以及培养基的组成，并且纳米纤维PCL可以作为生物学上优选的支架/底物，用于软骨细胞表型的增殖和维持。我们建议PCL纳米纤维结构可能是软骨组织工程的合适候选支架。

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