Assistant Professor, Department of Biochemistry, T.K.M. College of Arts and Science, kollam-5, kerala, India.
Department of Biochemistry, University of Kerala, Thiruvananthapuram, India.
J Biomater Sci Polym Ed. 2024 Dec;35(18):2845-2866. doi: 10.1080/09205063.2024.2391223. Epub 2024 Oct 21.
Cartilage tissue engineering (CTE) is a field of regenerative medicine focused on constructing ideal substitutes for injured cartilage by effectively combining cells, scaffolds, and stimulatory factors. CTE employing chondrocytes and biopolymer-based hydrogels has the potential to repair damaged cartilage. In this research, primary chondrocytes were extracted from the rib cartilage of rats and seeded on a hydrogel construct named HACF, which is made from hydroxyapatite, alginate, chitosan, and fucoidan. We then evaluated chondrogenesis on HACF cartilage construct. The results revealed that the primary chondrocytes were successfully isolated from rat rib cartilage by collagenase D digestion and HACF cartilage construct was effectively synthesized. Chondrocyte viability and its differentiation inside the scaffold HACF were determined by MTT assay, NRU assay, live/dead assay, DAPI nuclear staining, flow cytometry analysis (FCA), mRNA expression studies, and quantification of extracellular matrix components in the HACF scaffold. The findings indicated excellent chondrocyte viability within the HACF scaffold, with no noticeable changes in morphology. Apoptosis was not detected in the chondrocytes cultured on these hydrogels, as confirmed by DAPI staining, live/dead assay, and FCA. This demonstrates that the cells were capable of proliferating, dividing, multiplying, and maintaining their integrity on HACF scaffold. The results also showed more collagen deposition and glycosaminoglycan synthesis showing the good health of chondrocytes on the HACF construct. It indicates that HACF is an ideal scaffold supporting stable cartilage matrix production, highlighting its suitability for cartilage tissue engineering.
软骨组织工程(CTE)是再生医学领域的一个分支,专注于通过有效结合细胞、支架和刺激因子构建理想的受损软骨替代物。采用软骨细胞和基于生物聚合物的水凝胶的 CTE 具有修复受损软骨的潜力。在这项研究中,我们从大鼠肋软骨中提取原代软骨细胞,并将其接种在一种名为 HACF 的水凝胶构建体上,该构建体由羟基磷灰石、海藻酸钠、壳聚糖和褐藻糖胶组成。然后,我们评估了 HACF 软骨构建体上的软骨生成。结果表明,我们成功地通过胶原酶 D 消化从大鼠肋软骨中分离出原代软骨细胞,并有效地合成了 HACF 软骨构建体。通过 MTT 测定法、NRU 测定法、活/死测定法、DAPI 核染色、流式细胞术分析(FCA)、mRNA 表达研究以及 HACF 支架中细胞外基质成分的定量分析,确定了支架内软骨细胞的活力及其分化情况。结果表明,HACF 支架内的软骨细胞活力良好,形态无明显变化。DAPI 染色、活/死测定法和 FCA 证实,在这些水凝胶上培养的软骨细胞没有检测到凋亡。这表明细胞能够在 HACF 支架上增殖、分裂、繁殖并保持其完整性。结果还显示,更多的胶原蛋白沉积和糖胺聚糖合成表明 HACF 构建体上的软骨细胞健康状况良好。这表明 HACF 是一种支持稳定软骨基质生成的理想支架,突出了其在软骨组织工程中的适用性。