ABSTRACT

Purpose: Cartilage repair following trauma or degeneration is poor, making cell-based therapy an important avenue of treatment. Chondrocytes and mesenchymal stem cells have been extensively studied as potential candidates, although tendency toward hypertrophy and formation of mixed hyaline-fibrocartilage necessitates further optimization. Chondroprogenitors, isolated using fibronectin adhesion assay are reported to show reduced hypertrophy and enhanced chondrogenesis. Laminin, an essential component of extracellular matrix, has been shown to positively modulate chondrocyte proliferation, migration, and survival. The aim of our study was to evaluate the effect of laminin as a differential adhesion assay and obtain an enriched population of chondroprogenitors and assess its efficiency when compared to progenitors obtained via fibronectin.

Materials and methods: Chondrocytes were isolated from three osteoarthritic knee joints and subjected to fibronectin and laminin adhesion to obtain chondroprogenitors. After expansion in culture, they were assessed for differences in their biological characteristics based on growth kinetics, surface marker expression, gene expression for assessing markers of chondrogenesis and hypertrophy, and potential for tri-lineage differentiation.

Results: Our results showed that cells isolated by laminin and fibronectin both displayed comparable characteristics except in terms of proliferative potential (higher in laminin), gene expression of COL2A1 (lower in laminin) and trilineage potential where the laminin group showed higher osteogenic and adipogenic differentiation.

Conclusion: This was the first attempt to successfully isolate human articular cartilage derived chondroprogenitor clones using laminin, which retained stem cell like characteristics. Further evaluation to optimize this method will help enhance chondroprogenitor characteristics, for use in cartilage repair.

摘要

目的：外伤或退化后的软骨修复能力较差，因此细胞疗法成为重要的治疗途径。软骨细胞和间充质干细胞已被广泛研究作为潜在的候选者，尽管需要进一步优化肥大和形成混合透明纤维软骨的趋势。据报道，使用纤连蛋白粘附试验分离的软骨祖细胞显示出肥大减少和软骨形成增强。层粘连蛋白是细胞外基质的重要成分，已被证明可正向调节软骨细胞的增殖、迁移和存活。我们研究的目的是评估层粘连蛋白作为差异粘附试验的效果，并获得丰富的软骨祖细胞群，并与通过纤连蛋白获得的祖细胞相比评估其效率。

材料与方法：从三个骨关节炎膝关节分离软骨细胞，并进行纤连蛋白和层粘连蛋白粘附以获得软骨祖细胞。在培养扩增后，根据生长动力学、表面标记表达、用于评估软骨形成和肥大标记的基因表达以及三谱系分化的潜力，评估它们的生物学特性差异。

结果：我们的结果表明，通过层粘连蛋白和纤连蛋白分离的细胞都显示出相似的特征，除了在增殖潜力（层粘连蛋白中较高）、COL2A1 基因表达（层粘连蛋白中较低）和三系潜能方面，层粘连蛋白组显示出较高的成骨和脂肪形成分化.

结论：这是使用层粘连蛋白成功分离人关节软骨衍生的软骨祖细胞克隆的首次尝试，该克隆保留了干细胞样特征。进一步评估以优化该方法将有助于增强软骨祖细胞的特性，用于软骨修复。