Department of Veterinary Sciences, University of Parma, Italy.
Department of Veterinary Sciences, University of Parma, Italy.
Res Vet Sci. 2020 Dec;133:98-105. doi: 10.1016/j.rvsc.2020.08.017. Epub 2020 Sep 6.
In vitro studies have demonstrated that platelet lysate (PL) can serve as an alternative to platelet-rich plasma (PRP) to sustain chondrocyte proliferation and production of extracellular matrix components in chondrocytes. The present study aimed to evaluate the direct effects of PL on equine articular chondrocytes in vitro in order to provide a rationale for in vivo use of PL. An in vitro cell proliferation and de-differentiation model was used: primary articular chondrocytes isolated from horse articular cartilage were cultured at low density under adherent conditions to promote cell proliferation. Chondrocytes were cultured in serum-free medium, 10% foetal bovine serum (FBS) supplemented medium, or in the presence of alginate beads containing 5%, 10% and 20% PL. Cell proliferation and gene expression of relevant chondrocyte differentiation markers were investigated. The proliferative capacity of cultured chondrocytes, was sustained more effectively at certain concentrations of PL as compared to that with FBS. In addition, as opposed to FBS, PL, particularly at percentages of 5% and 10%, could maintain the gene expression pattern of relevant chondrocyte differentiation markers. In particular, 5% PL supplementation showed the best compromise between chondrocyte proliferation capacity and maintenance of differentiation. The results of the present study provide a rationale for using PL as an alternative to FBS for in vitro expansion of chondrocytes for matrix-assisted chondrocyte implantation, construction of 3D scaffolds for tissue engineering, and treatment of damaged articular cartilage.
体外研究表明,血小板裂解液(PL)可以替代富含血小板的血浆(PRP),以维持软骨细胞的增殖和细胞外基质成分的产生。本研究旨在评估 PL 对马关节软骨细胞的直接体外作用,为 PL 的体内应用提供依据。采用体外细胞增殖和去分化模型:从马关节软骨中分离出的原代关节软骨细胞在贴壁条件下以低密度培养,以促进细胞增殖。软骨细胞在无血清培养基、添加 10%胎牛血清(FBS)的培养基或含有 5%、10%和 20%PL 的藻酸盐珠中培养。研究了细胞增殖和相关软骨细胞分化标志物的基因表达。与 FBS 相比,PL 在某些浓度下更有效地维持培养软骨细胞的增殖能力。此外,与 FBS 相反,PL,特别是 5%和 10%,可以维持相关软骨细胞分化标志物的基因表达模式。特别是,5%PL 补充剂在维持软骨细胞增殖能力和分化之间表现出最佳的平衡。本研究结果为使用 PL 替代 FBS 进行体外扩增软骨细胞以用于基质辅助软骨细胞植入、组织工程 3D 支架构建以及治疗受损关节软骨提供了依据。