Vinod Elizabeth, Vinod Francis Deepak, Manickam Amirtham Soosai, Sathishkumar Solomon, Boopalan P R J V C
Department of Physiology, Christian Medical College, Vellore, India - 632002; Centre for Stem Cell Research, Christian Medical College, Vellore, India - 632002.
Department of Anatomy, Christian Medical College, Vellore, India - 632002.
Tissue Cell. 2019 Feb;56:107-113. doi: 10.1016/j.tice.2018.12.006. Epub 2019 Jan 2.
Limited self-restorative ability of the cartilage has necessitated the use of cell and tissue engineering based therapies. Recent advances in the isolation, expansion and characterization of articular cartilage derived chondroprogenitors(CPs) has gained popularity in its role for cartilage repair. Platelet rich plasma (PRP) is a reliable biological scaffold for in-vitro and in-vivo studies with reported therapeutic applications in cartilage and bone pathologies. The aim of this study was to evaluate whether human allogeneic PRP could serve as a biological scaffold for chondroprogenitors (CPs) in cartilage repair. CPs were isolated from the superficial layer of three osteoarthritic knee joints by fibronectin adhesion assay and characterized using flow cytometric analysis. Allogeneic citrated blood was harvested from three subjects to obtain PRP. CPs at a concentration of one million cells per ml were gelled with PRP using calcium chloride. The PRP-CP scaffolds were subjected for adipogeneic, osteogenic, chondrogeneic differentiation and processed for post differentiation-staining studies (Oil Red O, Von Kossa, Alcian blue staining), immunofluorescence (collagen II) and live dead assays (Calcein AM-Ethidium Homodimer). We show that PRP was able to sustain CP cell viability and differentiate towards adipogenic, osteogenic and chondrogenic lineage under appropriate culture conditions. We also noted positive extracellular matrix production in PRP-CP scaffolds cultured without chondrogenic supplementation. Our results suggest that PRP could be a promising bio-active scaffold due to its synergistic effect in supporting cell proliferation, maintaining cell viability and favoring extracellular matrix production. PRP can be used as biological scaffold for the delivery of CPs in cartilage healing.
软骨有限的自我修复能力使得基于细胞和组织工程的治疗方法成为必要。近年来，关节软骨来源的软骨祖细胞（CPs）在分离、扩增和特性鉴定方面取得的进展，使其在软骨修复中的作用日益受到关注。富血小板血浆（PRP）是一种可靠的生物支架，可用于体外和体内研究，在软骨和骨病理学方面具有报道的治疗应用。本研究的目的是评估人异体PRP是否可作为软骨修复中软骨祖细胞（CPs）的生物支架。通过纤连蛋白粘附试验从三个骨关节炎膝关节的表层分离CPs，并使用流式细胞术分析进行特性鉴定。从三名受试者采集异体枸橼酸盐血以获得PRP。将浓度为每毫升一百万个细胞的CPs与PRP用氯化钙凝胶化。对PRP-CP支架进行脂肪生成、成骨、软骨生成分化，并进行分化后染色研究（油红O、冯·科萨、阿尔辛蓝染色）、免疫荧光（胶原蛋白II）和活死检测（钙黄绿素AM-溴化乙锭二聚体）。我们表明，在适当的培养条件下，PRP能够维持CP细胞活力并向脂肪生成、成骨和软骨生成谱系分化。我们还注意到在无软骨生成补充剂培养的PRP-CP支架中有阳性细胞外基质产生。我们的结果表明，PRP因其在支持细胞增殖、维持细胞活力和促进细胞外基质产生方面的协同作用，可能是一种有前景的生物活性支架。PRP可作为软骨愈合中递送CPs的生物支架。
