Extracellular Matrix Laboratory, Department of Morphology, Institute of Biosciences, Universidade Estadual Paulista-UNESP, District of Rubião Júnior S/N, 18618-970, Botucatu, SP, Brazil.
Platelets. 2013;24(3):219-25. doi: 10.3109/09537104.2012.686255. Epub 2012 May 30.
Articular lesions are still a major challenge in orthopedics because of cartilage's poor healing properties. A major improvement in therapeutics was the development of autologous chondrocytes implantation (ACI), a biotechnology-derived technique that delivers healthy autologous chondrocytes after in vitro expansion. To obtain cartilage-like tissue, 3D scaffolds are essential to maintain chondrocyte differentiated status. Currently, bioactive 3D scaffolds are promising as they can deliver growth factors, cytokines, and hormones to the cells, giving them a boost to attach, proliferate, induce protein synthesis, and differentiate. Using mesenchymal stem cells (MSCs) differentiated into chondrocytes, one can avoid cartilage harvesting. Thus, we investigated the potential use of a platelet-lysate-based 3D bioactive scaffold to support chondrogenic differentiation and maintenance of MSCs. The MSCs from adult rabbit bone marrow (n = 5) were cultivated and characterized using three antibodies by flow cytometry. MSCs (1 × 10(5)) were than encapsulated inside 60 µl of a rabbit platelet-lysate clot scaffold and maintained in Dulbecco's Modified Eagle Medium Nutrient Mixture F-12 supplemented with chondrogenic inductors. After 21 days, the MSCs-seeded scaffolds were processed for histological analysis and stained with toluidine blue. This scaffold was able to maintain round-shaped cells, typical chondrocyte metachromatic extracellular matrix deposition, and isogenous group formation. Cells accumulated inside lacunae and cytoplasm lipid droplets were other observed typical chondrocyte features. In conclusion, the usage of a platelet-lysate bioactive scaffold, associated with a suitable chondrogenic culture medium, supports MSCs chondrogenesis. As such, it offers an alternative tool for cartilage engineering research and ACI.
关节病变仍然是骨科领域的一个主要挑战,因为软骨的愈合能力较差。治疗学的一个重大进展是开发自体软骨细胞移植(ACI),这是一种生物技术衍生的技术,可在体外扩增后提供健康的自体软骨细胞。为了获得类软骨组织,3D 支架对于维持软骨细胞分化状态至关重要。目前,生物活性 3D 支架具有很大的应用前景,因为它们可以向细胞输送生长因子、细胞因子和激素,为细胞提供附着、增殖、诱导蛋白质合成和分化的动力。使用分化为软骨细胞的间充质干细胞(MSCs),可以避免软骨采集。因此,我们研究了血小板裂解物 3D 生物活性支架支持软骨细胞分化和维持 MSCs 的潜力。从成年兔骨髓(n=5)中培养和鉴定 MSCs,使用三种抗体通过流式细胞术进行鉴定。然后将 1×10(5)个 MSCs 包裹在 60μl 的兔血小板裂解物凝块支架内,并在含有软骨诱导剂的 Dulbecco's Modified Eagle Medium Nutrient Mixture F-12 中维持。21 天后,对 MSCs 接种的支架进行组织学分析,并进行甲苯胺蓝染色。该支架能够维持圆形细胞、典型的软骨细胞异染细胞外基质沉积和同源群形成。细胞在陷窝内聚集,细胞质内的脂滴是观察到的其他典型软骨细胞特征。总之,使用血小板裂解物生物活性支架结合适当的软骨诱导培养基支持 MSCs 软骨生成。因此,它为软骨工程研究和 ACI 提供了一种替代工具。