ABSTRACT
INTRODUCTION: After injuries, the cartilage healing capacity is limited owing to its nature as a particular connective tissue without blood vessels, lymphatics, or nerves. The creation of artificial cartilage tissue mimics the biological properties of native cartilage and can reduce the need for donated tissue. Fibrin is a type of biodegradable scaffold that has great potential in tissue engineering applications. It can become good material for cell adhesion and proliferation in vitro. Therefore, this study aimed to create a cartilage tissue in vitro using umbilical cord-derived mesenchymal stem cells (UCMSC) and growth factor-rich fibrin (GRF) scaffolds. METHODS: UCMSCs were isolated and expanded, and platelet-rich plasma (PRP) preparations were performed following previously published protocols. PRP was activated (aPRP) by a 0.45-µm syringe filter to release growth factors inside the platelets. Each 2.105 of the UCMSCs were suspended in 2 ml of aPRP to make the mixture of MSC and PRP (MSC-PRP). Then, Ca2+ solution was added to this mixture to produce the fibril scaffold with UCMSCs inside. UCMSCs' adhesion and proliferation inside the scaffold were evaluated by observation under inverted microscopy, H-E staining, MTT assays, and scanning electron microscopy (SEM). The fibril structure containing UCMSCs was cultured, and chondrogenesis was induced using commercial chondrogenesis media for 21 days (iMSC-GRF). The differentiation in efficacy toward cartilage was evaluated based on the accumulation of aggrecan (acan), glycosaminoglycans (GAGs), and collagen type II (Col II). RESULTS: The results showed that we successfully created a cartilage tissue with some characteristics that mimic the properties of natural cartilage. The engineered cartilage tissue was positive with some cartilage protein, such as acan, GAG, and Coll II. In vitro cartilage presented some natural chondrocyte-like cells. The artificial cartilage tissue was positive for CD14, CD34, CD90, CD105, and HLA-DR and negative for CD44, CD45, and CD73. CONCLUSION: These results showed that using UCMSCs and growth factor-rich fibril from platelet-rich plasma was feasible to produce engineered cartilage tissue for further experiments or clinical usage.
ABSTRACT
INTRODUCTION: Articular cartilage is limited in self-repair following injuries due to avascular, lymphatic, and nerve absence. Recent treatments for cartilage injuries, such as physical therapy, anti-inflammatory medication, chondrocyte implantation, and joint replacement, still have limitations. This study aimed to evaluate the treatment efficacy of human umbilical cord-derived mesenchymal stem cell sheet (UCMSCS) transplantation in rat models of the osteochondral femoral head defect. METHODS: Models of osteochondral femoral head defect were produced in rats by drilling in order to reach the femoral bone tissue through the cartilage layer. Then, UCMSCS was implanted in the created cartilage lesion. The treatment efficacy was monitored by X-ray imaging. The cartilage regeneration was evaluated based on the hematoxylin and eosin staining, and proteoglycan accumulation was detected by staining Safranin O and Fast Green. The physiological, weight, or movement activity of rats were recorded during the treatment period. RESULTS: UCMSCS transplantation showed positive effects on the cartilage regeneration in osteochondral femoral head defect grade 4 (according to ICRS score/grade). Particularly, after 12 weeks of implantation of UCMSCS, the defect was filled with hyaline cartilage-like cells and accumulated a large density of proteoglycan. The osteochondral defect score significantly increased in the treated rats compared to the untreated rats (11.67 ± 0.6 and 9.67 ± 0.6, respectively) (p < 0.05). The histological score also increased in treated rats compared to untreated rats (21.33 ± 1.53 vs. 18.00 ± 1.00) (p < 0.0001). The accumulation of proteoglycan was higher in treated rats (20.50 ± 2.23) than untreated rats (5.38 ± 0.36) (p < 0.05). There was no change in the physiological activities between treated and untreated rats recorded during the study. CONCLUSION: MSCS transplantation could promote regeneration in advanced cartilage injury.
