Mesenchymal stem cells (MSCs) have the capacity for self-renewal and can form bone, fat, and cartilage. Alginate forms a viscous solution when dissolved in 0.9% saline and gels on contact with divalent cations. The viability and phenotype maintenance of chondrocytes in alginate beads have been well documented. However, little is known about the effect of microencapsulation in alginate on chondrogenesis of MSCs. In this study, human MSCs encapsulated in alginate beads were cultured in serum-free medium with the addition of transforming growth factor (TGF)beta1 (10 ng/mL), dexamethasone (10(-7) M), and ascorbate 2-phosphate (50 microg/mL). The MSCs in alginate assumed a rounded morphology with lacunae around them after 1 week in culture. Cell aggregates were observed at 2 weeks or longer in culture. Histological findings agreed with the clinical determination of hyaline cartilage, characterized by isolated cells with ground substance positive in Safranin-O staining and immunohistochemistry for collagen type II at the periphery of cells. Reverse transcriptase-polymerase chain reaction (RT-PCR) revealed the expression of COL2A1 and COL10A1, marker of chondrocytes and hypertrophy chondrocytes, respectively. These results indicate MSCs in alginate can form cartilage and the MSCs-alginate system represents a relevant model for the study of the molecular mechanisms involved in the chondrogenesis and endochondral ossification.
Mesenchymal stem cell (MSC)-based therapies may aid in the repair of articular cartilage defects. The purpose of this study was to investigate the effects of intraarticular injection of allogeneic MSCs in an in vivo anterior cruciate ligament transection (ACLT) model of osteoarthritis in rabbits. Allogeneic bone marrow-derived MSCs were isolated and cultured under hypoxia (1% O2). After 8 weeks following ACLT, MSCs suspended in hyaluronic acid (HA) were injected into the knees, and the contralateral knees were injected with HA alone. Additional controls consisted of a sham operation group as well as an untreated osteoarthritis group. The tissues were analyzed by macroscopic examination as well as histologic and immunohistochemical methods at 6 and 12 weeks post-transplantation. At 6 and 12 weeks, the joint surface showed less cartilage loss and surface abrasion after MSC injection as compared to the tissues receiving HA injection alone. Significantly better histological scores and cartilage content were observed with the MSC transplantation. Furthermore, engraftment of allogenic MSCs were evident in surface cartilage. Thus, injection of the allogeneic MSCs reduced the progression of osteoarthritis in vivo.
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