Chondrogenic Differentiation of Mesenchymal Stem Cells and Its Clinical Applications

Abstract

Tissue engineering has the potential to provide cartilaginous constructs capable of restoring the normal function of native articular cartilage following joint injury or degradation. One approach to functional tissue engineering of cartilage involves the in vitro cultivation of tissue constructs by using: (i) chondrogenic cells that can be selected, expanded, and transfected to overexpress the genes of interest, (ii) scaffolds that provide a defined three-dimensional structure for tissue development and biodegrade at a controlled rate. Understanding the functional potential of the cells and the signaling mechanisms underlying their differentiation should lead to innovative protocols for clinical orthopaedic interventions. A large number of growth factors and hormones have been implicated in the regulation of chondrocyte biology, relatively little is known about the intracellular signaling pathways involved. We have tried to define the roles of specific TGF- dependent signaling pathways involved in the regulation of chondrogenesis from human mesenchymal stem cells. Chondrogenesis induced by TGF-beta3 in alginate bead system was confirmed by examining cartilage specific type II collagen expression and aggrecan, whereas type I collagen expression was not affected by TGF-beta3. Type II collagen mRNA expression was expressed strongly during chondrogenesis and MEK inhibition (U0126) resulted in complete down-regulation of type II collagen. In contrast, aggrecan expression was detected in same level by treatment of U0126. These results strongly suggest that the ERK signaling cascade is involved in TGF-beta3 induced-chondrogenesis signaling pathways and a role of its pathway is necessary over a longer period to promote type II collagen expression. However, their end product properties in vivo have not been well known. In this study, an articular cartilage from chondrogenic MSCs with PLGA scaffolds (75:25 and 65:35) were made and analyzed its biochemical, histological and mechanical properties in vitro and in vivo. And also, we evaluated the cartilage formation in vivo through the injection of cell-thermosensitive gel complex, a newly developed injectable material. At 12 weeks after PLGA scaffolds containing chondrogenic MSCs transplantation, the separated rabbit distal femur showed a good gross articular cartilage appearance in the transplanted site. In indentation test, compare to the native articular cartilage, the engineered cartilage from two types of (75:25 and 65:35) achieved up to 30-60% in mechanical stiffness. And also, a new model for cartilage formation in bladder, at 14 weeks after injection, we could find out mass formation in the submucosal area grossly. H&E staining, alcian blue staining and other special staining confirmed the chondrogenic differentiation in the mass. These cell therapy technologies can provide the possibility of clinical applications for vesicoureteral reflux and reflux esophagitis, and urinary incontinence as well as articular cartilage regeneration.

Ontogeny of rat chondrocyte proliferation: studies in embryo, adult and osteoarthritic (OA) cartilage

“…Therefore, the induction of cell division in adult and OA cartilage remains an open subject. Recent studies point out the possibility of inducing chondrocyte proliferation by using novel techniques such as tissue engineering [37], mesenchymal stem cells [38], and dedifferentiated articular chondrocytes [39], in an attempt to restore the functional chondrocyte population in OA cartilage. These approaches shed more light on this subject.…”

Ex vivo expansion and pluripotential differentiation of cryopreserved human bone marrow mesenchymal stem cells

“…For chondrocytic differentiation in vitro of MSCs, it had been demonstrated that TGF-β was necessary for induction in vitro (Worster et al, 2001;Indrawattana et al, 2004;Lisignoli et al, 2005;Bai et al, 2004;Lee et al, 2004;Chen et al, 2003). In addition, other inducers were supplemented for chondrogenic induction, such as CDMP-1 (Bai et al, 2004), BMP (Indrawattana et al, 2004;Lee et al, 2004), GDF-5 (Lee et al, 2004), IGF-I (Worster et al, 2001;Zhang et al, 2004a;Indrawattana et al, 2004) or dexamethasone (Mackay et al, 1998). TGF-β induced-chondrogenesis was regulated through ERK1/2 signaling pathway.…”

PDGF, TGF-β, and FGF signaling is important for differentiation and growth of mesenchymal stem cells (MSCs): transcriptional profiling can identify markers and signaling pathways important in differentiation of MSCs into adipogenic, chondrogenic, and osteogenic lineages

“…Viability of these cells was checked by trypan blue dye exclusion test. Cells in differentiation medium were also stained with oil Red O (for adipocytes), von Kossa (for osteocytes), and alcian blue 39 (for chondrocytes) on day 14.…”