Chondrogenic differentiation of human mesenchymal stem cells in collagen type I hydrogels

Abstract

The chondrogenic differentiation of bone marrow-derived human mesenchymal stem cells (MSCs) in a collagen type I hydrogel, which is in clinical use for matrix-based autologous chondrocyte transplantation (ACT), was investigated. Collagen hydrogels with 2.5 x 10(5) MSCs/mL were fabricated and cultured for 3 weeks in a serum-free, defined, chondrogenic differentiation medium containing 10 ng/mL TGF-beta1 or 100 ng/mL BMP-2. Histochemistry revealed morphologically distinct, chondrocyte-like cells, surrounded by a sulfated proteoglycan-rich extracellular matrix in the TGF-beta1 and BMP-2 treated group, with more elongated cells seen in the BMP-2 treated group. Immunohistochemistry detected collagen type II (Col II) in the TGF-beta1 and BMP-2 treated group. Collagen type X (Col X) staining was positive in the TGF-beta1 but only very weak in the BMP-2 treated group. RT-PCR analyses revealed a specific chondrogenic differentiation with the expression of the cartilage specific marker genes Col II, Col X, and aggrecan (AGN) in the TGF-beta1 and the BMP-2 treated group, with earlier expression of these marker genes in the TGF-beta1 treated group. Interestingly, MSC-gels cultured in DMEM with 10% FBS (control) indicated few isolated chondrocyte-like cells but no expression of Col II or Col X could be detected. The results show, that MSCs cultured in a collagen type I hydrogel are able to undergo a distinct chondrogenic differentiation pathway, similar to that described for MSCs cultured in high-density pellet cultures. These findings are valuable in terms of ex vivo predifferentiation or in situ differentiation of MSCs in collagen hydrogels for articular cartilage repair.

Fibroblast Growth Factor-1 Is a Mesenchymal Stromal Cell-Secreted Factor Stimulating Proliferation of Osteoarthritic Chondrocytes in Co-Culture

“…For a long time, BMP-2 has been used as a strong stimulator for osteogenic differentiation of stem cells [35]. In other studies, however, BMP-2 has been applied to promote the chondrogenic differentiation of MSCs as well [24,26]. Meanwhile, it is also believed that BMP-2-induced chondrogenic differentiation of MSCs will eventually lead to hypertrophy and endochondral ossification [36,37].…”

“…In line with increased EdU incorporation in co-culture pellets, 1 cluster contained intracellular cell-cycle regulators, such as CCND1, -2, and -3. The fourth cluster contained two secreted growth factors FGF-1 and BMP-2, both of which are established modulators of chondrocyte proliferation and/or matrix production [24][25][26].…”

Hypoxic Conditions during Expansion Culture Prime Human Mesenchymal Stromal Precursor Cells for Chondrogenic Differentiation in Three-Dimensional Cultures

“…However, Nöth et al reported a contraction of up to 75% in the collagen gels, a process that could be results can be confirmed in in vivo studies, the application of MSCs will be facilitated. Our study does not prodisadvantageous for implantation purposes (33). We chose a low cell concentration for differentiation vide new insights regarding this aspect, but the gelatin hydrogels were found to be a suitable material for either due to the limited nutrient supply expected within the intervertebral disc tissue.…”

“…The gelatin hydrogels presented here as a model for the preparation of tion within a gelatin matrix. (33,54). However, Nöth et al reported a contraction of up to 75% in the collagen gels, a process that could be results can be confirmed in in vivo studies, the application of MSCs will be facilitated.…”

Comparison of Engineered Peptide-Glycosaminoglycan Microfibrous Hybrid Scaffolds for Potential Applications in Cartilage Tissue Regeneration

“…Attempts to mimic this environment by using hydrogel materials has shown promise in restoring functionally competent cartilage because of the inherent cross-linked hydrogel polymer networks which facilitate efficient transport of nutrients and actively form physical environments similar to that of articular cartilage [17]. Natural hydrogel polymers include chitosan, collagen, hyaluronic acid, gelatin and alginate, whereas synthetic polymers encompass poly(glycolic acid) (PGA), poly(lactic acid) (PLA), poly(ethylene oxide) (PEO), poly(vinyl alcohol) (PVA), poly(acrylic acid) (PAA) and their copolymers [18][19][20][21][22][23][24]. Typically, such hydrogel materials can be functionalized or assembled with specific functional groups to impart enhanced biocompatible properties.…”