Effects of transforming growth factor-β subtypes on in vitro cartilage production and mineralization of human bone marrow stromal-derived mesenchymal stem cells

Abstract

Human bone marrow stromal-derived mesenchymal stem cells (hBMSCs) will differentiate into chondrocytes in response to defined chondrogenic medium containing transforming growth factor-β (TGFβ). Results in the literature suggest that the three mammalian subtypes of TGFβ (TGFβ1, TGFβ2 and TGFβ3) provoke certain subtype-specific activities. Therefore, the aim of our study was to investigate whether the TGFβ subtypes affect chondrogenic differentiation of in vitro cultured hBMSCs differently. HBMSC pellets were cultured for 5 weeks in chondrogenic media containing either 2.5, 10 or 25 ng/ml of TGFβ1, TGFβ2 or TGFβ3. All TGFβ subtypes showed a comparable dose-response curve, with significantly less cartilage when 2.5 ng/ml was used and no differences between 10 and 25 ng/ml. Four donors with variable chondrogenic capacity were used to evaluate the effect of 10 ng/ml of either TGFβ subtype on cartilage formation. No significant TGFβ subtype-dependent differences were observed in the total amount of collagen or glycosaminoglycans. Cells from a donor with low chondrogenic capacity performed equally badly with all TGFβ subtypes, while a good donor overall performed well. After addition of β-glycerophosphate during the last 2 weeks of culture, the expression of hypertrophy markers was analysed and mineralization was demonstrated by alkaline phosphatase activity and alizarin red staining. No significant TGFβ subtype-dependent differences were observed in expression collagen type X or VEGF secretion. Nevertheless, pellets cultured with TGFβ1 had significantly less mineralization than pellets cultured with TGFβ3. In conclusion, this study suggests that TGFβ subtypes do affect terminal differentiation of in vitro cultured hBMSCs differently.

Growth Differentiation Factor-5 Enhances In Vitro Mesenchymal Stromal Cell Chondrogenesis and Hypertrophy

“…Mueller and Tuan performed an in-depth study investigating the contribution of TGF-b and dexamethasone, both constituents of the in vitro culture medium employed in the current study, concluding that the resultant MSC-derived chondrogenic pellets recapitulate EO in vitro [21]. More recently, Cals et al have demonstrated that individual TGF-b subtypes orchestrate in vitro terminal differentiation [52]. Exploitation of these observations has lead to the in vitro chondrogenic priming of MSCs, followed by subsequent implantation and natural ossification, as an improved method to engineer bone for regenerative applications [24,53].…”

Gene Delivery of TGF-β3 and BMP2 in an MSC-Laden Alginate Hydrogel for Articular Cartilage and Endochondral Bone Tissue Engineering

“…While it is perhaps unsurprising that the delivery of BMP2 appeared to support a more osteogenic phenotype 29,52 , the finding that pTGF-β3 delivery supported a similar phenotype was less expected. It is likely that the combination of the osteogenic stimulus provided by the nHA, with the overexpression of pTGF-β3 led to promotion of hypertrophic/osteogenic differentiation in the MSCs 53 . In contrast, nHA-mediated co-delivery of pTGF-β3 and pBMP2 resulted in suppressed calcification and collagen type X deposition, and promoted a more stable chondrogenic phenotype characterized by increased GAG and collagen type II production.…”

Silk fibroin/cartilage extracellular matrix scaffolds with sequential delivery of TGF-β3 for chondrogenic differentiation of adipose-derived stem cells

“…[14][15][16] Chondrocyte-related growth factors such as transforming growth factor β, 17 bone morphogenetic proteins, 18 and insulin-like growth factor 19 play a critical role in developing an appropriate microenvironment in cartilage tissue engineering. 20 However, the uncontrolled delivery of these factors may decrease their full reparative potential and have unwanted morphological effects. 21,22 To regulate the release of factors, loading microparticle containing factors into the scaffolds 23 and covalently binding the factors to the scaffolds have been used in recent research of chondrogenesis.…”