Synergistic interaction of hTGF-β3 with hBMP-6 promotes articular cartilage formation in chitosan scaffolds with hADSCs: implications for regenerative medicine

Abstract

Background Human TGF-β3 has been used in many studies to induce genes coding for typical cartilage matrix components and accelerate chondrogenic differentiation, making it the standard constituent in most cultivation media used for the assessment of chondrogenesis associated with various stem cell types on carrier matrices. However, in vivo data suggests that TGF-β3 and its other isoforms also induce endochondral and intramembranous osteogenesis in non-primate species to other mammals. Based on previously demonstrated improved articular cartilage induction by a using hTGF-β3 and hBMP-6 together on hADSC cultures and the interaction of TGF- β with matrix in vivo, the present study investigates the interaction of a chitosan scaffold as polyanionic polysaccharide with both growth factors. The study analyzes the difference between chondrogenic differentiation that leads to stable hyaline cartilage and the endochondral ossification route that ends in hypertrophy by extending the usual panel of investigated gene expression and stringent employment of quantitative PCR. Results By assessing the viability, proliferation, matrix formation and gene expression patterns it is shown that hTGF-β3 + hBMP-6 promotes improved hyaline articular cartilage formation in a chitosan scaffold in which ACAN with Col2A1 and not Col1A1 nor Col10A1 where highly expressed both at a transcriptional and translational level. Inversely, hTGF-β3 alone tended towards endochondral bone formation showing according protein and gene expression patterns. Conclusion These findings demonstrate that clinical therapies should consider using hTGF-β3 + hBMP-6 in articular cartilage regeneration therapies as the synergistic interaction of these morphogens seems to ensure and maintain proper hyaline articular cartilage matrix formation counteracting degeneration to fibrous tissue or ossification. These effects are produced by interaction of the growth factors with the polysaccharide matrix.

Differentiation Behaviour of Adipose-Derived Stromal Cells (ASCs) Seeded on Polyurethane-Fibrin Scaffolds In Vitro and In Vivo

“…There was an increase in gene expression of extracellular matrix proteins, such as collagen II and aggrecan, of the constructs treated with chondrogenic differentiation medium and growth factors; however, the increase was only significant for the collagen II expression in the BT and TI groups after 21 days (Figure 5). These results are consistent with the results of others, who described a stimulating effect on the chondrogenic differentiation of ASCs using various growth factor combinations [25,27,[29][30][31][32]55]. Therani et al [31] recently published a study on chondrogenic differentiation of human scalp ASCs using TGF-ß3 and BMP-6 and described effective chondrogenic differentiation with an increase in gene expression of SOX-9, aggrecan, and collagen II over a 14-day experimental period.…”

“…The capacity of ASCs to differentiate into the chondrogenic lineage is lower than that of BMSCs [25][26][27]. Various growth factors and growth factor combinations, including members of the TGF-ß family, as well as platelet-rich plasma (PRP) as an autologous growth factor source [44,53,54], are described in the literature to effectively stimulate ASCs towards a chondrogenic differentiation capacity similar to BMSCs [25,27,[29][30][31][32]55]. In the present in vitro study, different growth factor combinations containing TGF-ß3, BMP-6, and IGF, which are reported to have a beneficial effect on the chondrogenic differentiation potential of ASCs, were assessed.…”

“…Collagen X gene expression was significantly increased in all growth factor groups. Collagen X, a marker of hypertrophy, is an early indicator of hyaline cartilage formed during endochondral ossification and mineralization [27,30]. Ude et al [27] evaluated cartilage differentiation of ASCs and BMSCs with TGF-ß3 alone and with the combination of TGF-ß3 and BMP-6.…”

“…This limited chondrogenic potential of ASCs may be due to reduced expression of the TGF-ß receptor [25] and cell surface marker vascular cell adhesion molecule 1 (CD106) [29]. Thus, in the literature, considerations have been described to overcome this limitation [25,27,30,31]. A more effective stimulation towards chondrogenic differentiation of ASCs similar to BMSCs has been reported to be obtained by, e.g., high doses of BMP-6 [27,29], high concentrations of TGF-ß and IGF [27,32], and the combination of TGF-ß3 with BMP-6 [25,27,30,31].…”

ERK5 Transmits Shear Force Stimulation Signals of Chondrocyte Cultured in an Optimized in Vitro Shear Force Model

“…At the same time, mechanical load stimulation is also an important factor to promote the proliferation and differentiation of chondrocytes. According to reports [21] , one of the keys in cartilage tissue engineering is to nd the appropriate mechanical load, which can simulate joint movement, generate appropriate and coupled mechanical loads and biochemical stimulation to the cartilage, and promote the functional construction of engineered cartilage. For this reason, the cultivation method of using bioreactors as shear force providers has gradually been used widely in orthopedics researches [22] .…”