Induction of chondrogenesis of human placenta-derived mesenchymal stem cells via heparin-grafted human fibroblast derived matrix

Abstract

BackgroundFormation of mature and functional articular cartilage is still challenging in cartilage tissue engineering. This study investigates the potential of using heparin-grafted decellularized extracellular matrix (ECM) as a novel growth factor delivery platform towards human placenta-derived mesenchymal stem cells (hPMSCs) chondrogenic differentiation. Human fibroblast-derived extracellular matrix (hFDM) is naturally obtained from in vitro-cultured human lung fibroblasts via a mild decellularization process. hFDM was then conjugated with heparin via N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC) chemistry and subject to transforming growth factor (TGF)-β1 immobilization. Once heparin grafted-hFDM (hFDM-hep) and hPMSCs were co-embedded into collagen gel, they were examined for in vitro and in vivo chondrogenesis of hPMSCs for 4 weeks.ResultsWe identified heparin moieties on hFDM via toluidine blue O assay and Fourier transform infrared spectroscopy, respectively. We found out that collagen spheroids containing hFDM-hep and TGF-β1 exhibited a sustained release of growth factor for 28 days in vitro. Chondrogenesis of hPMSCs in vitro was supported by accumulated glycosaminoglycan (GAG) content and upregulated chondrogenic specific markers (collagen II, aggrecan, Sox9). Meanwhile, PKH26 - labeled hPMSCs incorporated collagen with either hFDM or hFDM-hep was pre-conditioned in a chondrogenic media for 3 days and subcutaneously implanted in the back of nude mice for 4 weeks. The implanted collagen spheroids containing both hPMSCs and hFDM-hep retained more viable hPMSCs and showed higher level of chondrogenic differentiation, based on immunostaining of collagen type II over collagen alone or Col/hFDM group. In addition, histological examination showed more positive signals of GAG via Safranin-O staining.ConclusionTGF-β1-immobilized hFDM-hep can provide an appropriate microenvironment for chondrogenic differentiation of hPMSCs in 3D collagen spheroid.Electronic supplementary materialThe online version of this article (10.1186/s40824-018-0121-2) contains supplementary material, which is available to authorized users.

Effects of latanoprost on the expression of TGF-β1 and Wnt / β-catenin signaling pathway in the choroid of form-deprivation myopia rats

“…With the onset of myopia with the increase in the rate, there are more and more serious complications caused by high myopia. The main complications of high myopia are scleral staphyloma, retinal hemorrhage, hole, atrophic degeneration, subretinal neovascularization, retinal detachment, glaucoma and cataract, which seriously affect human health (6). TGF -β 1 plays an important role in embryo development, cell differentiation, immune response and tissue regeneration Function (7,8), TGF -β 1 is one of the TGF -β subtypes, which is expressed in sclera, choroid and retina.…”

3D Spheroid Cultures of Stem Cells and Exosome Applications for Cartilage Repair

“…TGF-β1 enhances chondrocyte differentiation early on by modulating proliferation, increasing alkaline phosphatase activity, and increasing proteoglycan synthesis [ 58 ]. TGF-1 can be released in a sustained manner on human fibroblast-derived ECM–hep for up to 28 days, according to Noh et al [ 31 ] indicating that the interaction between heparin and TGF-1 is safe and effective. Some studies have found that incorporating heparin into hydrogels can improve the chondrocyte phenotype or promote the re-differentiation of dedifferentiated chondrocytes [ 59 ].…”

Effects of carrier solutions on the viability and efficacy of canine adipose-derived mesenchymal stem cells

“…cADMSCs stored in Hepa-Sal for 12 h showed the highest expression of both genes compared to other solutions. The higher chondrogenic ability of Hepa-Sal might be associated with the inherent nature of heparin as it ensures the secretion of endogenous growth factors such as transforming growth factor β1, vascular endothelial growth factor, and fibroblast growth factor from the cells, thereby promoting chondrogenesis [61,62]. Besides, several studies have shown that the incorporation of heparin in hydrogel can improve the re-differentiation of differentiated chondrocytes and chondrocyte phenotypes [63,64].…”