Hypoxic ADSCs-derived EVs Promote the Proliferation and Chondrogenic Differentiation of Cartilage Stem/progenitor Cells

2020 ◽  
Author(s):  
ke xue ◽  
Yongkang Jiang ◽  
Xiaodie Zhang ◽  
Jun Wu ◽  
Lin Qi ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Progenitor Cells ◽  
Extracellular Vesicles ◽  
Chondrogenic Differentiation ◽  
Cartilage Tissue Engineering ◽  
Biomechanical Analysis ◽  
Cartilage Tissue ◽  
Alginate Hydrogel ◽  
Cell Based Therapy

Abstract Background： Cartilage tissue engineering is a promising option for repairing cartilage defects caused by trauma, inflammation and osteoarthritis, although harvesting a large number of seeding cells with stable phenotypes remains a major challenge. Cartilage stem/progenitor cells (CSPCs) seem to be a promising cell source. Hypoxic extracellular vesicles secreted by mesenchymal stem cells may play a major role in cell-cell and tissue-tissue communication by transporting various RNAs and proteins in mesenchymal stem cell-based therapy. In the current study, we aimed to evaluate the effect of hypoxic adipose-derived stem cells (ADSCs)-derived extracellular vesicles (EVs) on CSPCs proliferation and differentiation. Methods: The characteristics of ADSCs-derived EVs were identified by and flow cytometric analysis. Proliferation, migration, and cartilage-related gene expression of CSPCs were measured with or without the presence of hypoxic ADSCs-derived EVs. The effect of ADSC-derived EVs on CSPCs were evaluated in alginate hydrogel culture, and SEM, histological staining, biochemical and biomechanical analysis were performed to evaluate the effect of hypoxic ADSCs-derived EVs on CSPCs in alginate hydrogel culture. Results: The results indicated that the majority of ADSC-derived EVs exhibited a round-shaped or cup-shaped morphology with a diameter of 40–1000 nm and expressed CD9, CD63, and CD81. CSPCs migration and proliferation were enhanced by hypoxic ADSCs-derived EVs, which also increased the expression of cartilage-related genes. The hypoxic ADSCs-derived EVs induced CSPCs to produce significantly more cartilage matrix and proteoglycan. Conclusions: The present study indicated that hypoxic ADSCs-derived EVs improved the proliferation and chondrogenic differentiation of CSPCs for cartilage tissue engineering.

Spidroin Striped Micropattern Promotes Chondrogenic Differentiation of Human Wharton’s Jelly Mesenchymal Stem Cells

2021 ◽  
Author(s):  
Anggraini Barlian ◽  
Dinda Hani’ah Arum Saputri ◽  
Adriel Hernando ◽  
Ekavianty Prajatelistia ◽  
Hutomo Tanoto
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Chondrogenic Differentiation ◽  
Spider Silk ◽  
Cartilage Tissue Engineering ◽  
Wharton’S Jelly ◽  
Cartilage Tissue ◽  
Type Ii ◽  
Wharton's Jelly

Abstract Cartilage tissue engineering, particularly micropattern, can influence the biophysical properties of mesenchymal stem cells (MSCs) leading to chondrogenesis. In this research, human Wharton’s jelly MSCs (hWJ-MSCs) were grown on a striped micropattern containing spider silk protein (spidroin) from Argiope appensa. This research aims to direct hWJ-MSCs chondrogenesis using micropattern made of spidroin bioink as opposed to fibronectin that often used as the gold standard. Cells were cultured on striped micropattern of 500 µm and 1000 µm width sizes without chondrogenic differentiation medium for 21 days. The immunocytochemistry result showed that spidroin contains RGD sequences and facilitates cell adhesion via integrin β1. Chondrogenesis was observed through the expression of glycosaminoglycan, type II collagen, and SOX9. The result on glycosaminoglycan content proved that 1000 µm was the optimal width to support chondrogenesis. Spidroin micropattern induced significantly higher expression of SOX9 mRNA on day-21 and SOX9 protein was located inside the nucleus starting from day-7. COL2A1 mRNA of spidroin micropattern groups was downregulated on day-21 and collagen type II protein was detected starting from day-14. These results showed that spidroin micropattern enhances chondrogenic markers while maintains long-term upregulation of SOX9, and therefore has the potential as a new method for cartilage tissue engineering.

scholarly journals Silencing Smad7 Potentiates BMP2-induced Chondrogenic Differentiation and Inhibits Endochondral Ossification in Human Synovial Derived Mesenchymal Stem Cells

2020 ◽  
Author(s):  
pengcheng xiao ◽  
Zhenglin Zhu ◽  
Chengcheng Du ◽  
Yongsheng Zeng ◽  
junyi Liao ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Chondrogenic Differentiation ◽  
Endochondral Ossification ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Cartilage Injuries

Abstract Background: Cartilage injuries pose formidable challenges for effective clinical management. Autologous stem cell-based therapies and transgene-enhanced cartilage tissue engineering may open new avenues for the treatment of cartilage injuries. Bone morphogenetic protein 2 (BMP2) is a promising chondrogenic growth factors for transgene-enhanced cartilage tissue engineering. However the BMP2 is failed to maintain a stable chondrogenic phenotype as it also induces robust endochondral ossification. Recently, human synovial derived mesenchymal stem cells (hSMSCs) arouse interested through the poor differentiation potential into osteogenic lineage. Smad7, a protein to antagonizes TGF-β/BMP signaling pathway has been discovered significant in the endochondral ossification. In the present study ,we further explore the effect of downregulate Smad7 in BMP2-induced chondrogenic differentiation of hSMSCs. Methods: hSMSCs were isolated from synovium of human knee joint through adhesion growth. In vitro and in vivo chondrogenic differentiation models of hSMSCs were constructed . Transgenes of BMP2, silencing Smad7 and Smad7 were expressed by adenoviral vectors. The osteogenic differentiation was detected by alkaline phosphatase staining, alizarin red staining. The chondrogenic differentiation was detected by alcian blue staining. Gene expression was determined by reverse transcription and quantitative real-time PCR (RT-qPCR), Immunofluorescence and immunohistochemistry. The subcutaneous stem cell implantation model was established and evaluated by micro-CT , h＆e staining, alcian blue staining and immunohistochemistry assay.Results: Compared to other MSCs, hSMSCs performed less of capability to osteogenic differentiation. But the occurrence of endochondral ossification is still inevasible during BMP2 induced cartilage formation. We found that silencing Smad7 enhanced the BMP2-induced chondrogenic differentiation of hSMSCs in vitro. Also, it leading to much less of hypertrophic differentiation. The subcutaneous stem cells implantation assays demonstrated silencing Smad7 potentiates BMP2-induced cartilage formation and inhibits endochondral ossification. Conclusion: This study strongly suggests that application of hSMSCs , cell scaffolds and silencing Smad7 can potentiate BMP2-induced chondrogenic differentiation and inhibit endochondral ossification. Thus, inhibit the expression of Smad7 in BMP2-induced hSMSCs differentiation may be a new strategy for cartilage tissue engineering.

scholarly journals HIF-1α as a Regulator of BMP2-Induced Chondrogenic Differentiation, Osteogenic Differentiation, and Endochondral Ossification in Stem Cells

2015 ◽  
Vol 36 (1) ◽  
pp. 44-60 ◽  
Author(s):  
Nian Zhou ◽  
Ning Hu ◽  
Jun-Yi Liao ◽  
Liang-Bo Lin ◽  
Chen Zhao ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Cartilage Repair ◽  
Chondrogenic Differentiation ◽  
Endochondral Ossification ◽  
Cartilage Tissue Engineering ◽  
Bone Morphogenetic Protein 2 ◽  
Cartilage Tissue ◽  
Cartilage Formation

Background/Aims: Joint cartilage defects are difficult to treat due to the limited self-repair capacities of cartilage. Cartilage tissue engineering based on stem cells and gene enhancement is a potential alternative for cartilage repair. Bone morphogenetic protein 2 (BMP2) has been shown to induce chondrogenic differentiation in mesenchymal stem cells (MSCs); however, maintaining the phenotypes of MSCs during cartilage repair since differentiation occurs along the endochondral ossification pathway. In this study, hypoxia inducible factor, or (HIF)-1α, was determined to be a regulator of BMP2-induced chondrogenic differentiation, osteogenic differentiation, and endochondral bone formation. Methods: BMP2 was used to induce chondrogenic and osteogenic differentiation in stem cells and fetal limb development. After HIF-1α was added to the inducing system, any changes in the differentiation markers were assessed. Results: HIF-1α was found to potentiate BMP2-induced Sox9 and the expression of chondrogenesis by downstream markers, and inhibit Runx2 and the expression of osteogenesis by downstream markers in vitro. In subcutaneous stem cell implantation studies, HIF-1α was shown to potentiate BMP2-induced cartilage formation and inhibit endochondral ossification during ectopic bone/cartilage formation. In the fetal limb culture, HIF-1α and BMP2 synergistically promoted the expansion of the proliferating chondrocyte zone and inhibited chondrocyte hypertrophy and endochondral ossification. Conclusion: The results of this study indicated that, when combined with BMP2, HIF-1α induced MSC differentiation could become a new method of maintaining cartilage phenotypes during cartilage tissue engineering.

scholarly journals Mesenchymal stem cells loaded on 3D-printed gradient poly(ε-caprolactone)/methacrylated alginate composite scaffolds for cartilage tissue engineering

2021 ◽  
Vol 8 (3) ◽  
Author(s):  
Yanyan Cao ◽  
Peng Cheng ◽  
Shengbo Sang ◽  
Chuan Xiang ◽  
Yang An ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Morphology ◽  
Chondrogenic Differentiation ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Composite Scaffolds ◽  
Individual Layer ◽  
3D Printed

Abstract Cartilage has limited self-repair ability due to its avascular, alymphatic and aneural features. The combination of three-dimensional (3D) printing and tissue engineering provides an up-and-coming approach to address this issue. Here, we designed and fabricated a tri-layered (superficial layer (SL), middle layer (ML) and deep layer (DL)) stratified scaffold, inspired by the architecture of collagen fibers in native cartilage tissue. The scaffold was composed of 3D printed depth-dependent gradient poly(ε-caprolactone) (PCL) impregnated with methacrylated alginate (ALMA), and its morphological analysis and mechanical properties were tested. To prove the feasibility of the composite scaffolds for cartilage regeneration, the viability, proliferation, collagen deposition and chondrogenic differentiation of embedded rat bone marrow mesenchymal stem cells (BMSCs) in the scaffolds were assessed by Live/dead assay, CCK-8, DNA content, cell morphology, immunofluorescence and real-time reverse transcription polymerase chain reaction. BMSCs-loaded gradient PCL/ALMA scaffolds showed excellent cell survival, cell proliferation, cell morphology, collagen II deposition and hopeful chondrogenic differentiation compared with three individual-layer scaffolds. Hence, our study demonstrates the potential use of the gradient PCL/ALMA construct for enhanced cartilage tissue engineering.

scholarly journals Cartilage Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells in Three-Dimensional Silica Nonwoven Fabrics

2018 ◽  
Vol 8 (8) ◽  
pp. 1398 ◽  
Author(s):  
Shohei Ishikawa ◽  
Kazutoshi Iijima ◽  
Kohei Sasaki ◽  
Mineo Hashizume ◽  
Masaaki Kawabe ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Bone Marrow ◽  
Chondrogenic Differentiation ◽  
Cartilage Tissue Engineering ◽  
Three Dimensional ◽  
Cartilage Tissue ◽  
Nonwoven Fabrics

In cartilage tissue engineering, three-dimensional (3D) scaffolds provide native extracellular matrix (ECM) environments that induce tissue ingrowth and ECM deposition for in vitro and in vivo tissue regeneration. In this report, we investigated 3D silica nonwoven fabrics (Cellbed®) as a scaffold for mesenchymal stem cells (MSCs) in cartilage tissue engineering applications. The unique, highly porous microstructure of 3D silica fabrics allows for immediate cell infiltration for tissue repair and orientation of cell–cell interaction. It is expected that the morphological similarity of silica fibers to that of fibrillar ECM contributes to the functionalization of cells. Human bone marrow-derived MSCs were cultured in 3D silica fabrics, and chondrogenic differentiation was induced by culture in chondrogenic differentiation medium. The characteristics of chondrogenic differentiation including cellular growth, ECM deposition of glycosaminoglycan and collagen, and gene expression were evaluated. Because of the highly interconnected network structure, stiffness, and permeability of the 3D silica fabrics, the level of chondrogenesis observed in MSCs seeded within was comparable to that observed in MSCs maintained on atelocollagen gels, which are widely used to study the chondrogenesis of MSCs in vitro and in vivo. These results indicated that 3D silica nonwoven fabrics are a promising scaffold for the regeneration of articular cartilage defects using MSCs, showing the particular importance of high elasticity.

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