scholarly journals Rapamycin-Induced Autophagy Promotes the Chondrogenic Differentiation of Synovium-Derived Mesenchymal Stem Cells in the Temporomandibular Joint in Response to IL-1β

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Wenjing Liu ◽  
Haiyun Luo ◽  
Ruolan Wang ◽  
Yiyuan Kang ◽  
Wenting Liao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Chondrogenic Differentiation ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Cartilage Defects ◽  
Inflammatory Microenvironment ◽  
Inflammatory Conditions ◽  
Cartilage Injuries ◽  
Important Regulator

Cartilage defects in temporomandibular disorders (TMD) lead to chronic pain and seldom heal. Synovium-derived mesenchymal stem cells (SMSCs) exhibit superior chondrogenesis and have become promising seed cells for cartilage tissue engineering. However, local inflammatory conditions that affect the repair of articular cartilage by SMSCs present a challenge, and the specific mechanism through which the function remains unclear. Thus, it is important to explore the chondrogenesis of SMSCs under inflammatory conditions of TMD such that they can be used more effectively in clinical treatment. In this study, we obtained SMSCs from TMD patients with severe cartilage injuries. In response to stimulation with IL-1β, which is well known as one of the most prevalent cytokines in TMD, MMP13 expression increased, while that of SOX9, aggrecan, and collagen II decreased during chondrogenic differentiation. At the same time, IL-1β upregulated the expression of mTOR and decreased the ratio of LC3-II/LC3-I and the formation of autophagosomes. Further study revealed that rapamycin pretreatment promoted the migration of SMSCs and the expression of chondrogenesis-related markers in the presence of IL-1β by inducing autophagy. 3-Benzyl-5-((2-nitrophenoxy)methyl)-dihydrofuran-2(3H)-one (3BDO), a new activator of mTOR, inhibited autophagy and increased the expression of p-GSK3βser9 and β-catenin, simulating the effect of IL-1β stimulation. Furthermore, rapamycin reduced the expression of mTOR, whereas the promotion of LC3-II/LC3-I was blocked by the GSK3β inhibitor TWS119. Taken together, these results indicate that rapamycin enhances the chondrogenesis of SMSCs by inducing autophagy, and GSK3β may be an important regulator in the process of rapamycin-induced autophagy. Thus, inducing autophagy may be a useful approach in the chondrogenic differentiation of SMSCs in the inflammatory microenvironment and may represent a novel TMD treatment.

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 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 Chondrogenic Differentiation of Mesenchymal Stem Cells in Three-Dimensional Chitosan Film Culture

2017 ◽  
Vol 26 (3) ◽  
pp. 417-427 ◽  
Author(s):  
Tsai-Jung Lu ◽  
Fang-Yao Chiu ◽  
Hsiao-Ying Chiu ◽  
Ming-Chau Chang ◽  
Shih-Chieh Hung
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Chondrogenic Differentiation ◽  
Chitosan Film ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Cartilage Defects ◽  
Induction Medium ◽  
Alcian Blue Staining ◽  
Film Culture

Articular cartilage has a very limited capacity for self-repair, and mesenchymal stem cells (MSCs) have the potential to treat cartilage defects and osteoarthritis. However, in-depth mechanistic studies regarding their applications are required. Here we demonstrated the use of chitosan film culture for promoting chondrogenic differentiation of MSCs. We found that MSCs formed spheres 2 days after seeding on dishes coated with chitosan. When MSCs were induced in a chondrogenic induction medium on chitosan films, the size of the spheres continuously increased for up to 21 days. Alcian blue staining and immunohistochemistry demonstrated the expression of chondrogenic proteins, including aggrecan, type II collagen, and type X collagen at 14 and 21 days of differentiation. Importantly, chitosan, with a medium molecular weight (size: 190–310 kDa), was more suitable than other sizes for inducing chondrogenic differentiation of MSCs in terms of sphere size and expression of chondrogenic proteins and endochondral markers. We identified that the mechanistic target of rapamycin (mTOR) signaling and its downstream S6 kinase (S6K)/S6 were activated in chitosan film culture compared to that of monolayer culture. The activation of mTOR/S6K was continuously upregulated from days 2 to 7 of differentiation. Furthermore, we found that mTOR/S6K signaling was required for chondrogenic differentiation of MSCs in chitosan film culture through rapamycin treatment and mTOR knockdown. In conclusion, we showed the suitability of chitosan film culture for promoting chondrogenic differentiation of MSCs and its potential in the development of new strategies in cartilage tissue engineering.

scholarly journals Protective Effects of a Hyaluronan-Binding Peptide (P15-1) on Mesenchymal Stem Cells in an Inflammatory Environment

2021 ◽  
Vol 22 (13) ◽  
pp. 7058
Author(s):  
Thorsten Kirsch ◽  
Fenglin Zhang ◽  
Olivia Braender-Carr ◽  
Mary K. Cowman
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Chondrogenic Differentiation ◽  
Therapeutic Strategy ◽  
Cartilage Defects ◽  
Mrna Levels ◽  
Protective Effects ◽  
Human Bmscs ◽  
Hyaluronan Binding

Mesenchymal stem cells (MSCs) obtained from various sources, including bone marrow, have been proposed as a therapeutic strategy for the improvement of tissue repair/regeneration, including the repair of cartilage defects or lesions. Often the highly inflammatory environment after injury or during diseases, however, greatly diminishes the therapeutic and reparative effectiveness of MSCs. Therefore, the identification of novel factors that can protect MSCs against an inflammatory environment may enhance the effectiveness of these cells in repairing tissues, such as articular cartilage. In this study, we investigated whether a peptide (P15-1) that binds to hyaluronan (HA), a major component of the extracellular matrix of cartilage, protects bone-marrow-derived MSCs (BMSCs) in an inflammatory environment. The results showed that P15-1 reduced the mRNA levels of catabolic and inflammatory markers in interleukin-1beta (IL-1β)-treated human BMSCs. In addition, P15-1 enhanced the attachment of BMSCs to HA-coated tissue culture dishes and stimulated the chondrogenic differentiation of the multipotential murine C3H/10T1/2 MSC line in a micromass culture. In conclusion, our findings suggest that P15-1 may increase the capacity of BMSCs to repair cartilage via the protection of these cells in an inflammatory environment and the stimulation of their attachment to an HA-containing matrix and chondrogenic differentiation.

Effects of Passaging on the Migration Response of Synovium-Derived Stem Cells to an Applied DC Electric Field

2011 ◽  
Author(s):  
Andrea R. Tan ◽  
Elena Alegre-Aguarón ◽  
Divya N. Dujari ◽  
Sonal R. Sampat ◽  
J. Chloë Bulinski ◽  
...  
Keyword(s):  
Stem Cells ◽  
Growth Factor ◽  
Electric Field ◽  
Cartilage Tissue Engineering ◽  
Joint Space ◽  
Cartilage Tissue ◽  
Cartilage Defects ◽  
Cell Sources ◽  
Dc Electric Field ◽  
Migration Response

Strategies for cartilage tissue engineering and repair have recently focused on cell sources from the surrounding joint tissue as an alternative to chondrocytes. Synovium-derived stem cells (SDSCs) are found in the intimal layer of the synovium, the thin overlying capsule surrounding the joint space [1] and have been found to exhibit a greater chondrogenic potential than stem cells from other origins such as bone marrow stem cells or adipose derived stem cells [2–4]. Under directed cues, these cells have been shown to be capable of migrating from the synovium membrane into articular cartilage defects, though the mechanism behind such movement is unclear. As a first step, we have previously shown that SDSCs expanded in 2D monolayer culture in a growth factor cocktail of TGF-β1, FGF, and PDGF-ββ exhibit directed cathodal migration with perpendicular alignment when under the influence of an applied DC electric field [5]. As cellular behavior and response to an external stimulus can change with exposure to growth factors and passage number, we look here to characterize the effects of passaging on the migration response of SDSCs to an applied electric field. We hypothesize that if these cells develop more chondrocyte-like characteristics with growth factor passaging, their response will mimic that which has previously been reported for chondrocytes, notably directed cathodal (negative pole) migration and perpendicular realignment of the long axis to the direction of applied field [6].

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