Expression of Collagen Type II and Osteocalcin Genes in Mesenchymal Stem Cells from Rats Treated with Lead acetate II

Background: Lead is one of the sustainable metals with devastating effects on many tissues. This study, examined the adverse effect of lead poisoning on the gene expression of collagen type II and osteocalcin by mesenchymal stem cells (MSCs) cultured in chondrogenic and osteogenic media, respectively. Methods: We used 18 male Wistar rats, divided in 3 groups. In addition to libitum feed as the control, treatment I and treatment II groups were fed by distilled water, distilled water with a dose of 50 ppm lead acetate II and distilled water with a dose of 100 ppm lead acetate II, respectively, over a 2-month period. The MSCs of rat femur were isolated in DMEM medium. After the second passage, the media were replaced separately with chondrogenic and osteogenic media over another 21 days. Then, Collagen Type II and Osteocalcin genes expression were investigated by real time PCR. Results: Collagen Type II and Osteocalcin genes expression in treatments I and II groups showed meaningful decreases compared with that of the control group. Also, the concentration of collagen type II in treatment II group in chondrogenic medium was significantly reduced compared with Osteocalcin concentration in osteogenic medium. Conclusion: We found that poisoning with lead and its accumulation at doses of 50 and 100 ppm in femoral bone marrow of rats decreased the expression of the collagen type II and osteocalcin genes in MSCs and in the chondrogenic and osteogenic media, respectively.

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Auricular Cartilage Regeneration with Adipose-Derived Stem Cells in Rabbits

Mediators of Inflammation ◽

10.1155/2018/4267158 ◽

2018 ◽

Vol 2018 ◽

pp. 1-8 ◽

Cited By ~ 2

Author(s):

Se-Joon Oh ◽

Hee-Young Park ◽

Kyung-Un Choi ◽

Sung-Won Choi ◽

Sung-Dong Kim ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Cartilage Defect ◽

Collagen Type ◽

Cartilage Regeneration ◽

Control Group ◽

Adipose Derived Stem Cells ◽

Type Ii ◽

Collagen Type Ii ◽

Auricular Cartilage

Tissue engineering cell-based therapy using induced pluripotent stem cells and adipose-derived stem cells (ASCs) may be promising tools for therapeutic applications in tissue engineering because of their abundance, relatively easy harvesting, and high proliferation potential. The purpose of this study was to investigate whether ASCs can promote the auricular cartilage regeneration in the rabbit. In order to assess their differentiation ability, ASCs were injected into the midportion of a surgically created auricular cartilage defect in the rabbit. Control group was injected with normal saline. After 1 month, the resected auricles were examined histopathologically and immunohistochemically. The expression of collagen type II and transforming growth factor-β1 (TGF-β1) were analyzed by quantitative polymerase chain reaction. Histopathology showed islands of new cartilage formation at the site of the surgically induced defect in the ASC group. Furthermore, Masson’s trichrome staining and immunohistochemistry for S-100 showed numerous positive chondroblasts. The expression of collagen type II and TGF-β1 were significantly higher in the ASCs than in the control group. In conclusion, ASCs have regenerative effects on the auricular cartilage defect of the rabbit. These effects would be expected to contribute significantly to the regeneration of damaged cartilage tissue in vivo.

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Activin and Nodal Are Not Suitable Alternatives to TGFβ for Chondrogenic Differentiation of Mesenchymal Stem Cells

Cartilage ◽

10.1177/1947603516667585 ◽

2016 ◽

Vol 8 (4) ◽

pp. 432-438 ◽

Cited By ~ 1

Author(s):

Laurie M. G. de Kroon ◽

Esmeralda N. Blaney Davidson ◽

Roberto Narcisi ◽

Eric Farrell ◽

Peter M. van der Kraan ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Chondrogenic Differentiation ◽

Collagen Type ◽

Transforming Growth Factor ◽

Terminal Differentiation ◽

Transforming Growth Factor Β ◽

Type Ii ◽

Collagen Type Ii ◽

Protein Levels

Objective Previously, we demonstrated the importance of transforming growth factor-β (TGFβ)-activated SMAD2/3 signaling in chondrogenesis of bone marrow–derived mesenchymal stem cells (BMSCs). However, TGFβ also signals via the SMAD1/5/9 pathway, which is known to induce terminal differentiation of BMSCs. In this study, we investigated whether other SMAD2/3-activating ligands, Activin and Nodal, can induce chondrogenic differentiation of BMSCs without inducing terminal differentiation. Design Activation of SMAD2/3 signaling and chondrogenesis were evaluated in human BMSCs ( N = 3 donors) stimulated with TGFβ, Activin, or Nodal. SMAD2/3 activation was assessed by determining phosphorylated-SMAD2 (pSMAD2) protein levels and SMAD2/3-target gene expression of SERPINE1. Chondrogenesis was determined by ACAN and COL2A1 transcript analysis and histological examination of proteoglycans and collagen type II. Results Both Activin and TGFβ enhanced pSMAD2 and SERPINE1 expression compared to the control condition without growth factors, demonstrating activated SMAD2/3 signaling. pSMAD2 and SERPINE1 had a higher level of expression following stimulation with TGFβ than with Activin, while Nodal did not activate SMAD2/3 signaling. Of the 3 ligands tested, only TGFβ induced chondrogenic differentiation as shown by strongly increased transcript levels of ACAN and COL2A1 and positive histological staining of proteoglycans and collagen type II. Conclusions Even with concentrations up to 25 times higher than that of TGFβ, Activin and Nodal do not induce chondrogenic differentiation of BMSCs; thus, neither of the 2 ligands is an interesting alternative candidate for TGFβ to induce chondrogenesis without terminal differentiation. To obtain stable cartilage formation by BMSCs, future studies should decipher how TGFβ-induced terminal differentiation can be prevented.

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The Antihypertensive Drug Nifedipine Modulates the Metabolism of Chondrocytes and Human Bone Marrow-Derived Mesenchymal Stem Cells

10.21203/rs.2.10478/v1 ◽

2019 ◽

Author(s):

Ilona Uzieliene ◽

Eiva Bernotiene ◽

Greta Urbonaite ◽

Jaroslav Denkovskij ◽

Edvardas Bagdonas ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Antihypertensive Drug ◽

Mitochondrial Respiration ◽

Collagen Type ◽

Cell Types ◽

Cartilage Tissue ◽

Type Ii ◽

Collagen Type Ii

Abstract Aging is associated with the development of various chronic diseases, in which both hypertension and osteoarthritis (OA) are dominant. Currently, there is no effective treatment for OA, whereas hypertension is often treated using L-type voltage-operated calcium channel (VOCC) blocking drugs, nifedipine being among the most classical ones. Although nifedipine together with other VOCC inhibitors plays an important role in people wellbeing, there are unresolved questions on its possible effect on cartilage tissue homeostasis and the development of OA. Due to that, the aim of this study was to analyse the effects of nifedipine on metabolic processes in human chondrocytes and bone marrow mesenchymal stem cells (BMMSCs). To analyze whether those events were mediated specifically through VOCC, agonist BayK8644 was used. Our results demonstrate that nifedipine downregulated chondrocyte proliferation rate as well as mitochondrial respiration and ATP production (Agilent Seahorse) in both cell types. Analysis of cartilage explant histological sections by electron microscopy also suggested that part of mitochondria lose their activity in response to nifedipine.However, switch of energetic metabolic pathway towards glycolytic was observed only in chondrocytes. Stimulation with either nifedipine or BayK8644 resulted in elevated production of collagen type II and proteoglycans in micromass cultures under chondrogenic condition, although the effects of VOCC inhibitor Bay8466 were less expressed. Nitric oxide (NO) activity, as measured by flow cytometry, was upregulated by nifedipine in BMMSCs and particularly chondrocytes, suggesting that NO at least in part may account for the effects of nifedipine on metabolism in both tested cell types.Taken together, we conclude that antihypertensive drug nifedipine inhibits mitochondrial respiration in both chondrocytes and BMMSCs and that these effects may be associated with increased NO accumulation and pro-inflammatory activity. Glycolytic capacity was enhanced only in chondrocytes, suggesting that these cells have the capacity to switch from oxidative phosphorylation to glycolysis and alter their metabolic activity in response to VOCC inhibition. Finally, nifedipine stimulated production of collagen type II and proteoglycans in both cell types, implying its potentially beneficial anabolic effects on articular cartilage. These results highlight a potential link between consumption of antihypertensive drugs and cartilage health

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Chondrogenic differentiation of bovine bone marrow mesenchymal stem cells (MSCs) in different hydrogels: Influence of collagen type II extracellular matrix on MSC chondrogenesis

Biotechnology and Bioengineering ◽

10.1002/bit.20828 ◽

2006 ◽

Vol 93 (6) ◽

pp. 1152-1163 ◽

Cited By ~ 329

Author(s):

Darko Bosnakovski ◽

Morimichi Mizuno ◽

Gonhyung Kim ◽

Satoshi Takagi ◽

Masahiro Okumura ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Extracellular Matrix ◽

Mesenchymal Stem Cells ◽

Chondrogenic Differentiation ◽

Collagen Type ◽

Bovine Bone ◽

Type Ii ◽

Collagen Type Ii ◽

Marrow Mesenchymal Stem Cells

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P76 Major osteochondral defect (ICRS4 > 10CM2) treated with autologus bone marrow mesenchymal stem cells transplanted in a collagen type II extracellular matrix

Osteoarthritis and Cartilage ◽

10.1016/s1063-4584(07)61431-9 ◽

2007 ◽

Vol 15 ◽

pp. B107 ◽

Cited By ~ 1

Author(s):

R. Mardones ◽

R. Martinez ◽

M.J. Paredes ◽

L. Valenzuela ◽

M. Orrego

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Extracellular Matrix ◽

Mesenchymal Stem Cells ◽

Collagen Type ◽

Osteochondral Defect ◽

Type Ii ◽

Collagen Type Ii ◽

Marrow Mesenchymal Stem Cells

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Integrin α10β1-Selected Mesenchymal Stem Cells Mitigate the Progression of Osteoarthritis in an Equine Talar Impact Model

The American Journal of Sports Medicine ◽

10.1177/0363546519899087 ◽

2020 ◽

Vol 48 (3) ◽

pp. 612-623

Author(s):

Michelle L. Delco ◽

Margaret Goodale ◽

Jan F. Talts ◽

Sarah L. Pownder ◽

Matthew F. Koff ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Synovial Fluid ◽

Prostaglandin E2 ◽

Collagen Type ◽

Type Ii ◽

Collagen Type Ii ◽

Posttraumatic Osteoarthritis ◽

India Ink ◽

Joint Trauma

Background: Early intervention with mesenchymal stem cells (MSCs) after articular trauma has the potential to limit progression of focal lesions and prevent ongoing cartilage degeneration by modulating the joint environment and/or contributing to repair. Integrin α10β1 is the main collagen type II binding receptor on chondrocytes, and MSCs that are selected for high expression of the α10 subunit have improved chondrogenic potential. The ability of α10β1-selected (integrin α10high) MSCs to protect cartilage after injury has not been investigated. Purpose: To investigate integrin α10high MSCs to prevent posttraumatic osteoarthritis in an equine model of impact-induced talar injury. Study Design: Controlled laboratory study. Methods: Focal cartilage injuries were created on the tali of horses (2-5 years, n = 8) by using an impacting device equipped to measure impact stress. Joints were treated with 20 × 106 allogenic adipose-derived α10high MSCs or saline vehicle (control) 4 days after injury. Synovial fluid was collected serially and analyzed for protein content, cell counts, markers of inflammation (prostaglandin E2, tumor necrosis factor α) and collagen homeostasis (procollagen II C-propeptide, collagen type II cleavage product), and glycosaminoglycan content. Second-look arthroscopy was performed at 6 weeks, and horses were euthanized at 6 months. Joints were imaged with radiographs and quantitative 3-T magnetic resonance imaging. Postmortem examinations were performed, and India ink was applied to the talar articular surface to identify areas of cartilage fibrillation. Synovial membrane and osteochondral histology was performed, and immunohistochemistry was used to assess type I and II collagen and lubricin. A mixed effect model with Tukey post hoc and linear contrasts or paired t tests were used, as appropriate. Results: Integrin α10high MSC-treated joints had less subchondral bone sclerosis on radiographs ( P = .04) and histology ( P = .006) and less cartilage fibrillation ( P = .04) as compared with control joints. On gross pathology, less India ink adhered to impact sites in treated joints than in controls, which may be explained by the finding of more prominent lubricin immunostaining in treated joints. Prostaglandin E2 concentration in synovial fluid and mononuclear cell synovial infiltrate were increased in treated joints, suggesting possible immunomodulation by integrin α10high MSCs. Conclusion: Intra-articular administration of integrin α10high MSCs is safe, and evidence suggests that the cells mitigate the effects of joint trauma. Clinical Relevance: This preclinical study indicates that intra-articular therapy with integrin α10high MSCs after joint trauma may be protective against posttraumatic osteoarthritis.

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