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Chondrogenic effects of exogenous retinoic acid or a retinoic acid receptor antagonist (LE135) on equine chondrocytes and bone marrow-derived mesenchymal stem cells in monolayer culture.
Abstract: To determine effects of various concentrations of retinoic acid (RA) or a synthetic RA receptor antagonist (LE135) on equine chondrocytes or bone marrow-derived equine mesenchymal stem cells (BMDMSCs) in monolayer cultures. Methods: Articular cartilage and BMDMSCs from 5 clinically normal horses. Methods: Monolayers of chondrocytes cultured in standard media and of BMDMSCs cultured in chondrogenic media were treated with RA at concentrations of 0, 0.1, 1, or 10 μM or LE135 at concentrations of 0, 0.1, 1, or 10 μM on day 0. On days 7 and 14, samples were analyzed for DNA concentration, chondrocyte morphology or features consistent with chondrogenesis (ie, chondral morphology [scored from 0 to 4]), and gene expression of collagen type Ia (CI), collagen type II (CII), and aggrecan. Results: Chondrocytes treated with RA had more mature chondral morphology (range of median scores, 3.0 to 4.0) than did untreated controls (range of median scores, 0.5 to 0.5). Chondrocytes treated with LE135 did not sustain chondrocyte morphology. All BMDMSCs had evidence of chondral morphology or high CII:CI ratio. Retinoic acid (1 or 10 μM) or LE135 (10 μM) treatment decreased DNA content of BMDMSC cultures. At 0.1 and 1 μM concentrations, LE135 weakly but significantly increased chondral morphology scores, compared with untreated controls, but lack of aggrecan expression and lack of increased CII:CI ratio, compared with that of controls, did not affect chondrogenesis. Conclusions: RA promoted maturation and hypertrophy in chondrocytes but not BMDMSCs in monolayer cultures. Deficiency or blockade of RA may prevent hypertrophy and maturation of differentiated chondrocytes.
Publication Date: 2011-07-07 PubMed ID: 21728848DOI: 10.2460/ajvr.72.7.884Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- N.I.H.
- Extramural
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research article investigates the effect of retinoic acid or a synthetically-created antagonist on equine chondrocytes (cartilage cells) and bone marrow-derived mesenchymal stem cells. The study found that retinoic acid promoted maturation and growth in chondrocytes but not in stem cells.
Research Methods
The methods of the experiment involved:
- Using articular cartilage and mesenchymal stem cells taken from the bone marrow of five healthy horses.
- Chondrocytes were cultured in a standard medium while mesenchymal stem cells were cultured in a chondrogenic (cartilage-forming) medium.
- The cultured cells were then exposed to either retinoic acid or the LE135 antagonist at varying concentrations.
- Cells were observed and evaluated on day 7 and 14 for DNA concentration, cell morphology and specific gene expression.
Results
The results of the experiment indicated:
- Chondrocytes treated with retinoic acid exhibited more mature cartilage cell morphology compared to untreated controls.
- Chondrocytes treated with the LE135 antagonist did not maintain the typical cartilage cell shape.
- All mesenchymal stem cells exhibited evidence of cartilage cell formation or had a high collagen type II to collagen type I ratio, which is indicative of a shift towards a cartilage cell type.
- Either retinoic acid or the LE135 antagonist resulted in decreased DNA content in the mesenchymal stem cell cultures.
- While the LE135 antagonist managed to slightly increase scores for cartilage cell formation, it did not trigger aggrecan expression and therefore did not affect the process of cartilage formation.
Conclusion
The conclusion drawn from the study is that retinoic acid promotes cell growth and development in chondrocytes, but not in mesenchymal stem cells. In fact, it was determined that either a deficiency or blockage of retinoic acid could inhibit the maturation and hypertrophy of differentiated chondrocytes.
Cite This Article
APA
Henderson SE, Santangelo KS, Bertone AL.
(2011).
Chondrogenic effects of exogenous retinoic acid or a retinoic acid receptor antagonist (LE135) on equine chondrocytes and bone marrow-derived mesenchymal stem cells in monolayer culture.
Am J Vet Res, 72(7), 884-892.
https://doi.org/10.2460/ajvr.72.7.884 Publication
Researcher Affiliations
- Comparative Orthopedic Molecular Medicine and Applied Research Laboratory, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210, USA.
MeSH Terms
- Aggrecans / genetics
- Aggrecans / metabolism
- Animals
- Bone Marrow Cells / cytology
- Bone Marrow Cells / drug effects
- Bone Marrow Cells / metabolism
- Cartilage, Articular / cytology
- Cartilage, Articular / drug effects
- Cell Culture Techniques
- Cell Differentiation
- Chondrocytes / cytology
- Chondrocytes / drug effects
- Chondrogenesis
- Collagen Type I / genetics
- Collagen Type I / metabolism
- Collagen Type II / genetics
- Collagen Type II / metabolism
- Dibenzazepines / metabolism
- Gene Expression Regulation
- Horses / anatomy & histology
- Horses / physiology
- Mesenchymal Stem Cells / cytology
- Mesenchymal Stem Cells / drug effects
- Mesenchymal Stem Cells / metabolism
- Receptors, Retinoic Acid / antagonists & inhibitors
- Receptors, Retinoic Acid / genetics
- Tretinoin / metabolism
- Tretinoin / pharmacology
Grant Funding
- K08 AR049201 / NIAMS NIH HHS
Citations
This article has been cited 6 times.- Hu B, Zou X, Yu Y, Jiang Y, Xu H. METTL3 promotes SMSCs chondrogenic differentiation by targeting the MMP3, MMP13, and GATA3. Regen Ther 2023 Mar;22:148-159.
- Reisbig NA, Pinnell E, Scheuerman L, Hussein H, Bertone AL. Synovium extra cellular matrices seeded with transduced mesenchymal stem cells stimulate chondrocyte maturation in vitro and cartilage healing in clinically-induced rat-knee lesions in vivo. PLoS One 2019;14(3):e0212664.
- Prosser A, Scotchford C, Roberts G, Grant D, Sottile V. Integrated Multi-Assay Culture Model for Stem Cell Chondrogenic Differentiation. Int J Mol Sci 2019 Feb 22;20(4).
- Jie Z, Liang Y, Yi P, Tang H, Soong L, Cong Y, Zhang K, Sun J. Retinoic Acid Regulates Immune Responses by Promoting IL-22 and Modulating S100 Proteins in Viral Hepatitis. J Immunol 2017 May 1;198(9):3448-3460.
- Ham O, Lee CY, Kim R, Lee J, Oh S, Lee MY, Kim J, Hwang KC, Maeng LS, Chang W. Therapeutic Potential of Differentiated Mesenchymal Stem Cells for Treatment of Osteoarthritis. Int J Mol Sci 2015 Jul 2;16(7):14961-78.
- Song H, Chang W, Song BW, Hwang KC. Specific differentiation of mesenchymal stem cells by small molecules. Am J Stem Cells 2012;1(1):22-30.
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