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Optimizing Tenogenic Differentiation of Equine Adipose-Derived Mesenchymal Stem Cells (eq-ASC) Using TGFB3 Along with BMP Antagonists
OBJECTIVE: Tendon repair strategies usually are accompanied by pathological mineralization and scar tissue formation that increases the risk of re-injuries. This study aimed to establish an efficient tendon regeneration method simultaneously with a reduced risk of ectopic bone formation. MATERIALS A...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royan Institute
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9428478/ https://www.ncbi.nlm.nih.gov/pubmed/36043405 http://dx.doi.org/10.22074/cellj.2022.7892 |
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author | Shojaee, Asiyeh Ejeian, Fatemeh Parham, Abbas Nasr-Esfahani, Mohammad Hossein |
author_facet | Shojaee, Asiyeh Ejeian, Fatemeh Parham, Abbas Nasr-Esfahani, Mohammad Hossein |
author_sort | Shojaee, Asiyeh |
collection | PubMed |
description | OBJECTIVE: Tendon repair strategies usually are accompanied by pathological mineralization and scar tissue formation that increases the risk of re-injuries. This study aimed to establish an efficient tendon regeneration method simultaneously with a reduced risk of ectopic bone formation. MATERIALS AND METHODS: In this experimental study, tenogenic differentiation was induced through transforming growth factor- β3 (TGFB3) treatment in combination with the inhibiting concentrations of bone morphogenetic proteins (BMP) antagonists, gremlin-2 (GREM2), and a Wnt inhibitor, namely sclerostin (SOST). The procedure’s efficacy was evaluated using real-time polymerase chain reaction (qPCR) for expression analysis of tenogenic markers and osteo- chondrogenic marker genes. The expression level of two tenogenic markers, SCX and MKX, was also evaluated by immunocytochemistry. Sirius Red staining was performed to examine the amounts of collagen fibers. Moreover, to investigate the impact of the substrate on tenogenic differentiation, the nanofibrous scaffolds that highly resemble tendon extracellular matrix was employed. RESULTS: Aggregated features formed in spontaneous normal culture conditions followed by up-regulation of tenogenic and osteogenic marker genes, including SCX, MKX, COL1A1, RUNX2, and CTNNB1. TGFB3 treatment exaggerated morphological changes and markedly amplified tenogenic differentiation in a shorter period of time. Along with TGFB3 treatment, inhibition of BMPs by GREM2 and SOST delayed migratory events to some extent and dramatically reduced osteo-chondrogenic markers synergistically. Nanofibrous scaffolds increased tenogenic markers while declining the expression of osteo-chondrogenic genes. CONCLUSION: These findings revealed an appropriate in vitro potential of spontaneous tenogenic differentiation of eq- ASCs that can be improved by simultaneous activation of TGFB and inhibition of osteoinductive signaling pathways. |
format | Online Article Text |
id | pubmed-9428478 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Royan Institute |
record_format | MEDLINE/PubMed |
spelling | pubmed-94284782022-09-03 Optimizing Tenogenic Differentiation of Equine Adipose-Derived Mesenchymal Stem Cells (eq-ASC) Using TGFB3 Along with BMP Antagonists Shojaee, Asiyeh Ejeian, Fatemeh Parham, Abbas Nasr-Esfahani, Mohammad Hossein Cell J Original Article OBJECTIVE: Tendon repair strategies usually are accompanied by pathological mineralization and scar tissue formation that increases the risk of re-injuries. This study aimed to establish an efficient tendon regeneration method simultaneously with a reduced risk of ectopic bone formation. MATERIALS AND METHODS: In this experimental study, tenogenic differentiation was induced through transforming growth factor- β3 (TGFB3) treatment in combination with the inhibiting concentrations of bone morphogenetic proteins (BMP) antagonists, gremlin-2 (GREM2), and a Wnt inhibitor, namely sclerostin (SOST). The procedure’s efficacy was evaluated using real-time polymerase chain reaction (qPCR) for expression analysis of tenogenic markers and osteo- chondrogenic marker genes. The expression level of two tenogenic markers, SCX and MKX, was also evaluated by immunocytochemistry. Sirius Red staining was performed to examine the amounts of collagen fibers. Moreover, to investigate the impact of the substrate on tenogenic differentiation, the nanofibrous scaffolds that highly resemble tendon extracellular matrix was employed. RESULTS: Aggregated features formed in spontaneous normal culture conditions followed by up-regulation of tenogenic and osteogenic marker genes, including SCX, MKX, COL1A1, RUNX2, and CTNNB1. TGFB3 treatment exaggerated morphological changes and markedly amplified tenogenic differentiation in a shorter period of time. Along with TGFB3 treatment, inhibition of BMPs by GREM2 and SOST delayed migratory events to some extent and dramatically reduced osteo-chondrogenic markers synergistically. Nanofibrous scaffolds increased tenogenic markers while declining the expression of osteo-chondrogenic genes. CONCLUSION: These findings revealed an appropriate in vitro potential of spontaneous tenogenic differentiation of eq- ASCs that can be improved by simultaneous activation of TGFB and inhibition of osteoinductive signaling pathways. Royan Institute 2022-07 2022-07-27 /pmc/articles/PMC9428478/ /pubmed/36043405 http://dx.doi.org/10.22074/cellj.2022.7892 Text en Any use, distribution, reproduction or abstract of this publication in any medium, with the exception of commercial purposes, is permitted provided the original work is properly cited. https://creativecommons.org/licenses/by-nc/3.0/This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial 3.0 (CC BY-NC 3.0) License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Shojaee, Asiyeh Ejeian, Fatemeh Parham, Abbas Nasr-Esfahani, Mohammad Hossein Optimizing Tenogenic Differentiation of Equine Adipose-Derived Mesenchymal Stem Cells (eq-ASC) Using TGFB3 Along with BMP Antagonists |
title | Optimizing Tenogenic Differentiation of Equine Adipose-Derived
Mesenchymal Stem Cells (eq-ASC) Using TGFB3 Along
with BMP Antagonists |
title_full | Optimizing Tenogenic Differentiation of Equine Adipose-Derived
Mesenchymal Stem Cells (eq-ASC) Using TGFB3 Along
with BMP Antagonists |
title_fullStr | Optimizing Tenogenic Differentiation of Equine Adipose-Derived
Mesenchymal Stem Cells (eq-ASC) Using TGFB3 Along
with BMP Antagonists |
title_full_unstemmed | Optimizing Tenogenic Differentiation of Equine Adipose-Derived
Mesenchymal Stem Cells (eq-ASC) Using TGFB3 Along
with BMP Antagonists |
title_short | Optimizing Tenogenic Differentiation of Equine Adipose-Derived
Mesenchymal Stem Cells (eq-ASC) Using TGFB3 Along
with BMP Antagonists |
title_sort | optimizing tenogenic differentiation of equine adipose-derived
mesenchymal stem cells (eq-asc) using tgfb3 along
with bmp antagonists |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9428478/ https://www.ncbi.nlm.nih.gov/pubmed/36043405 http://dx.doi.org/10.22074/cellj.2022.7892 |
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