Modulating the oxidative environment during mesenchymal stem cells chondrogenesis with serum increases collagen accumulation in agarose culture.
Abstract: Chondrogenesis of mesenchymal stem cells (MSCs) is induced in culture conditions that have been associated with oxidative stress, although the extent to which the oxidative environment affects differentiation and extracellular matrix (ECM) accumulation is not known. The objectives of this study were to evaluate the oxidative environment during MSCs chondrogenesis in conventional serum-free medium, and the effect of serum-supplementation on intracellular reactive oxygen species (ROS) and chondrogenesis. Young adult equine MSCs were seeded into agarose and cultured in chondrogenic medium, with or without 5% fetal bovine serum (FBS), for up to 15 days. Samples were evaluated for intracellular ROS, the antioxidant glutathione, ECM and gene expression measures of chondrogenesis, and carbonylation as an indicator of oxidative damage. Intracellular ROS increased with time in culture, and was lower in medium supplemented with FBS. Glutathione decreased ∼12-fold during early chondrogenesis (p < 0.0001), and was not affected by FBS (p = 0.25). After 15 days of culture, FBS supplementation increased hydroxyproline accumulation ∼80% (p = 0.0002); otherwise, measures of chondrogenesis were largely unaffected. Protein carbonylation in chondrogenic MSCs cultures was not significantly different between serum-free and FBS cultures (p = 0.72). Supplementation with adult equine serum increased hydroxyproline accumulation by 45% over serum-free culture (p = 0.0006). In conclusion, this study characterized changes in the oxidative environment during MSC chondrogenesis, and suggested that lowering ROS may be an effective approach to increase collagen accumulation. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:506-514, 2018.
© 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.
Publication Date: 2017-06-26 PubMed ID: 28548680DOI: 10.1002/jor.23618Google Scholar: Lookup
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- Journal Article
Summary
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The research article focused on the effects of oxidative environment on the chondrogenesis, differentiation, and accumulation of extracellular matrix of mesenchymal stem cells. It found that reducing oxidative stress through serum-supplementation could enhance collagen accumulation.
Objective of the Study
- The researchers aimed to investigate the oxidative environment during the chondrogenesis of mesenchymal stem cells (MSCs) in conventional serum-free medium. They also aimed to explore the impact of serum-supplementation on intra-cellular reactive oxygen species (ROS) and chondrogenesis.
Methodology
- MSCs from young adult equine were placed in agarose and cultured in chondrogenic medium, either with or without 5% fetal bovine serum (FBS), for up to 15 days.
- The samples were then evaluated for intra-cellular ROS, the antioxidant glutathione, extracellular matrix (ECM), and the gene expression measures of chondrogenesis.
- The samples were also checked for protein carbonylation as an indicator of oxidative damage.
Findings
- Throughout the duration of culture, intra-cellular ROS increased and was seen to be lower in medium supplemented with FBS.
- Glutathione decreased ∼12-fold during the early stages of chondrogenesis (p < 0.0001), and FBS supplementation did not affect this change (p = 0.25).
- FBS supplementation, post 15 days of culture, resulted in an ∼80% increase in hydroxyproline accumulation (p = 0.0002).
- With the exception of this, the chondrogenesis measurements were largely unaffected.
- Protein carbonylation in chondrogenic MSCs cultures had no significant difference between serum-free and FBS cultures (p = 0.72).
- Supplementation with adult equine serum also increased hydroxyproline accumulation by 45% as compared to serum-free culture (p = 0.0006).
Conclusion
- The study helped understand the changes in the oxidative environment during MSC chondrogenesis.
- The results suggest that reducing ROS could be effective in increasing collagen accumulation. This might aid in developing improved methods for stem cell treatment and research.
Cite This Article
APA
Tangtrongsup S, Kisiday JD.
(2017).
Modulating the oxidative environment during mesenchymal stem cells chondrogenesis with serum increases collagen accumulation in agarose culture.
J Orthop Res, 36(1), 506-514.
https://doi.org/10.1002/jor.23618 Publication
Researcher Affiliations
- Department of Clinical Sciences, Orthopaedic Research Center, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins 80523, Colorado.
- Department of Clinical Sciences, Orthopaedic Research Center, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins 80523, Colorado.
MeSH Terms
- Animals
- Cell Differentiation
- Cells, Cultured
- Chondrogenesis
- Collagen / metabolism
- Culture Media
- Extracellular Matrix / metabolism
- Glutathione / analysis
- Horses
- Mesenchymal Stem Cells / cytology
- Protein Carbonylation
- Reactive Oxygen Species / metabolism
- Sepharose
Citations
This article has been cited 9 times.- Liang R, Yang X, Yew PYM, Sugiarto S, Zhu Q, Zhao J, Loh XJ, Zheng L, Kai D. PLA-lignin nanofibers as antioxidant biomaterials for cartilage regeneration and osteoarthritis treatment.. J Nanobiotechnology 2022 Jul 16;20(1):327.
- Kitching M, Inguva S, Ramani M, Gao Y, Marsili E, Cahill P. Biosynthesis of Gold Nanoparticles by Vascular Cells in vitro.. Front Microbiol 2022;13:813511.
- Komsa-Penkova R, Stavreva G, Belemezova K, Kyurkchiev S, Todinova S, Altankov G. Mesenchymal Stem-Cell Remodeling of Adsorbed Type-I Collagen-The Effect of Collagen Oxidation.. Int J Mol Sci 2022 Mar 11;23(6).
- Salvador-Clavell R, Martín de Llano JJ, Milián L, Oliver M, Mata M, Carda C, Sancho-Tello M. Chondrogenic Potential of Human Dental Pulp Stem Cells Cultured as Microtissues.. Stem Cells Int 2021;2021:7843798.
- Chijimatsu R, Miwa S, Okamura G, Miyahara J, Tachibana N, Ishikura H, Higuchi J, Maenohara Y, Tsuji S, Sameshima S, Takagi K, Nakazato K, Kawaguchi K, Yamagami R, Inui H, Taketomi S, Tanaka S, Saito T. Divergence in chondrogenic potential between in vitro and in vivo of adipose- and synovial-stem cells from mouse and human.. Stem Cell Res Ther 2021 Jul 15;12(1):405.
- Salm A, Krishnan SR, Collu M, Danton O, Hamburger M, Leonti M, Almanza G, Gertsch J. Phylobioactive hotspots in plant resources used to treat Chagas disease.. iScience 2021 Apr 23;24(4):102310.
- Yan W, Diao S, Fan Z. The role and mechanism of mitochondrial functions and energy metabolism in the function regulation of the mesenchymal stem cells.. Stem Cell Res Ther 2021 Feb 17;12(1):140.
- Tangtrongsup S, Kisiday JD. Differential Effects of the Antioxidants N-Acetylcysteine and Pyrrolidine Dithiocarbamate on Mesenchymal Stem Cell Chondrogenesis.. Cell Mol Bioeng 2019 Apr;12(2):153-163.
- Kisiday JD, Schwartz JA, Tangtrongsup S, Goodrich LR, Grande DA. Culture Conditions that Support Expansion and Chondrogenesis of Middle-Aged Rat Mesenchymal Stem Cells.. Cartilage 2020 Jul;11(3):364-373.
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