Enhanced chondrogenic differentiation of equine bone marrow-derived mesenchymal stem cells in zirconia microwell substrata.
Abstract: In human cartilage tissue engineering, three-dimensional zirconia substrata have the potential advantage of producing many uniform cell clusters of controlled size without xenobiotic material, allowing easy clinical application. The objective of this study was to evaluate the possibility of using zirconia porous three-dimensional microwell substrata for chondrogenic differentiation of equine bone marrow-derived mesenchymal stem cells (BMMSCs) in vitro. In regular medium, 8 × 10, 2 × 10, and 5 × 10 equine BMMSCs from five thoroughbred horses were cultured on zirconia microwell substrata for 4 days to allow formation of clusters. The medium was replaced by chondrogenic culture medium. After chondrogenic culture for 7, 14 and 21 days, analysis of collagen type II alpha 1 gene (COL2A1) gene expression and observation of chondrogenic aggregates by scanning electron microscopy (SEM) were performed. SEM showed size-controlled cell clusters and increasing extracellular matrix over time when using 5 × 10 cells. The expression of COL2A1 on day 7 and 14 with 5 × 10 cells was significantly higher than that of conventional pellet culture with 2 × 10 cells. Histological evaluation by immunohistochemical staining for type II collagen (ColII) was performed after chondrogenic culture for 7 days. The clusters showed wide distribution of ColII. The results suggest that the zirconia substrata have the potential to enhance the chondrogenic differentiation of equine BMMSCs, allowing effective equine cartilage tissue engineering without xenobiotic materials.
Copyright © 2019 Elsevier Ltd. All rights reserved.
Publication Date: 2019-07-10 PubMed ID: 31352283DOI: 10.1016/j.rvsc.2019.07.005Google Scholar: Lookup
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- Journal Article
Summary
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This study explored the effectiveness of zirconia microwell substrata in enhancing the chondrogenic (cartilage-forming) differentiation of stem cells derived from horse bone marrow. The research demonstrated that these substrata can potentially be used for effective, xenobiotic-free equine cartilage tissue engineering.
Objective and Methodology
- The goal of the study was to investigate the suitability of utilizing zirconia porous 3D microwell substrata to enhance chondrogenic differentiation of horse-derived bone marrow mesenchymal stem cells (BMMSCs) in vitro.
- The researchers cultured these BMMSCs on zirconia microwell substrata to allow the formation of homogeneous cell clusters.
- The conventional cell culture medium was then replaced by chondrogenic culture medium to encourage cartilage differentiation.
Analysis and Results
- The researchers performed a gene expression analysis on the COL2A1 gene – which is integral to the formation of cartilage – after chondrogenic cultures were maintained for 7, 14 and 21 days.
- Scanning Electron Microscopy (SEM) was used to observe the chondrogenic aggregates. Over time, SEM demonstrated an increase in the extracellular matrix (which plays a key role in tissue functionality and structure) and size-controlled cell clusters when 5 × 10 cells were used.
- Immunohistochemical staining was carried out for type II collagen after chondrogenic culture for 7 days. The results showed a wide distribution of type II collagen, which indicates successful chondrogenic differentiation.
- The experiments culminated in the conclusion that zirconia substrata can potentially enhance the chondrogenic differentiation of horse-derived BMMSCs.
<li COL2A1 expression at day 7 and 14 were notably higher when using 5 × 10 cells than conventional pellet culture with 2 × 10 cells.
Implications
- The findings present a strong case for the potential use of zirconia substrata in equine cartilage tissue engineering. This technique does not involve any xenobiotic substances, which are foreign to the body and can incite an immune response.
- Using a naturally occurring, biocompatible material like zirconia offers a promising direction for cell-based therapeutic strategies for both equine and potentially human cartilage repair.
Cite This Article
APA
Inui T, Haneda S, Sasaki M, Furuoka H, Ito M, Yanagawa M, Hiyama M, Tabata Y, Sasaki N.
(2019).
Enhanced chondrogenic differentiation of equine bone marrow-derived mesenchymal stem cells in zirconia microwell substrata.
Res Vet Sci, 125, 345-350.
https://doi.org/10.1016/j.rvsc.2019.07.005 Publication
Researcher Affiliations
- Department of Clinical Veterinary Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro-shi, Hokkaido 080-8555, Japan.
- Department of Clinical Veterinary Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro-shi, Hokkaido 080-8555, Japan.
- Department of Clinical Veterinary Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro-shi, Hokkaido 080-8555, Japan.
- Department of Clinical Veterinary Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro-shi, Hokkaido 080-8555, Japan.
- Department of Clinical Veterinary Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro-shi, Hokkaido 080-8555, Japan.
- Department of Clinical Veterinary Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro-shi, Hokkaido 080-8555, Japan.
- Department of Clinical Veterinary Science, Yamaguchi University, Yoshida, Yamaguchi city, Yamaguchi 753-8515, Japan.
- Department of Regeneration Science and Engeneering Lab. of Biomaterials, Kyoto University, Kawahara-cho, Seigoin, Sakyo-ku, Kyoto-city, Kyoto 606-8507, Japan.
- Department of Clinical Veterinary Science, Yamaguchi University, Yoshida, Yamaguchi city, Yamaguchi 753-8515, Japan. Electronic address: nsasaki@yamaguchi-u.ac.jp.
MeSH Terms
- Animals
- Cell Differentiation
- Chondrogenesis
- Horses
- Mesenchymal Stem Cells / cytology
- Mesenchymal Stem Cells / metabolism
- Zirconium / chemistry
Citations
This article has been cited 4 times.- Moriwaki T, Tani H, Haga K, Tohyama S. Protocol for production of homogeneous iPSC spheroids and microtissues using the suction technique. STAR Protoc 2025 Jun 20;6(2):103891.
- Nunes FC, Santos SIP, Colnago LA, Hammer P, Ferreira JA, Ambrósio CE, Pallone EMJA. Impact of ZrO(2) Content on the Formation of Sr-Enriched Phosphates in Al(2)O(3)/ZrO(2) Nanocomposites for Bone Tissue Engineering. Materials (Basel) 2024 Apr 19;17(8).
- Moriwaki T, Tani H, Haga K, Morita-Umei Y, Soma Y, Umei TC, Sekine O, Takatsuna K, Kishino Y, Kanazawa H, Fujita J, Fukuda K, Tohyama S, Ieda M. Scalable production of homogeneous cardiac organoids derived from human pluripotent stem cells. Cell Rep Methods 2023 Dec 18;3(12):100666.
- Vidane AS, Nunes FC, Ferreira JA, Fukumasu H, Freitas SH, Pallone EM, Ambrósio CE. Biocompatibility and interaction of porous alumina-zirconia scaffolds with adipose-derived mesenchymal stem cells for bone tissue regeneration. Heliyon 2023 Sep;9(9):e20128.
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