Tenogenic induction of human adipose-derived stem cells by soluble tendon extracellular matrix: composition and transcriptomic analyses
Abstract Background Tendon healing is clinically challenging largely due to its inferior regenerative capacity. We have previously prepared a soluble, DNA-free, urea-extracted bovine tendon-derived extracellular matrix (tECM) that exhibits strong pro-tenogenic bioactivity on human adipose-derived st...
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| Format: | Article |
| Language: | English |
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BMC
2022-07-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-022-03038-0 |
| _version_ | 1818162736770580480 |
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| author | Ying Rao Chenxian Zhu Hoi Ching Suen Shuting Huang Jinyue Liao Dai Fei Elmer Ker Rocky S. Tuan Dan Wang |
| author_facet | Ying Rao Chenxian Zhu Hoi Ching Suen Shuting Huang Jinyue Liao Dai Fei Elmer Ker Rocky S. Tuan Dan Wang |
| author_sort | Ying Rao |
| collection | DOAJ |
| description | Abstract Background Tendon healing is clinically challenging largely due to its inferior regenerative capacity. We have previously prepared a soluble, DNA-free, urea-extracted bovine tendon-derived extracellular matrix (tECM) that exhibits strong pro-tenogenic bioactivity on human adipose-derived stem cells (hASCs). In this study, we aimed to elucidate the mechanism of tECM bioactivity via characterization of tECM protein composition and comparison of transcriptomic profiles of hASC cultures treated with tECM versus collagen type I (Col1) as a control ECM component. Methods The protein composition of tECM was characterized by SDS-PAGE, hydroxyproline assay, and proteomics analysis. To investigate tECM pro-tenogenic bioactivity and mechanism of action, differentiation of tECM-treated hASC cultures was compared to serum control medium or Col1-treated groups, as assessed via immunofluorescence for tenogenic markers and RNA Sequencing (RNA-Seq). Results Urea-extracted tECM yielded consistent protein composition, including collagens (20% w/w) and at least 17 non-collagenous proteins (< 100 kDa) based on MS analysis. Compared to current literature, tECM included key tendon ECM components that are functionally involved in tendon regeneration, as well as those that are involved in similar principal Gene Ontology (GO) functions (ECM-receptor interaction and collagen formation) and signaling pathways (ECM-receptor interaction and focal adhesion). When used as a cell culture supplement, tECM enhanced hASC proliferation and tenogenic differentiation compared to the Col1 and FBS treatment groups based on immunostaining of tenogenesis-associated markers. Furthermore, RNA-Seq analysis revealed a total of 584 genes differentially expressed among the three culture groups. Specifically, Col1-treated hASCs predominantly exhibited expression of genes and pathways related to ECM-associated processes, while tECM-treated hASCs expressed a mixture of ECM- and cell activity-associated processes, which may explain in part the enhanced proliferation and tenogenic differentiation of tECM-treated hASCs. Conclusions Our findings showed that urea-extracted tECM contained 20% w/w collagens and is significantly enriched with other non-collagenous tendon ECM components. Compared to Col1 treatment, tECM supplementation enhanced hASC proliferation and tenogenic differentiation as well as induced distinct gene expression profiles. These findings provide insights into the potential mechanism of the pro-tenogenic bioactivity of tECM and support the development of future tECM-based approaches for tendon repair. |
| first_indexed | 2024-12-11T16:38:25Z |
| format | Article |
| id | doaj.art-d23f72a4ac794411b796f4898c75df97 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-11T16:38:25Z |
| publishDate | 2022-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-d23f72a4ac794411b796f4898c75df972022-12-22T00:58:23ZengBMCStem Cell Research & Therapy1757-65122022-07-0113112110.1186/s13287-022-03038-0Tenogenic induction of human adipose-derived stem cells by soluble tendon extracellular matrix: composition and transcriptomic analysesYing Rao0Chenxian Zhu1Hoi Ching Suen2Shuting Huang3Jinyue Liao4Dai Fei Elmer Ker5Rocky S. Tuan6Dan Wang7Institute for Tissue Engineering and Regenerative Medicine, The Chinese University of Hong KongInstitute for Tissue Engineering and Regenerative Medicine, The Chinese University of Hong KongSchool of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong KongInstitute for Tissue Engineering and Regenerative Medicine, The Chinese University of Hong KongSchool of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong KongInstitute for Tissue Engineering and Regenerative Medicine, The Chinese University of Hong KongInstitute for Tissue Engineering and Regenerative Medicine, The Chinese University of Hong KongInstitute for Tissue Engineering and Regenerative Medicine, The Chinese University of Hong KongAbstract Background Tendon healing is clinically challenging largely due to its inferior regenerative capacity. We have previously prepared a soluble, DNA-free, urea-extracted bovine tendon-derived extracellular matrix (tECM) that exhibits strong pro-tenogenic bioactivity on human adipose-derived stem cells (hASCs). In this study, we aimed to elucidate the mechanism of tECM bioactivity via characterization of tECM protein composition and comparison of transcriptomic profiles of hASC cultures treated with tECM versus collagen type I (Col1) as a control ECM component. Methods The protein composition of tECM was characterized by SDS-PAGE, hydroxyproline assay, and proteomics analysis. To investigate tECM pro-tenogenic bioactivity and mechanism of action, differentiation of tECM-treated hASC cultures was compared to serum control medium or Col1-treated groups, as assessed via immunofluorescence for tenogenic markers and RNA Sequencing (RNA-Seq). Results Urea-extracted tECM yielded consistent protein composition, including collagens (20% w/w) and at least 17 non-collagenous proteins (< 100 kDa) based on MS analysis. Compared to current literature, tECM included key tendon ECM components that are functionally involved in tendon regeneration, as well as those that are involved in similar principal Gene Ontology (GO) functions (ECM-receptor interaction and collagen formation) and signaling pathways (ECM-receptor interaction and focal adhesion). When used as a cell culture supplement, tECM enhanced hASC proliferation and tenogenic differentiation compared to the Col1 and FBS treatment groups based on immunostaining of tenogenesis-associated markers. Furthermore, RNA-Seq analysis revealed a total of 584 genes differentially expressed among the three culture groups. Specifically, Col1-treated hASCs predominantly exhibited expression of genes and pathways related to ECM-associated processes, while tECM-treated hASCs expressed a mixture of ECM- and cell activity-associated processes, which may explain in part the enhanced proliferation and tenogenic differentiation of tECM-treated hASCs. Conclusions Our findings showed that urea-extracted tECM contained 20% w/w collagens and is significantly enriched with other non-collagenous tendon ECM components. Compared to Col1 treatment, tECM supplementation enhanced hASC proliferation and tenogenic differentiation as well as induced distinct gene expression profiles. These findings provide insights into the potential mechanism of the pro-tenogenic bioactivity of tECM and support the development of future tECM-based approaches for tendon repair.https://doi.org/10.1186/s13287-022-03038-0TendonExtracellular matrixAdipose-derived stem cellsMass spectrometryRNA sequencingBioinformatics |
| spellingShingle | Ying Rao Chenxian Zhu Hoi Ching Suen Shuting Huang Jinyue Liao Dai Fei Elmer Ker Rocky S. Tuan Dan Wang Tenogenic induction of human adipose-derived stem cells by soluble tendon extracellular matrix: composition and transcriptomic analyses Stem Cell Research & Therapy Tendon Extracellular matrix Adipose-derived stem cells Mass spectrometry RNA sequencing Bioinformatics |
| title | Tenogenic induction of human adipose-derived stem cells by soluble tendon extracellular matrix: composition and transcriptomic analyses |
| title_full | Tenogenic induction of human adipose-derived stem cells by soluble tendon extracellular matrix: composition and transcriptomic analyses |
| title_fullStr | Tenogenic induction of human adipose-derived stem cells by soluble tendon extracellular matrix: composition and transcriptomic analyses |
| title_full_unstemmed | Tenogenic induction of human adipose-derived stem cells by soluble tendon extracellular matrix: composition and transcriptomic analyses |
| title_short | Tenogenic induction of human adipose-derived stem cells by soluble tendon extracellular matrix: composition and transcriptomic analyses |
| title_sort | tenogenic induction of human adipose derived stem cells by soluble tendon extracellular matrix composition and transcriptomic analyses |
| topic | Tendon Extracellular matrix Adipose-derived stem cells Mass spectrometry RNA sequencing Bioinformatics |
| url | https://doi.org/10.1186/s13287-022-03038-0 |
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