Potential of Soluble Decellularized Extracellular Matrix for Musculoskeletal Tissue Engineering – Comparison of Various Mesenchymal Tissues
BackgroundIt is well studied that preparations of decellularized extracellular matrix (ECM) obtained from mesenchymal tissues can function as biological scaffolds to regenerate injured musculoskeletal tissues. Previously, we reported that soluble decellularized ECMs derived from meniscal tissue demo...
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Frontiers Media S.A.
2020-11-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2020.581972/full |
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| author | Hiroto Hanai George Jacob George Jacob Shinichi Nakagawa Rocky S. Tuan Rocky S. Tuan Norimasa Nakamura Norimasa Nakamura Norimasa Nakamura Kazunori Shimomura |
| author_facet | Hiroto Hanai George Jacob George Jacob Shinichi Nakagawa Rocky S. Tuan Rocky S. Tuan Norimasa Nakamura Norimasa Nakamura Norimasa Nakamura Kazunori Shimomura |
| author_sort | Hiroto Hanai |
| collection | DOAJ |
| description | BackgroundIt is well studied that preparations of decellularized extracellular matrix (ECM) obtained from mesenchymal tissues can function as biological scaffolds to regenerate injured musculoskeletal tissues. Previously, we reported that soluble decellularized ECMs derived from meniscal tissue demonstrated excellent biocompatibility and produced meniscal regenerate with native meniscal anatomy and biochemical characteristics. We therefore hypothesized that decellularized mesenchymal tissue ECMs from various mesenchymal tissues should exhibit tissue-specific bioactivity. The purpose of this study was to test this hypothesis using porcine tissues, for potential applications in musculoskeletal tissue engineering.MethodsNine types of porcine tissue, including cartilage, meniscus, ligament, tendon, muscle, synovium, fat pad, fat, and bone, were decellularized using established methods and solubilized. Although the current trend is to develop tissue specific decellularization protocols, we selected a simple standard protocol across all tissues using Triton X-100 and DNase/RNase after mincing to compare the outcome. The content of sulfated glycosaminoglycan (sGAG) and hydroxyproline were quantified to determine the biochemical composition of each tissue. Along with the concentration of several growth factors, known to be involved in tissue repair and/or maturation, including bFGF, IGF-1, VEGF, and TGF-β1. The effect of soluble ECMs on cell differentiation was explored by combining them with 3D collagen scaffold culturing human synovium derived mesenchymal stem cells (hSMSCs).ResultsThe decellularization of each tissue was performed and confirmed both histologically [hematoxylin and eosin (H&E) and 4’,6-diamidino-2-phenylindole (DAPI) staining] and on the basis of dsDNA quantification. The content of hydroxyproline of each tissue was relatively unchanged during the decellularization process when comparing the native and decellularized tissue. Cartilage and meniscus exhibited a significant decrease in sGAG content. The content of hydroxyproline in meniscus-derived ECM was the highest when compared with other tissues, while sGAG content in cartilage was the highest. Interestingly, a tissue-specific composition of most of the growth factors was measured in each soluble decellularized ECM and specific differentiation potential was particularly evident in cartilage, ligament and bone derived ECMs.ConclusionIn this study, soluble decellularized ECMs exhibited differences based on their tissue of origin and the present results are important going forward in the field of musculoskeletal regeneration therapy. |
| first_indexed | 2024-12-12T18:45:42Z |
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| last_indexed | 2024-12-12T18:45:42Z |
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| spelling | doaj.art-d94111f71655413d89be2425485e66562022-12-22T00:15:32ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2020-11-01810.3389/fcell.2020.581972581972Potential of Soluble Decellularized Extracellular Matrix for Musculoskeletal Tissue Engineering – Comparison of Various Mesenchymal TissuesHiroto Hanai0George Jacob1George Jacob2Shinichi Nakagawa3Rocky S. Tuan4Rocky S. Tuan5Norimasa Nakamura6Norimasa Nakamura7Norimasa Nakamura8Kazunori Shimomura9Department of Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, JapanDepartment of Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, JapanDepartment of Orthopaedics, Tejasvini Hospital, Mangalore, IndiaDepartment of Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, JapanCenter for Cellular and Molecular Engineering, Department of Orthopaedic Surgery, University of Pittsburgh, Pittsburgh, PA, United StatesInstitute for Tissue Engineering and Regenerative Medicine, The Chinese University of Hong Kong, Hong Kong, ChinaDepartment of Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, JapanInstitute for Medical Science in Sports, Osaka Health Science University, Osaka, JapanGlobal Center for Medical Engineering and Informatics, Osaka University, Suita, JapanDepartment of Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, JapanBackgroundIt is well studied that preparations of decellularized extracellular matrix (ECM) obtained from mesenchymal tissues can function as biological scaffolds to regenerate injured musculoskeletal tissues. Previously, we reported that soluble decellularized ECMs derived from meniscal tissue demonstrated excellent biocompatibility and produced meniscal regenerate with native meniscal anatomy and biochemical characteristics. We therefore hypothesized that decellularized mesenchymal tissue ECMs from various mesenchymal tissues should exhibit tissue-specific bioactivity. The purpose of this study was to test this hypothesis using porcine tissues, for potential applications in musculoskeletal tissue engineering.MethodsNine types of porcine tissue, including cartilage, meniscus, ligament, tendon, muscle, synovium, fat pad, fat, and bone, were decellularized using established methods and solubilized. Although the current trend is to develop tissue specific decellularization protocols, we selected a simple standard protocol across all tissues using Triton X-100 and DNase/RNase after mincing to compare the outcome. The content of sulfated glycosaminoglycan (sGAG) and hydroxyproline were quantified to determine the biochemical composition of each tissue. Along with the concentration of several growth factors, known to be involved in tissue repair and/or maturation, including bFGF, IGF-1, VEGF, and TGF-β1. The effect of soluble ECMs on cell differentiation was explored by combining them with 3D collagen scaffold culturing human synovium derived mesenchymal stem cells (hSMSCs).ResultsThe decellularization of each tissue was performed and confirmed both histologically [hematoxylin and eosin (H&E) and 4’,6-diamidino-2-phenylindole (DAPI) staining] and on the basis of dsDNA quantification. The content of hydroxyproline of each tissue was relatively unchanged during the decellularization process when comparing the native and decellularized tissue. Cartilage and meniscus exhibited a significant decrease in sGAG content. The content of hydroxyproline in meniscus-derived ECM was the highest when compared with other tissues, while sGAG content in cartilage was the highest. Interestingly, a tissue-specific composition of most of the growth factors was measured in each soluble decellularized ECM and specific differentiation potential was particularly evident in cartilage, ligament and bone derived ECMs.ConclusionIn this study, soluble decellularized ECMs exhibited differences based on their tissue of origin and the present results are important going forward in the field of musculoskeletal regeneration therapy.https://www.frontiersin.org/articles/10.3389/fcell.2020.581972/fulldecellularized extracellular matrixsoluble factorgrowth factormesenchymal tissuetissue engineering |
| spellingShingle | Hiroto Hanai George Jacob George Jacob Shinichi Nakagawa Rocky S. Tuan Rocky S. Tuan Norimasa Nakamura Norimasa Nakamura Norimasa Nakamura Kazunori Shimomura Potential of Soluble Decellularized Extracellular Matrix for Musculoskeletal Tissue Engineering – Comparison of Various Mesenchymal Tissues Frontiers in Cell and Developmental Biology decellularized extracellular matrix soluble factor growth factor mesenchymal tissue tissue engineering |
| title | Potential of Soluble Decellularized Extracellular Matrix for Musculoskeletal Tissue Engineering – Comparison of Various Mesenchymal Tissues |
| title_full | Potential of Soluble Decellularized Extracellular Matrix for Musculoskeletal Tissue Engineering – Comparison of Various Mesenchymal Tissues |
| title_fullStr | Potential of Soluble Decellularized Extracellular Matrix for Musculoskeletal Tissue Engineering – Comparison of Various Mesenchymal Tissues |
| title_full_unstemmed | Potential of Soluble Decellularized Extracellular Matrix for Musculoskeletal Tissue Engineering – Comparison of Various Mesenchymal Tissues |
| title_short | Potential of Soluble Decellularized Extracellular Matrix for Musculoskeletal Tissue Engineering – Comparison of Various Mesenchymal Tissues |
| title_sort | potential of soluble decellularized extracellular matrix for musculoskeletal tissue engineering comparison of various mesenchymal tissues |
| topic | decellularized extracellular matrix soluble factor growth factor mesenchymal tissue tissue engineering |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2020.581972/full |
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