Gelatin methacryloyl (GelMA) loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells(ADSCs) conditioned medium promotes wound healing and vascular regeneration in aged skin
Abstract Background Aging skin is characterized by a disturbed structure and lack of blood supply, which makes it difficult to heal once injured. ADSCs secrete large amounts of cytokines, which promote wound healing and vascular regeneration through paracrine secretion, and the number of cytokines c...
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| Format: | Article |
| Language: | English |
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American Association for the Advancement of Science (AAAS)
2023-02-01
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| Series: | Biomaterials Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40824-023-00352-3 |
| _version_ | 1797279810018344960 |
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| author | Shiyi Li Jiachen Sun Jinxiu Yang Yi Yang Hongfan Ding Boya Yu Kui Ma Minliang Chen |
| author_facet | Shiyi Li Jiachen Sun Jinxiu Yang Yi Yang Hongfan Ding Boya Yu Kui Ma Minliang Chen |
| author_sort | Shiyi Li |
| collection | DOAJ |
| description | Abstract Background Aging skin is characterized by a disturbed structure and lack of blood supply, which makes it difficult to heal once injured. ADSCs secrete large amounts of cytokines, which promote wound healing and vascular regeneration through paracrine secretion, and the number of cytokines can be elevated by hypoxic pretreating. However, the components of ADSCs are difficult to retain in wounds. Gelatin methacrylate (GelMA) is a photopolymerizable hydrogel synthesized from gelatin and has recently emerged as a potentially attractive material for tissue engineering applications. GelMA loaded with concentrated hypoxic pretreated ADSCs conditioned medium could provide a new method of treating wounds in aged skin. Methods Primary ADSCs were isolated from human adipose tissue and characterized by flow cytometry and differentiation test. ADSCs in passages 4-6 were pretreated in the hypoxic and normoxic environments to collect conditioned medium, the conditioned medium was then concentrated to prepare concentrated ADSCs conditioned medium(cADSC-CM)(the one collected from ADSCs under hypoxia was called hypo-CM ,and the one from normoxia was called nor-CM). The concentration of cytokines was detected. After treated with cADSC-CM, the abilities of proliferation, migration, and tube formation of human umbilical vascular endothelial cells (HUVECs) were assayed, and Akt/mTOR and MAPK signal pathway was detected using western blotting. GelMA+hypo-CM hydrogel was prepared, and a comprehensive evaluation of morphology, protein release efficiency, degradation rate, mechanical properties, and rheology properties were performed. Full-thickness skin wounds were created on the backs of 20-month-old mice. After surgery, GelMA, GelMA+F12, GelMA+hypo-CM, and GelMA+nor-CM were applied to the wound surface respectively. H&E, Masson, and immunohistochemistry staining were performed, and a laser Doppler perfusion imager was used to evaluate the blood perfusion. The student’s t-test was used for analysis between two groups and a one-way analysis of variance (ANOVA) was used for analysis among multi groups. Results Our results revealed that 1) wounds in aged skin healed more slowly than that in young skin and exhibited poorer perfusion; 2) hypoxic pretreated ADSCs secreted more cytokines including VEGF by activating HIF1α; 3) hypo-CM promoted proliferation and migration of HUVECs through VEGF/Akt/mTOR and MAPK signal pathway; 4) GelMA-hypoCM accelerated wound healing and angiogenesis in aged skin in vivo. Conclusion GelMA loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells conditioned medium could accelerate wound healing in aged skin by promoting angiogenesis. |
| first_indexed | 2024-03-07T16:31:25Z |
| format | Article |
| id | doaj.art-4b4fef607bb2412c8b57f27f8a55a5b1 |
| institution | Directory Open Access Journal |
| issn | 2055-7124 |
| language | English |
| last_indexed | 2024-03-07T16:31:25Z |
| publishDate | 2023-02-01 |
| publisher | American Association for the Advancement of Science (AAAS) |
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| series | Biomaterials Research |
| spelling | doaj.art-4b4fef607bb2412c8b57f27f8a55a5b12024-03-03T10:11:38ZengAmerican Association for the Advancement of Science (AAAS)Biomaterials Research2055-71242023-02-0127111710.1186/s40824-023-00352-3Gelatin methacryloyl (GelMA) loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells(ADSCs) conditioned medium promotes wound healing and vascular regeneration in aged skinShiyi Li0Jiachen Sun1Jinxiu Yang2Yi Yang3Hongfan Ding4Boya Yu5Kui Ma6Minliang Chen7Senior Department of Burns and Plastic Surgery, The Fourth Medical Center of Chinese PLA General HospitalSenior Department of Burns and Plastic Surgery, The Fourth Medical Center of Chinese PLA General HospitalSenior Department of Burns and Plastic Surgery, The Fourth Medical Center of Chinese PLA General HospitalSenior Department of Burns and Plastic Surgery, The Fourth Medical Center of Chinese PLA General HospitalSenior Department of Burns and Plastic Surgery, The Fourth Medical Center of Chinese PLA General HospitalSenior Department of Burns and Plastic Surgery, The Fourth Medical Center of Chinese PLA General HospitalResearch Center for Tissue Repair and Regeneration Affiliated to the Medical Innovation Research Division, Chinese PLA General HospitalSenior Department of Burns and Plastic Surgery, The Fourth Medical Center of Chinese PLA General HospitalAbstract Background Aging skin is characterized by a disturbed structure and lack of blood supply, which makes it difficult to heal once injured. ADSCs secrete large amounts of cytokines, which promote wound healing and vascular regeneration through paracrine secretion, and the number of cytokines can be elevated by hypoxic pretreating. However, the components of ADSCs are difficult to retain in wounds. Gelatin methacrylate (GelMA) is a photopolymerizable hydrogel synthesized from gelatin and has recently emerged as a potentially attractive material for tissue engineering applications. GelMA loaded with concentrated hypoxic pretreated ADSCs conditioned medium could provide a new method of treating wounds in aged skin. Methods Primary ADSCs were isolated from human adipose tissue and characterized by flow cytometry and differentiation test. ADSCs in passages 4-6 were pretreated in the hypoxic and normoxic environments to collect conditioned medium, the conditioned medium was then concentrated to prepare concentrated ADSCs conditioned medium(cADSC-CM)(the one collected from ADSCs under hypoxia was called hypo-CM ,and the one from normoxia was called nor-CM). The concentration of cytokines was detected. After treated with cADSC-CM, the abilities of proliferation, migration, and tube formation of human umbilical vascular endothelial cells (HUVECs) were assayed, and Akt/mTOR and MAPK signal pathway was detected using western blotting. GelMA+hypo-CM hydrogel was prepared, and a comprehensive evaluation of morphology, protein release efficiency, degradation rate, mechanical properties, and rheology properties were performed. Full-thickness skin wounds were created on the backs of 20-month-old mice. After surgery, GelMA, GelMA+F12, GelMA+hypo-CM, and GelMA+nor-CM were applied to the wound surface respectively. H&E, Masson, and immunohistochemistry staining were performed, and a laser Doppler perfusion imager was used to evaluate the blood perfusion. The student’s t-test was used for analysis between two groups and a one-way analysis of variance (ANOVA) was used for analysis among multi groups. Results Our results revealed that 1) wounds in aged skin healed more slowly than that in young skin and exhibited poorer perfusion; 2) hypoxic pretreated ADSCs secreted more cytokines including VEGF by activating HIF1α; 3) hypo-CM promoted proliferation and migration of HUVECs through VEGF/Akt/mTOR and MAPK signal pathway; 4) GelMA-hypoCM accelerated wound healing and angiogenesis in aged skin in vivo. Conclusion GelMA loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells conditioned medium could accelerate wound healing in aged skin by promoting angiogenesis.https://doi.org/10.1186/s40824-023-00352-3Aged woundAngiogenesisGelMA hydrogelADSCsConditioned mediumHypoxia |
| spellingShingle | Shiyi Li Jiachen Sun Jinxiu Yang Yi Yang Hongfan Ding Boya Yu Kui Ma Minliang Chen Gelatin methacryloyl (GelMA) loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells(ADSCs) conditioned medium promotes wound healing and vascular regeneration in aged skin Biomaterials Research Aged wound Angiogenesis GelMA hydrogel ADSCs Conditioned medium Hypoxia |
| title | Gelatin methacryloyl (GelMA) loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells(ADSCs) conditioned medium promotes wound healing and vascular regeneration in aged skin |
| title_full | Gelatin methacryloyl (GelMA) loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells(ADSCs) conditioned medium promotes wound healing and vascular regeneration in aged skin |
| title_fullStr | Gelatin methacryloyl (GelMA) loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells(ADSCs) conditioned medium promotes wound healing and vascular regeneration in aged skin |
| title_full_unstemmed | Gelatin methacryloyl (GelMA) loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells(ADSCs) conditioned medium promotes wound healing and vascular regeneration in aged skin |
| title_short | Gelatin methacryloyl (GelMA) loaded with concentrated hypoxic pretreated adipose-derived mesenchymal stem cells(ADSCs) conditioned medium promotes wound healing and vascular regeneration in aged skin |
| title_sort | gelatin methacryloyl gelma loaded with concentrated hypoxic pretreated adipose derived mesenchymal stem cells adscs conditioned medium promotes wound healing and vascular regeneration in aged skin |
| topic | Aged wound Angiogenesis GelMA hydrogel ADSCs Conditioned medium Hypoxia |
| url | https://doi.org/10.1186/s40824-023-00352-3 |
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