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Feasibility of repairing skin defects by VEGF165 gene-modified iPS-HFSCs seeded on a 3D printed scaffold containing astragalus polysaccharide |
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| Authors: | Weibin Du Jintao Hu Xiaolong Huang Zhenwei Wang Huateng Zhou Yadong Yang Huahui Hu Rongliang Chen Fuxiang Shen Renfu Quan |
| Affiliation: | 1. Research Institute of Orthopedics, The Affiliated Jiangnan Hospital of Zhejiang Chinese Medical University, Hangzhou, China;2. Orthopedics and Traumatology Department, Hangzhou Traditional Chinese Medicine Hospital Affiliated to Zhejiang Chinese Medical University, Hangzhou, China Contribution: Data curation (equal), Methodology (equal), Validation (equal), Writing - original draft (equal);3. Research Institute of Orthopedics, The Affiliated Jiangnan Hospital of Zhejiang Chinese Medical University, Hangzhou, China Hangzhou Xiaoshan Hospital of Traditional Chinese Medicine, Hangzhou, China Contribution: Resources (supporting), Validation (supporting);4. Research Institute of Orthopedics, The Affiliated Jiangnan Hospital of Zhejiang Chinese Medical University, Hangzhou, China Hangzhou Xiaoshan Hospital of Traditional Chinese Medicine, Hangzhou, China Contribution: Conceptualization (equal), Formal analysis (equal), Writing - original draft (equal);5. Research Institute of Orthopedics, The Affiliated Jiangnan Hospital of Zhejiang Chinese Medical University, Hangzhou, China Hangzhou Xiaoshan Hospital of Traditional Chinese Medicine, Hangzhou, China Contribution: Data curation (supporting), Formal analysis (supporting), Resources (supporting);6. School of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, China Contribution: Conceptualization (supporting), Methodology (supporting), Software (supporting);7. Research Institute of Orthopedics, The Affiliated Jiangnan Hospital of Zhejiang Chinese Medical University, Hangzhou, China Hangzhou Xiaoshan Hospital of Traditional Chinese Medicine, Hangzhou, China Contribution: Formal analysis (supporting), Validation (supporting), Visualization (supporting);8. Research Institute of Orthopedics, The Affiliated Jiangnan Hospital of Zhejiang Chinese Medical University, Hangzhou, China Hangzhou Xiaoshan Hospital of Traditional Chinese Medicine, Hangzhou, China Contribution: Conceptualization (supporting), Data curation (supporting), Formal analysis (supporting);9. Research Institute of Orthopedics, The Affiliated Jiangnan Hospital of Zhejiang Chinese Medical University, Hangzhou, China Hangzhou Xiaoshan Hospital of Traditional Chinese Medicine, Hangzhou, China Contribution: Supervision (supporting), Writing - review & editing (supporting) |
| Abstract: | The preparation of biodegradable scaffolds loaded with cells and cytokine is a feature of tissue-engineered skin. IPSCs-based tissue-engineered skin treatment for wound repair is worth exploring. Healthy human skin fibroblasts were collected and reprogrammed into iPSCs. After gene modification and induction, CK19+/Integrinβ1+/CD200+ VEGF165 gene-modified iPS-HFSCsGFP were obtained and identified by a combination of immunofluorescence and RT-qPCR. Astragalus polysaccharide-containing 3D printed degradable scaffolds were prepared and co-cultured with VEGF165 gene-modified iPS-HFSCsGFP, and the biocompatibility and spatial structure of the tissue-engineered skin was analysed by cell counting kit-8 (CCK8) assay and scanning electron microscopy. Finally, the tissue-engineered skin was transplanted onto the dorsal trauma of nude mice, and the effect of tissue-engineered skin on the regenerative repair of total skin defects was evaluated by a combination of histology, immunohistochemistry, immunofluorescence, RT-qPCR, and in vivo three-dimensional reconstruction under two-photon microscopy. CK19+/Integrinβ1+/CD200+ VEGF165 gene-modified iPS-HFSCsGFP, close to the morphology and phenotype of human-derived hair follicle stem cells, were obtained. The surface of the prepared 3D printed degradable scaffold containing 200 μg/mL astragalus polysaccharide was enriched with honeycomb-like meshwork, which was more conducive to the proliferation of the resulting cells. After tissue-engineered skin transplantation, combined assays showed that it promoted early vascularization, collagen and hair follicle regeneration and accelerated wound repair. VEGF165 gene-modified iPS-HFSCsGFP compounded with 3D printed degradable scaffolds containing 200 μg/mL astragalus polysaccharide can directly and indirectly participate in vascular, collagen, and hair follicle regeneration in the skin, achieving more complete structural and functional skin regenerative repair. |
| Keywords: | 3D printed degradable scaffold astragalus polysaccharide hair follicle stem cells induced pluripotent stem cells regeneration and repair skin defect |