Feasibility of repairing skin defects by VEGF165 gene-modified iPS-HFSCs seeded on a 3D printed scaffold containing astragalus polysaccharide

Weibin Du; Jintao Hu; Xiaolong Huang; Zhenwei Wang; Huateng Zhou; Yadong Yang; Huahui Hu; Rongliang Chen; Fuxiang Shen; Renfu Quan

doi:10.1111/jcmm.17800

Feasibility of repairing skin defects by VEGF₁₆₅ gene-modified iPS-HFSCs seeded on a 3D printed scaffold containing astragalus polysaccharide

J Cell Mol Med. 2023 Aug;27(15):2136-2149. doi: 10.1111/jcmm.17800. Epub 2023 Jun 1.

Authors

Weibin Du^{1

2}, Jintao Hu³, Xiaolong Huang^{1

2}, Zhenwei Wang^{1

2}, Huateng Zhou^{1

2}, Yadong Yang⁴, Huahui Hu^{1

2}, Rongliang Chen^{1

2}, Fuxiang Shen^{1

2}, Renfu Quan^{1

2}

Affiliations

¹ Research Institute of Orthopedics, The Affiliated Jiangnan Hospital of Zhejiang Chinese Medical University, Hangzhou, China.
² Hangzhou Xiaoshan Hospital of Traditional Chinese Medicine, Hangzhou, China.
³ Orthopedics and Traumatology Department, Hangzhou Traditional Chinese Medicine Hospital Affiliated to Zhejiang Chinese Medical University, Hangzhou, China.
⁴ School of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, China.

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-HFSCs^GFP 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-HFSCs^GFP , 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-HFSCs^GFP , 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-HFSCs^GFP 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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Collagen
Feasibility Studies
Humans
Mice
Mice, Nude
Polysaccharides / pharmacology
Printing, Three-Dimensional
Skin Transplantation* / methods
Tissue Engineering / methods
Tissue Scaffolds / chemistry
Vascular Endothelial Growth Factor A* / genetics

Substances

Vascular Endothelial Growth Factor A
Collagen
Polysaccharides