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Featured researches published by Changcan Shi.


ACS Applied Materials & Interfaces | 2017

CAGW Peptide- and PEG-Modified Gene Carrier for Selective Gene Delivery and Promotion of Angiogenesis in HUVECs in Vivo

Jing Yang; Xuefang Hao; Qian Li; Mary Akpanyung; Abdelilah Nejjari; Agnaldo Luis Neve; Xiangkui Ren; Jintang Guo; Yakai Feng; Changcan Shi; Wencheng Zhang

Gene therapy is a promising strategy for angiogenesis, but developing gene carriers with low cytotoxicity and high gene delivery efficiency in vivo is a key issue. In the present study, we synthesized the CAGW peptide- and poly(ethylene glycol) (PEG)-modified amphiphilic copolymers. CAGW peptide serves as a targeting ligand for endothelial cells (ECs). Different amounts of CAGW peptide were effectively conjugated to the amphiphilic copolymer via heterofunctional poly(ethylene glycol). These CAG- and PEG-modified copolymers could form nanoparticles (NPs) by self-assembly method and were used as gene carriers for the pEGFP-ZNF580 (pZNF580) plasmid. CAGW and PEG modification coordinately improved the hemocompatibility and cytocompatibility of NPs. The results of cellular uptake showed significantly enhanced internalization efficiency of pZNF580 after CAGW modification. Gene expression at mRNA and protein levels demonstrated that EC-targeted NPs possessed high gene delivery efficiency, especially the NPs with higher content of CAGW peptide (1.16 wt %). Furthermore, in vitro and in vivo vascularization assays also showed outstanding vascularization ability of human umbilical vein endothelial cells treated by the NP/pZNF580 complexes. This study demonstrates that the CAGW peptide-modified NP is a promising candidate for gene therapy in angiogenesis.


Journal of Materials Chemistry B | 2017

Mixed micelles obtained by co-assembling comb-like and grafting copolymers as gene carriers for efficient gene delivery and expression in endothelial cells

Qian Li; Xuefang Hao; Juan Lv; Xiangkui Ren; Kunyu Zhang; Ihsan Ullah; Yakai Feng; Changcan Shi; Wencheng Zhang

Gene delivery can enhance the endothelialization of biomaterial surfaces. However, the lack of efficient target function is still the major concern that hinders the clinical application of gene therapy. With the aim to develop a specific targeting gene carrier for endothelial cells (ECs), the Cys-Arg-Glu-Asp-Val-Trp (CREDVW) peptide was linked to the comb-like copolymer of poly(lactide-co-3(S)-methyl-morpholine-2,5-dione)-poly(poly(ethylene glycol) monomethacrylate) (PLMD-PPEGMA) to form the CREDVW modified copolymer PLMD-PPEGMA-CREDVW, which could enhance the special recognition of ECs. Mixed micelles were then prepared by co-assembling this comb-like copolymer and the amphiphilic grafting copolymer poly(lactide-co-3(S)-methyl-morpholine-2,5-dione)-g-polyethylenimine (PLMD-g-PEI). These mixed micelles with the CREDVW-functional peptide exhibited good pEGFP-ZNF580 (pDNA) binding ability and could condense it into complexes with proper size and positive zeta potential. The MTT results demonstrated the low cytotoxicity of the CREDVW-modified mixed micelle/pDNA complexes. The internalization efficiency of the CREDVW-modified complexes with targeting function was about two times higher than the dysfunctional CREVDW-modified complexes. Besides, the transfection efficiency of these complexes was more pronounced, compared to the control group, PEI(10 kDa)/pDNA, as detected by means of in vitro transfection studies. Western blot analysis demonstrated relatively high protein levels in the transfected cells by CREDVW-modified mixed micelle/pDNA complexes, up to 75%, in comparison to the control group (26%). In addition, the cell migration ability was significantly improved as demonstrated by the wound healing assay. These results indicated that the mixed micelles could act as an active targeting gene carrier, having both tunable gene transfection efficiency and low cytotoxicity, which are beneficial for the endothelialization of biomaterial surface.


Macromolecular Rapid Communications | 2016

Multitargeting Gene Delivery Systems for Enhancing the Transfection of Endothelial Cells

Jing Yang; Qian Li; Xiao Yang; Yakai Feng; Xiangkui Ren; Changcan Shi; Wencheng Zhang

Gene therapy demonstrates promising prospects on cardiovascular diseases. However, nonviral gene delivery system has relatively low transfection efficiency, especially for endothelial cells (ECs). Herein, typical cell-penetrating peptide (TAT), nuclear localization signals (NLSs), and REDV functional peptide have been used to prepare multitargeting complexes. These complexes exhibit higher transfection efficiency owing to the targeting sequences of REDV and NLSs as well as the cell-penetrating function of TAT. The multifunction of the complexes provides high cell uptake, endo/lysosomal escape, and nucleus accumulation of the encapsulated DNA. Thus these multitargeting complexes can provide a potential platform for gene delivery, especially for EC transfection.


Polymers | 2017

CAGW Peptide Modified Biodegradable Cationic Copolymer for Effective Gene Delivery

Xinghong Duo; Jun Wang; Qian Li; Agnaldo Luis Neve; Mary Akpanyung; Abdelilah Nejjari; Zaidi Ali; Yakai Feng; Wencheng Zhang; Changcan Shi

In recent years, gene therapy has become a promising technology to enhance endothelialization of artificial vascular grafts. The ideal gene therapy requires a gene carrier with low cytotoxicity and high transfection efficiency. In this paper, we prepared a biodegradable cationic copolymer poly(d,l-lactide-co-glycolide)-graft-PEI (PLGA-g-PEI), grafted Cys-Ala-Gly-Trp (CAGW) peptide onto this copolymer via the thiol-ene Click-reaction, and then prepared micelles by a self-assembly method. pEGFP-ZNF580 plasmids (pDNA) were condensed by these micelles via electrostatic interaction to form gene complexes. The CAGW peptide enables these gene complexes with special recognition for endothelial cells, which could enhance their transfection. As a gene carrier system, the PLGA-g-PEI-g-CAGW/pDNA gene complexes were evaluated and the results showed that they had suitable diameter and zeta potential for cellular uptake, and exhibited low cytotoxicity and high transfection efficiency for EA.hy926 cells.


Journal of Materials Chemistry B | 2017

Multi-targeting peptides for gene carriers with high transfection efficiency

Jing Zhao; Qian Li; Xuefang Hao; Xiangkui Ren; Jintang Guo; Yakai Feng; Changcan Shi

Non-viral gene carriers for gene therapy have been developed for many years. But the gene transfection is generally limited by deficient cellular uptake, low endo/lysosome escape, and weak nuclear translocation. Some targeting peptides have been conjugated onto gene carriers for highly efficient gene delivery. These targeting carriers can overcome some of these limitations to efficiently deliver therapeutic genes into desired cells. In this review, we will summarize the recent development of multi-targeting peptide immobilized non-viral gene carriers for efficient gene therapy, especially for the targeting and suppression of tumor cells, and the transfection and proliferation of endothelial cells. The peptide functionalization of gene carriers is a promising strategy to promote the elimination of solid tumors and the rapid endothelialization of artificial blood vessels.


Journal of Nanobiotechnology | 2018

Oligohistidine and targeting peptide functionalized TAT-NLS for enhancing cellular uptake and promoting angiogenesis in vivo

Qian Li; Xuefang Hao; Syed Saqib Ali Zaidi; Jintang Guo; Xiangkui Ren; Changcan Shi; Wencheng Zhang; Yakai Feng

BackgroundGene therapy has been developed and used in medical treatment for many years, especially for the enhancement of endothelialization and angiogenesis. But slow endosomal escape rate is still one of the major barriers to successful gene delivery. In order to evaluate whether introducing oligohistidine (Hn) sequence into gene carriers can promote endosomal escape and gene transfection or not, we designed and synthesized Arg-Glu-Asp-Val (REDV) peptide functionalized TAT-NLS-Hn (TAT: typical cell-penetrating peptide, NLS: nuclear localization signals, Hn: oligohistidine sequence, n: 4, 8 and 12) peptides with different Hn sequence lengths. pEGFP-ZNF580 (pZNF580) was condensed by these peptides to form gene complexes, which were used to transfect human umbilical vein endothelial cells (HUVECs).ResultsMTT assay showed that the gene complexes exhibited low cytotoxicity for HUVECs. The results of cellular uptake and co-localization ratio demonstrated that the gene complexes prepared from TAT-NLS-Hn with long Hn sequence (n = 12) benefited for high internalization efficiency of pZNF580. In addition, the results of western blot analysis and PCR assay of REDV-TAT-NLS-H12/pZNF580 complexes showed significantly enhanced gene expression at protein and mRNA level. Wound healing assay and transwell migration assay also confirmed the improved proliferation and migration ability of the transfected HUVECs by these complexes. Furthermore, the in vitro and in vivo angiogenesis assay illustrated that these complexes could promote the tube formation ability of HUVECs.ConclusionThe above results indicated that the delivery efficiency of pZNF580 and its expression could be enhanced by introducing Hn sequence into gene carriers. The Hn sequence in REDV-TAT-NLS-Hn is beneficial for high gene transfection. These REDV and Hn functionalized TAT-NLS peptides are promising gene carriers in gene therapy.


RSC Advances | 2017

Electrospun PCL-PIBMD/SF blend scaffolds with plasmid complexes for endothelial cell proliferation

Lingchuang Bai; Qian Li; Xinghong Duo; Xuefang Hao; Wencheng Zhang; Changcan Shi; Jintang Guo; Xiangkui Ren; Yakai Feng

Tissue engineering scaffolds with gene delivery function play an important role in DNA-based vascular tissue engineering. In the present work, we used biodegradable polyester–polydepsipeptide, silk fibroin (SF) and gene complexes to prepare electrospun scaffolds encapsulating gene complexes in order to enhance the proliferation of endothelial cells. A series of nanofibrous scaffolds with different properties including fiber diameter, hydrophilicity, porosity and mechanical properties were prepared by electrospinning technology with adjusting the weight ratio of poly(e-caprolactone)-b-poly(isobutyl-morpholine-2,5-dione) (PCL-PIBMD) and SF. PCL-PIBMD/SF blend scaffolds were optimized to obtain the scaffolds with a weight ratio of 90/10 to have superior mechanical performance and good biocompatibility. pEGFP-ZNF580 plasmid (pZNF580) complexes were electrosprayed onto these PCL-PIBMD/SF blend scaffolds to promote the proliferation of endothelial cells. In order to maintain the stability and integrity of plasmid complexes loaded in scaffolds, the composite scaffolds were fabricated by alternatively layer-by-layer electrospinning and electrospraying techniques. These composite scaffolds showed obviously low platelet adhesion and good histocompatibility. They could effectively enhance the adhesion, spreading and proliferation of human umbilical vein endothelial cells. These results indicated that the composite scaffolds could serve as an attractive platform to deliver therapeutic genes for vascular tissue engineering.


Journal of Materials Chemistry B | 2018

Red-blood-cell-mimetic gene delivery systems for long circulation and high transfection efficiency in ECs

Xuefang Hao; Qian Li; Huaning Wang; Khan Muhammad; Jintang Guo; Xiangkui Ren; Changcan Shi; Shihai Xia; Wencheng Zhang; Yakai Feng

Recently, the red blood cell (RBC) membrane has been used as a mimetic nanocoating for nanoparticles for drug delivery systems to promote their biocompatibility. In the present study, the nano-sized RBC membrane was coated on the surface of gene complexes through electrostatic interactions to prepare biomimetic gene delivery systems so as to improve their biocompatibility and prolong their circulation time in vivo. The structure of the biomimetic gene delivery systems was determined by transmission electron microscopy (TEM) and confocal laser scanning microscopy (CLSM). They exhibited low cytotoxicity and high transfection efficiency in endothelial cells (ECs), which could improve the migration ability of ECs. Besides, the biomimetic gene delivery systems exhibited strong immune evasion and long in vivo circulation time. The phagocytic rate of these biomimetic gene delivery systems reduced 52% compared with that of the PLGA-PEI/pZNF580 control group (without RBC membrane modification). Their circulation time in vivo was more than 2 times higher than that of the control group. Consequently, we provide a simple method for the preparation of camouflaged gene delivery systems, which can further facilitate the development of a gene delivery platform for the therapy of vascular diseases via enhancing EC transfection. This strategy will open up a new avenue for gene delivery systems by RBC membrane camouflage.


Journal of Materials Chemistry B | 2018

POSS-cored and peptide functionalized ternary gene delivery systems with enhanced endosomal escape ability for efficient intracellular delivery of plasmid DNA

Xuefang Hao; Qian Li; Hasnain Ali; Syed Saqib Ali Zaidi; Jintang Guo; Xiangkui Ren; Changcan Shi; Shihai Xia; Wencheng Zhang; Yakai Feng

Biocompatibility, stability and high efficiency profiles are critical points for promoting the practical applications of gene delivery systems. The incorporation of cell-penetrating peptides (CPPs), REDV, and a nuclear localization signal (NLS) peptide sequence has been considered to be a promising strategy for developing efficient gene carriers to transfect vascular endothelial cells (ECs). However, these integrated multifunctional peptide carriers are usually limited by their inefficient targeting function and weak endosomal escape ability. Aiming to develop more efficient gene carriers, the integrated multifunctional REDV-G-TAT-G-NLS-C sequence was conjugated to polyhedral oligomeric silsesquioxane (POSS) by heterobifunctional poly(ethylene glycol) in the current study. This star-shaped polymer carrier complexed with the pZNF580 plasmid to form gene complexes, and then the histidine-rich peptide of REDV-TAT-NLS-H12 (TP-H12) was incorporated into their surface to obtain ternary gene delivery systems with enhanced endosomal escape ability. These ternary gene delivery systems exhibited low cytotoxicity towards ECs and possessed high REDV-mediated cellular uptake, excellent internalization efficiency, rapid endosomal escape and high nucleus translocation capacity. The endosomal escape of the ternary complexes was improved due to the pH buffering capacity of the histidine residue in TP-H12 and the optimized macropinocytosis internalization pathway. Moreover, these CPP-based ternary gene delivery systems have high gene delivery efficiency and could improve the migration of ECs as demonstrated by gene expression and transwell assay. These systems may serve as a promising candidate for gene delivery and transfection in ECs, which is advantageous for EC migration and endothelialization on the biomaterial surface.


Biomaterials Science | 2017

Star-shaped copolymer grafted PEI and REDV as a gene carrier to improve migration of endothelial cells

Juan Lv; Xuefang Hao; Qian Li; Mary Akpanyung; Abdelilah Nejjari; Agnaldo Luis Neve; Xiangkui Ren; Yakai Feng; Changcan Shi; Wencheng Zhang

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Bin Liang

First Affiliated Hospital of Wenzhou Medical University

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