Publications by authors named "Changcan Shi"

36 Publications

Controlled release of KGF-2 for regulation of wound healing by KGF-2 complexed with "lotus seedpod surface-like" porous microspheres.

J Mater Chem B 2021 05;9(19):4039-4049

Department of Intensive Care, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.

Keratinocyte growth factor-2 (KGF-2) can regulate the proliferation and differentiation of keratinocyte, which plays a remarkable role in maintaining normal tissue structure and promoting wound healing. As an effective strategy, KGF-2 solution is widely used in the treatment of wounds in clinical applications. However, KGF-2 in solution cannot achieve sustained release, which results in drug loss and unnecessary waste. Polysaccharide hemostasis microspheres (PHMs) are an ideal drug loading platform due to their special "lotus seedpod surface-like" morphology and structure. Herein, to realize the controllable release of KGF-2, PHMs loaded with KGF-2 ([email protected]) were prepared. It was found that the bioavailability of KGF-2 was improved greatly. Most importantly, [email protected] can reduce inflammation and accelerate the wound healing process due to the controlled release of KGF-2. [email protected] might be a potential alternative strategy for wound healing in future clinical applications.
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http://dx.doi.org/10.1039/d1tb00148eDOI Listing
May 2021

Hydrodynamic cavitation: A feasible approach to intensify the emulsion cross-linking process for chitosan nanoparticle synthesis.

Ultrason Sonochem 2021 Jun 20;74:105551. Epub 2021 Apr 20.

School of Biological and Chemical Engineering, Guangxi University of Science and Technology, Liuzhou 545006, China; Guangxi Key Laboratory of Green Processing of Sugar Resources, Liuzhou 545006, China.

Chitosan nanoparticles (NPs) exhibit great potential in drug-controlled release systems. A controlled hydrodynamic cavitation (HC) technique was developed to intensify the emulsion crosslinking process for the synthesis of chitosan NPs. Experiments were performed using a circular venturi and under varying operating conditions, i.e., types of oil, addition mode of glutaraldehyde (Glu) solution, inlet pressure (P), and rheological properties of chitosan solution. Palm oil was more appropriate for use as the oil phase for the HC-intensified process than the other oil types. The addition mode of water-in-oil (W/O) emulsion containing Glu (with Span 80) was more favorable than the other modes for obtaining a narrow distribution of chitosan NPs. The minimum size of NPs with polydispersity index of 0.342 was 286.5 nm, and the maximum production yield (P) could reach 47.26%. A positive correlation was found between the size of NPs and the droplet size of W/O emulsion containing chitosan at increasing P. Particle size, size distribution, and the formation of NPs were greatly dependent on the rheological properties of the chitosan solution. Fourier transform infrared spectroscopy (FTIR) analysis indicated that the molecular structure of palm oil was unaffected by HC-induced effects. Compared with ultrasonic horn, stirring-based, and conventional drop-by-drop processes, the application of HC to intensify the emulsion crosslinking process allowed the preparation of a finer and a narrower distribution of chitosan NPs in a more energy-efficient manner. The novel route developed in this work is a viable option for chitosan NP synthesis.
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http://dx.doi.org/10.1016/j.ultsonch.2021.105551DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8091060PMC
June 2021

Fabricating poly(vinyl alcohol)/gelatin composite sponges with high absorbency and water-triggered expansion for noncompressible hemorrhage and wound healing.

J Mater Chem B 2021 02 26;9(6):1568-1582. Epub 2021 Jan 26.

Wenzhou Institute of Biomaterials and Engineering, Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou, Zhejiang 325011, China.

It is challenging for traditional hemostatic sponges to control massive and noncompressible hemorrhages in the military field and accidental trauma. In this work, a series of highly fluid-absorbent composite sponges with rapid expansion ability based on norbornene anhydride-modified poly(vinyl alcohol) and gelatin ([email protected]) were developed by a foaming technique, chemical and physical crosslinking reactions and lyophilization. The prepared [email protected] exhibited a 3500% maximum water absorption ratio with a fast water absorption speed, which was suitable for blood component concentration. Owing to its interconnected macroporous structure, robust mechanical strength and high resilience, the compressed sponge could rapidly re-expand to more than 10 times its volume in response to water and blood. Moreover, due to the synergistic effect of the PVA-based sponge and gelatin, [email protected] could obviously shorten the hemostasis time and reduce blood loss in SD rat liver defect noncompressible hemorrhage models, and exhibited better wound healing effects in a full-thickness skin defect model than commercial sponges. These results suggest that [email protected] is a potential candidate for controlling noncompressible hemorrhage and promoting wound healing.
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http://dx.doi.org/10.1039/d0tb02480eDOI Listing
February 2021

Liquiritigenin-Loaded Submicron Emulsion Protects Against Doxorubicin-Induced Cardiotoxicity via Antioxidant, Anti-Inflammatory, and Anti-Apoptotic Activity.

Int J Nanomedicine 2020 17;15:1101-1115. Epub 2020 Feb 17.

School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210046, People's Republic of China.

Background: The clinical use of doxorubicin (DOX) is severely limited due to its cardiotoxicity. Thus, there is a need for prophylactic and treatment strategies against DOX-induced cardiotoxicity.

Purpose: The purpose of this study was to develop a liquiritigenin-loaded submicron emulsion (Lq-SE) with enhanced oral bioavailability and to explore its efficacy against DOX-induced cardiotoxicity.

Methods: Lq-SE was prepared using high-pressure homogenization and characterized using several analytical techniques. The formulation was optimized by central composite design response surface methodology (CCD-RSM). In vivo pharmacokinetic studies, biochemical analyses, reactive oxygen species (ROS) assays, histopathologic assays, and Western blot analyses were performed.

Results: Each Lq-SE droplet had a mean particle size of 221.7 ± 5.80 nm, a polydispersity index (PDI) of 0.106 ± 0.068 and a zeta potential of -28.23 ± 0.42 mV. The area under the curve (AUC) of Lq-SE was 595% higher than that of liquiritigenin (Lq). Lq-SE decreased the release of serum cardiac enzymes and ameliorated histopathological changes in the hearts of DOX-challenged mice. Lq-SE significantly reduced oxidative stress by adjusting the levels of ROS, increasing the activity of antioxidative enzymes and inhibiting the protein expression of NOX4 and NOX2. Furthermore, Lq-SE significantly improved the inflammatory response through the mitogen-activated protein kinase (MAPK)/nuclear factor-κB (NF-κB) signalling pathway and induced cardiomyocyte apoptosis.

Conclusion: Lq-SE could be used as an effective cardioprotective agent against DOX in chemotherapy to enable better treatment outcomes.
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http://dx.doi.org/10.2147/IJN.S235832DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7034974PMC
June 2020

Polysaccharide Based Hemostatic Strategy for Ultrarapid Hemostasis.

Macromol Biosci 2020 04 18;20(4):e1900370. Epub 2020 Feb 18.

Wenzhou Institute of Biomaterials and Engineering, Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou, Zhejiang, 325011, China.

Bleeding complications usually cause significant morbidity and mortality in civilian and military populations. In clinical application, hemostatic sponges, gauzes, hydrogel, and bandages are widely used as the traditional effective hemostatic products for hemorrhage. However, the traditional hemostatic devices or agents cannot meet the requirement for treatment of massive bleeding. Therefore, the excellent hemostatic performance of hemostatic products are of great significance for saving lives. Natural polysaccharides, as the main chemical component, have been widely used in the preparation of hemostasis due to their perfect biocompatibility and biodegradability. Polysaccharide based hemostatic products are available in variety of forms, such as, hydrogel, sponges, gauze and microspheres. The purpose of the present review is to report the research progress on polysaccharide hemostatic products and technology.
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http://dx.doi.org/10.1002/mabi.201900370DOI Listing
April 2020

Polysaccharide-Based Lotus Seedpod Surface-Like Porous Microsphere with Precise and Controllable Micromorphology for Ultrarapid Hemostasis.

ACS Appl Mater Interfaces 2019 Dec 5;11(50):46558-46571. Epub 2019 Dec 5.

Institute of Chemistry and Biochemistry , Free University of Berlin , Takustrasse 3 , Berlin 14195 , Germany.

Rapid water absorption rate has become a bottleneck that limits ultrarapid hemostatic performance of hemostatic microspheres. Herein, we reported a "lotus seedpod surface-like" polysaccharide hemostatic microsphere (PHM) with "macropits on surface" morphology and "micropores in macropits" structure. Unique macropits on surface can promote the water absorption rate because they are advantageous to quickly guide blood into the micropores. Special micropores are internally connected with each other, which endows PHM with high water absorption ratio. During the process of blood entering the micropores from micropits, the pore size decreases gradually. In this way, blood clotting factors could be rapidly concentrated. PHM showed the highest water absorption rate (40.7 mL/s/cm) and rapid hemostatic property in vivo (hemostatic time shortened from 210 to 45 s). Lotus seedpod surface-like PHMs are believed to have further clinical application as an effective hemostasis.
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http://dx.doi.org/10.1021/acsami.9b17543DOI Listing
December 2019

Peptide-immobilized starch/PEG sponge with rapid shape recovery and dual-function for both uncontrolled and noncompressible hemorrhage.

Acta Biomater 2019 11 23;99:220-235. Epub 2019 Aug 23.

Wenzhou Institute of Biomaterials and Engineering, Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou, Zhejiang 325011, China. Electronic address:

It is challenging for traditional hemostatic sponges to meet the clinic demand for both uncontrolled and noncompressible hemorrhage. With the aim to develop a rapid shape recovery material with both active and passive hemostatic performance, a dual-functional hemostatic sponge (TRAP-Sp) with a macroporous structure and good mechanical properties for controlling massive and noncompressible hemorrhage was prepared by chemically immobilizing thrombin-receptor-agonist-peptide (TRAP) onto a starch/polyethylene glycol (PEG) sponge. The TRAP2-Sp1 showed the best hemostatic performance among all samples in both rat artery uncontrollable hemorrhage and liver defect noncompressible hemorrhage models. When analyzing the hemostatic mechanism of TRAP-Sp, the high water absorption capacity of the sponge contributed to absorbing plasma, concentrating blood cells, and enhancing blood coagulation. After absorbing water, the shape-fixed TRAP-Sp with sufficient mechanical strength and high resilience can rapidly expand and apply pressure to the wound. TRAP immobilized on the sponge could activate the adhered platelets in an active pathway. Additionally, evaluation of cytotoxicity, hemolysis, and histology further highlighted the adequate biocompatibility of TRAP-Sp. With excellent hemostatic performance and biosafety, this sponge could be a potential candidate as a topical hemostatic agent for uncontrolled and noncompressible hemorrhage. STATEMENT OF SIGNIFICANCE: There is a need for innovative hemostatic materials for both uncontrolled and noncompressible hemorrhage. This manuscript describes a rapid shape recovery hemostatic sponge with both active and passive hemostatic performances synthesized by foaming technique, cross-linking reaction, and chemical immobilization of thrombin-receptor-agonist-peptide (TRAP). On contact with blood, the shape-fixed sponge can not only rapidly recover its original shape and concentrate platelets and RBCs but also activate the adhered platelets efficiently. The dual-functional sponge has excellent hemostatic efficacy in rat femoral artery hemorrhage and can control noncompressible hemorrhage in penetrating liver wound. Thus, we believe that this sponge could be a potential candidate as a topical hemostatic agent for uncontrolled and noncompressible hemorrhage.
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http://dx.doi.org/10.1016/j.actbio.2019.08.039DOI Listing
November 2019

Poly(lactide-co-glycolide) grafted hyaluronic acid-based electrospun fibrous hemostatic fragments as a sustainable anti-infection and immunoregulation material.

J Mater Chem B 2019 08;7(32):4997-5010

School of Ophthalmology & Optometry, Eye Hospital, School of Biomedical Engineering, Wenzhou Medical University, Wenzhou, Zhejiang 325027, China. and Wenzhou Institute of Biomaterials and Engineering, Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou, Zhejiang 325011, China and Engineering Research Center of Clinical Functional Materials and Diagnosis & Treatment Devices of Zhejiang Province, Wenzhou Institute of Biomaterials and Engineering, Wenzhou, Zhejiang 325011, China.

Poly(lactide-co-glycolide) (PLGA) copolymers are promising synthetic materials in the biomedical field. However, in wound management, their hydrophobic properties limit their further application because of their poor adhesion to the surface of moist wounds. Furthermore, the lack of hemostatic materials with sustainable anti-infection and immunoregulation functions remains a highly significant clinical problem, as commercially available hemostatic products, such as Arista™, Celox™ and QuikClot™, do not have sufficient infection prevention and immunoregulation properties. Herein, we employ electrospinning, ammonia dissociation and surface grafting techniques to develop a series of PLGA-based hemostatic materials, including a PLGA electrospun fibrous membrane, PLGA-NH2 fibrous particles and PLGA-hyaluronic acid fibrous fragments (PLGA-HA FFs). Notably, we load azithromycin on the PLGA-HA FFs to endow them with anti-infection and immunoregulation properties. The hemostatic mechanism analysis demonstrates that the PLGA-HA FFs show superior hemostasis performance compared to traditional gauzes. The results show that the PLGA-HA FFs can act as a versatile platform with high encapsulation of azithromycin (83.03% ± 2.81%) and rapid hemostasis (28 ± 2 s) as well as prominent cytocompatibility towards L929 cells, RAW 264.7 cells and red blood cells. We believe that the current research proposes a possible strategy to synthesize materials that achieve not only safe and effective hemostasis, but also have anti-infection and immunoregulation properties for the development of further hemostatic products.
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http://dx.doi.org/10.1039/c9tb00659aDOI Listing
August 2019

Fabricating antimicrobial peptide-immobilized starch sponges for hemorrhage control and antibacterial treatment.

Carbohydr Polym 2019 Oct 20;222:115012. Epub 2019 Jun 20.

Wenzhou Institute of Biomaterials and Engineering, Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou, Zhejiang 325011, China. Electronic address:

It is important to control immediate hemorrhage and prevent infection simultaneously in the wound management. However, most of hemostatic materials are associated with low efficiency of hemostasis, poor biocompatibility and lack of antimicrobial properties. A kind of starch-based macroporous sponges (KR-Sps) immobilized covalently with antimicrobial peptide KR12 using highly efficient thiol-ene photo click reaction were developed. The physical properties of these sponges could be fine-tuned by varying the ratio of modified starch/HS-PEG-SH and the polymer concentration. The in vitro and vivo results demonstrated that KR-Sps induced thrombosis, shortened clotting time and reduced the blood loss at bleeding site. Besides, KR12 immobilized sponge exhibited inherent antimicrobial properties against Gram (+) and (-) bacteria and methicillin-resistant Staphylococcus aureus (MRSA), which could maintain at least 5 days. Therefore, KR-Sps were believed to be an excellent candidate as hemostatic and antimicrobial product for the intraoperative wound management.
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http://dx.doi.org/10.1016/j.carbpol.2019.115012DOI Listing
October 2019

Idarubicin-loaded methoxy poly(ethylene glycol)--poly(l-lactide-co-glycolide) nanoparticles for enhancing cellular uptake and promoting antileukemia activity.

Int J Nanomedicine 2019 11;14:543-556. Epub 2019 Jan 11.

Laboratory of Internal Medicine, The First Affiliated Hospital of Wenzhou Medical University, Nanbaixiang, Ouhai District, Wenzhou, Zhejiang 325000, China,

Purpose: Nanoparticle (NP)-based drug delivery approaches have tremendous potential for enhancing treatment efficacy and decreasing doses of chemotherapeutics. Idarubicin (IDA) is one of the most common chemotherapeutic drugs used in the treatment of acute myeloid leukemia (AML). However, severe side effects and drug resistance markedly limit the application of IDA.

Methods: In this study, we encapsulated IDA in polymeric NPs and validated their antileukemia activity in vitro and in vivo.

Results: NPs with an average diameter of 84 nm was assembled from a methoxy poly(ethylene glycol)--poly(l-lactide-co-glycolide) (mPEG-PLGA). After loading of IDA, IDA-loaded mPEG-PLGA NPs (IDA/mPEG-PLGA NPs) were formed. The in vitro release data showed that the IDA/mPEG-PLGA NPs have excellent sustained release property. IDA/mPEG-PLGA NPs had exhibited the lower IC than pure IDA. Moreover, IDA/mPEG-PLGA NPs in the same concentration substantially induced apoptosis than did pure IDA. Most importantly, IDA/MPEG-PLGA NPs significantly decreased the infiltration of leukemia blasts and improved the overall survival of MLL-AF9-induced murine leukemia compared with free IDA. However, the blank NPs were nontoxic to normal cultured cells in vitro, suggesting that NPs were the safe carrier.

Conclusion: Our data suggest that IDA/mPEG-PLGA NPs might be a suitable carrier to encapsulate IDA. Low dose of IDA/mPEG-PLGA NPs can be used as a conventional dosage for antileukemia therapy to reduce side effect and improve survival.
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http://dx.doi.org/10.2147/IJN.S190027DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6333394PMC
February 2019

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

J Mater Chem B 2018 Oct 6;6(37):5975-5985. Epub 2018 Sep 6.

School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Tianjin 300350, China.

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.
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http://dx.doi.org/10.1039/c8tb01789aDOI Listing
October 2018

Tannic Acid Cross-linked Polysaccharide-Based Multifunctional Hemostatic Microparticles for the Regulation of Rapid Wound Healing.

Macromol Biosci 2018 11 20;18(11):e1800209. Epub 2018 Sep 20.

Wenzhou Institute of Biomaterials and Engineering, Ningbo Institute of Industrial Tehcnology (CNITECH), Chinese Academy of Sciences (CAS), Wenzhou, Zhejiang, 325011, China.

Hemostatic microparticles (HMs) have been widely used in surgery. To improve the comprehensive performance of HMs, multifunctional HMs named HM and HM ' are prepared from starch, carboxymethyl chitosan, hyaluronic acid, and tannic acid. Herein, tannic acid is used as an effective cross-linker. A 3D network structure for cell growth and wound repair can be formed by secondary cross-linking. Through synergistic effect of these natural materials, the process of wound healing can be regulated controllably. HM and HM ' have the ability of rapid hemostasis. Moreover, HM ' shows excellent properties in antibacteria and wound healing acceleration. Blood clotting time treated with different HMs is shortened obviously from 436.8 s to 126 s. Compared with Celox, HM and HM ' exhibited better broad spectrum antibacterial activity against both Escherichia coli and Staphylococcus aureus. Notably, the wound can be repaired rapidly by HM ' in 14 days. These multifunctional HMs might have an important prospect in clinical application.
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http://dx.doi.org/10.1002/mabi.201800209DOI Listing
November 2018

W Modified Polymeric Micelles with Different Hydrophobic Cores for Efficient Gene Delivery and Capillary-like Tube Formation.

ACS Biomater Sci Eng 2018 Aug 3;4(8):2870-2878. Epub 2018 Jul 3.

School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Tianjin 300350, China.

Recently, polymeric micelles with different biodegradable hydrophobic cores, such as poly(lactide--glycolide) (PLGA) and poly(lactide--3(S)-methyl-morpholine-2,5-dione) (PLMD), have been used for gene delivery. The biodegradable hydrophobic cores should play an important role in gene delivery. However, little research has focused on selectively promoting proliferation and migration of endothelial cells (ECs) as well as vascularization by altering hydrophobic cores of polymeric micelles. Herein, we prepared two kinds of W peptide (selective adhesion for ECs) modified micelles with PLGA and PLMD as hydrophobic cores, respectively, and poly(ethylene glycol) (PEG) and polyethylenimine (PEI) as mixed hydrophilic shell. Their ability of condensing pEGFP-ZNF580 (pZNF580) to form gene complexes was proved by agarose gel electrophoresis assay. MTT results showed that the relative cell viability of the micelles with PLMD cores was higher than control groups and the micelles with PLGA cores. The cellular uptake ability of these W modified gene complexes was higher than the complexes without W target function. A similar trend was also found in transfection tests in vitro, which further demonstrated the effect of W peptide and different hydrophobic cores on gene delivery. The number of migrated cells treated by the gene complexes with PLGA cores was 82 (nontarget group) and 115 (target group), whereas the complexes with PLMD cores was 88 (nontarget group) and 120 (target group). Capillary-like tube formation of W peptide modified complexes with PLMD core group was much higher (about 6 times) than the PEI(10 kDa)/pZNF580 group. These results demonstrated that transfection efficiency, cell proliferation, migration, and vascularization could be promoted by altering hydrophobic cores and W modification.
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http://dx.doi.org/10.1021/acsbiomaterials.8b00529DOI Listing
August 2018

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

J Mater Chem B 2018 Jul 20;6(25):4251-4263. Epub 2018 Jun 20.

School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Tianjin 300350, China.

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-H (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.
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http://dx.doi.org/10.1039/c8tb00786aDOI Listing
July 2018

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

J Nanobiotechnology 2018 Mar 26;16(1):29. Epub 2018 Mar 26.

School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, China.

Background: Gene 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 (H) 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-H (TAT: typical cell-penetrating peptide, NLS: nuclear localization signals, H: oligohistidine sequence, n: 4, 8 and 12) peptides with different H 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).

Results: MTT 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-H with long H 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-H/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.

Conclusion: The above results indicated that the delivery efficiency of pZNF580 and its expression could be enhanced by introducing H sequence into gene carriers. The H sequence in REDV-TAT-NLS-H is beneficial for high gene transfection. These REDV and H functionalized TAT-NLS peptides are promising gene carriers in gene therapy.
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http://dx.doi.org/10.1186/s12951-018-0358-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5870920PMC
March 2018

Core/Shell Gene Carriers with Different Lengths of PLGA Chains to Transfect Endothelial Cells.

Langmuir 2017 11 8;33(46):13315-13325. Epub 2017 Nov 8.

Department of Physiology and Pathophysiology, Logistics University of Chinese People's Armed Police Force , Tianjin 300162, China.

In order to improve the transfection efficiency and reduce the cytotoxicity of gene carriers, many strategies have been used to develop novel gene carriers. In this study, five complex micelles (MSP(2 k), MSP(4 k), MSP(6 k), MSP(8 k), and MSP(10 k)) were prepared from methoxy-poly(ethylene glycol)-b-poly(d,l-lactide-co-glycolide) (mPEG-b-PLGA) and sorbitol-poly(d,l-lactide-co-glycolide)-graft-PEI (sorbitol-PLGA-g-PEI, where the designed molecular weights of PLGA chains were 2 kDa, 4 kDa, 6 kDa, 8 kDa, and 10 kDa, respectively) copolymers by a self-assembly method, and the mass ratio of mPEG-b-PLGA to sorbitol-PLGA-g-PEI was 1/3. These complex micelles and their gene complexes had appropriate sizes and zeta potentials, and pEGFP-ZNF580 (pDNA) could be efficiently internalized into EA.hy926 cells by their gene complexes (MSP(2 k)/pDNA, MSP(4 k)/pDNA, MSP(6 k)/pDNA, MSP(8 k)/pDNA, and MSP(10 k)/pDNA). The MTT assay results demonstrated that the gene complexes had low cytotoxicity in vitro. When the hydrophobic PLGA chain increased above 6 kDa, the gene complexes showed higher performance than that prepared from short hydrophobic chains. Moreover, the relative ZNF580 protein expression levels in MSP(6 k)/pDNA, MSP(8 k)/pDNA, and MSP(10 k)/pDNA) groups were 79.6%, 71.2%, and 73%, respectively. These gene complexes could promote the transfection of endothelial cells, while providing important information and insight for the design of new and effective gene carriers to promote the proliferation and migration of endothelial cells.
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http://dx.doi.org/10.1021/acs.langmuir.7b02934DOI Listing
November 2017

Multi-targeting peptides for gene carriers with high transfection efficiency.

J Mater Chem B 2017 Oct 4;5(40):8035-8051. Epub 2017 Oct 4.

School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Tianjin 300350, China.

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.
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http://dx.doi.org/10.1039/c7tb02012kDOI Listing
October 2017

Design and development of polysaccharide hemostatic materials and their hemostatic mechanism.

Biomater Sci 2017 Nov;5(12):2357-2368

School of Chemical Engineering and Technology, Collaborative Innovation Center of Chemical Science and Chemical Engineering (Tianjin), Tianjin University, Tianjin 300072, China.

The formation of stable blood clots or hemostasis is essential to prevent major blood loss and death from excessive bleeding. However, the body's own coagulation process is not able to accomplish timely hemostasis without the assistance of hemostatic agents. For developing novel topical hemostatic agents, tissue adhesives and sealants, it is necessary to understand the coagulation process and the hemostasis mechanism of different materials. Among hemostatic materials, polysaccharides are naturally derived polymers having excellent biodegradable and biocompatible properties. This review provides an overview of polysaccharide-based hemostatic materials and agents, including their advantages and drawbacks in hemostatic applications. Furthermore, polysaccharide-based hemostatic materials with anti-microbial and healing functions are also introduced.
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http://dx.doi.org/10.1039/c7bm00554gDOI Listing
November 2017

Multifunctional Gene Carriers with Enhanced Specific Penetration and Nucleus Accumulation to Promote Neovascularization of HUVECs in Vivo.

ACS Appl Mater Interfaces 2017 Oct 5;9(41):35613-35627. Epub 2017 Oct 5.

Department of Physiology and Pathophysiology, Logistics University of Chinese People's Armed Police Force , Tianjin 300309, China.

Recently, gene therapy has attracted much attention, especially for the treatment of vascular disease. However, it is still challenging to develop the gene carriers with high biocompatibility as well as highly efficient gene delivery to overcome multiple barriers. Herein, a frequently used cell-penetrating peptide PKKKRKV (TAT) was selected as a functional sequence of the gene carrier with distinctive cell-penetrating ability. REDV peptide with selectively targeting function for endothelial cells (ECs) and nuclear localization signals (NLS) were integrated with this TAT peptide to obtain a highly efficient gene delivery system with ECs specificity and nucleus accumulation capacity. Besides, the glycine sequences with different repeat numbers were inserted into the above integrated peptide. These glycine sequences acted as a flexible spacer arm to exert the targeting, cell-penetrating, and nucleus accumulation functions of each functional peptide. Three tandem peptides REDV-G-TAT-G-NLS (m = 0, 1, and 4) complexed with pZNF580 plasmid to form gene complexes. The results of hemocompatibility and cytocompatibility indicated that these peptides and gene complexes were nontoxic and biocompatible. The internalization efficiency and mechanism of these gene complexes were investigated. The internalization efficiency was improved as the introduction of targeting REDV and glycine sequence, and the REDV-G-TAT-G-NLS/pZNF580 (TP-G4/pZNF580) complexes showed the highest cellular uptake among the gene complexes. The TP-G4/pZNF580 complexes also presented significantly higher internalization efficiency (∼1.36 times) in human umbilical vein endothelial cells (HUVECs) than human umbilical artery smooth muscle cells. TP-G4/pZNF580 complexes substantially promoted the expression of pZNF580 by confocal live cell imaging, gene delivery efficiency, and HUVECs migration assay. The in vitro and in vivo revascularization ability of transfected HUVECs was further enhanced obviously. In conclusion, these multifunctional REDV-G-TAT-G-NLS peptides offer a promising and efficacious delivery option for neovascularization to treat vascular diseases.
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http://dx.doi.org/10.1021/acsami.7b11615DOI Listing
October 2017

Bioreducible, hydrolytically degradable and targeting polymers for gene delivery.

J Mater Chem B 2017 May 5;5(18):3253-3276. Epub 2017 Apr 5.

School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Tianjin 300350, China.

Recently, synthetic gene carriers have been intensively developed owing to their promising application in gene therapy and considered as a suitable alternative to viral vectors because of several benefits. But cationic polymers still face some problems like low transfection efficiency, cytotoxicity, and poor cell recognition and internalization. The emerging engineered and smart polymers can respond to some changes in the biological environment like pH change, ionic strength change and redox potential, which is beneficial for cellular uptake. Redox-sensitive disulfide based and hydrolytically degradable cationic polymers serve as gene carriers with excellent transfection efficiency and good biocompatibility owing to degradation in the cytoplasm. Additionally, biodegradable polymeric micelles with cell-targeting function are recently emerging gene carriers, especially for the transfection of endothelial cells. In this review, some strategies for gene carriers based on these bioreducible and hydrolytically degradable polymers will be illustrated.
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http://dx.doi.org/10.1039/c7tb00275kDOI Listing
May 2017

CAGW Peptide Modified Biodegradable Cationic Copolymer for Effective Gene Delivery.

Polymers (Basel) 2017 Apr 28;9(5). Epub 2017 Apr 28.

Institute of Biomaterials and Engineering, Wenzhou Medical University, Wenzhou 325011, Zhejiang, China.

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--glycolide)-graft-PEI (PLGA--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--PEI--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.
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http://dx.doi.org/10.3390/polym9050158DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6432137PMC
April 2017

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

J Mater Chem B 2017 Feb 9;5(8):1673-1687. Epub 2017 Feb 9.

School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Tianjin 300350, China.

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.
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http://dx.doi.org/10.1039/c6tb02212jDOI Listing
February 2017

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

ACS Appl Mater Interfaces 2017 Feb 24;9(5):4485-4497. Epub 2017 Jan 24.

Department of Physiology and Pathophysiology, Logistics University of Chinese People's Armed Police Force , Tianjin 300162, China.

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.
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http://dx.doi.org/10.1021/acsami.6b14769DOI Listing
February 2017

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

Biomater Sci 2017 Feb;5(3):511-522

Department of Physiology and Pathophysiology, Logistics University of Chinese People's Armed Police Force, Tianjin 300162, China.

In this work, a biodegradable star-shaped copolymer poly(lactide-co-3(S)-methyl-morpholine-2,5-dione) (Star-(PLMD)) was synthesized via ring-opening polymerization (ROP), and subsequently a gene carrier Star-PLMD-g-PEI-g-PEG-CREDVW was prepared by grafting polyethyleneimine (PEI), polyethylene glycol (PEG) and targeting peptide REDV onto Star-(PLMD). This gene carrier could form stable micelles to condense pEGFP-ZNF580 through electrostatic interaction. The resulting complexes were biocompatible and showed high efficiency in gene delivery. In addition, these complexes exhibited high selectivity for endothelial cells (ECs), high transfection efficiency and enhanced migration of ECs. The protein level of ZNF580 expression was significantly high (up to 85%), while the control group was only 51%. This combination of degradability, targeting ligand and star-structure strategy exhibits a significant advantage in transfection efficiency and migration of ECs.
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http://dx.doi.org/10.1039/c6bm00856aDOI Listing
February 2017

Co-self-assembly of cationic microparticles to deliver pEGFP-ZNF580 for promoting the transfection and migration of endothelial cells.

Int J Nanomedicine 2017;12:137-149. Epub 2016 Dec 20.

Department of Physiology and Pathophysiology, Logistics University of Chinese People's Armed Police Force, Tianjin, People's Republic of China.

The gene transfection efficiency of polyethylenimine (PEI) varies with its molecular weight. Usually, high molecular weight of PEI means high gene transfection, as well as high cytotoxicity in gene delivery in vivo. In order to enhance the transfection efficiency and reduce the cytotoxicity of PEI-based gene carriers, a novel cationic gene carrier was developed by co-self-assembly of cationic copolymers. First, a star-shaped copolymer poly(3(S)-methyl-morpholine-2,5-dione-co-lactide) (P(MMD-co-LA)) was synthesized using D-sorbitol as an initiator, and the cationic copolymer (P(MMD-co-LA)-g-PEI) was obtained after grafting low-molecular weight PEI. Then, by co-self-assembly of this cationic copolymer and a diblock copolymer methoxy-poly(ethylene glycol) (mPEG)-b-P(MMD-co-LA), microparticles (MPs) were formed. The core of MPs consisted of a biodegradable block of P(MMD-co-LA), and the shell was formed by mPEG and PEI blocks. Finally, after condensation of pEGFP-ZNF580 by these MPs, the plasmids were protected from enzymatic hydrolysis effectively. The result indicated that pEGFP-ZNF580-loaded MP complexes were suitable for cellular uptake and gene transfection. When the mass ratio of mPEG-b-P(MMD-co-LA) to P(MMD-co-LA)-g-PEI reached 3/1, the cytotoxicity of the complexes was very low at low concentration (20 μg mL). Additionally, pEGFP-ZNF580 could be transported into endothelial cells (ECs) effectively via the complexes of MPs/pEGFP-ZNF580. Wound-healing assay showed that the transfected ECs recovered in 24 h. Cationic MPs designed in the present study could be used as an applicable gene carrier for the endothelialization of artificial blood vessels.
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http://dx.doi.org/10.2147/IJN.S107593DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5191575PMC
February 2017

Multitargeting Gene Delivery Systems for Enhancing the Transfection of Endothelial Cells.

Macromol Rapid Commun 2016 Dec 28;37(23):1926-1931. Epub 2016 Sep 28.

Department of Physiology and Pathophysiology, Logistics University of Chinese People's Armed Police Force, Tianjin, 300162, China.

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.
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http://dx.doi.org/10.1002/marc.201600345DOI Listing
December 2016

PEI-g-P(LA-co-MMD)-g-PEI nanoparticles as gene carriers to promote the proliferation of human vascular endothelial cells.

J Control Release 2015 Sep 19;213:e89-90. Epub 2015 Aug 19.

School of Chemical Engineering and Technology, Collaborative Innovation Center of Chemical Science and Chemical Engineering (Tianjin), Tianjin University, Tianjin 300072, China; Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072, China. Electronic address:

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http://dx.doi.org/10.1016/j.jconrel.2015.05.149DOI Listing
September 2015

REDV Peptide Conjugated Nanoparticles/pZNF580 Complexes for Actively Targeting Human Vascular Endothelial Cells.

ACS Appl Mater Interfaces 2015 Sep 3;7(36):20389-99. Epub 2015 Sep 3.

School of Chemical Engineering and Technology, Tianjin University , Tianjin 300072, China.

Herein, we demonstrate that the REDV peptide modified nanoparticles (NPs) can serve as a kind of active targeting gene carrier to condensate pZNF580 for specific promotion of the proliferation of endothelial cells (ECs). First, we synthesized a series of biodegradable amphiphilic copolymers by ring-opening polymerization reaction and graft modification with REDV peptide. Second, we prepared active targeting NPs via self-assembly of the amphiphilic copolymers using nanoprecipitation technology. After condensation with negatively charged pZNF580, the REDV peptide modified NPs/pZNF580 complexes were formed finally. Due to the binding affinity toward ECs of the specific peptide, these REDV peptide modified NPs/pZNF580 complexes could be recognized and adhered specifically by ECs in the coculture system of ECs and human artery smooth muscle cells (SMCs) in vitro. After expression of ZNF580, as the key protein to promote the proliferation of ECs, the relative ZNF580 protein level increased from 15.7% to 34.8%. The specificity in actively targeting ECs of the REDV peptide conjugated NPs/pZNF580 complexes was still retained in the coculture system. These findings in the present study could facilitate the development of actively targeting gene carriers for the endothelialization of artificial blood vessels.
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http://dx.doi.org/10.1021/acsami.5b06286DOI Listing
September 2015

Surface tailoring for selective endothelialization and platelet inhibition via a combination of SI-ATRP and click chemistry using Cys-Ala-Gly-peptide.

Acta Biomater 2015 Jul 1;20:69-81. Epub 2015 Apr 1.

Department of Physiology and Pathophysiology, Longistics University of Chinese People's Armed Police Force, Tianjin 300162, China. Electronic address:

Surface tailoring is an attractive approach to enhancing selective endothelialization, which is a prerequisite for current vascular prosthesis applications. Here, we modified polycarbonate urethane (PCU) surface with both poly(ethylene glycol) and Cys-Ala-Gly-peptide (CAG) for the purpose of creating a hydrophilic surface with targeting adhesion of endothelial cells (ECs). In the first step, PCU-film surface was grafted with poly(ethylene glycol) methacrylate (PEGMA) to covalently tether hydrophilic polymer brushes via surface initiated atom transfer radical polymerization (SI-ATRP), followed by grafting of an active monomer pentafluorophenyl methacrylate (PFMA) by a second ATRP. The postpolymerization modification of the terminal reactive groups with allyl amine molecules created pendant allyl groups, which were subsequently functionalized with cysteine terminated CAG-peptide via photo-initiated thiol-ene click chemistry. The functionalized surfaces were characterized by water contact angle and XPS analysis. The growth and proliferation of human ECs or human umbilical arterial smooth muscle cells on the functionalized surfaces were investigated for 1, 3 and 7 day/s. The results indicated that these peptide functionalized surfaces exhibited enhanced EC adhesion, growth and proliferation. Furthermore, they suppressed platelet adhesion in contact with platelet-rich plasma for 2h. Therefore, these surfaces with EC targeting ligand could be an effective anti-thrombogenic platform for vascular tissue engineering application.
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http://dx.doi.org/10.1016/j.actbio.2015.03.032DOI Listing
July 2015

Nanoparticles complexed with gene vectors to promote proliferation of human vascular endothelial cells.

Adv Healthc Mater 2015 Jun 9;4(8):1225-35. Epub 2015 Mar 9.

School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China.

Amphiphilic block copolymers containing biodegradable hydrophobic segments of depsipeptide based copolymers have been synthesized and explored as gene carriers for enhancing proliferation of endothelial cells in vitro. These polymers form nanoparticles (NPs) with positive charges on their surface, which could condense recombinant plasmids of enhanced green fluorescent protein plasmid and ZNF580 gene (pEGFP-ZNF580) and protect them against DNase I. ZNF580 gene is efficiently transported into EA.hy926 cells to promote their proliferation, whereby the transfection efficiency of NPs/pEGFP-ZNF580 is approximately similar to that of Lipofectamine 2000. These results indicate that the NPs might have potential as a carrier for pEGFP-ZNF580, which could support endothelialization of cardiovascular implants.
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http://dx.doi.org/10.1002/adhm.201400817DOI Listing
June 2015