The effect of hirudin modification of silk fibroin on cell growth and antithrombogenicity

Wang, Qiongyu; Tu, Fangfang; Liu, Yunfei; Zhang, Yujin; Li, Helei; Zhao, Kang; Yin, Yin; Wang, Jiannan

doi:10.1016/j.msec.2017.02.035

Cited by 20 publications

(11 citation statements)

References 39 publications

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“…Silk fibroin is a natural protein synthesized and secreted by silkworms and spiders. Bombyx mori silk fibroin has been extensively used for biomaterials because it is readily obtainable on large quantities ad displays good biocompatibility and biodegradability, resulting in practicable cell adhesion and cell spreading, high proliferative and anti-thrombosis activities, and potent ability to induce tissue repair [1,2,3,4,5,6,7]. Wild silkworm species such as Antheraea yamamai , Antheraea pernyi , Antheraea mylitta , Antheraea assama , and Philosamia ricini secrete another type of silk fibroin with arginine-glycine-aspartate (RGD) motif repeats in the protein chain [8,9,10,11,12], which is absent in mulberry ( B. mori ) silk fibroin [13].…”

Section: Introductionmentioning

confidence: 99%

An RGD-Containing Peptide Derived from Wild Silkworm Silk Fibroin Promotes Cell Adhesion and Spreading

Zhao

Wang

et al. 2018

Polymers

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Arginine-Glycine-Aspartate (RGD) tripeptide can promote cell adhesion when present in the amino acid of proteins such as fibronectin. In order to demonstrate the bioactivity of an RGD-containing silk protein, a gene encoding the RGD motif-containing peptide GSGAGGRGDGGYGSGSS (–RGD–) derived from nonmulberry silk was designed and cloned, then multimerised and inserted into a commercial pGEX expression vector for recombinant expression of (–RGD–)n peptides. Herein, we focus on two glutathione-S-transferase (GST)-tagged fusion proteins, GST–(–RGD–)4 and GST–(–RGD–)8, which were expressed in Escherichia coli BL21, purified by GST affinity chromatography, and analyzed with sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry (MS). Target peptides (–RGD–)4 and (–RGD–)8 (6.03 and 11.5 kDa) were cleaved from the GST-tag by thrombin digestion, as verified with MS and SDS-PAGE. Isoelectric point analysis confirmed that target peptides were expressed and released in accordance with the original design. Target peptides self-assembled into a mainly α-helical structure, as determined by circular dichroism spectroscopy. Furthermore, (–RGD–)4 and (–RGD–)8 modified mulberry silk fibroin films were more effective for rapid cell adhesion, spreading and proliferative activity of L929 cells than some chemically synthesized RGD peptides modified and mulberry silk lacking the RGD motif.

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Section: Introductionmentioning

confidence: 99%

An RGD-Containing Peptide Derived from Wild Silkworm Silk Fibroin Promotes Cell Adhesion and Spreading

Zhao

Wang

et al. 2018

Polymers

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“…Regenerated silk fibroin possesses better blood compatibility in comparison with poly-tetrafluoroethylene and Dacron [27]. Certainly, scholars have put forth more effort to improve the antithrombogenicity of small-caliber vascular transplantation by modification with heparin [28,29] or hirudin [27,30]. Here, the SFTSs with a fabric design in the medium showed remarkable antithrombotic properties, and satisfactory resistance and resilience to tissue oppression or blood pressure, owing to their excellent mechanical properties [17,18].…”

Section: Resultsmentioning

confidence: 99%

Steady-State Behavior and Endothelialization of a Silk-Based Small-Caliber Scaffold In Vivo Transplantation

Sun

Tian

et al. 2019

Polymers

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A silk-based small-caliber tubular scaffold (SFTS), which is fabricated using a regenerated silk fibroin porous scaffold embedding a silk fabric core layer, has been proved to possess good cell compatibility and mechanical properties in vitro. In this study, the endothelialization ability and the steady-state blood flow of SFTSs were evaluated in vivo by implanting and replacing a common carotid artery in a rabbit. The results of the color doppler ultrasound and angiographies showed that the blood flow was circulated in the grafts without aneurysmal dilations or significant stenoses at any time point, and ran stronger and close to the autologous blood vessel from one month after implantation. The SFTSs presented an initial tridimensionality without being distorted or squashed. SEM and immunohistochemistry results showed that a clear and discontinuous endodermis appeared after one month of implantation; when implanted for three months, an endothelial layer fully covered the inner surface of SFTSs. RT-PCR results indicated that the gene expression level of CD31 in SFTSs was 45.8% and 75.3% by that of autologous blood vessels at 3 months and 12 months, respectively. The VEGF gene showed a high expression level that continued to increase after implantation.

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“…Increasing evidence validates that patients with cardiovascular disease can be treated and controlled with hirudin [ 35 – 37 ]. Researchers has found that hirudin contributed to suppressing growth of fibroblasts [ 38 ], and to eliminating ROS accumulation [ 39 ]. Our results demonstrated that hirudin decreased viability of Ang II-induced myocardial fibroblasts, depressed ROS accumulation, LDH activity and MDA content, and enhanced SOD activity.…”

Section: Discussionmentioning

confidence: 99%

Hirudin Protects Ang II-Induced Myocardial Fibroblasts Fibrosis by Inhibiting the Extracellular Signal-Regulated Kinase1/2 (ERK1/2) Pathway

Wang

Jin

2018

Med Sci Monit

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BackgroundMyocardial fibrosis is closely related to all types of cardiovascular diseases. Hirudin is widely used in the prevention and treatment of cardiovascular diseases and cancers. In this study, we examined the potential role(s) and mechanism of hirudin in angiotensin II (Ang II)-induced myocardial fibrosis.Material/MethodsThe viability of myocardial fibroblasts, and reactive oxygen species (ROS) rates were measured respectively using cell counting kit-8 (CCK-8) and flow cytometry. Malondialdehyde (MDA) content, the activities of lactate dehydrogenase (LDH), and superoxide dismutase (SOD) were detected by the respective kits. The mRNA and protein levels of fibrosis-related factors were separately assessed by qRT-PCR and western blot.ResultsOur data revealed that hirudin suppressed the viability of myocardial fibroblasts, and that it relieved the proliferation induced by Ang II in a dose-dependent manner. We also found that hirudin reduced ROS production, LDH activity, and MDA content; however, it enhanced SOD activity. Moreover, while hirudin significantly downregulated the levels of matrix metalloproteinase-2 (MMP-2), MMP-9, fibronectin (FN), transforming growth factor beta 1 (TGF-β1), collagen-I (COL-I), and COL-III, it upregulated the expression level of tissue inhibitor of metalloproteinases-2 (TIMP-2). Furthermore, phosphorylated extracellular signal-regulated kinase1/2 (p-ERK1/2) was decreased by hirudin, compared to the Ang-II group.ConclusionsHirudin depressed Ang II-induced myocardial fibroblasts via inhibiting oxidative stress, regulating fibrosis-related factors, and repressing the ERK1/2 pathway.

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The effect of hirudin modification of silk fibroin on cell growth and antithrombogenicity

Cited by 20 publications

References 39 publications

An RGD-Containing Peptide Derived from Wild Silkworm Silk Fibroin Promotes Cell Adhesion and Spreading

An RGD-Containing Peptide Derived from Wild Silkworm Silk Fibroin Promotes Cell Adhesion and Spreading

Steady-State Behavior and Endothelialization of a Silk-Based Small-Caliber Scaffold In Vivo Transplantation

Hirudin Protects Ang II-Induced Myocardial Fibroblasts Fibrosis by Inhibiting the Extracellular Signal-Regulated Kinase1/2 (ERK1/2) Pathway

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