Bioactivity of silk fibroin peptides on vascular endothelial cells

Abstract

To determine the contribution of non-repetitive domains to the bioactivity of the heavy chain in silk fibroin (SF) macromolecules, a gene motif f(1) encoding this fragment and its multimers (f(4) and f(8)) were biosynthesized from Escherichia coli BL21. Based on the positive application potential of SF materials for the vascular tissue engineering, this study focused on examining the active response of these polypeptides to vascular endothelial cells. Biosynthetic polypeptides F(1), F(4), and F(8) were separately grafted onto the surfaces of bioinert polyethylene terephthalate (PET) films, resulting in remarkable improvements in the spread and proliferation of human umbilical vein endothelial cells (HUVECs). Using the same grafting dose, the activity of cells on polypeptide-modified PET films enhanced with the increase of the molecular weight of those grafted polypeptides from F(1) to F(8). Meanwhile, the growth of cells on the surface of the alkaline-treated PET film was improved, indicating that the hydrophilicity of the surface material had influence on the growth of HUVECs. Moreover, on surfaces with the same water contact angle, the spread and proliferation activity of cells on PET films were significantly lower than those on polypeptide-modified PET films.

Graphical abstract

Keywords

silk fibroin heavy chain / non-repetitive fragment / cytoactive / endothelial cell

References

[1] Xu J J, Wang Y N, Ding M Y, . Sequence-structure characterization of recombinant polypeptides derived from silk fibroin heavy chain.Materials Science and Engineering C, 2020, 111: 110831

[2] Huang H Y, Tian Z F, Yi H G, . Designing and cloning of the gene sequence encoding silk fibroin amorphous domain.Journal of Donghua University (English Edition), 2012, 29(6): 489–492

[3] Song G Z, Zheng C D, Liu Y F, . In vitro extracellular matrix deposition by vascular smooth muscle cells grown in fibroin scaffolds, and the regulation of TGF-β1.Materials & Design, 2021, 199: 109428

[4] Alessandrino A, Chiarini A, Biagiotti M, . Three-layered silk fibroin tubular scaffold for the repair and regeneration of small caliber blood vessels: from design to in vivo pilot tests.Frontiers in Bioengineering and Biotechnology, 2019, 7: 356

[5] Li H L, Wang Y N, Sun X L, . Steady-state behavior and endothelialization of a silk-based small-caliber scaffold in vivo transplantation.Polymers, 2019, 11(8): 1303

[6] Bai S M, Han H Y, Huang X W, . Silk scaffolds with tunable mechanical capability for cell differentiation.Acta Biomaterialia, 2015, 20: 22–31

[7] Yamada H, Igarashi Y, Takasu Y, . Identification of fibroin-derived peptides enhancing the proliferation of cultured human skin fibroblasts.Biomaterials, 2004, 25(3): 467–472

[8] Kim E D, Bayaraa T, Shin E J, . Fibroin-derived peptides stimulate glucose transport in normal and insulin-resistant 3T3-L1 adipocytes.Biological & Pharmaceutical Bulletin, 2009, 32(3): 427–433

[9] Machado R, Azevedo-Silva J, Correia C, . High level expression and facile purification of recombinant silk-elastin-like polymers in auto induction shake flask cultures.AMB Express, 2013, 3(1): 11

[10] An B, DesRochers T M, Qin G, . The influence of specific binding of collagen–silk chimeras to silk biomaterials on hMSC behavior.Biomaterials, 2013, 34(2): 402–412

[11] Kuang H Z, Wang Y, Shi Y, . Construction and performance evaluation of Hep/silk–PLCL composite nanofiber small-caliber artificial blood vessel graft.Biomaterials, 2020, 259: 120288

[12] Li H L, Song G Z, Tian W, . Motility and function of smooth muscle cells in a silk small-caliber tubular scaffold after replacement of rabbit common carotid artery.Materials Science and Engineering C, 2020, 114: 110977

[13] Xu S, Li Q T, Pan H T, . Tubular silk fibroin/gelatin-tyramine hydrogel with controllable layer structure and its potential application for tissue engineering.ACS Biomaterials Science & Engineering, 2020, 6(12): 6896–6905

[14] Yang G Q, Wu M Y, Yi H G, . Biosynthesis and characterization of a non-repetitive polypeptide derived from silk fibroin heavy chain.Materials Science and Engineering C, 2016, 59: 278–285

[15] Zhao H R, Yang Y X, Yi H G, . Biosynthesis of a potentially functional polypeptide derived from silk fibroin.Bio-Medical Materials and Engineering, 2014, 24(6): 2057–2064

[16] Wang J N, Yan S Q, Lu C D, . Biosynthesis and characterization of typical fibroin crystalline polypeptides of silkworm Bombyx mori.Materials Science and Engineering C, 2009, 29(4): 1321–1325

[17] Shi P G, Zhang L, Tian W, . Preparation and anticoagulant activity of functionalised silk fibroin.Chemical Engineering Science, 2019, 199: 240–248

[18] Jaziri H, Mokhtar S, Kyosev Y, . Influence of fatigue stress on the radial strength of polymeric braided vascular stents.Polymers for Advanced Technologies, 2022, 33(2): 627–637

[19] Jirofti N, Mohebbi-Kalhori D, Samimi A, . Fabrication and characterization of a novel compliant small-diameter PET/PU/PCL triad-hybrid vascular graft.Biomedical Materials, 2020, 15(5): 055004

[20] Van de Voorde B, Benmeridja L, Giol E D, . Potential of poly(alkylene terephthalate)s to control endothelial cell adhesion and viability.Materials Science and Engineering C, 2021, 129: 112378

[21] Giol E D, Van Vlierberghe S, Unger R E, . Biomimetic strategy towards gelatin coatings on PET.Effect of protocol on coating stability and cell-interactive properties. Journal of Materials Chemistry B: Materials for Biology and Medicine, 2019, 7(8): 1258–1269

[22] Tzoneva R, Seifert B, Albrecht W, . Poly(ether imide) membranes: studies on the effect of surface modification and protein pre-adsorption on endothelial cell adhesion, growth and function.Journal of Biomaterials Science: Polymer Edition, 2008, 19(7): 837–852

[23] Huang Z S, Bi L, Zhang Z Y, . Effects of dimethylolpropionic acid modification on the characteristics of polyethylene terephthalate fibers.Molecular Medicine Reports, 2012, 6(4): 709–715

[24] Jiang Y C, Wang H A, Wang X F, . Surface modification with hydrophilic and heparin-loaded coating for endothelialization and anticoagulation promotion of vascular scaffold.International Journal of Biological Macromolecules, 2022, 219: 1146–1154

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