Protein-coated resorbable synthetic polymeric nanofibrous membranes are promising for the fabrication of advanced skin substitutes. We fabricated electrospun polylactic acid and poly(lactide-co-glycolic acid) nanofibrous membranes and coated them with fibrin or collagen I. Fibronectin was attached to a fibrin or collagen nanocoating, in order further to enhance the cell adhesion and spreading. Fibrin regularly formed a coating around individual nanofibers in the membranes, and also formed a thin noncontinuous nanofibrous mesh on top of the membranes. Collagen also coated most of the fibers of the membrane and randomly created a soft gel on the membrane surface. Fibronectin predominantly adsorbed onto a thin fibrin mesh or a collagen gel, and formed a thin nanofibrous structure. Fibrin nanocoating greatly improved the attachment, spreading, and proliferation of human dermal fibroblasts, whereas collagen nanocoating had a positive influence on the behavior of human HaCaT keratinocytes. In addition, fibrin stimulated the fibroblasts to synthesize fibronectin and to deposit it as an extracellular matrix. Fibrin coating also showed a tendency to improve the ultimate tensile strength of the nanofibrous membranes. Fibronectin attached to fibrin or to a collagen coating further enhanced the adhesion, spreading, and proliferation of both cell types.

Department of Anatomy and Biomechanics Faculty of Physical Education and Sport Charles University

Department of Biomaterials and Tissue Engineering Institute of Physiology Czech Academy of Sciences

Department of Biomaterials and Tissue Engineering Institute of Physiology Czech Academy of Sciences; 2nd Faculty of Medicine Charles University Prague

Department of Biomaterials and Tissue Engineering Institute of Physiology Czech Academy of Sciences; Department of Anatomy and Biomechanics Faculty of Physical Education and Sport Charles University

Department of Chemistry and Physics of Surfaces and Biointerfaces Institute of Macromolecular Chemistry

Department of Composites and Carbon Materials Institute of Rock Structure and Mechanics Czech Academy of Sciences Prague Czech Republic

InStar Technologies Liberec

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Eisenbud D, Huang NF, Luke S, Silberklang M. Skin substitutes and wound healing: current status and challenges. Wounds. 2004;16(1):2–17.

McMillan JR, Akiyama M, Tanaka M, et al. Small-diameter porous poly(ε-caprolactone) films enhance adhesion and growth of human cultured epidermal keratinocyte and dermal fibroblast cells. Tissue Eng. 2007;13(4):789–798. PubMed

Sun L, Stout DA, Webster TJ. The nano-effect: improving the long-term prognosis for musculoskeletal implants. J Long Term Eff Med Implants. 2012;22(3):195–209. PubMed

Bacakova L, Filova E, Parizek M, Ruml T, Svorcik V. Modulation of cell adhesion, proliferation and differentiation on materials designed for body implants. Biotechnol Adv. 2011;29(6):739–767. PubMed

Groeber F, Holeiter M, Hampel M, Hinderer S, Schenke-Layland K. Skin tissue engineering: in vivo and in vitro applications. Adv Drug Deliv Rev. 2011;63(4–5):352–366. PubMed

Kai D, Liow SS, Loh XJ. Biodegradable polymers for electrospinning: towards biomedical applications. Mater Sci Eng C Mater Biol Appl. 2014;45:659–670. PubMed

Hoveizi E, Nabiuni M, Parivar K, Rajabi-Zeleti S, Tavakol S. Functionalisation and surface modification of electrospun polylactic acid scaffold for tissue engineering. Cell Biol Int. 2014;38(1):41–49. PubMed

Bacakova M, Lopot F, Hadraba D, et al. Effects of fiber density and plasma modification of nanofibrous membranes on the adhesion and growth of HaCaT keratinocytes. J Biomater Appl. 2015;29(6):837–853. PubMed

Bacakova M, Musilkova J, Riedel T, et al. The potential applications of fibrin-coated electrospun polylactide nanofibers in skin tissue engineering. Int J Nanomedicine. 2016;11:771–789. PubMed PMC

Rajangam T, An SS. Fibrinogen and fibrin based micro and nano scaffolds incorporated with drugs, proteins, cells and genes for therapeutic biomedical applications. Int J Nanomedicine. 2013;8:3641–3662. PubMed PMC

O’Toole EA. Extracellular matrix and keratinocyte migration. Clin Exp Dermatol. 2001;26(6):525–530. PubMed

Auxenfans C, Fradette J, Lequeux C, et al. Evolution of three dimensional skin equivalent models reconstructed in vitro by tissue engineering. Eur J Dermatol. 2009;19(2):107–113. PubMed

Nuutila K, Peura M, Suomela S, et al. Recombinant human collagen III gel for transplantation of autologous skin cells in porcine full-thickness wounds. J Tissue Eng Regen Med. 2015;9(12):1386–1393. PubMed

Sarkar SD, Farrugia BL, Dargaville TR, Dhara S. Chitosan-collagen scaffolds with nano/microfibrous architecture for skin tissue engineering. J Biomed Mater Res A. 2013;101(12):3482–3492. PubMed

Peh P, Lim NS, Blocki A, et al. Simultaneous delivery of highly diverse bioactive compounds from blend electrospun fibers for skin wound healing. Bioconjug Chem. 2015;26(7):1348–1358. PubMed

Niiyama H, Kuroyanagi Y. Development of novel wound dressing composed of hyaluronic acid and collagen sponge containing epidermal growth factor and vitamin C derivative. J Artif Organs. 2014;17(1):81–87. PubMed

Butler CE, Orgill DP. Simultaneous in vivo regeneration of neodermis, epidermis, and basement membrane. Adv Biochem Eng Biotechnol. 2005;94:23–41. PubMed

Wang F, Wang M, She Z, et al. Collagen/chitosan based two- compartment and bi-functional dermal scaffolds for skin regeneration. Mater Sci Eng C Mater Biol Appl. 2015;52:155–162. PubMed

Anish S. Skin substitutes in dermatology. Indian J Dermatol Venereol Leprol. 2015;81(2):175–178. PubMed

Debels H, Hamdi M, Abberton K, Morrison W. Dermal matrices and bioengineered skin substitutes: a critical review of current options. Plast Reconstr Surg Glob Open. 2015;3(1):e284. PubMed PMC

Mutsaers SE, Bishop JE, McGrouther G, Laurent GJ. Mechanisms of tissue repair: from wound healing to fibrosis. Int J Biochem Cell Biol. 1997;29(1):5–17. PubMed

Laurens N, Koolwijk P, de Maat MP. Fibrin structure and wound healing. J Thromb Haemost. 2006;4(5):932–939. PubMed

Gorodetsky R, Clark RA, An J, et al. Fibrin microbeads (FMB) as biodegradable carriers for culturing cells and for accelerating wound healing. J Invest Dermatol. 1999;112(6):866–872. PubMed

Fabris G, Trombelli L, Schincaglia GP, Cavallini R, Calura G, del Senno L. Effects of a fibrin-fibronectin sealing system on proliferation and type I collagen synthesis of human PDL fibroblasts in vitro. J Clin Periodontol. 1998;25(1):11–14. PubMed

Ahmed TA, Dare EV, Hincke M. Fibrin: a versatile scaffold for tissue engineering applications. Tissue Eng Part B Rev. 2008;14(2):199–215. PubMed

Braziulis E, Biedermann T, Hartmann-Fritsch F, et al. Skingineering I: engineering porcine dermo-epidermal skin analogues for autologous transplantation in a large animal model. Pediatr Surg Int. 2011;27(3):241–247. PubMed

Mazzone L, Pontiggia L, Reichmann E, Ochsenbein-Kölble N, Moehrlen U, Meuli M. Experimental tissue engineering of fetal skin. Pediatr Surg Int. 2014;30(12):1241–1247. PubMed

Monteiro IP, Gabriel D, Timko BP, et al. A two-component pre-seeded dermal-epidermal scaffold. Acta Biomater. 2014;10(12):4928–4938. PubMed PMC

de la Puente P, Ludena D, Fernandez A, Aranda JL, Varela G, Iglesias J. Autologous fibrin scaffolds cultured dermal fibroblasts and enriched with encapsulated bFGF for tissue engineering. J Biomed Mater Res A. 2011;99(4):648–654. PubMed

Sivan U, Jayakumar K, Krishnan LK. Constitution of fibrin-based niche for in vitro differentiation of adipose-derived mesenchymal stem cells to keratinocytes. Biores Open Access. 2014;3(6):339–347. PubMed PMC

Kranz H, Ubrich N, Maincent P, Bodmeier R. Physicomechanical properties of biodegradable poly(D,L-lactide) and poly(D,L-lactide-co-glycolide) films in the dry and wet states. J Pharm Sci. 2000;89(12):1558–1566. PubMed

Lück M, Pistel KF, Li YX, Blunk T, Müller RH, Kissel T. Plasma protein adsorption on biodegradable microspheres consisting of poly(D,L-lactide-co-glycolide), poly(L-lactide) or ABA triblock copolymers containing poly(oxyethylene): influence of production method and polymer composition. J Control Release. 1998;55(2–3):107–120. PubMed

Ishaug SL, Yaszemski MJ, Bizios R, Mikos AG. Osteoblast function on synthetic biodegradable polymers. J Biomed Mater Res. 1994;28(12):1445–1453. PubMed

Tsai WB, Chen CH, Chen JF, Chang KY. The effects of types of degradable polymers on porcine chondrocyte adhesion, proliferation and gene expression. J Mater Sci Mater Med. 2006;17(4):337–343. PubMed

Riedel T, Brynda E, Dyr JE, Houska M. Controlled preparation of thin fibrin films immobilized at solid surfaces. J Biomed Mater Res A. 2009;88(2):437–447. PubMed

Boukamp P, Petrussevska RT, Breitkreutz D, Hornung J, Markham A, Fusenig NE. Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell Biol. 1988;106(3):761–771. PubMed PMC

Hornig-Do HT, von Kleist-Retzow JC, Lanz K, et al. Human epidermal keratinocytes accumulate superoxide due to low activity of Mn-SOD, leading to mitochondrial functional impairment. J Invest Dermatol. 2007;127(5):1084–1093. PubMed

Rho KS, Jeong L, Lee G, et al. Electrospinning of collagen nanofibers: effects on the behavior of normal human keratinocytes and early-stage wound healing. Biomaterials. 2006;27(8):1452–1461. PubMed

Fu X, Xu M, Liu J, Qi Y, Li S, Wang H. Regulation of migratory activity of human keratinocytes by topography of multiscale collagen-containing nanofibrous matrices. Biomaterials. 2014;35(5):1496–1506. PubMed PMC

Mahjour SB, Fu X, Yang X, Fong J, Sefat F, Wang H. Rapid creation of skin substitutes from human skin cells and biomimetic nanofibers for acute full-thickness wound repair. Burns. 2015;41(8):1764–1774. PubMed PMC

Mazlyzam AL, Aminuddin BS, Fuzina NH, et al. Reconstruction of living bilayer human skin equivalent utilizing human fibrin as a scaffold. Burns. 2007;33(3):355–363. PubMed

Nair RP, Joseph J, Harikrishnan VS, Krishnan VK, Krishnan L. Contribution of fibroblasts to the mechanical stability of in vitro engineered dermal-like tissue through extracellular matrix deposition. Biores Open Access. 2014;3(5):217–225. PubMed PMC

Sese N, Cole M, Tawil B. Proliferation of human keratinocytes and cocultured human keratinocytes and fibroblasts in three-dimensional fibrin constructs. Tissue Eng Part A. 2011;17(3–4):429–437. PubMed

Cox S, Cole M, Tawil B. Behavior of human dermal fibroblasts in three-dimensional fibrin clots: dependence on fibrinogen and thrombin concentration. Tissue Eng. 2004;10(5–6):942–954. PubMed

Gugerell A, Schossleitner K, Wolbank S, et al. High thrombin concentrations in fibrin sealants induce apoptosis in human keratinocytes. J Biomed Mater Res A. 2012;100(5):1239–1247. PubMed

Kubo M, Van de Water L, Plantefaber LC, et al. Fibrinogen and fibrin are anti-adhesive for keratinocytes: a mechanism for fibrin eschar slough during wound repair. J Invest Dermatol. 2001;117(6):1369–1381. PubMed

Weiss E, Yamaguchi Y, Falabella A, Crane S, Tokuda Y, Falanga V. Un-cross-linked fibrin substrates inhibit keratinocyte spreading and replication: correction with fibronectin and factor XIII cross-linking. J Cell Physiol. 1998;174(1):58–65. PubMed

Koivisto L, Larjava K, Häkkinen L, Uitto VJ, Heino J, Larjava H. Different integrins mediate cell spreading, haptotaxis and lateral migration of HaCaT keratinocytes on fibronectin. Cell Adhes Commun. 1999;7(3):245–257. PubMed

Trombetta-Esilva J, Eadie EP, Zhang Y, Norris RA, Borg TK, Bradshaw AD. The effects of age and the expression of SPARC on extracellular matrix production by cardiac fibroblasts in 3-D cultures. PLoS One. 2013;8(11):e79715. PubMed PMC

Tuan TL, Song A, Chang S, Younai S, Nimni ME. In vitro fibroplasia: matrix contraction, cell growth, and collagen production of fibroblasts cultured in fibrin gels. Exp Cell Res. 1996;223(1):127–134. PubMed

Marchisio PC, Cancedda R, De Luca M. Structural and functional studies of integrin receptors in cultured human keratinocytes. Cell Differ Dev. 1990;32(3):355–359. PubMed

Staatz WD, Fok KF, Zutter MM, Adams SP, Rodriguez BA, Santoro SA. Identification of a tetrapeptide recognition sequence for the α2β1 integrin in collagen. J Biol Chem. 1991;266(12):7363–7367. PubMed

Scharffetter-Kochanek K, Klein CE, Heinen G, et al. Migration of a human keratinocyte cell line (HACAT) to interstitial collagen type I is mediated by the α2β1-integrin receptor. J Invest Dermatol. 1992;98(1):3–11. PubMed

Engler A, Bacakova L, Newman C, Hategan A, Griffin M, Discher D. Substrate compliance versus ligand density in cell on gel responses. Biophys J. 2004;86(1 Pt 1):617–628. PubMed PMC

Eastwood M, Porter R, Khan U, McGrouther G, Brown R. Quantitative analysis of collagen gel contractile forces generated by dermal fibroblasts and the relationship to cell morphology. J Cell Physiol. 1996;166(1):33–42. PubMed

Agis H, Collins A, Taut AD, et al. Cell population kinetics of collagen scaffolds in ex vivo oral wound repair. PLoS One. 2014;9(11):e112680. PubMed PMC

Mateos-Timoneda MA, Castano O, Planell JA, Engel E. Effect of structure, topography and chemistry on fibroblast adhesion and morphology. J Mater Sci Mater Med. 2014;25(7):1781–1787. PubMed

Ruoslahti E, Pierschbacher MD. New perspectives in cell adhesion: RGD and integrins. Science. 1987;238(4826):491–497. PubMed

Shive MS, Anderson JM. Biodegradation and biocompatibility of PLA and PLGA microspheres. Adv Drug Deliv Rev. 1997;28(1):5–24. PubMed

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