Dermal injuries and chronic wounds usually regenerate with scar formation. Successful treatment without scarring might be achieved by pre-seeding a wound dressing with cells. We aimed to prepare a wound dressing fabricated from sodium carboxymethylcellulose (Hcel® NaT), combined with fibrin and seeded with dermal fibroblasts in vitro. We fabricated the Hcel® NaT in a porous and homogeneous form (P form and H form, respectively) differing in structural morphology and in the degree of substitution of hydroxyl groups. Each form of Hcel® NaT was functionalized with two morphologically different fibrin structures to improve cell adhesion and proliferation, estimated by an MTS assay. Fibrin functionalization of the Hcel® NaT strongly enhanced colonization of the material with human dermal fibroblasts. Moreover, the type of fibrin structures influenced the ability of the cells to adhere to the material and proliferate on it. The fibrin mesh filling the void spaces between cellulose fibers better supported cell attachment and subsequent proliferation than the fibrin coating, which only enwrapped individual cellulose fibers. On the fibrin mesh, the cell proliferation activity on day 3 was higher on the H form than on the P form of Hcel® NaT, while on the fibrin coating, the cell proliferation on day 7 was higher on the P form. The Hcel® NaT wound dressing functionalized with fibrin, especially when in the form of a mesh, can accelerate wound healing by supporting fibroblast adhesion and proliferation.

Department of Biomaterials and Tissue Engineering Institute of Physiology of the Czech Academy of Sciences 14220 Prague Czech Republic

Holzbecher Ltd Bleaching and Dyeing Plant in Zlic 55203 Zlic Czech Republic

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Bacakova L., Novotna K., Sopuch T., Havelka P. Cell interaction with cellulose-based scaffolds for tissue engineering—A review. In: Mondal I.H., editor. Cellulose and Cellulose Composites: Modification, Characterization and Applications. Nova Science Publishers, Inc.; New York, NY, USA: 2015. pp. 341–375.

Kalia S., Dufresne A., Cherian B.M., Kaith B.S., Averous L., Njuguna J., Nassiopoulos E. Cellulose-Based Bio- and Nanocomposites: A Review. Int. J. Polym. Sci. 2011;2011:837875. doi: 10.1155/2011/837875. DOI

Klemm D., Heublein B., Fink H.P., Bohn A. Cellulose: Fascinating biopolymer and sustainable raw material. Angew. Chem. Int. Ed. 2005;44:3358–3393. doi: 10.1002/anie.200460587. PubMed DOI

Coseri S. Cellulose: To depolymerize... or not to? Biotechnol. Adv. 2017;35:251–266. doi: 10.1016/j.biotechadv.2017.01.002. PubMed DOI

Ohta S., Nishiyama T., Sakoda M., Machioka K., Fuke M., Ichimura S., Inagaki F., Shimizu A., Hasegawa K., Kokudo N., et al. Development of carboxymethyl cellulose nonwoven sheet as a novel hemostatic agent. J. Biosci. Bioeng. 2015;119:718–723. doi: 10.1016/j.jbiosc.2014.10.026. PubMed DOI

Ramli N.A., Wong T.W. Sodium carboxymethylcellulose scaffolds and their physicochemical effects on partial thickness wound healing. Int. J. Pharm. 2011;403:73–82. doi: 10.1016/j.ijpharm.2010.10.023. PubMed DOI

Wong T.W., Ramli N.A. Carboxymethylcellulose film for bacterial wound infection control and healing. Carbohydr. Polym. 2014;112:367–375. doi: 10.1016/j.carbpol.2014.06.002. PubMed DOI

Mutsaers S.E., Bishop J.E., McGrouther G., Laurent G.J. Mechanisms of tissue repair: From wound healing to fibrosis. Int. J. Biochem. Cell Biol. 1997;29:5–17. doi: 10.1016/S1357-2725(96)00115-X. PubMed DOI

Tracy L.E., Minasian R.A., Caterson E.J. Extracellular Matrix and Dermal Fibroblast Function in the Healing Wound. Adv. Wound Care. 2016;5:119–136. doi: 10.1089/wound.2014.0561. PubMed DOI PMC

Wright K.A., Nadire K.B., Busto P., Tubo R., McPherson J.M., Wentworth B.M. Alternative delivery of keratinocytes using a polyurethane membrane and the implications for its use in the treatment of full-thickness burn injury. Burns. 1998;24:7–17. doi: 10.1016/S0305-4179(97)00075-2. PubMed DOI

van den Bogaerdt A.J., Ulrich M.M., van Galen M.J., Reijnen L., Verkerk M., Pieper J., Lamme E.N., Middelkoop E. Upside-down transfer of porcine keratinocytes from a porous, synthetic dressing to experimental full-thickness wounds. Wound Repair Regen. 2004;12:225–234. doi: 10.1111/j.1067-1927.2004.012115.x. PubMed DOI

Johnen C., Steffen I., Beichelt D., Braeutigam K., Witascheck T., Toman N., Moser V., Ottomann C., Hartmann B., Gerlach J.C. Culture of subconfluent human fibroblasts and keratinocytes using biodegradable transfer membranes. Burns. 2008;34:655–663. doi: 10.1016/j.burns.2007.08.023. PubMed DOI

Mohamad N., Loh E.Y.X., Fauzi M.B., Ng M.H., Mohd Amin M.C.I. In vivo evaluation of bacterial cellulose/acrylic acid wound dressing hydrogel containing keratinocytes and fibroblasts for burn wounds. Drug Deliv. Transl. Res. 2018:1–9. doi: 10.1007/s13346-017-0475-3. PubMed DOI

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;63:352–366. doi: 10.1016/j.addr.2011.01.005. PubMed DOI

Laurens N., Koolwijk P., de Maat M.P. Fibrin structure and wound healing. J. Thromb. Haemost. 2006;4:932–939. doi: 10.1111/j.1538-7836.2006.01861.x. PubMed DOI

Reinke J.M., Sorg H. Wound repair and regeneration. Eur. Surg. Res. 2012;49:35–43. doi: 10.1159/000339613. PubMed DOI

Rajangam T., An S.S. Fibrinogen and fibrin based micro and nano scaffolds incorporated with drugs, proteins, cells and genes for therapeutic biomedical applications. Int. J. Nanomed. 2013;8:3641–3662. doi: 10.2147/IJN.S43945. PubMed DOI PMC

Bacakova M., Musilkova J., Riedel T., Stranska D., Brynda E., Zaloudkova M., Bacakova L. The potential applications of fibrin-coated electrospun polylactide nanofibers in skin tissue engineering. Int. J. Nanomed. 2016;11:771–789. doi: 10.2147/IJN.S99317. PubMed DOI PMC

Bacakova M., Pajorova J., Stranska D., Hadraba D., Lopot F., Riedel T., Brynda E., Zaloudkova M., Bacakova L. Protein nanocoatings on synthetic polymeric nanofibrous membranes designed as carriers for skin cells. Int. J. Nanomed. 2017;12:1143–1160. doi: 10.2147/IJN.S121299. PubMed DOI PMC

Pajorova J., Bacakova M., Musilkova J., Broz A., Hadraba D., Lopot F., Bacakova L. Morphology of a fibrin nanocoating influences dermal fibroblast behavior. Int. J. Nanomed. 2018;13:3367–3380. doi: 10.2147/IJN.S162644. PubMed DOI PMC

Melander A., Vuorinen T. Determination of the degree of polymerisation of carboxymethyl cellulose by size exclusion chromatography. Carbohydr. Polym. 2001;46:227–233. doi: 10.1016/S0144-8617(00)00303-9. DOI

Riedel T., Brynda E., Dyr J.E., Houska M. Controlled preparation of thin fibrin films immobilized at solid surfaces. J. Biomed. Mater. Res. A. 2009;88:437–447. doi: 10.1002/jbm.a.31755. PubMed DOI

Tamer T.M., Collins M.N., Valachova K., Hassan M.A., Omer A.M., Mohy-Eldin M.S., Svik K., Jurcik R., Ondruska L., Biro C., et al. MitoQ Loaded Chitosan-Hyaluronan Composite Membranes for Wound Healing. Materials. 2018;11:569. doi: 10.3390/ma11040569. PubMed DOI PMC

Zulkifli F.H., Hussain F.S., Rasad M.S., Mohd Yusoff M. Nanostructured materials from hydroxyethyl cellulose for skin tissue engineering. Carbohydr. Polym. 2014;114:238–245. doi: 10.1016/j.carbpol.2014.08.019. PubMed DOI

Song S.H., Kim J.E., Lee Y.J., Kwak M.H., Sung G.Y., Kwon S.H., Son H.J., Lee H.S., Jung Y.J., Hwang D.Y. Cellulose film regenerated from Styela clava tunics have biodegradability, toxicity and biocompatibility in the skin of SD rats. J. Mater. Sci. Mater. Med. 2014;25:1519–1530. doi: 10.1007/s10856-014-5182-8. PubMed DOI

Mohseni M., Shamloo A., Aghababaei Z., Vossoughi M., Moravvej H. Antimicrobial Wound Dressing Containing Silver Sulfadiazine with High Biocompatibility: In Vitro Study. Artif. Organs. 2016;40:765–773. doi: 10.1111/aor.12682. PubMed DOI

Novotna K., Havelka P., Sopuch T., Kolarova K., Vosmanska V., Lisa V., Svorcik V., Bacakova L. Cellulose-based materials as scaffolds for tissue engineering. Cellulose. 2013;20:2263–2278. doi: 10.1007/s10570-013-0006-4. DOI

Broussard K.C., Powers J.G. Wound dressings: Selecting the most appropriate type. Am. J. Clin. Dermatol. 2013;14:449–459. doi: 10.1007/s40257-013-0046-4. PubMed DOI

Ahmadi H., Williams G. Permanent scarring in a partial thickness scald burn dressed with Biobrane. J. Plast. Reconstr. Aesthet. Surg. 2009;62:697–698. doi: 10.1016/j.bjps.2008.01.014. PubMed DOI

Boateng J.S., Matthews K.H., Stevens H.N., Eccleston G.M. Wound healing dressings and drug delivery systems: A review. J. Pharm. Sci. 2008;97:2892–2923. doi: 10.1002/jps.21210. PubMed DOI

Rodriguez-Merchan E.C. Local fibrin glue and chitosan-based dressings in haemophilia surgery. Blood Coagul. Fibrinolysis. 2012;23:473–476. doi: 10.1097/MBC.0b013e3283555379. PubMed DOI

Yildirim S., Ozener H.O., Dogan B., Kuru B. Effect of topically applied hyaluronic acid on pain and palatal epithelial wound healing: An examiner-masked, randomized, controlled clinical trial. J. Periodontol. 2018;89:36–45. doi: 10.1902/jop.2017.170105. PubMed DOI

Bacakova L., Filova E., Kubies D., Machova L., Proks V., Malinova V., Lisa V., Rypacek F. Adhesion and growth of vascular smooth muscle cells in cultures on bioactive RGD peptide-carrying polylactides. J. Mater. Sci. Mater. Med. 2007;18:1317–1323. doi: 10.1007/s10856-006-0074-1. PubMed DOI

Hopp I., Michelmore A., Smith L.E., Robinson D.E., Bachhuka A., Mierczynska A., Vasilev K. The influence of substrate stiffness gradients on primary human dermal fibroblasts. Biomaterials. 2013;34:5070–5077. doi: 10.1016/j.biomaterials.2013.03.075. PubMed DOI

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;86:617–628. doi: 10.1016/S0006-3495(04)74140-5. PubMed DOI PMC

Carr M.E., Jr., Hermans J. Size and density of fibrin fibers from turbidity. Macromolecules. 1978;11:46–50. doi: 10.1021/ma60061a009. PubMed DOI

Riedelova-Reicheltova Z., Brynda E., Riedel T. Fibrin nanostructures for biomedical applications. Physiol. Res. 2016;65:S263–S272. PubMed

Becker J.C., Domschke W., Pohle T. Biological in vitro effects of fibrin glue: Fibroblast proliferation, expression and binding of growth factors. Scand. J. Gastroenterol. 2004;39:927–932. doi: 10.1080/00365520410003371. PubMed DOI

Gsib O., Duval J.L., Goczkowski M., Deneufchatel M., Fichet O., Larreta-Garde V., Bencherif S.A., Egles C. Evaluation of Fibrin-Based Interpenetrating Polymer Networks as Potential Biomaterials for Tissue Engineering. Nanomaterials. 2017;7:436. doi: 10.3390/nano7120436. PubMed DOI PMC

Stevens M.M., George J.H. Exploring and engineering the cell surface interface. Science. 2005;310:1135–1138. doi: 10.1126/science.1106587. PubMed DOI

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;29:739–767. doi: 10.1016/j.biotechadv.2011.06.004. PubMed DOI

Mazlyzam A.L., Aminuddin B.S., Fuzina N.H., Norhayati M.M., Fauziah O., Isa M.R., Saim L., Ruszymah B.H. Reconstruction of living bilayer human skin equivalent utilizing human fibrin as a scaffold. Burns. 2007;33:355–363. doi: 10.1016/j.burns.2006.08.022. PubMed DOI

Dvorankova B., Smetana K., Jr., Vacik J., Jelinkova M. Cultivation of keratinocytes on poly HEMA and their migration after inversion. Folia Biol. 1996;42:83–86. PubMed

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