Abstract
The present study reports construction of wound dressing materials from degradable natural polymers such as hydroxy derivatives of carboxylic acids (PHAs) and 3-hydroxybutyrate/4-hydroxybutyrate [P(3HB/4HB)] as copolymer. The developed polymer films and electrospun membranes were evaluated for its wound healing properties with Grafts—elastic nonwoven membranes carrying fibroblast cells derived from adipose tissue multipotent mesenchymal stem cells. The efficacy of nonwoven membranes of P(3HB/4HB) carrying the culture of allogenic fibroblasts was assessed against model skin defects in Wistar rats. The morphological, histological and molecular studies revealed the presence of fibroblasts on dressing materials which facilitated wound healing, vascularization and regeneration. Further it was also observed that cells secreted extracellular matrix proteins which formed a layer on the surface of membranes and promoted the migration of epidermal cells from the neighboring tissues surrounding the wound. The wounds under the P(3HB/4HB) membrane carrying cells healed 1.4 times faster than the wounds under the cell-free membrane and 3.5 times faster than the wounds healing under the eschar (control).The complete wound healing process was achieved at Day 14. Thus the study highlights the importance of nonwoven membranes developed from degradable P(3HB/4HB) polymers in reducing inflammation, enhancing angiogenic properties of skin and facilitating better wound healing process.
Similar content being viewed by others
Explore related subjects
Discover the latest articles, news and stories from top researchers in related subjects.References
MacNeil S. Progress and opportunities for tissue-engineered skin. Nature. 2007;445:874–80.
MacNeil S. Biomaterials for tissue engineering of skin. Mater Today. 2008;11:26–35.
Canto’n I, Cole DM, Kemp EH, Watson PF, Chunthapong J, Ryan AJ, MacNeil S, Haycock JW. Development of a 3D human in vitro skin co-culture model for detecting irritants in real-time. Biotechnol Bioeng. 2010;106:794–803.
Deshpande P, Ramachandran C, Sangwan VS, MacNeil S. Cultivation of Limbal Epithelial Cells on Electrospun Poly(lactide-co-glycolide) Scaffolds for Delivery to the Cornea. In: Wright B, Connon CJ, eds. Corneal Regenerative Medicine, Methods and Protocols. New York: Humana Press; 2013. p. 179–85.
Seland H, Gusafson C-J, Johnson H, Junker JPE, Kratz G. Transplantation of acellular dermis and keratinocytes cultured on porous biodegradable microcarriers into full-thickness skin injuries on athymic rats. Burns. 2011;37:99–108.
Metcalfe AD, Ferguson MWJ. Tissue engineering of replacement skin: the crossroads of biomaterials, wound healing, embryonic development, stem cells and regeneration. JR Soc Interface. 2007;4:413–37.
Walker MNM, Wright KT, Fuller HR, MacNeil S, Johnson WEB. Mesenchymal stem cell-conditioned medium accelerates skin wound healing: an in vitro study of fibroblast and keratinocyte scratch assays. Exp Cell Res. 2010;316:1271 –81.
Walker NG, Mistry AR, Smith LE, Eves PC, Tsaknakis G, Forster S, Watt SM, MacNeil S. A chemically defined carrier for the delivery of human mesenchymal stem/stromal cells to skin wounds. Tissue Eng. Part C. 2012;18:143–55.
Blais M, Parenteau-Bareil R, Cadau S, Berthod F. Concise review: tissue-engineered skin and nerve regeneration in burn treatment. Stem Cells Transl Med. 2013;2:545–51.
Kim SS, Gwak SJ, Choi CY, Kim BS. Skin regeneration using keratinocytes and dermal fibroblasts cultured on biodegradable microspherical polymer scaffolds. J Biomed Mater Res B Appl Biomater. 2005;75:369–77.
Liu P, Deng Z, Han S, Liu T, Wen N, Lu W, Geng X, Huang S, Jin Y. Tissue-engineered skin containing mesenchymal stem cells improves burn wounds. Artif Organs. 2008;32:925–31.
Yang EK, Seo YK, Youn HH, Lee DH, Park SN, Park JK. Tissue engineered artificial skin composed of dermis and epidermis. Artif Organs. 2000;24:7–17.
MacNeil S, Shepherd J, Smith L. Production of Tissue-Engineered Skin and Oral Mucosa for Clinical and Experimental Use. In: Haycock JW, ed. 3D Cell Culture, Methods and Protocols. New York: Humana Press; 2011. p. 129–53.
Sharma K, Bullock A, Ralston D, MacNeil S. Development of a one-step approach for the reconstruction of full thickness skin defects using minced split thickness skin grafts and biodegradable synthetic scaffolds as a dermal substitute. Burns. 2014;40:957–65.
Sill TJ, von Recum HA. Electrospinning: applications in drug delivery and tissue engineering. Biomaterials. 2008;29:1989–2006.
Rošic R, Kocbek P, Pelipenko J, Kristl J, Baumgartner S. Nanofibers and their biomedical use. Acta Pharm. 2013;63:295–304.
Ramalingman M, Seeram R. Nano-featured scaffolds for tissue engineering: a review of spinning methodologies. Tissue Eng. 2006;12:437–70.
Williams SF, Martin DP. Applications of PHAs in Medicine and Pharmacy. In: Steinbüchel A, ed. Series of Biopolymers in 10 vol. 4. VCY Verlag GmbH: Willey; 2002. p. 91–121.
Chen G-Q, Wu Q. The application of polyhydroxyalkanoates as tissue engineering materials. Biomaterials. 2005;26:6565–78.
Hazer B, Steinbüchel A. Increased diversification of polyhydroxyalkanoates by modification reactions for industrial and medical applications. Appl Microbiol Biotechnol. 2007;74:1–12.
Martin DP, Badhwar A, Shah DV, Rizk S, Eldridge SN, Gagne DH, Ganatra A, Darois RE, Williams SF, Tai H-C, Scott JR. Characterization of poly-4-hydroxybutyrate mesh for hernia repair applications. J Surg Res. 2013;184:766–73.
Saad B, Hirt TD, Welti M, Uhlschmid GK, Neuenschwander P, Suter UW. Development of degradable polyesterurerethans for medical applications: in vitro and in vivo evaluations. J Biomed Mater Res. 1997;36:65–74.
Tezcaner A, Bugra K, Hasirci V. Retinal pigment epithelium cell culture on surface modified poly(hydroxybutyrate-co-hydroxyvalerate) thin films. Biomaterials. 2003;24:4573–83.
Liu P, Deng Z, Han S, Liu T, Wen N, Lu W, Geng X, Huang S, Jin Y. Tissue-engineered skin containing mesenchymal stem cells improves burn wounds. Artif Organs. 2008;32:925–31.
Lucchesi C, Ferreira BMP, Duek EAR, Santos AR Jr., Joazeiro PP. Increased response of Vero cells to PHBV matrices treated by plasma. J Mater Sci-Mater M. 2008;19:635–43.
Köse GT, Kokusuz F, Özkul A, Soysal Y, Ozdemir T, Yildiz C, Hasirci V. Tissue engineered cartilage on collagen and PHBV matrices. Biomaterials. 2005;26:5187–97.
Qu XH, Wu Q, Liang J, Qu X, Wang SG, Chen GQ. Enhanced vascular-related cellular affinity on surface modified copolyesters of 3-hydroxybutyrate and 3-hydroxyhexanoate (PHBHHx). Biomaterials. 2005;26:6991–7001.
Rentsch C, Rentsch B, Breier A, Hofmann A, Manthey S, Scharnweber D, Biewener A, Zwipp H. Evaluation of the osteogenic potential and vascularization of 3D poly(3)hydroxybutyrate scaffolds subcutaneously implanted in nude rats. J Biomed Mater Res A. 2010;92:185–95.
Wang L, Wang ZH, Shen CY, You ML, Xiao JF, Chen GQ. Differentiation of human bone marrow mesenchymal stem cells grown in terpolyesters of 3-hydroxyalkanoates scaffolds into nerve cells. Biomaterials. 2010;31:1691–98.
You M, Peng G, Li J, Ma P, Wang Z, Shu W, Peng S, Chen GQ. Chondrogenic differentiation of human bone marrow mesenchymal stem cells on polyhydroxyalkanoate (PHA) scaffolds coated with PHA granule binding protein PhaP fused with RGD peptide. Biomaterials. 2011;32:2305–13.
Shishatskaya EI, Volova TG. A comparative investigation of biodegradable polyhydroxyalkanoate films as matrices for in vitro cell cultures. J Mater Sci-Mater M. 2004;15:915–23.
Volova TG, Shishatskaya EI. Results of biomedical studies of PHAs produced in Institute of Biophysics SB RAS and Siberian Federal University. Chapter № 21 in the book «Polyhydroxyalkanoates (PHA): Biosynthesis, Industrial Production and Applications in Medicine». USA: Nova Scienses Publ. Inc. NY; 2014. p. 273–330.
Shishatskaya EI, Volova TG, Efremov SN, Puzyr AP, Mogilnaya OA. Tissue response to the implantation of biodegradable polyhydroxyalkanoate sutures. J Mater Sci-Mater М. 2004;15:719–28.
Shishatskaya EI, Chlusov IA, Volova TG. A hybrid PHA-hydroxyapatite composite for biomedical application: production and investigation. J Biomater Sci Polym Ed. 2006;17:481–98.
Shishatskaya EI, Voinova ON, Goreva AV, Mogilnaya OA, Volova TG. Biocompatability of polyhydroxybutyrate Microspheres: in vitro and in vivo evaluation. Journal of Siberian Federal University. Biology. 2008;1:66–77.
Shishatskaya EI, Kamendov IV, Starosvetsky SI, Vinnik YS, Markelova NM, Shageev AA, Khorzhevsky VA, Peryanova OV, Shumiliva AA. An in vivo study of osteoplastic properties of resorbable poly-3-hydroxybutyrate in models of segmental osteotomy and chronic osteomyelitis. Artif Cells Nanomed Biotechnol. 2014;42:344–55.
Volova TG, Shishatskaya EI, Sinskey AJ. Degradable Polymers: Production, Properties and Applications. USA: Nova Science Pub., Inc. NY; 2013.
Goreva AV, Shishatskaya EI, Kuzmina AM, Volova TG, Sinskey AJ. Microparticles prepared from biodegradable polyhydroxyalkanoates as matrix for encapsulation of cytostatic drug. J Mat Sci-Mater M. 2013;24:1905–15.
Eke G, Kuzmina AM, Goreva AV, Shishatskaya EI, Hasirci N, Hasirci V. in vitro and transdermal penetration of PHBV micro/nanoparticles. J Mater Sci-Mater M. 2014;25:1471–81.
Volova TG, Shishatskaya EI, Nikolaeva ED, Sinskey AJ. in vivo study of 2D PHA matrices of different chemical compositions: tissue reactions and biodegradations. Mater Sci Technol. 2014;30:549–57.
Ji Y, Li XT, Chen GQ. Interactions between a poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) terpolyester and human keratinocytes. Biomaterials. 2008;29:3807–14.
Volova TG, Kiselev EG, Shishatskaya EI, Zhila NO, Boyandin AN, Syrvacheva DA, Vinogradova ON, Kalacheva GS, Vasiliev AD, Peterson IV. Cell growth and accumulation of polyhydroxyalkanoates from CO2 and H2 of a hydrogen-oxidizing bacterium, Cupriaviduseutrophus B-10646. Bioresour Technol. 2013;146:215–22.
Volova TG and Shishatskaya EI. RF Patent No. 2439143.
Volova T, Goncharov D, Sukovatyi A, Shabanov A, Nikolaeva E, Shishatskaya E. Electrospinning of polyhydroxyalkanoate fibrous scaffolds: effects on electrospinning parameters on structure and properties. J BiomaterSciPolym Ed. 2014;25:370–93.
Kaelble DH. Dispersion-polar surface tension properties of organic solids. J Adhes. 1970;2:66–81.
Jelvani S, Pazokian H, MoradiFarisar S. Effect of CO2pulsed laser irradiation on improving the biocompatibility of a polyethersulfone film. J Phys: Conference Ser. 2013;414:012021
Kang YO, Yoon IS, Lee SY, Kim DD, Lee SJ, Park WH, Hudson SM. Chitosan-coated poly(vinyl alcohol) nanofibers for wound dressings. J Biomed Mater Res B Appl Biomater. 2010;92:568–76.
Franco RA, Min YK, Yang HM, Lee BT. Fabrication and biocompatibility of novel bilayer scaffold for skin tissue engineering applications. J Biomater Appl. 2013;27:605–15.
Jin G, Prabhakaran MP, Kai D, Sathesh KA, Arunachalam KD, Ramakrishna S. Tissue engineered plant extracts as nanofibrous wound dressing. Biomaterials. 2013;34:724–34.
Pezeshki-Modaress M, Mirzadeh H, Zandi M. Gelatin-gag electrospunnanofibrous scaffold for skin tissue engineering: Fabrication and modeling of process parameters. Mater Sci Eng C. 2015;48:704–712.
Blackwood KA, McKean R, Canton I, Freeman CO, Franklin KL, Cole D, Brook I, Farthing P, Rimmer S, Haycock JW, Ryan AJ, MacNeil S. Development of biodegradable electrospun scaffolds for dermal replacement. Biomaterials. 2008;29:3091–104.
Shishatskaya EI, Volova TG, Gordeev SA, Puzyr AP. Degradation of P(3HB) and P(3HB-co-3HV) in biological media. J BiomaterSci Polymer Ed. 2005;16:643–57.
Li X-T, Zhang Y, Chen G-Q. Nanofibrouspolyhydroxyalkanoate matrices as cell growth supporting materials. Biomaterials. 2008;29:3720–28.
Sombatmankhong K, Sanchavanakit N, Pavasant P, Supaphol P. Bone scaffolds from electrospun fiber mats of poly(3-hydroxybutyrate), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and their blend. Polymer (Guildf). 2007;48:1419–27.
Kim Y-J, Bae H-I, Kwon OK, Choi M-S. Three-dimensional gastric cancer cell culture using nanofiber scaffold for chemosensitivity test. Int J BiolMacromol. 2009;45:65–71.
Zhou J, Peng S-W, Wang Y-Y, Zheng S-B, Wang Y, Chen G-Q. The use of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) scaffolds for tarsal repair in eyelid reconstruction in the rat. Biomaterials. 2010;31:7512–18.
Murueva AV, Shershneva AM, Shishatskaya EI, Volova TG. The Use of Polymeric Microcarriers Loaded with Anti-Inflammatory Substances in the Therapy of Experimental Skin Wounds. Bull ExpBiol Med, Pharmacology and Toxicology. 2014;157:597–602.
Potapov IV, Il’inskii OM, Kurenkova LG, Sevast’yanov VI, Egorova VA, Zaidenov VA, Rasulov MF, Onishchenko NA. ElastoPHB® Membrane Systems with Immobilized Bone Marrow Stromal Cells Optimize Conditions for Regeneration of Damaged Tissue. Cell TechnolBiol Med. 2005;3:132–37. (in Russian)
Volova TG, Sevastianov VI, Shishatskaya EI. Matrix of polyhydroxyalkanoates for cell and tissue engineering. In: Shumakov VI, ed. Polyhydroxyalkanoates – Biodegradable Polymers for Medicine. Krasnoyarsk: Platinum; 2006. p. 209–16. (in Russian)
Acknowledgments
The study was supported by the RFBR/KRSTSF (Project “The scientific bases of construction and application of biocompatible resorbable biomaterials and hybride tissue engineered systems for tissue formation repairing defects of skin”) – research and development of wound dressings; State budget allocated to the fundamental research at the Russian Academy of Sciences (project No 01201351505) –polymer synthesis.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Rights and permissions
About this article
Cite this article
Shishatskaya, E.I., Nikolaeva, E.D., Vinogradova, O.N. et al. Experimental wound dressings of degradable PHA for skin defect repair. J Mater Sci: Mater Med 27, 165 (2016). https://doi.org/10.1007/s10856-016-5776-4
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10856-016-5776-4