In vivo evaluation of bacterial cellulose/acrylic acid wound dressing hydrogel containing keratinocytes and fibroblasts for burn wounds

  • Najwa Mohamad
  • Evelyn Yun Xi Loh
  • Mh Busra Fauzi
  • Min Hwei Ng
  • Mohd Cairul Iqbal Mohd Amin
Original Article


The healing of wounds, including those from burns, currently exerts a burden on healthcare systems worldwide. Hydrogels are widely used as wound dressings and in the field of tissue engineering. The popularity of bacterial cellulose-based hydrogels has increased owing to their biocompatibility. Previous study demonstrated that bacterial cellulose/acrylic acid (BC/AA) hydrogel increased the healing rate of burn wound. This in vivo study using athymic mice has extended the use of BC/AA hydrogel by the addition of human epidermal keratinocytes and human dermal fibroblasts. The results showed that hydrogel loaded with cells produces the greatest acceleration on burn wound healing, followed by treatment with hydrogel alone, compared with the untreated group. The percentage wound reduction on day 13 in the mice treated with hydrogel loaded with cells (77.34 ± 6.21%) was significantly higher than that in the control-treated mice (64.79 ± 6.84%). Histological analysis, the expression of collagen type I via immunohistochemistry, and transmission electron microscopy indicated a greater deposition of collagen in the mice treated with hydrogel loaded with cells than in the mice administered other treatments. Therefore, the BC/AA hydrogel has promising application as a wound dressing and a cell carrier.


Cell carrier Tissue regeneration Skin injury Wound healing 



This work was supported by funding from Universiti Kebangsaan Malaysia (GP-K007818 and DIP-2015-026).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethics approval and consent to participate

All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000 (5). Informed consent was obtained from all patients for being included in the study.

All institutional and national guidelines for the care and use of laboratory animals were followed.


  1. 1.
    Sen S, Greenhalgh D, Palmieri T. Review of burn injury research for the year 2009. J Burn Care Res. 2010;31(6):836–48. Scholar
  2. 2.
    Fagenholz PJ, Sheridan RL, Harris NS, Pelletier AJ, Camargo CA. National study of emergency department visits for burn injuries, 1993 to 2004. J Burn Care Res. 2007;28(5):681–90. Scholar
  3. 3.
    Gibran NS, Wiechman S, Meyer W, Edelman L, Fauerbach J, Gibbons L, et al. Summary of the 2012 ABA Burn Quality Consensus Conference. J Burn Care Res. 2013;34(4):361–85. Scholar
  4. 4.
    Oryan A, Jalili M, Kamali A. Tissue engineering in burn wound healing: current modalities and future directions. Int Clin Pathol J. 2017;4(1):00085.CrossRefGoogle Scholar
  5. 5.
    Rowan MP, Cancio LC, Elster EA, Burmeister DM, Rose LF, Natesan S, et al. Burn wound healing and treatment: review and advancements. Crit Care. 2015;19(1):243. Scholar
  6. 6.
    Kumar RJ, Kimble RM, Boots R, Pegg SP. Treatment of partial-thickness burns: a prospective, randomized trial using Transcyte. ANZ J Surg. 2004;74(8):622–6. Scholar
  7. 7.
    Wojtowicz AM, Oliveira S, Carlson MW, Zawadzka A, Rousseau CF, Baksh D. The importance of both fibroblasts and keratinocytes in a bilayered living cellular construct used in wound healing. Wound Repair Regen. 2014;22(2):246–55. Scholar
  8. 8.
    Werner S, Krieg T, Smola H. Keratinocyte-fibroblast interactions in wound healing. J Invest Dermatol. 2007;127(5):998–1008. Scholar
  9. 9.
    Hassan A, Halim AS, Hilmi ABMA. Bilayer engineered skin substitute for wound repair in an irradiation-impeded healing model on rat. Adv Wound Care (New Rochelle). 2015;4(5):312–20.
  10. 10.
    Hoffman AS. Hydrogels for biomedical applications. Adv Drug Deliv Rev. 2002;54(1):3–12.
  11. 11.
    Takei T, Nakahara H, Ijima H, Kawakami K. Synthesis of a chitosan derivative soluble at neutral pH and gellable by freeze thawing, and its application in wound care. Acta Biomater. 2012;8(2):686–93. Scholar
  12. 12.
    Balakrishnan B, Mohanty M, Umashankar PR, Jayakrishnan A. Evaluation of an in situ forming hydrogel wound dressing based on oxidized alginate and gelatin. Biomaterials. 2005;26(32):6335–42. Scholar
  13. 13.
    Drury JL, Mooney DJ. Hydrogels for tissue engineering: scaffold design variables and applications. Biomaterials. 2003;24(24):4337–51. Scholar
  14. 14.
    Sun G, Zhang X, Shen Y, Sebastian R, Dickinson LE, Talbot KF, et al. Dextran hydrogel scaffolds enhance angiogenic responses and promote complete skin regeneration during burn wound healing. Proc Natl Acad Sci. 2011;108(52):20976–81. Scholar
  15. 15.
    Varkey M, Ding J, Tredget EE, Wound-Healing-Research-Group. The effect of keratinocytes on the biomechanical characteristics and pore microstructure of tissue engineered skin using deep dermal fibroblasts. Biomaterials. 2014;35(36):9591–8. Scholar
  16. 16.
    Cacicedo ML, Castro MC, Servetas I, Bosnea L, Boura K, Tsafrakidou P, et al. Progress in bacterial cellulose matrices for biotechnological applications. Bioresour Technol. 2016;213:172–80. Scholar
  17. 17.
    Fu L, Zhang J, Yang G. Present status and applications of bacterial cellulose-based materials for skin tissue repair. Carbohydr Polym. 2013;92(2):1432–42. Scholar
  18. 18.
    Wippermann J, Schumann D, Klemm D, Kosmehl H, Salehi-Gelani S, Wahlers T. Preliminary results of small arterial substitute performed with a new cylindrical biomaterial composed of bacterial cellulose. Eur J Vasc Endovasc. 2009;37(5):592–6. Scholar
  19. 19.
    Czaja W, Krystynowicz A, Bielecki S, Brown RM. Microbial cellulose—the natural power to heal wounds. Biomaterials. 2006;27(2):145–51. Scholar
  20. 20.
    Kwak MH, Kim JE, Go J, Koh EK, Song SH, Son HJ, et al. Bacterial cellulose membrane produced by Acetobacter sp. A10 for burn wound dressing applications. Carbohydr Polym. 2015;122:387–98.
  21. 21.
    Muangman P, Opasanon S, Suwanchot S, Thangthed O. Efficiency of microbial cellulose dressing in partial-thickness burn wounds. J Am Coll Certif Wound Spec. 2011;3(1):16–9.
  22. 22.
    Klemm D, Schumann D, Udhardt U, Marsch S. Bacterial synthesized cellulose—artificial blood vessels for microsurgery. Prog Polym Sci. 2001;26(9):1561–603. Scholar
  23. 23.
    Mohamad N, Mohd Amin MC, Pandey M, Ahmad N, Rajab NF. Bacterial cellulose/acrylic acid hydrogel synthesized via electron beam irradiation: accelerated burn wound healing in an animal model. Carbohydr Polym. 2014;114:312–20. Scholar
  24. 24.
    Mohamad N, Buang F, Mat Lazim A, Ahmad N, Martin C, Mohd Amin MC. Characterization and biocompatibility evaluation of bacterial cellulose-based wound dressing hydrogel: effect of electron beam irradiation doses and concentration of acrylic acid. J Biomed Mater Res B Appl Biomater. 2017;105(8):2553–64. Scholar
  25. 25.
    Seet WT, Manira M, Khairul Anuar K, Chua KH, Ahmad Irfan AW, Ng MH, et al. Shelf-life evaluation of bilayered human skin equivalent, MyDerm™. PLoS One. 2012;7:e40978.
  26. 26.
    Loo Y, Wong YC, Cai EZ, Ang CH, Raju A, Lakshmanan A, et al. Ultrashort peptide nanofibrous hydrogels for the acceleration of healing of burn wounds. Biomaterials. 2014;35:1–10.
  27. 27.
    Busra MF, Chowdhury SR, Ismail F, Saim A, Idrus RH. Tissue-engineered skin substitute enhances wound healing after radiation therapy. Adv Skin Wound Care. 2016;29(3):120–9. Scholar
  28. 28.
    Idrus RB, Rameli MA, Low KC, Law JX, Chua KH, Latiff MB, et al. Full-thickness skin wound healing using autologous keratinocytes and dermal fibroblasts with fibrin: bilayered versus single-layered substitute. Adv Skin Wound Care. 2014;27(4):171–80. Scholar
  29. 29.
    Rogers AA, Walmsley RS, Rippon MG, Bowler PG. Adsorption of serum-derived proteins by primary dressings: implications for dressing adhesion to wounds. J Wound Care. 2016;8:403–6.
  30. 30.
    Qiu Y, Qiu L, Cui J, Wei Q. Bacterial cellulose and bacterial cellulose-vaccarin membranes for wound healing. Mater Sci Eng C Mater Biol Appl. 2016;59:303–9. Scholar
  31. 31.
    Li J, Chen J, Kirsner R. Pathophysiology of acute wound healing. Clin Dermatol. 2007;25(1):9–18. Scholar
  32. 32.
    Arai M, Matsuzaki T, Ihara S. Wound closure on the neonatal rat skin I. The modulation of the thickness of epidermis at the closing incisional wounds. CellBio. 2013;2(4):248–56. Scholar
  33. 33.
    Boateng JS, Matthews KH, Stevens HN, Eccleston GM. Wound healing dressings and drug delivery systems: a review. J Pharm Sci. 2008;97(8):2892–923. Scholar
  34. 34.
    Lorenti A. Wound healing: from epidermis culture to tissue engineering. CellBio. 2012;1(02):17–29. Scholar
  35. 35.
    Madaghiele M, Demitri C, Sannino A, Ambrosio L. Polymeric hydrogels for burn wound care: advanced skin wound dressings and regenerative templates. Burns Trauma. 2014;2(4):153–61. Scholar
  36. 36.
    Candi E, Schmidt R, Melino G. The cornified envelope: a model of cell death in the skin. Nat Rev Mol Cell Biol. 2005;6(4):328–40. Scholar
  37. 37.
    Smiley AK, Klingenberg JM, Boyce ST, Supp DM. Keratin expression in cultured skin substitutes suggests that the hyperproliferative phenotype observed in vitro is normalized after grafting. Burns. 2006;32(2):135–8. Scholar
  38. 38.
    Byrne C, Tainsky M, Fuchs E. Programming gene expression in developing epidermis. Development. 1994;120(9):2369–83.PubMedGoogle Scholar
  39. 39.
    Kumar V, Cotran RZ, Robbins SL. Basic pathology. 7th ed. Philadelphia: Saunders; 2003. p. 873.Google Scholar
  40. 40.
    Gurtner GC, Werner S, Barrandon Y, Longaker MT. Wound repair and regeneration. Nature. 2008;453(7193):314–21. Scholar
  41. 41.
    Darby I, Skalli O, Gabbiani G. Alpha-smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab Investig. 1990;63(1):21–9.PubMedGoogle Scholar
  42. 42.
    Desmouliere A, Guyot C, Gabbiani G. The stroma reaction myofibroblast: a key player in the control of tumor cell behavior. Int J Dev Biol. 2004;48(5–6):509–17. Scholar
  43. 43.
    Serini G, Gabbiani G. Mechanisms of myofibroblast activity and phenotypic modulation. Exp Cell Res. 1999;250(2):273–83. Scholar
  44. 44.
    Darby IA, Laverdet B, Bonte F, Desmouliere A. Fibroblasts and myofibroblasts in wound healing. Clin Cosmet Investig Dermatol. 2014;7:301–11. Scholar
  45. 45.
    Desmouliere A, Redard M, Darby I, Gabbiani G. Apoptosis mediates the decrease in cellularity during the transition between granulation tissue and scar. Am J Pathol. 1995;146(1):56–66.PubMedPubMedCentralGoogle Scholar
  46. 46.
    Brandner JM, Haftek M, Niessen CM. Adherens junctions, desmosomes and tight junctions in epidermal barrier function. Open Dermatol J. 2010;4:14–20.Google Scholar

Copyright information

© Controlled Release Society 2018

Authors and Affiliations

  • Najwa Mohamad
    • 1
  • Evelyn Yun Xi Loh
    • 1
  • Mh Busra Fauzi
    • 2
  • Min Hwei Ng
    • 2
  • Mohd Cairul Iqbal Mohd Amin
    • 1
  1. 1.Faculty of PharmacyUniversiti Kebangsaan MalaysiaKuala LumpurMalaysia
  2. 2.Tissue Engineering CentreUniversiti Kebangsaan Malaysia Medical CentreKuala LumpurMalaysia

Personalised recommendations