Repositing honey incorporated electrospun nanofiber membranes to provide anti-oxidant, anti-bacterial and anti-inflammatory microenvironment for wound regeneration

  • Ripon Sarkar
  • Aritri Ghosh
  • Ananya Barui
  • Pallab Datta
Tissue Engineering Constructs and Cell Substrates Original Research
Part of the following topical collections:
  1. Tissue Engineering Constructs and Cell Substrates


Topical application of honey for tissue regeneration, has recently regained attention in clinical practice with controlled studies affirming its efficacy and indicating its role in regeneration over repair. Parallely, to overcome difficulties of applying raw honey, several product development studies like nanofibrous matrices have been reported. However, one approach concentrated on achieving highest possible honey loading in the nanofiber membranes while other studies have found that only specific honey dilutions result in differential cellular responses on wound healing and re-epithelization. From these results, it can be suggested that high honey loading provides optimum external microenvironment, low-loaded membranes could provide a more conducive internal microenvironment for tissue regeneration. With this hypothesis, this paper sought to evaluate ability of low-honey loaded nanofibers to modulate the anti-oxidant, anti-biofilm and anti-inflammatory properties which are important to be maintained in wound micro-environment. A loading-dependent reduction of biofilm formation and anti-oxidant activity was noted in different concentration ranges investigated. After scratch assay, a certain honey loading (0.5%) afforded the maximum re-epithelization. Since there is lack of methods to determine anti-inflammatory properties of nanofiber membranes during epithelial healing process, we performed anti-inflammatory assessment of nano-fibers by evaluating the expressions of pro-inflammatory markers-Cycloxygenase-2 (COX-2) and Interleukin-6 (IL-6) and to confirm the optimized concentration. Considering the role of COX-2 and IL-6, the novel methodology used in this study can also be developed as an assay for anti-inflammatory matrices for wound healing.

List of Abbreviation






Polymorphonuclear luekocyte




Platelet derived growth factor


Vascular endothelial growth factor


Transforming growth factor


Extra cellular matrix


3-[4,5-dimethylthiazol-2-yl]-diphenyltetrazolium bromide




Fetal bovine serum


Poly vinyl alcohol


Scanning Electron Microscopy


3-[4,5-dimethylthiazol-2-yl]-diphenyltetrazolium bromide




fetal bovine serum


Poly vinyl alcohol


Scanning Electron Microscopy


Dulbecco’s modified Eagle’s medium


Differential interference contrast





AB would like to acknowledge SERB, Govt. of India financial assistance through Fast Track project numbers SB/FTP/ETA/265-2012 and DST INSPIRE faculty award to PD via IFA2012-LSBM-48. Assistance from TEQIP Phase II to IIEST Shibpur for procurement of Inverted Florescence microscope and TEQIP COE for AFM is also acknowledged.

Compliance with ethical standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

10856_2018_6038_MOESM1_ESM.docx (397 kb)
Supplementary Information(DOCX 397 kb)


  1. 1.
    Rose LF, Chan RK. The burn wound microenvironment. Adv Wound Care. 2016;5:106–18.CrossRefGoogle Scholar
  2. 2.
    Frykberg RG, Banks J. Challenges in the treatment of chronic wounds. Adv Wound Care. 2015;4:560–82. Scholar
  3. 3.
    Kruse CR, Nuutila K, Lee CCY, Kiwanuka E, Singh M, Caterson EJ, et al. The external microenvironment of healing skin wounds. Wound Repair Regen. 2015;23:456–64.CrossRefGoogle Scholar
  4. 4.
    Schultz GS, Davidson JM, Kirsner RS, Herman IM. Dynamic reciprocity in the wound microenvironment. Wound Repair Regen. 2012;19:134–48.CrossRefGoogle Scholar
  5. 5.
    Braund R, Hook S, Medlicott NJ. The role of tropical growth factors in chroninc wounds. Curr Drug Deliv U Arab EMIR. 2007;4:195–204.CrossRefGoogle Scholar
  6. 6.
    Telgenhoff D, Shroot B. Cellular senescence mechanisms in chronic wound healing. Cell Death Differ. 2005;12:695–8. Scholar
  7. 7.
    Muller M, Trocme C, Lardy B, Morel F, Halimi S, Benhamou PY. Matrix metalloproteinases and diabetic foot ulcers: the ratio of MMP-1 to TIMP-1 is a predictor of wound healing. Diabet Med. 2008;25:419–26. Scholar
  8. 8.
    Andreu V, Mendoza G, Arruebo M, Irusta S. Smart dressings based on nanostructured fibers containing natural origin antimicrobial, anti-inflammatory, and regenerative compounds. Mater. 2015;8:5154–93.CrossRefGoogle Scholar
  9. 9.
    Dhall S, Do D, Garcia M, Wijesinghe DS, Brandon A, Kim J, Sanchez A, Lyubovitsky J, Gallagher S, Nothnagel EA, Chalfant CE, Patel RP, Schiller N, Martins-Green M, Appanna VD. A novel model of chronic wounds: importance of redox imbalance and biofilm-forming bacteria for establishment of chronicity. PLoS ONE 2014;9(10):e109848.Google Scholar
  10. 10.
    Saikaly SK, Khachemoune A. Honey and wound healing: an update. Am J Clin Dermatol. 2017;18:237–51. Scholar
  11. 11.
    Tian X, Yi L-J, Ma L, Zhang L, Song G-M, Wang Y. Effects of honey dressing for the treatment of DFUs: a systematic review. Int. J Nurs Sci. 2014;1:224–31. Scholar
  12. 12.
    Sarkar S, Mukhopadhyay A, Chaudhary A, Rajput M, Pawar HS, Mukherjee R. et al. Therapeutic interfaces of honey in diabetic wound pathology. Wound Med. 2017;18:21–32. Scholar
  13. 13.
    Cooper R. Honey in wound care: antibacterial properties. GMS Krankenhhyg. Interdiszip. German Medical Science GMS Publishing House; 2007;2:Doc51. Scholar
  14. 14.
    Maleki H, Gharehaghaji AA, Dijkstra PJ. A novel honey-based nanofibrous scaffold for wound dressing application. J Appl Polym Sci. 2013;127:4086–92.CrossRefGoogle Scholar
  15. 15.
    Barnthip N. Preparation of honey-gelatin nanofibers as the prototype of wound-healing and covering materials by electrospinning process. J Bionanoscience. 2015;9:475–9.CrossRefGoogle Scholar
  16. 16.
    Turaga U, Singh V, Gibson A, Maharubin S, Korzeniewski C, Presley S, et al. Preparation and characterization of honey-treated PVA nanowebs. AATCC J Res. 2016;3:25–31.CrossRefGoogle Scholar
  17. 17.
    Arslan A, Simşek M, Aldemir SD, Kazaroğlu N, Gümüşderelioğlu M. Honey based PET or PET/chitosan fibrous wound dressings: effect of honey on electrospinning process. J Biomater Sci Polym Ed. 2014;10:999–1012.CrossRefGoogle Scholar
  18. 18.
    Sarhan WA, Azzazy HME, El-Sherbiny IM. Honey/chitosan nanofiber wound dressing enriched with allium sativum and cleome droserifolia: enhanced antimicrobial and wound healing activity. ACS Appl Mater Interfaces. 2016;8:6379–90.CrossRefGoogle Scholar
  19. 19.
    Rieger KA, Birch NP, Schiffman JD. Designing electrospun nanofiber mats to promote wound healing – a review. J Mater Chem B. 2013;1:4531. Scholar
  20. 20.
    Vellayappan MV, Jaganathan SK, Manikandan A. Nanomaterials as a game changer in the management and treatment of diabetic foot ulcers. RSC Adv. 2016;6:114859–78. Scholar
  21. 21.
    Abrigo M, McArthur SL, Kingshott P. Electrospun nanofibers as dressings for chronic wound care: Advances, challenges, and future prospects. Macromol Biosci. 2014;14:772–92.CrossRefGoogle Scholar
  22. 22.
    Sarhan WA, Azzazy HME. High concentration honey chitosan electrospun nanofibers: Biocompatibility and antibacterial effects. Carbohydr Polym. 2015;122:135–43. Scholar
  23. 23.
    Chaudhary A, Bag S, Barui A, Banerjee P, Chatterjee J. Honey dilution impact on in vitro wound healing: Normoxic and hypoxic condition. Wound Repair Regen. 2017;23:412–22. Scholar
  24. 24.
    Chaudhary A, Bag S, Mandal M, Krishna Karri SP, Barui A, Rajput M, et al. Modulating prime molecular expressions and in vitro wound healing rate in keratinocyte (HaCaT) population under characteristic honey dilutions. J Ethnopharmacol. 2015;166:211–9.CrossRefGoogle Scholar
  25. 25.
    Rajput M, Bhandaru N, Anura A, Pal M, Pal B, Paul RR, et al. Differential behavior of normal and fibrotic fibroblasts under the synergistic influence of micropillar topography and the rigidity of honey/silk-fibroin substrates. ACS Biomater Sci Eng. 2016;2:1528–39.CrossRefGoogle Scholar
  26. 26.
    Rajput M, Bhandaru N, Barui A, Chaudhary A, Paul RR, Mukherjee R, et al. Nano-patterned honey incorporated silk fibroin membranes for improving cellular compatibility. RSC Adv [Internet]. 2014;4:44674–88. Scholar
  27. 27.
    Destaye AG, Lin C-K, Lee C-K. Glutaraldehyde Vapor Cross-linked Nanofibrous PVA Mat with in situ formed silver nanoparticles. ACS Appl Mater Interfaces [Internet]. 2013;5:4745–52. Scholar
  28. 28.
    Clauss M, Tafin UF, Betrisey B, Garderen N van, Trampuz A, Ilchmann T, et al. Influence of physico-chemical material characteristics on Staphylococcal biofilm formation – a qualitative and quantitative in vitro analysis of five different calcium phosphate bone grafts. Eur Cells Mater. 2014;28:39–50.CrossRefGoogle Scholar
  29. 29.
    Topuz F, Uyar T. Electrospinning of gelatin with tunable fiber morphology from round to flat/ribbon. Mater Sci Eng C. 2017;80:371–8. Scholar
  30. 30.
    Megelski S, Stephens JS, Bruce Chase D, Rabolt JF. Micro- and nanostructured surface morphology on electrospun polymer fibers. Macromolecules. 2002;35:8456–66.CrossRefGoogle Scholar
  31. 31.
    Tang C, Saquing CD, Harding JR, Khan SA. In situ cross-linking of electrospun poly(vinyl alcohol) nanofibers. Macromolecules. 2010;43:630–7.CrossRefGoogle Scholar
  32. 32.
    Koombhongse S, Liu W, Reneker DH. Flat polymer ribbons and other shapes by electrospinning. J Polym Sci Part B Polym Phys. 2001;39:2598–606. Scholar
  33. 33.
    Ramakrishna S, Fujihara K, Teo W-E, Lim T-C, Zuwei M. Electrospinning process. An Introdroduction to electrospining nanofibers. Singapore: World Scientific Publishing Co; 2005.CrossRefGoogle Scholar
  34. 34.
    Sarhan WA, Azzazy HME, El-Sherbiny IM. The effect of increasing honey concentration on the properties of the honey/polyvinyl alcohol/chitosan nanofibers. Mater Sci Eng C. 2016;67:276–84. Scholar
  35. 35.
    Khan MQ, Lee H, Khatri Z, Kharaghani D, Khatri M, Ishikawa T. et al. Fabrication and characterization of nanofibers of honey/poly(1,4-cyclohexane dimethylene isosorbide trephthalate) by electrospinning. Mater Sci Eng C. 2017;81:247–51. Scholar
  36. 36.
    Wu M-C, Liao H-C, Chou Y, Hsu C-P, Yen W-C, Chuang C-M, et al. Manipulation of nanoscale phase separation and optical properties of P3HT/PMMA polymer blends for photoluminescent electron beam resist. J Phys Chem B. 2010;114:10277–84. Scholar
  37. 37.
    Tanaka K, Yoon J-S, Takahara A, Kajiyama T. Ultrathinning-induced surface phase separation of Polystyrene/Poly(vinyl methyl ether) blend film. Macromolecule. 1995;28:934–8. Scholar
  38. 38.
    Svečnjak L, Biliškov N, Bubalo D, Barišić D. Application of infrared spectroscopy in honey analysis. Agric Conspec Sci. 2011;76:191–5. Scholar
  39. 39.
    Anjos O, Campos MG, Ruiz PC, Antunes P. Application of FTIR-ATR spectroscopy to the quantification of sugar in honey. Food Chem. 2015;169:218–23.CrossRefGoogle Scholar
  40. 40.
    Turaga U, Singh V, Behrens R, Korzeniewski C, Jinka S, Smith E, et al. Breathability of standalone poly(vinyl alcohol) nanofiber webs. Ind Eng Chem Res. 2014;53:6951–8. Scholar
  41. 41.
    Balaji A, Jaganathan SK, Ismail AF, Rajasekar R. Fabrication and hemocompatibility assessment of novel polyurethane-based bio-nanofibrous dressing loaded with honey and Carica papaya extract for the management of burn injuries. Int J Nanomed NZ. 2016;11:4339–55.CrossRefGoogle Scholar
  42. 42.
    Wang T, Zhu XK, Xue XT, Wu DY. Hydrogel sheets of chitosan, honey and gelatin as burn wound dressings. Carbohydr Polym. 2012;88:75–83. Scholar
  43. 43.
    Molan P, Rhodes T. Honey: a biologic wound dressing. Wounds. 2015;27:141–51.Google Scholar
  44. 44.
    Azzazy HME-S, Sarhan WAA. Bio-compatible apitherapeutic nanofibers. 2015. Scholar
  45. 45.
    Raynaud A, Ghezali L, Gloaguen V, Liagre B, Quero F, Petit JM. Honey-induced macrophage stimulation: AP-1 and NF-??B activation and cytokine production are unrelated to LPS content of honey. Int Immunopharmacol. 2013;17:874–9. Scholar
  46. 46.
    Cooper RA, Molan PC, Harding KG. Antibacterial activity of honey against strains of Staphylococcus aureus from infected wounds. J R Soc Med. 1999;92:283–5. Scholar
  47. 47.
    Rodriguez-Saona LE, Allendorf ME. Use of FTIR for rapid authentication and detection of adulteration of food. Annu Rev Food Sci Technol. 2011;2:467–83.CrossRefGoogle Scholar
  48. 48.
    Svečnjak L, Bubalo D, Baranović G, Novosel H. Optimization of FTIR-ATR spectroscopy for botanical authentication of unifloral honey types and melissopalynological data prediction. Eur Food Res Technol. 2015;240:1101–15. Scholar
  49. 49.
    Xu H, Ma L, Shi H, Gao C, Han C. Chitosan–hyaluronic acid hybrid film as a novel wound dressing: in vitro and in vivo studies. Polym Adv Technol. 2007;18:869–75. Scholar
  50. 50.
    Asran AS, Razghandi K, Aggarwal N, Michler GH, Groth T. Nanofibers from blends of polyvinyl alcohol and polyhydroxy butyrate as potential scaffold material for tissue engineering of skin. Biomacromolecules. 2010;11:3413–21. Scholar
  51. 51.
    Kamoun EA, Kenawy E-RS, Tamer TM, El-Meligy MA, Mohy Eldin MS. Poly (vinyl alcohol)-alginate physically crosslinked hydrogel membranes for wound dressing applications: characterization and bio-evaluation. Arab J Chem. 2015;8:38–47. Scholar
  52. 52.
    Bryan N, Ahswin H, Smart N, Bayon Y, Wohlert S, Hunt JA. Reactive oxygen species (ROS)-a family of fate deciding molecules pivotal in constructive inflammation and wound healing. Eur Cells Mater. 2012;24:249–65.CrossRefGoogle Scholar
  53. 53.
    Dunnill C, Patton T, Brennan J, Barrett J, Dryden M, Cooke J, et al. Reactive oxygen species (ROS) and wound healing: the functional role of ROS and emerging ROS-modulating technologies for augmentation of the healing process. Int Wound J. 2017;14:89–96. Scholar
  54. 54.
    Wagener FADTG, Carels CE, Lundvig DMS. Targeting the redox balance in inflammatory skin conditions. Int J Mol Sci. 2013;14:9126–67. Scholar
  55. 55.
    Kurahashi T, Fujii J. Roles of antioxidative enzymes in wound healing. J Dev Biol. 2015;3:57–70.CrossRefGoogle Scholar
  56. 56.
    Zhao G, Usui ML, Lippman SI, James GA, Stewart PS, Fleckman P, et al. Biofilms and inflammation in chronic wounds. Adavnces Wound Care. 2013;2:389–99.CrossRefGoogle Scholar
  57. 57.
    Omar A, Wright JB, Schultz G, Burrell R, Nadworny P. Microbial biofilms and chronic wounds. Microorganisms. Switzerland; 2017;5:9.Google Scholar
  58. 58.
    Hall-Stoodley L, Costerton JW, Stoodley P. Bacterial biofilms: from the Natural environment to infectious diseases. Nat Rev Micro. 2004;2:95–108. Scholar
  59. 59.
    Elliott CG, Forbes TL, Leask A, Hamilton DW. Inflammatory microenvironment and tumor necrosis factor alpha as modulators of periostin and CCN2 expression in human non-healing skin wounds and dermal fibroblasts. Matrix Biol. 2015;43:71–84.CrossRefGoogle Scholar
  60. 60.
    Landén NX, Li D, Ståhle M. Transition from inflammation to proliferation: a critical step during wound healing. Cell Mol Life Sci. 2016;73:3861–85.CrossRefGoogle Scholar
  61. 61.
    Shah JMY, Omar E, Pai DR, Sood S. Cellular events and biomarkers of wound healing. Indian J Plast Surg. 2012;45:220–8. Scholar
  62. 62.
    Kessler-Becker D, Krieg T, Eckes B. Expression of pro-inflammatory markers by human dermal fibroblasts in a three-dimensional culture model is mediated by an autocrine interleukin-1 loop. Biochem J. 2004;379:351–8. Scholar
  63. 63.
    Chin GC, Diegelmann RF, Schultz GS. Cellular and molecular regulation of wound healing. In: Falabella AF, Kirsne RS, editors. Wound Heal. Boca Raton, USA: Taylor & Francis Group; 2005. pp. 17–37.Google Scholar
  64. 64.
    Nooh HZ, Nermeen MN-E. The dual anti-inflammatory and antioxidant activities of natural honey promote cell proliferation and neural regeneration in a rat model of colitis. Acta Histochem. 2016;118:588–95.CrossRefGoogle Scholar
  65. 65.
    McCarty SM, Percival SL. Proteases and delayed wound healing. Adv Wound Care. 2013;2:438–47. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Ripon Sarkar
    • 1
  • Aritri Ghosh
    • 1
  • Ananya Barui
    • 1
  • Pallab Datta
    • 1
  1. 1.Centre for Healthcare Science and TechnologyIndian Institute of Engineering Science and Technology ShibpurHowrahIndia

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