Biological Trace Element Research

, Volume 168, Issue 1, pp 169–180 | Cite as

Boron and Poloxamer (F68 and F127) Containing Hydrogel Formulation for Burn Wound Healing

  • Selami Demirci
  • Ayşegül Doğan
  • Emre Karakuş
  • Zekai Halıcı
  • Atila Topçu
  • Elif Demirci
  • Fikrettin SahinEmail author


Burn injuries, the most common and destructive forms of wounds, are generally accompanied with life-threatening infections, inflammation, reduced angiogenesis, inadequate extracellular matrix production, and lack of growth factor stimulation. In the current study, a new antimicrobial carbopol-based hydrogel formulated with boron and pluronic block copolymers was evaluated for its healing activity using in vitro cell culture techniques and an experimental burn model. Cell viability, gene expression, and wound healing assays showed that gel formulation increased wound healing potential. In vitro tube-like structure formation and histopathological examinations revealed that gel not only increased wound closure by fibroblastic cell activity, but also induced vascularization process. Moreover, gel formulation exerted remarkable antimicrobial effects against bacteria, yeast, and fungi. Migration, angiogenesis, and contraction-related protein expressions including collagen, α-smooth muscle actin, transforming growth factor-β1, vimentin, and vascular endothelial growth factor were considerably enhanced in gel-treated groups. Macrophage-specific antigen showed an oscillating expression at the burn wounds, indicating the role of initial macrophage migration to the wound site and reduced inflammation phase. This is the first study indicating that boron containing hydrogel is able to heal burn wounds effectively. The formulation promoted burn wound healing via complex mechanisms including stimulation of cell migration, growth factor expression, inflammatory response, and vascularization.


Boron Burn Pluronic Silver sulfadiazine Wound healing 



This study was supported by Yeditepe University and by grants from the Scientific and Technological Research Council of Turkey (TUBITAK, project code: 112 M495). The current formulation has a patent application to the Turkish patent institute with the application number of 2013/04209.

Conflict of Interest

The authors have no conflict of interest.

Ethical Approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

Supplementary material

12011_2015_338_Fig5_ESM.gif (38 kb)
Supplementary Figure 1

a Schematic diagram of the whole experimental plan. Cell viability analysis of NIH 3T3 cells treated with different concentrations of b NaB (1–500 μg/ml), c F68 (5–20 μg/ml), and d F127 (5-20 μg/ml). e NaB and gel formulation stimulated NIH 3T3 cell migration after 36 h incubation period (objective ×4). f Tube formation of HUVEC cells treated with NaB, F68, and F127 (alone or in combination) after 24 h (objective ×4). Black arrows indicate defined concentrations for NaB (15 μg/ml), F68 (10 μg/ml), and F127 (10 μg/ml).Scale bar, 400 μm. Data represent the mean values ± SD per sample from three separate experiments performed in triplicate. Comparisons were performed by ANOVA (Tukey’s post hoc). * P < 0.05: comparison with control group. Control: growth medium-treated NIH 3T3 cells (for b, c, d, and e sections) or HUVECs (for f section). F68: 10 μg/ml pluronic F68; F127: 10 μg/ml pluronic F127; F68-F127: combination of 10 μg/ml pluronic F68 and F127; NaB: 15 μg/ml sodium pentaborate pentahydrate; Gel: combination of 15 μg/ml NaB, 10 μg/ml pluronic F68 and F127 containing growth medium (GIF 37 kb)

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High-resolution image (TIFF 1,721 kb)
12011_2015_338_Fig6_ESM.gif (60 kb)
Supplementary Figure 2

Immunocytochemical analysis of NIH 3T3 cells treated with different reagents for 24 h. Gel formulation resulted in remarkable increase in Col I and VIM expression levels. On the other hand, α-SMA was weakly expressed in NaB group. DAPI used for nucleus staining. Scale bar, 50 μm. Control: growth medium-treated NIH 3T3 cells. F68: 10 μg/ml pluronic F68; F127: 10 μg/ml pluronic F127; F68-F127: combination of 10 μg/ml pluronic F68 and F127; NaB: 15 μg/ml sodium pentaborate pentahydrate; Gel: combination of 15 μg/ml NaB, 10 μg/ml pluronic F68 and F127 containing growth medium. α-SMA: alpha-smooth muscle actin, Col I: collagen type I, and VIM: vimentin (GIF 60 kb)

12011_2015_338_MOESM2_ESM.tif (6.4 mb)
High-resolution image (TIFF 6,575 kb)
12011_2015_338_Fig7_ESM.gif (49 kb)
Supplementary Figure 3

a Macroscopic images of burn wounds at days 3, 7, and 14. b Macroscopic evaluation of deep second-degree burn wound. c Hematoxylin and eosin staining of second-degree burn wound (GIF 48 kb)

12011_2015_338_MOESM3_ESM.tif (761 kb)
High-resolution image (TIFF 761 kb)


  1. 1.
    Bayat M, Vasheghani MM, Razavi N, Taheri S, Rakhshan M (2005) Effect of low-level laser therapy on the healing of second-degree burns in rats: a histological and microbiological study. J Photochem Photobiol B 78(2):171–177CrossRefPubMedGoogle Scholar
  2. 2.
    Alemdaroğlu C, Değim Z, Çelebi N, Zor F, Öztürk S, Erdoğan D (2006) An investigation on burn wound healing in rats with chitosan gel formulation containing epidermal growth factor. Burns 32(3):319–327CrossRefPubMedGoogle Scholar
  3. 3.
    Church D, Elsayed S, Reid O, Winston B, Lindsay R (2006) Burn wound infections. Clin Microbiol Rev 19(2):403–434PubMedCentralCrossRefPubMedGoogle Scholar
  4. 4.
    Dalal ND, Lee RC (2008) Treatment of burn injury by cellular repair. J Craniofac Surg 19(4):903–906CrossRefPubMedGoogle Scholar
  5. 5.
    Lehnhardt M, Jafari HJ, Druecke D, Steinstraesser L, Steinau H, Klatte W, Schwake R, Homann H (2005) A qualitative and quantitative analysis of protein loss in human burn wounds. Burns 31(2):159–167CrossRefPubMedGoogle Scholar
  6. 6.
    Fox CL Jr, Lasker SE, Winfield JM, Mersheimer WL (1954) Albumin, potassium, sodium, and chloride redistribution and erythrocyte loss after surgical trauma and extensive burns. Ann Surg 140(4):524–532PubMedCentralCrossRefPubMedGoogle Scholar
  7. 7.
    Atiyeh BS, Hayek SN, Gunn SW (2005) New technologies for burn wound closure and healing-review of the literature. Burns 31(8):944–956CrossRefPubMedGoogle Scholar
  8. 8.
    Silver S, Phung LT, Silver G (2006) Silver as biocides in burn and wound dressings and bacterial resistance to silver compounds. J Ind Microbiol Biotechnol 33(7):627–634CrossRefPubMedGoogle Scholar
  9. 9.
    Burd A, Kwok CH, Hung SC, Chan HS, Gu H, Lam WK, Huang L (2007) A comparative study of the cytotoxicity of silver‐based dressings in monolayer cell, tissue explant, and animal models. Wound Repair Regen 15(1):94–104CrossRefPubMedGoogle Scholar
  10. 10.
    Benderdour M, Van Bui T, Hess K, Dicko A, Belleville F, Dousset B (2000) Effects of boron derivatives on extracellular matrix formation. J Trace Elem Med Biol 14(3):168–173CrossRefPubMedGoogle Scholar
  11. 11.
    Nielsen FH, Penland JG (1999) Boron supplementation of peri‐menopausal women affects boron metabolism and indices associated with macromineral metabolism, hormonal status and immune function. J Trace Elem Exp Med 12(3):251–261CrossRefGoogle Scholar
  12. 12.
    Gorustovich AA, Steimetz T, Nielsen FH, Guglielmotti MB (2008) Histomorphometric study of alveolar bone healing in rats fed a boron‐deficient diet. Anat Rec 291(4):441–447CrossRefGoogle Scholar
  13. 13.
    Blech M, Martin C, Borrelly J, Hartemann P (1990) Treatment of deep wounds with loss of tissue. Value of a 3 percent boric acid solution. Presse medicale 19(22):1050–1052Google Scholar
  14. 14.
    Chebassier N, Ouijja EH, Viegas I, Dreno B (2004) Stimulatory effect of boron and manganese salts on keratinocyte migration. Acta Derm Venereol 84(3):191–194CrossRefPubMedGoogle Scholar
  15. 15.
    Doğan A, Yalvaç ME, Şahin F, Kabanov AV, Palotás A, Rizvanov AA (2012) Differentiation of human stem cells is promoted by amphiphilic pluronic block copolymers. Int J Nanomedicine 7:4849–4860PubMedCentralPubMedGoogle Scholar
  16. 16.
    Kabanov AV, Batrakova EV, Sriadibhatla S, Yang Z, Kelly DL, Alakov VY (2005) Polymer genomics: shifting the gene and drug delivery paradigms. J Control Release 101(1):259–271CrossRefPubMedGoogle Scholar
  17. 17.
    Sahay G, Gautam V, Luxenhofer R, Kabanov AV (2010) The utilization of pathogen-like cellular trafficking by single chain block copolymer. Biomaterials 31(7):1757–1764PubMedCentralCrossRefPubMedGoogle Scholar
  18. 18.
    Khalil EA, Afifi FU, Al-Hussaini M (2007) Evaluation of the wound healing effect of some Jordanian traditional medicinal plants formulated in Pluronic F127 using mice (Mus musculus). J Ethnopharmacol 109(1):104–112CrossRefPubMedGoogle Scholar
  19. 19.
    Choi JS, Yoo HS (2010) Pluronic/chitosan hydrogels containing epidermal growth factor with wound‐adhesive and photo‐crosslinkable properties. J Biomed Mater Res A 95(2):564–573CrossRefPubMedGoogle Scholar
  20. 20.
    Faulkner DM, Sutton ST, Hesford JD, Faulkner BC, Major DA, Hellewell TB, Laughon MM, Rodeheaver GT, Edlich RF (1997) A new stable pluronic® F68 gel carrier for antibiotics in contaminated wound treatment. Am J Emerg Med 15(1):20–24CrossRefPubMedGoogle Scholar
  21. 21.
    Erukova VY, Krylova OO, Antonenko YN, Melik-Nubarov NS (2000) Effect of ethylene oxide and propylene oxide block copolymers on the permeability of bilayer lipid membranes to small solutes including doxorubicin. Biochim Biophys Acta 1468(1):73–86CrossRefPubMedGoogle Scholar
  22. 22.
    Murhammer DW, Goochee CF (1990) Sparged animal cell bioreactors: mechanism of cell damage and Pluronic F-68 protection. Biotechnol Prog 6(5):391–397CrossRefPubMedGoogle Scholar
  23. 23.
    Kant V, Gopal A, Kumar D, Gopalkrishnan A, Pathak NN, Kurade NP, Tandan SK, Kumar D (2013) Topical pluronic F-127 gel application enhances cutaneous wound healing in rats. Acta Histochemica 116(1):5–13CrossRefPubMedGoogle Scholar
  24. 24.
    Yuhua S, Ligen L, Jiake C, Tongzhu S (2012) Effect of Poloxamer 188 on deepening of deep second-degree burn wounds in the early stage. Burns 38(1):95–101CrossRefPubMedGoogle Scholar
  25. 25.
    Nalbandian RM, Henry RL, Balko KW, Adams DV, Neuman NR (1987) Pluronic F‐127 gel preparation as an artificial skin in the treatment of third‐degree burns in pigs. J Biomed Mater Res A 21(9):1135–1148CrossRefGoogle Scholar
  26. 26.
    Lai C-C, Huang F-M, Yang H-W, Chan Y, Huang M-S, Chou M-Y, Chang Y-C (2001) Antimicrobial activity of four root canal sealers against endodontic pathogens. Clin Oral Investig 5(4):236–239CrossRefPubMedGoogle Scholar
  27. 27.
    Kalaycı S, Demirci S, Sahin F (2014) Determination of antimicrobial properties of picaridin and DEET against a broad range of microorganisms. World J Microbiol Biotechnol 30(2):407–411CrossRefPubMedGoogle Scholar
  28. 28.
    Walter M, Wright KT, Fuller H, MacNeil S, Johnson WEB (2010) Mesenchymal stem cell-conditioned medium accelerates skin wound healing: an in vitro study of fibroblast and keratinocyte scratch assays. Exp Cell Res 316(7):1271–1281CrossRefPubMedGoogle Scholar
  29. 29.
    Tang L, Ma X, Tian Q, Cheng Y, Yao H, Liu Z, Qu X, Han X (2013) Inhibition of angiogenesis and invasion by DMBT is mediated by downregulation of VEGF and MMP-9 through Akt pathway in MDA-MB-231 breast cancer cells. Food Chem Toxicol 56:204–213CrossRefPubMedGoogle Scholar
  30. 30.
    Vorauer-Uhl K, Fürnschlief E, Wagner A, Ferko B, Katinger H (2001) Topically applied liposome encapsulated superoxide dismutase reduces postburn wound size and edema formation. Eur J Pharm Sci 14(1):63–67CrossRefPubMedGoogle Scholar
  31. 31.
    Xu J, Liu X, Chen J, Zacharek A, Cui X, Savant-Bhonsale S, Liu Z, Chopp M (2009) Simvastatin enhances bone marrow stromal cell differentiation into endothelial cells via notch signaling pathway. Am J Physiol Cell Physiol 296(3):C535–C543PubMedCentralCrossRefPubMedGoogle Scholar
  32. 32.
    Mohajeri D, Mesgari M, Doustar Y, Nazeri M (2011) Comparison of the effect of normal saline and silver sulfadiazine on healing of skin burn wounds in rats: a histopathological study. Middle East J Sci Res 10(1):08–14Google Scholar
  33. 33.
    Poon VK, Burd A (2004) In vitro cytotoxicity of silver: implication for clinical wound care. Burns 30(2):140–147CrossRefPubMedGoogle Scholar
  34. 34.
    Nzietchueng RM, Dousset B, Franck P, Benderdour M, Nabet P, Hess K (2002) Mechanisms implicated in the effects of boron on wound healing. J Trace Elem Med Biol 16(4):239–244CrossRefPubMedGoogle Scholar
  35. 35.
    Doğan A, Demirci S, Çağlayan AB, Kılıç E, Günal MY, Uslu Ü, Cumbul A, Şahin F (2014) Sodium pentaborate pentahydrate and pluronic containing hydrogel increases cutaneous wound healing in vitro and in vivo. Biol Trace Elem Res 162(1–3):72–79PubMedGoogle Scholar
  36. 36.
    Gümüstekin K, Seven B, Karabulut N, Aktas Ö, Gürsan N, Aslan S, Keles M, Varoglu E, Dane S (2004) Effects of sleep deprivation, nicotine, and selenium on wound healing in rats. Int J Neurosci 114(11):1433–1442CrossRefPubMedGoogle Scholar
  37. 37.
    Wetzler C, Kämpfer H, Stallmeyer B, Pfeilschifter J, Frank S (2000) Large and sustained induction of chemokines during impaired wound healing in the genetically diabetic mouse: prolonged persistence of neutrophils and macrophages during the late phase of repair. J Invest Dermatol 115(2):245–253CrossRefPubMedGoogle Scholar
  38. 38.
    Sidhu GS, Mani H, Gaddipati JP, Singh AK, Seth P, Banaudha KK, Patnaik GK, Maheshwari RK (1999) Curcumin enhances wound healing in streptozotocin induced diabetic rats and genetically diabetic mice. Wound Repair Regen 7(5):362–374CrossRefPubMedGoogle Scholar
  39. 39.
    Lee Y-H, Chang J-J, Chien C-T, Yang M-C, Chien H-F (2012) Antioxidant sol–gel improves cutaneous wound healing in streptozotocin-induced diabetic rats. Exp Diabetes Res 2012:504693. doi: 10.1155/2012/504693 PubMedCentralPubMedGoogle Scholar
  40. 40.
    Zhu H, Ka B, Murad F (2007) Nitric oxide accelerates the recovery from burn wounds. World J Surg 31:624–631CrossRefPubMedGoogle Scholar
  41. 41.
    Tonnesen MG, Feng X, Clark RA (2000) Angiogenesis in wound healing. J Investig Dermatol Symp Proc 5(1):40–46CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Selami Demirci
    • 1
  • Ayşegül Doğan
    • 1
  • Emre Karakuş
    • 2
  • Zekai Halıcı
    • 3
  • Atila Topçu
    • 3
  • Elif Demirci
    • 4
  • Fikrettin Sahin
    • 1
    Email author
  1. 1.Department of Genetics and Bioengineering, Faculty of Engineering and ArchitectureYeditepe UniversityIstanbulTurkey
  2. 2.Department of Pharmacology and ToxicologyAtaturk University School of Veterinary MedicineErzurumTurkey
  3. 3.Department of Pharmacology, Faculty of MedicineAtaturk UniversityErzurumTurkey
  4. 4.Department of Pathology, Medical FacultyAtaturk UniversityErzurumTurkey

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