Healing of Skin Wounds in Rats: Influence of Low-Level Laser Therapy, Alone or Combined with Natural Products

Diana Milanez Ávila Dias Maciel, Sheila Canevese Rahal, Carlos Eduardo Fonseca Alves, Ivan Felismino Charas dos Santos, Alexandre Botelho de Abreu Sampaio, Laís Rosa Nagai, Barbara Wagner Duarte Ferraz de Camargo, Gabriela Cunha Salewski

Abstract


Background: A great variety of natural products have been evaluated for the skin wound healing due to anti-inflammatory, antioxidant and antibacterial activities, and procollagen synthesis, of which may be mentioned the honey and propolis. In turn, low-level laser therapy (LLLT) is considered an important tool in the treatment of skin wounds, because of ability to raise the ATP production, to stimulate the microcirculation, and formation of new blood vessels. Therefore,this study aimed to assess the influence of LLLT, alone or combined with natural products, in the healing of excisional skin wounds in rats.

Materials, Methods & Results:  One hundred twenty-six male Wistar rats, aged 14 weeks, were randomly divided into seven groups (n = 18 per group) according to wound treatments: G1 (negative control): 0.9% saline solution; G2 (positive control): allantoin + zinc oxide ointment; G3: LLLT; G4: mixture of honey + 5% propolis hydroalcoholic solution; G5: LLLT + honey + hydroalcoholic solution of 5% propolis; G6: LLLT + honey in natura; G7: LLLT + 5% propolis hydroalcoholic solution. Six rats from each group were euthanized on the 7th, 14th, and 21th postoperative days. Macroscopic and histological evaluations of wound healing were performed. A decrease in wound area occurred in all groups, but in general G4 had the higher reduction and G1 and G3 had the lower reductions. These findings corresponded to the percentage of wound contraction, since the largest contraction was G4 and the lowest contractions were G1 and G3. Histological analysis showed no significant difference among groups on the 7th postoperative day. On postoperative day 14, a significant decrease in hemorrhage occurred between G1xG4, G1xG5 and G1xG7, and a significant decrease in congestion between G1xG7 and G2xG5. In addition, intensity of the inflammatory infiltrate showed a significant reduction between G1xG7, G2xG7 and G3xG7. On postoperative day 21, a significant changed from mixed to mononuclear inflammatory infiltrate was observed between G1xG3, G1xG4, G1xG6, G1xG7, G2xG3, and G2xG6. Regarding organization of fibroblasts, there were differences between G1xG6 and G1xG7. 


Discussion: Spite the red laser has been associated to in increase fibroblast proliferation, G3 showed inadequate decrease in wound area and percentage of contraction. Histological examination showed no differences among groups in the inflammatory phase. However, in the proliferative and remodeling phases around days 14 and 21 were found some differences among groups, which may be related to the actions of the natural products, or laser. Studies have shown that honey acts as a repair and anti-inflammatory agent in skin woundthat may be associated to lower hemorrhage in G4 and G4 compared to G1 on day 14. Propolis is related to reduction of free radicals and increasing the amount of collagen, which may have contributed for quality improvement of the inflammatory infiltrate in G4 and G7 compared to G1, and fibroblast organization in G7 at postoperative day 21. In conclusion, macroscopically the mixture of honey + 5% propolis hydroalcoholic solution was the most effective in reducing wound area and increasing wound contraction. However, based on the parameters evaluated histologically, may be highlighted the evolution of the treatments with LLLT + honey, as well as LLLT + 5% propolis hydroalcoholic solution.

Keywords: honey, propolis, biomodulation, wound, healing.


Full Text:

PDF

References


Andrade F.S.S.D., Clark R.M & Ferreira M.L. 2014. Effects of low-level laser therapy on wound healing. Revista do Colégio Brasileiro de Cirurgiões. 41(2): 129-133.

Al-Watban F.A.H. & Andres B.L. 2001. Laser photons and pharmacological treatments in wound healing. Laser Therapy. 12: 1-9.

Bayat M., Azari A. & Galmohammadi M.G. 2010. Effects of 780nm low-level laser therapy with a pulsed gallium aluminum arsenide laser on the healing of a surgically induced open skin wound of rat. Photomedicine and Laser Surgery. 28(4): 465-470.

Benhanifia M.B., Boukraâ L., Hammoudi S.M., Sulaiman S.A. & Manivannan L. 2011. Recent patents on topical application of honey in wound and burn management. Recent Patents on Inflammation & Allergy Drug Discovery. 5(1): 81-86

Bergman A., Yanai J., Weiss J., Bell D. & David M.P. 1983. Acceleration of wound healing by topical application of honey. American Journal of Surgery. 145(3): 374-376.

Chaves M.E., Araújo A.R., Piancastelli A.C. & Pinotti M. 2014. Effects of low-power light therapy on wound healing: LASER x LED. Anais Brasileiros de Dermatologia. 89(4): 616-623.

Garraud O., Hozzein W.N. & Badr G. 2017. Wound healing: time to look for intelligent, ‘natural’ immunological approaches? BMC Immunology. 18(Suppl 1): 39-65.

Gonçalves R.V., Novaes R.D., Matta S.L.P., Benevides G.P., Faria F.R. & Pinto M.V.M. 2010. Comparative study of the effects of galliumaluminium- arsenide laser photobiomodulation and healing oil on skin wound in Wistar rats: a histomorphometric study. Photomedicine and Laser Surgery. 28(5): 597-602.

Güngörmüs M. & Akyol U. 2009. The effect of gallium-aluminum arsenide 808nm low-level laser therapy on healing of skin incisions made using a diode laser. Photomedicine and Laser Surgery. 27(6): 895-899.

Ibrahim N., Wong S.K., Mohamed I.N., Mohamed N., Chin K.Y., Ima-Nirwana S. & Shuid A.N. 2018. Wound healing properties of selected natural products. International Journal of Environmental Research and Public Health. 15(11): 1-23.

Kotian S., Bhat K., Pai S., Nayak J., Souza A., Gourisheti K. & Padma D. 2018. The role of natural medicines on wound healing: a biomechanical, histological, biochemical and molecular study. Ethiopian Journal of Science. 28(6): 759-770.

Lima F.J.C., Barbosa F.T. & Sousa-Rodrigues C.F. 2014. Use alone or in combination of red and infrared laser in skin wounds. Journal of Lasers in Medical Sciences. 5(2): 51-57.

Lima F.J., Oliveira Neto O.B., Barbosa F.T., Nascimento Galvão A.M., Ramos F.W., Lima C.C. & Sousa Rodrigues C.F. 2016. Is there a protocol in experimental skin wounds in rats using low-level diode laser therapy (LLDLT) combining or not red and infrared wavelengths? Systematic review. Lasers in Medical Science. 31(4): 779-787.

Mani F., Damasceno H.C., Novelli E.L., Martins E.A. & Sforcin J.M. 2006. Propolis: Effect of different concentrations, extracts and intake period on seric biochemical variables. Journal of Ethnopharmacology. 105(1-2): 95-98.

Majtan J. 2014. Honey: An immunomodulator in wound healing. Wound Repair and Regeneration. 22(2): 187-192.

Martinotti S. & Ranzato E. 2018. Honey, wound repair and regenerative medicine. Journal of Functional Biomaterials. 9(2): 1-7.

Medrado A.P., Soares A.P., Santos E.T., Reis S.R.A. & Andrade Z.A. 2008. Influence of laser photobiomodulation upon connective tissue remodeling during wound healing. Journal of Photochemistry and Photobiology B: Biology. 92(3): 144-152.

Meo S.A., Al-Asiri S.A., Mahesar A.L. & Ansari M.J. 2017. Role of honey in modern medicine. Saudi Journal of Biological Sciences. 24(5): 975-978.

Molan P.C. 1999. The role of honey in the management of wounds. Journal of Wound Care. 8(8): 415-418.

Ito E.H., Araújo W.L.P., Shinohara A.J., Barros D.C.B., Camilli M.P. & Orsi R.O. 2018. Physicochemical characteristics of honeys of Apis mellifera L. from the Polo cuesta region, Sao Paulo, Brazil. Boletim de Indústria Animal. 75: 1-9.

Oryan A., Alemzadeh E. & Moshiri A. 2018. Potential role of propolis in wound healing: Biological properties and therapeutic activities. Biomedicine & Pharmacotherapy. 98: 469-483.

Pasupuleti V.R., Sammugam L., Ramesh N. & Gan S.H. 2017. Honey, propolis, and royal jelly: a comprehensive review of their biological actions and health benefits. Oxidative Medicine and Cellular Longevity. 21p. doi: 10.1155/2017/1259510.

Pereira R.F. & Bártolo P.J. 2016. Traditional therapies for skin wound healing. Advances in Wound Care. 5(5): 208-229.

Rocha Junior A.M., Vieira B.J., Andrade L.C.F. & Aarestrup F.M. 2007. Effects of low-level laser therapy on the progress of wound healing in humans: the contribution of in vitro and in vivo experimental studies. Jornal Vascular Brasileiro. 6(3): 258-266.

Takzaree N., Hadjiakhondi A., Hassanzadeh G., Rouini M.R. & Manayi A. 2016. Synergistic effect of honey and propolis on cutaneous wound healing in rats. Acta Medica Iranica. 54(4): 233-239.

Zabaiou N., Fouache A., Trousson A., Baron S., Zellagui A., Lahouel M. & Lobaccaro J.A. 2017. Biological properties of propolis extracts: something new from an ancient product. Chemistry and Physics of Lipids. 207(Pt B): 214-222.




DOI: https://doi.org/10.22456/1679-9216.99845

Copyright (c) 2020 Diana Milanez Ávila Dias Maciel, Sheila Canevese Rahal

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.