Evaluation of glycine as a monotherapy and in combination with neomycin in promoting wound healing in rabbits
DOI:
https://doi.org/10.30539/1pfjs828Keywords:
Glycine, neomycin, wound healing, Staphylococcus aureus, combinationAbstract
Background: Skin wounds infected with Staphylococcus aureus pose a significant challenge in veterinary medicine due to their delay in healing and impairment of tissue repair. Therefore, this study aims to evaluate the therapeutic effect of neomycin and glycine, both individually and in combination, on accelerating the healing of infected skin wounds experimentally. Methodology: The study included two parts: in vitro and in vivo. In the in vitro part, the minimum inhibitory concentration (MIC) for both neomycin and glycine were determined, and a checkerboard test was performed to evaluate the interaction between them. In the animal study, twenty adult rabbits were randomly divided into four groups, five rabbits for each: a positive control group (untreated), a neomycin ointment group (5% in white petrolatum), a glycine ointment group (2.5% in white petrolatum), and a combination group treated with neomycin (2.5%) + glycine (2%) formulated in a white petrolatum base. Surgical wounds measuring 2 cm² were created on the backs of the rabbits, and then all of them were injected with a bacterial suspension containing S. aureus. The wound area was assessed using photographs and analyzed using ImageJ software on the days 7, 14, and 21 after treatment. Results: Neomycin exhibited a higher inhibitory activity against S. aureus than glycine. The interaction test between the two compounds also showed a fractional inhibitory concentration index (FICI) value of 2, indicating neither a synergistic nor an antagonistic interaction (no interaction). In contrast, animal results showed that all treatment groups had a significant reduction in wound area compared to the control group, with the combination group (neomycin + glycine) achieving the fastest healing rates during the observation period. Conclusions: The combination of neomycin and glycine improves wound healing in vivo, although no synergistic interaction between the two compounds was found in the in vitro study. This suggests that the positive effect in animals may be related to local physiological mechanisms related to the inflammatory response and accelerated tissue regeneration.
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References
CLSI. (2016). M100: Performance standards for antimicrobial susceptibility testing (26th ed.). Wayne, PA: Clinical and Laboratory Standards Institute. https://webstore.ansi.org/standards/clsi/clsim100s26
Hassan, M. A., et al. (2024). Evaluation of Xylazine–Ketamine Anesthesia in Rabbits Undergoing Tendon Surgery: A Prospective Randomized Controlled Study. New Valley Veterinary Journal, 4(1). https://doi.org/10.21608/nvvj.2023.247402.1038
Franca, A., Gaio, V., Lopes, N., & Melo, L. D. R. (2021). Virulence factors in coagulase-negative staphylococci. Pathogens, 10(2), 170. https://doi.org/10.3390/pathogens10020170
Hong, S., Su, S., Gao, Q., Chen, M., Xiao, L., Cui, R., et al. (2023). Enhancement of β-lactam-mediated killing of Gram-negative bacteria by lysine hydrochloride. Microbiology Spectrum, 11(4), e01198-23. https://doi.org/10.1128/spectrum.01198-23
Kožár, M., Hamilton, H., & Koščová, J. (2018). Types of wounds and the prevalence of bacterial contamination of wounds in the clinical practice of small animals. Folia Veterinaria, 62(4), 39. https://doi.org/10.2478/fv-2018-0036
Maali, Y., Badiou, C., Martins-Simões, P., Hodille, E., Bes, M., Andenesch, F., et al. (2018). Understanding the virulence of Staphylococcus pseudintermedius: A major role of pore-forming toxins. Frontiers in Cellular and Infection Microbiology, 8, 221. https://doi.org/10.3389/fcimb.2018.00221
Ibrahim, O. M. S. (2012). Evaluation of antibacterial activity of Capparis spinosa (Al-Kabara) and Aloe vera extracts against bacterial isolates from skin wound infections in vitro and in vivo. Kufa Journal for Veterinary Medical Sciences, 3(2), 23–35. https://doi.org/10.36326/kjvs/2012/v3i23938
Mingeot-Leclercq, M. P., & Tulkens, P. M. (1999). Aminoglycosides: Nephrotoxicity. Antimicrobial Agents and Chemotherapy, 43(5), 1003–1012. https://doi.org/10.1128/AAC.43.5.1003
Moazen, J., Riyahi, F., & Hallaj, B. (2022). Characterization of virulence genes and antibiotic resistance of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible Staphylococcus aureus (MSSA) isolates in ICU and non-ICU wards. Trends in Medical Sciences, 2(2), e129037. https://doi.org/10.5812/tms-129037
Pomba, C., Rantala, M., Greko, C., Baptiste, K. E., Catry, B., van Duijkeren, E., … et al. (2017). Public health risk of antimicrobial resistance transfer from companion animals. Journal of Antimicrobial Chemotherapy, 72(4), 957–968. https://doi.org/10.1093/jac/dkw481
Razak, M. A., Begum, P. S., Viswanath, B., & Rajagopal, S. (2017). Multifaceted beneficial effect of nonessential amino acid glycine. Oxidative Medicine and Cellular Longevity, 2017, 1716701. https://doi.org/10.1155/2017/1716701
Stokes, J. M., Lopatkin, A. J., Lobritz, M. A., & Collins, J. J. (2019). Bacterial metabolism and antibiotic efficacy. Cell Metabolism, 30(2), 251–259. https://doi.org/10.1016/j.cmet.2019.06.009
Tasneem, U., Mehmood, K., Majid, M., Ullah, S. R., & Andleeb, S. (2022). Methicillin-resistant Staphylococcus aureus: A brief review of virulence and resistance. Journal of the Pakistan Medical Association, 72(3), 509–515. https://doi.org/10.47391/JPMA.0504
Wilkinson, H. N., & Hardman, M. J. (2020). Wound healing: Cellular mechanisms and pathological outcomes. Open Biology, 10(9), 200223. https://doi.org/10.1098/rsob.200223
Wu, G. (2010). Functional amino acids in growth, reproduction, and health. Advances in Nutrition, 1(1), 31–37. https://doi.org/10.3945/an.110.1008
Wu, G., Bazer, F. W., Dai, Z., Li, D., Wang, J., & Wu, Z. (2013). Functional amino acids in nutrition and health. Amino Acids, 45(3), 407–417. https://doi.org/10.1007/s00726-013-1500-6
Windahl, U., Bengtsson, B., Nyman, A.K., & Holst, B.S. (2015). The Distribution of Pathogens and Their Antimicrobial Susceptibility Patterns among Canine SurgicalWound Infections in Sweden in Relation to Different Risk Factors. Acta Veterinaria Scandinavica, 57, 11. https://doi.org/10.1186/s13028-015-0102-6
Pomba, C., Rantala, M., Greko, C., Baptiste, K. E., Catry, B., van Duijkeren, E., … et al. (2017). Public health risk of antimicrobial resistance transfer from companion animals. Journal of Antimicrobial Chemotherapy, 72(4), 957–968.
Zhong, Z., Zhao, Y., & Zhang, J. (2020). Protective role of glycine in wound healing and tissue regeneration. Frontiers in Physiology, 11, 1267. https://doi.org/10.3389/fphys.2020.01267
Zhao, X. L., Chen, Z. G., Yang, T. C., Jiang, M., Wang, J., Cheng, Z. X., Yang, M. J., Zhu, J. X., Zhang, T. T., Li, H., Peng, B., & Peng, X. X. (2021). Glutamine promotes antibiotic uptake to kill multidrug-resistant uropathogenic bacteria. Science Translational Medicine, 13(619), eabj0716. https://doi.org/10.1126/scitranslmed.abj0716
Cheng, Z. X., Guo, C., Chen, Z. G., ….et al. (2019). Glycine, serine and threonine metabolism confounds efficacy of complement-mediated killing. Nature Communications, 10(1), 3325. https://doi.org/10.1038/s41467-019-11129-5
Ye, J. Z., Lin, X. M., Cheng, Z. X., et al. (2018). Identification and efficacy of glycine, serine and threonine metabolism in potentiating kanamycin-mediated killing of Edwardsiella piscicida. Journal of Proteomics, 183, 34–44. https://doi.org/10.1016/j.jprot.2018.05.006
Kou, T. S., Wu, J. H., Chen, X. W., Chen, Z. G., Zheng, J., & Peng, B. (2022). Exogenous glycine promotes oxidation of glutathione and restores sensitivity of bacterial pathogens to serum-induced cell death. Redox Biology, 58, 102512. https://doi.org/10.1016/j.redox.2022.102512
Lescat, M., Schuetz, A., & Bernier, A. (2019). Preparation and use of Alamar Blue reagent in microbiological assays. Journal of Microbiological Methods, 160, 76–81. https://doi.org/10.1016/j.mimet.2019.03.003
Gurtner, G. C., Werner, S., Barrandon, Y., & Longaker, M. T. (2008). Wound repair and regeneration. Nature, 453(7193), 314–321. https://doi.org/10.1038/nature07039
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