Evaluation of antibiotic activity of methicillin in healing of full-thickness infected wounds with sensitized methicillin resistant Staphylococcus aureus in presence of HAMLET

Document Type : Original Article


1 Young Researchers and Elite Club, Urmia Branch, Islamic Azad University, Urmia, Iran

2 Department of Surgery and Diagnostic Imaging, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran


Objective(s): The novel healing choices for handling of infections due to multidrug resistant Staphylococcus aureus are reguired. HAMLET has been reported to be able to sensitize bacterial pathogens to traditional antimicrobial agents. The aim was to assess wound healing activity of methicillin in presence of HAMLET in methicillin resistant S. aureus (MRSA) infected wounds.
Materials and Methods: Fifty male rats were randomized into five groups of ten animals each. In CONTROL group, 0.1 ml sterile saline 0.9% solution was added to the wounds with no infection.  In MRSA group, the wounds were infected with MRSA and only treated with 0.1 ml the sterile saline (0.9%) solution. In MRSA/HAMLET group, infected wounds were cured with HAMLET (100 µg). In group MRSA/ Met, animals with infected wounds were cured with 0.1 ml local use of 1 mg/ml methicillin. In MRSA/Met/HAMLET group, animals with infected wounds were cured with local use of 0.1 ml solution of methicillin (1 mg/ml) and HAMLET (100 µg). All test formulations were used for ten consecutive days, twice a day, beginning from first treatment.
Results: Microbiological examination, planimetric, histological and quantitative morphometric studies, immunohistochemical staining for angiogenesis, determination of hydroxyproline levels and RT-PCR for Caspase 3, Bcl-2 and p53 showed that there was significant difference between animals in MRSA/Met/ HAMLET group compared to other groups (P<0.05).
Conclusion: HAMLET could make methicillin beneficial for handling of MRSA infected wounds and had the prospective effect to consider this harmless agent for local application.


Main Subjects

Srinivas RB, Kiran KRR, Naidu VG, Madhusudhana K, Agwane SB, Ramakrishna S, et al. Evaluation of antimicrobial, antioxidant and wound healing potentials of Holoptelea integrifolia. J Ethnopharmacol. 2008;115:249-256.
Khan HA, Ahmad A, Mehboob R. Nosocomial infections and their control strategies. Asian Pac J Trop Biomed 2015;5:509-514.
Calfee DP. Methicillin-resistant Staphylococcus aureus and vancomycin resistant enterococci, and other Gram-positives in healthcare. Curr Opin Infect Dis 2012; 25:385-394.
Gould IM, David MZ, Esposito S, Garau J, Lina G, Mazzei T, Peters G. New insights into meticillin-resistant Staphylococcus aureus (MRSA) pathogenesis, treatment and resistance. Int J Antimicrob Agents 2012; 39:96–104.
Klevens RM, Morrison MA, Nadle J, Petit S, Gershman K, Ray S, et al. Active bacterial core surveillance (ABCs) MRSA investigators. Invasive methicillin-resistant Staphylococcus aureus infections in the United States. JAMA 2007; 298:1763-1771.
Fischbach MA, Walsh CT. Antibiotics for emerging pathogens. Science 2007; 325:1089–1093.
Braine T. Race against time to develop new antibiotics. Bull World Health Organ 2011; 89:88–89.
Alanis AJ. Resistance to antibiotics: are we in the post-antibiotic era? Arch Med Res 2005; 36:697–705.
Marks LR, Clementi EA, Hakansson AP. The human milk protein-lipid complex HAMLET sensitizes bacterial pathogens to traditional antimicrobial agents. PLoS One 2012;7(8):e43514.
Marks LR, Clementi EA, Hakansson AP. Sensitization of Staphylococcus aureus to methicillin and other antibiotics in vitro and in vivo in the presence of HAMLET. PLoS One 2013;8:e63158
Svensson M, Hakansson A, Mossberg AK, Linse S, Svanborg C. Conversion of alpha-lactalbumin to a protein inducing apoptosis. Proc Natl Acad Sci USA 2000; 97: 4221–4226.
Kumar MS, Kirubanandan, S, Sripriya R, Sehgal PK. Triphala promotes healing of infected full-thickness dermal wound. J Surg Res 2008;144:94-101.
Qiu Z, Kwon AH, Kamiyama Y. Effects of plasma fibronectin on the healing of full-thickness skin wounds in streptozotocin-induced diabetic rats. J Surg Res 2007;138:64-70.
Peters EJ, Lipsky BA. Diagnosis and management of infection in the diabetic foot. Med Clin North Am. 2013; 97:911-946
Pattanayak SP, Sunita P. Wound healing, anti-microbial and antioxidant potential of Dendrophthoe falcata (L.f) Ettingsh. J Ethnopharmacol 2008; 120:241-247.
Dogan S, Demirer S, Kepenekci I, Erkek B, Kiziltay A, Hasirci N, et al. Epidermal growth factor containing wound closure enhances wound healing in non-diabetic and diabetic rats. Int Wound J 2009; 6:107–115.
Martin J, Zenilman M, Lazarus GS. Molecular microbiology: new dimensions for cutaneous biology and wound healing. J Invest Dermatol 2010; 130:38-48.
Berridge MJ. Cell Stress, Inflammatory Responses and Cell Death. Cell Signal Biol  2014 ;1:1-30.
Harper D, Young A, McNaught CE. The physiology of wound healing. Surgery (Oxford) 2014; 32:445-450.
Rodero M, Khosrotehrani K. Skin wound healing modulation by macrophages. Int J Clin Exp Pathol 2010; 3:643–653.
Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007; 35:495-516.
Wu YS, Chen SN. Apoptotic cell: linkage of inflammation and wound healing. Front Pharmacol 2014;5:1.
Bhan S, Mitra R, Arya AK, Pandey HP, Tripathi K. A Study on evaluation of apoptosis and expression of Bcl-2-Related marker in wound healing of streptozotocin-induced diabetic rats. ISRN Dermatol 7(2013), Article ID 739054, 6 pages.
Hockenbery D, Nunez G, Milliman C, Schreiber RD, Korsmeyer SJ. Bcl-2 is an inner itochondrialmembrane protein that blocks programmed cell death. Nature 1990; 348:334–336.
Jean SS, Hsueh PR. Current review of antimicrobial treatment of nosocomial pneumonia caused by multidrug-resistant pathogens. Expert Opin Pharmacother 2011;12: 2145–2148.
Rose WE, Poppens PT. Impact of biofilm on the in vitro activity of vancomycin alone and in combination with tigecycline and rifampicin against Staphylococcus aureus. J Antimicrob Chemother.  2009; 63:485–488.
Tang HJ, Chen CC, Ko WC, Yu WL, Chiang SR, Chuang YC. In vitro efficacy of antimicrobial agents against high-inoculum or biofilm-embedded methicillin resistant Staphylococcus aureus with vancomycin minimal inhibitory concentrations equal to 2 mug/ml (VA2-MRSA). Int J Antimicrob Agents 2011; 38:46–51.