Abstract
This study was carried out to evaluate the effectiveness of the triple combination of biologically synthesized silver nanoparticles, anti-microbial peptides and visible blue light against pathogenic clinical methicillin resistance Staphylococcus aureus (MRSA) in vivo after inducing full thickness skin injury in the initially induced diabetic rats. This experiment of in vivo study was for evaluation the role of the combinations in promoting wound healing after wound inducing infection using pathogenic MRSA (0.25ml from 1.5 ×108 cfu/ml) in diabetic rats. 108 diabetic rats (aged 8-10 weeks weighing 200-250g) were divided equally into nine groups, (treatment started after the appearance of infection signs). Group A: Negative control group, group B: Positive control group, Group C: infected treated with AgNPs 8µg/ml, group D: infected treated with Gramicidin 16µg/ml, group E: infected treated with blue light for 1 hour, group F: infected treated with combination of AgNPs 0.25µg/ml with blue light for 1 hour, group G: infected treated with combination of Gramicidin 8 µg/ml and blue light for 1 hour, group H: infected treated with combination of AgNPs and Gramicidin 0.25µg/ml, 0.5 µg/ml respectively, group I : infected treated with triple combination of AgNPs ,Gramicidin 0.25µg/ml, 0.5µg/ml respectively and blue light for 1 hour. All groups treated topically daily for three days. Wound healing was assessed by measuring wound contraction at 7, 14 and 21 days post wound induction. Viable bacterial count at 21 day post wound induction. Results were according to data analysis, healing rate of treated rat with the triple combination was significantly (p < 0.001) faster (within 7 days) than treated rat with other double combination and each agent alone. All symptoms returned to near normal values at the end of treatment. In conclusion the topical therapy of the triple combination of AgNPs, visible blue light and antimicrobial peptides promote wound healing process in diabetic infected wound in a short period of time. A new approach can be used to combat serious infections caused by MRSA by combining AgNPs, blue light, and antibiotics.
Highlights
الخلاصة: أجريت هذه الدراسة لتقييم فعالية التركيبة الدوائية الثلاثية المكونة من دقائق الفضة النانوية المصنعة حيويا والببتيدات المضادة للجراثيم والضوء الأزرق المرئي ضد عزلة المكورات العنقودية الذهبية المقاومة للمثيسيلين الممرضة ) )methicillin resistance Staphylococcus aureus (MRSA)داخل الجسم الحي بعد إحداث جرح لجميع طبقات الجلد في الجرذان المصابة مسبقا بداء السكري. اجريت هذه التجربة داخل الجسم الحي لتقييم دور التركيبات في تعزيز التئام الجروح بعد احداث الجرح المصاب باستخدام ) )MRSAالممرضة ) 0.25مليلتر من )1.5 × 108 cfu/mlفي الجرذان المصابة بداء
Blue light is recently attracting increasing attention as a novel phototherapy-based antimicrobial agent that has significant antimicrobial activity against a broad range of bacterial and fungal pathogens with less chance to resistance development compared to antibiotics [12,13]
All wound surfaces post-wounding at the time of MRSA challenge with obvious signs of infection
Summary
The S. aureus (MRSA) was obtained from Al-Hilla Surgical Hospital, Iraq. Hinton broth and the suspension was centrifuged at 1000 rpm for 15 min. - Induction of infection After creation of the full thickness skin incision, 10 μl of the prepared bacterial isolate (MRSA) at concentration of 1.5×108 CFU/mL was spread along the created wound and let to interfere within the wound and left the wound opened. - Group (B): infected wound treated with glycerine topically daily with AgNPs for three days. - Group (C): infected wound treated with AgNPs alone topically at conc.8μg/ml daily and for three days. - Group (E): infected wound treated with blue light exposure alone for 1 hour daily and for three days - Group (F): infected wound treated with combination of AgNPs topically at conc. - Group (H): infected wound treated with combination of AgNPs and Gramicidin topically at conc. - Group (I): infected wound treated with triple combination of AgNPs and Gramicidin topically at conc. Statistical analysis was applied by two ways ANOVA with least significant differences (LSD) to compare groups means .Probability level P0.05 and to test the significant differences between the averages of the traits and considered statistically significant applying the SAS method [22]
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