Doxycycline-Coated Silicone Breast Implants are Non-Toxic and Reduce Biofilm Formation In Vitro
Mitchell J. Skinner, B.S.1, Jennifer E. Baker, MD1, Ryan M. Boudreau, MD1, Aaron P. Seitz, MD1, Michael J. Edwards, MD1, Erich Gulbins, MD, PhD1,2, Ryan M. Gobble, MD1.
1University of Cincinnati, College of Medicine, Department of Surgery, Cincinnati, OH, USA, 2University of Duisburg-Essen, University Hospital Essen, Department of Molecular Biology, Duisburg, Germany.
Infection and capsular contracture are two common complications after silicone breast implantation. It is estimated that approximately 0.5-3% of augmentations and reconstructions will develop an infectious complication, and 20-60% of implant pockets are positive for bacteria at time of revision surgery. Staphylococcus aureus and Pseudomonas aeruginosa are the predominant species found in breast implant infections. We have developed a novel method of coating silicone implants previously shown to prevent implant infection in vitro. We hypothesize that this coated silicone will be non-toxic to fibroblasts and will prevent biofilm formation in vitro
Silicone Coating: Pieces of silicone breast implants were coated with doxycycline or ethanol (vehicle control) using our novel method resulting in an evaporative induced molecular crystal film on the surface.
Neutral Red Uptake: L929 mouse fibroblasts were seeded into a 96-well plate and incubated at 37C for 24 hours. Prior media was removed and media that had been incubated with doxycycline-coated and ethanol-coated silicone breast implants for 24 hours was added and incubated for 24 hours. Prior media was removed, and neutral red medium was added and incubated for 3 hours. Wells were washed, and neutral red desorb was added and shaken for 10 minutes. Absorbance was obtained and analyzed.
Fibroblast Toxicity: Doxycycline-coated and ethanol-coated implants were placed into a 24-well plate and seeded with L929 mouse fibroblast cells. The plate was incubated at 37C for 24 hours. Implants were removed and fixed with 2.5% gluteraldehyde for 2 hours, washed with PBS, stained 1% osmium tetroxide for 2 hours, washed with PBS, and allowed to air dry. Samples were sputter coated with silver for 5 seconds and viewed under scanning electron microscopy (SEM). Biofilm Evaluation: Doxycycline-coated and ethanol-coated implants were placed into a 24-well plate and seeded with 1000 cfu of methicillin-resistant S. aureus (MRSA) or P. aeruginosa and incubated for 24 hours at 37C. Implants were processed the same as Fibroblast toxicity and viewed under SEM.
Doxycycline-coated silicone implants demonstrated 92% viability of fibroblast based on a neutral red uptake assay demonstrating nontoxicity. SEM of doxycycline-coated implants after incubation with fibroblasts demonstrated similar viability and adhesion as an ethanol-coated control (Figure 1). Doxycycline-coated silicone implants prevented biofilm formation of MRSA (Figure 2) and P. aeruginosa (Figure 3) as compared to ethanol-coated control.
Surgical site infections are a dreaded complication often leading to biofilm formation and the development of capsular contracture. We have now demonstrated that doxycycline-coated silicone implants are non-toxic and prevent biofilm formation in vitro. Doxycycline-coated implants may be useful to prevent both bacterial infections and capsular contracture.
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