Abstract
Urinary tract infections are among the most common bacterial infections in humans. Moreover, they are highly recurrent and increasingly often resistant to antibiotics. The antimicrobial properties of the amniotic membrane (AM), the innermost layer of fetal membranes, have been briefly reported in the literature, however, the results of published studies are often inconsistent and unclear; moreover, its effect on uropathogenic bacteria has not yet been investigated. Further, there is no data in the literature about the effect of AM preparation and storage on its antimicrobial properties. To examine the impact of several preparation procedures on the antimicrobial properties of AM, we prepared patches and homogenates of fresh (fAM) and cryopreserved (cAM) human AM and tested them on 14 selected Gram-positive and Gram-negative uropathogenic bacteria. By employing novel antimicrobial efficiency assays we showed that fAM and cAM homogenates have broad-spectrum antimicrobial activity against all here tested uropathogenic bacteria, except for Serratia marcescens. Moreover, they had a potent effect also on the multiple-resistant clinical strains of uropathogenic Escherichia coli. Interestingly, the patches of fAM and cAM had no antimicrobial effect on any of the tested strains. We therefore prepared and stored AM patches according to the standard procedure for clinical use in ophthalmology, which includes the cryopreservation of antibiotic-treated AM, and performed antimicrobial efficiency assays. Our findings suggest that the ultrastructure of AM patches could enable the retention of added antibiotics. In addition, we also prepared gentamicin-resistant uropathogenic E. coli strains, which confirmed that the antimicrobial effect of antibiotic-treated AM patches can be attributed to the antibiotic alone. To summarize, here we describe novel protocols for preparation and storage of AM to ensure the preservation of its antimicrobial factors. Moreover, we describe the mechanism of AM retention of antibiotics, based on which the AM could potentially be used as a drug delivery vehicle in future clinically applicable approaches.
Highlights
Amniotic membrane, the innermost layer of fetal membranes, surrounds the developing fetus and forms the amniotic cavity
All plates in all assays were overgrown with bacteria, showing that fresh amniotic membrane (fAM) and cryopreserved AM (cAM) patches have no inhibitory effect on the growth of any here tested strain (Figures 1, 2A–H,J,L)
Our study demonstrated for the first time that amniotic membrane (AM) has a powerful broadspectrum antimicrobial effect on Gram-positive and Gramnegative bacteria
Summary
The innermost layer of fetal membranes, surrounds the developing fetus and forms the amniotic cavity. The AM contributes to prevention of intrauterine infection, which is critical for proper fetal development (King et al, 2007; PrabhuDas et al, 2015). AM has already been recognized as an antimicrobial agent in dermatology for treating chronic wounds (Mohammadi et al, 2013; ElHeneidy et al, 2016; Sant’Anna et al, 2016). There are two completed clinical trials and 1 clinical trial in running, registered on the NIH Clinical Trials website, which are studying the antimicrobial potential of AM or AM-derived cells in dental medicine (chronic periodontitis) and dermatology (burns, complex wounds). None of the clinical studies are testing the antimicrobial effect of the AM in the field of urology (ClinicalTrials.gov)
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