Disinfection of Human Amniotic Membrane Using a Hydrodynamic System with Ozonated Water

Human amniotic membrane (hAM) has a great potential in regenerative medicine as it can be disinfected, preserved, and stored. The present study evaluated a hydrodynamic system that uses ozonated water for disinfecting hAM and assessed possible morphological alterations using histological analysis and scanning electron microscopy (SEM). hAM fragments were experimentally contaminated with Staphylococcus aureus (10(3) CFU/mL) and submitted to disinfection processes for 5, 10 and 15 minutes (1.95, 3.9 and 5.85 mg/cm(2) (O-3), respectively). The results of the water microbiological analysis showed that no microbial growth was detected in all water samples. The hAM surface monitoring with swab method and the analysis of fragment inoculated directly into the culture media revealed that no microbial growth was detected after 10 and 15 minutes. The SEM and histological analysis of hAM revealed that morphological alterations in the epithelium began after 10 minutes 3.9 mg/cm(2) (O-3) of the disinfection process, and significant changes in the epithelial cells were observed after 15 minutes (5.85 mg/cm(2) (O-3)). These findings indicates that the structural integrity of hAM was maintained during the process and suggest that the technique is a promising system to disinfect this biological material.