The purpose of this study was to determine the expression of natural antimicrobials in primary cultured amnion epithelial cells and to examine their regulation by interleukin-1 beta (IL-1beta).
Primary amnion epithelial cells were cultured from samples that were obtained at prelabor cesarean section (n = 12) and stimulated with IL-1beta. Natural antimicrobial messenger RNA expression was determined by real-time quantitative polymerase chain reaction, and protein was measured by enzyme-linked immunosorbent assay. Data was analyzed by analysis of variance.
Primary amnion epithelial cells express messenger RNA for human beta defensin (HBD) 1 to 3, secretory leukocyte protease inhibitor and elafin, but not HBD4. IL-1beta 10 ng/mL stimulates HBD2 messenger RNA in a biphasic pattern, with a 51-fold increase at 6 hours and a 67-fold at 12 hours (P < .001). HBD2 protein production is significantly increased by 24 hours (P < .05).
The amnion produces potent natural antimicrobials that may help protect the pregnancy from infection. HBD2 production is dramatically upregulated by the labor-associated inflammatory cytokine IL-1beta.
"The results of Hsp-60 and Hsp-70 secretion by CHD and AM compartments is consistent with other antimicrobial human bdefensins involved in the amniotic infection response. It has been reported that human amniotic epithelium cells stimulated with IL- 1b, E. coli LPS, and prostaglandin induced an increase in the secretion of b-defensin type 1 and 2 after 12 h of incubation  . Recently, evidence has shown an increase by 1.5-fold of Hsp-70 secretion in patients with intra-amniotic infection . "
"In general, there are some natural antimicrobials produced in amniotic fluid during pregnancy and the epithelial layer consists of human beta-defensins 1-3 (HBD), elafin and secretory leukocyte protease inhibitor (SLPI), among that HBD-2 is a strong antibiotic and is expressed in response to IL-1 in amniotic epithelial cells (Stock et al., 2007; King et al., 2007). "
[Show abstract][Hide abstract] ABSTRACT: The present study investigated the antimicrobial activity of human amniotic and chorionic membranes against some common bacterial and fungal pathogens. The findings clearly demonstrated the antimicrobial effect of both the amniotic and chorionic membranes against the tested bacterial and fungal pathogens at different dilutions by their maximum and minimum inhibitory zones. The maximum inhibition zone was measured in amniotic membrane compared to chorionic membrane in both the bacterial and fungal activity plates. While assessing the four different dilutions (5X10 5 , 5X10 6 , 5X10 7 and 5X10 8), the similar diameter of inhibition zone was observed in 1.5X10 5 and 1.5X10 6 dilutions. The study clearly confirmed the antimicrobial activity effect of both amniotic and chorionic membranes against several bacterial and fungal pathogens in which maximum activity was recorded by amniotic membrane.
"To date, hAM has clinical indication in ophthalmology and is being tested in clinical trials in dermatology and soft tissue surgery. Many beneficial properties of hAM, including anti-bacterial, anti-viral, anti-inflammatory, analgesic, anti-fibrotic, anatomical and vapor barrier properties have been reported and it reportedly facilitates wound healing, re-epithelialization and reduction of scarring (Kjaergaard et al. 1999; Li et al. 2005; Stock et al. 2007; Parolini et al. 2008; Tao and Fan 2009; Insausti et al. 2010). These pleiotropic functions are related in part to its capacity to synthesize and release biologically active substances, including cytokines and signaling molecules (Parolini et al. 2008; Insausti et al. 2010). "
[Show abstract][Hide abstract] ABSTRACT: In the past 20 years, human amniotic membrane (hAM) has become widely recommended as an ophthalmic surgical patch, and as a substrate for stem cell tissue equivalents for ocular surface reconstruction. HAM reduces ocular surface scarring and inflammation, and enhances epithelialization. In addition, it shows limited immunogenicity and some anti-microbial properties. Thanks to these properties, hAM has been also used in wound healing, especially for burns and ulcers. Since its first clinical applications, hAM has been used for other indications, such as oral and maxillofacial, earnose- throat, gynaecological and orthopaedic surgeries. This review will describe: (i) past and current clinical uses of hAM; (ii) accepted processing methods, including preparation, preservation, sterilization and de-epithelialization and their impact on the properties of hAM, especially growth factor release, cell viability and immunological properties; (iii) its applications in tissue engineering, namely as scaffold or carrier of biological molecules. Economical aspects are presented at the end of this review. Existing patents are reported for each section and existing marketed products are listed. Patents, products and ongoing clinical trials, were identified by electronic searches on the internet in January 2013. In conclusion, in view of published data, hAM seems to have real market potential in regenerative medicine, in particular in emerging fields such as oral and maxillofacial, ear-nose-throat, gynaecological and orthopaedic surgery.
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