tissue-processing-isolation-primary-fibroblasts-from-human

Tissue Processing and Isolation of Primary Fibroblasts from the Human Vagina

Pelvic organ prolapse is a disorder that seriously impacts the quality of life of women. It occurs when muscles and ligaments weaken and cause pelvic ...

Massachusetts General Hospital, Vincent Center for Reproductive Biology

The objective of the study was to evaluate the upper genital tract (UGT) presence of vaginal bacterial species using sensitive molecular methods capable of detecting fastidious bacterial vaginosis (BV)-associated bacteria.

Colonization of the upper genital tract by vaginal bacterial species in nonpregnant women.

Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN) is a systemic disease that can be associated with debilitating acute and chronic complications across multiple organ systems. As patients with acute SJS/TEN are often treated in a burn intensive care unit (BICU), we surveyed burn centers across the United States to determine their approach to the care of these patients. The goal of our study was to identify best practices and possible variations in the care of patients with acute SJS/TEN. We demonstrate that the method of diagnosis, use of systemic therapies, and involvement of subspecialists varied significantly between burn centers. Beyond supportive care provided to every patient, our data highlights a lack of standardization in the acute care of patients with SJS/TEN. A comprehensive guideline for the care of patients with acute SJS/TEN is indicated.

Burn unit care of Stevens Johnson syndrome/toxic epidermal necrolysis: A survey.

Cervicovaginal microbiota play a critical role in women's health and reproductive outcomes. Despite being one of the simplest commensal bacterial communities in the human body, we are only beginning to appreciate its complex dynamic nature and important role in host immune modulation. In this review, we discuss the "optimal" cervicovaginal bacterial community composition, the impact of microbiota on gynecologic and obstetric outcomes, and the hurdles to developing a deeper mechanistic understanding of the function of the cervicovaginal microbiome. We then describe efforts to durably alter microbial composition in this compartment by promotion of Lactobacillus colonization with probiotics, modulation of vaginal pH, hormonal administration, and the eradication of pathogenic bacteria with antibiotics. Finally, we draw on lessons learned from the deeply investigated gut microbiome to suggest future avenues of research into host-pathogen interactions in the female genital tract.

Cervicovaginal Microbiota and Reproductive Health: The Virtue of Simplicity.

Inflammatory bowel diseases (IBD) affect 200-400 people per 100,000 in the United States, about half of whom are women. We aimed to define the prevalence of vulvovaginal symptoms and association with IBD activity in a large cohort of women.

Association Between Vulvovaginal Discomfort and Activity of Inflammatory Bowel Diseases.

Half of postmenopausal women experience genitourinary syndrome of menopause, for which many use lubricating vaginal products. The effect of vaginal products on uropathogenic and commensal vaginal bacteria is poorly understood. We evaluated the effect of five common vaginal products (KY Jelly, Replens Silky Smooth lubricant, coconut oil, Replens Long-Lasting moisturizer or Trimo-San) on growth and viability of Escherichia coli and Lactobacillus crispatus. Bacteria were co-cultured products alone and in the presence of both vaginal epithelial cells and selected products. Bacterial growth was compared between conditions using an unpaired t-test or ANOVA, as appropriate. All products except for coconut oil significantly inhibited growth of laboratory and clinical strains of Escherichia coli (p < 0.02). Only two products (Replens Long-Lasting moisturizer and Trimo-San) significantly inhibited growth of Lactobacillus crispatus (p < 0.01), while the product Replens Silky Smooth stimulated growth (p < 0.01). Co-culture of selected products in the presence of vaginal epithelial cells eliminated the inhibitory effects of the products on E. coli. In conclusion, in vitro exposure to vaginal moisturizing and lubricating products inhibited growth of Escherichia coli, though the inhibition was mitigated by the presence of vaginal epithelial cells. Lactobacillus crispatus demonstrated less growth inhibition than Escherichia coli.

Effect of commercial vaginal products on the growth of uropathogenic and commensal vaginal bacteria.

Urinary tract infections (UTIs) are one of the most common infections in women. The only proven preventive strategy for recurrent UTIs is prophylactic antibiotics. Given growing antibiotic resistance, the use of probiotics has been proposed as an alternative to antibiotics. Herein, we discuss the current evidence to support the possibility that exogenous lactobacilli may limit the pathogenicity of uropathogens such as E. coli. Probiotics appear to have a significant potential in prevention of recurrent UTI, however, additional data are needed to understand how they can be effectively used in clinical practice.

Understanding the potential of lactobacilli in recurrent UTI prevention.

Half of all postmenopausal women report symptoms of vulvar, vaginal, or urinary discomfort with substantial impact on sexual function and quality of life; underlying mechanisms leading to symptoms are poorly understood.

Association between postmenopausal vulvovaginal discomfort, vaginal microbiota, and mucosal inflammation.

Consequences of long-term B cell depletion with rituximab are not well understood. We describe inflammatory vaginitis as a potential side effect of long-term rituximab treatment, distinct from previously described vulvovaginal pyoderma gangrenosum.

Inflammatory vaginitis in women on long-term rituximab treatment for autoimmune disorders.

We compared the effect of commercial vaginal douching products on Lactobacillus crispatus, L. jensenii, L. gasseri, L. iners, E. coli, and immortalized vaginal epithelial cells (VK2). All studied douching products (vinegar, iodine and baking soda based) induced epithelial cell death, and all inhibited growth of E. coli. Co-culture of vaginal epithelial cells with any of the lactobacilli immediately following exposure to douching products resulted in a trend to less human cell death. However, co-culture of epithelial cells with L. iners was associated with higher production of IL6 and IL8, and lower IL1RA regardless of presence or type of douching solution. Co-culture with L. crispatus or L. jensenii decreased IL6 production in the absence of douches, but increased IL6 production after exposure to vinegar. Douching products may be associated with epithelial disruption and inflammation, and may reduce the anti-inflammatory effects of beneficial lactobacilli.

Impact of vaginal douching products on vaginal Lactobacillus, Escherichia coli and epithelial immune responses.

Bacterial vaginosis (BV), the overgrowth of diverse anaerobic bacteria in the vagina, is the most common cause of vaginal symptoms worldwide. BV frequently recurs after antibiotic therapy, and the best probiotic treatments only result in transient changes from BV-associated states to "optimal" communities dominated by a single species of Lactobacillus. Therefore, additional treatment strategies are needed to durably alter vaginal microbiota composition for patients with BV. Vaginal microbiota transplantation (VMT), the transfer of vaginal fluid from a healthy person with an optimal vaginal microbiota to a recipient with BV, has been proposed as one such alternative. However, VMT carries potential risks, necessitating strict safety precautions. Here, we present an FDA-approved donor screening protocol and detailed methodology for donation collection, storage, screening, and analysis of VMT material. We find that Lactobacillus viability is maintained for over six months in donated material stored at - 80 °C without glycerol or other cryoprotectants. We further show that species-specific quantitative PCR for L. crispatus and L. iners can be used as a rapid initial screening strategy to identify potential donors with optimal vaginal microbiomes. Together, this work lays the foundation for designing safe, reproducible trials of VMT as a treatment for BV.

Caroline Mitchell

Department of Obstetrics,
Gynecology and Reproductive Biology,
Department of Obstetrics, Gynecology and Reproductive Biology

Massachusetts General Hospital, Vincent Center for Reproductive Biology

Tissue Processing and Isolation of Primary Fibroblasts from the Human Vagina

Caroline Mitchell

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Obstetrics,Gynecology and Reproductive Biology,Department of Obstetrics, Gynecology and Reproductive Biology

Massachusetts General Hospital, Vincent Center for Reproductive Biology

Tissue Processing and Isolation of Primary Fibroblasts from the Human Vagina

Department of Obstetrics,
Gynecology and Reproductive Biology,
Department of Obstetrics, Gynecology and Reproductive Biology