Biotherapeutic agents and vaginal health

Treatment of vaginal infection requires different drugs although the recurrence rate post treatment remains high due to adverse effects on the beneficial microbiota. Thus, there are clear clinical advantages for the use of biotherapeutic agents (prebiotics and/or probiotics) for treating these infections. Pre‐ and probiotic beneficial effects can be delivered topically or systemically. In general, both approaches have the potential to optimize, maintain and restore the ecology of the vaginal ecosystem. Specific carbohydrates provide a therapeutic approach for controlling infections by stimulating the growth of the indigenous lactobacilli but inhibiting the growth and adhesion of pathogens to the vaginal epithelial cells. Overall, little evidence exists to promote the prevention or treatment of vaginal disease with prebiotic carbohydrates in formulations such as pessaries, creams or douches. However, recent reports have promoted prebiotic applications in ecosystems other than the gut and include the mouth, skin and vagina. This review focuses on the utilization of pre‐ and probiotics for vaginal health.     

Human vaginal epithelial multilayer tissue culture

Summary Fragments of normal human adult vagina, when explanted onto glass slides gave rise to outgrowing sheets of pure epithelium, which had microscopic morphological features in common with normal vaginal epithelium. Infrequent fibroblast contamination was observed. Proliferating epithelial cells formedmultilayers of stratified squamous epithelium and demonstrated a progressive decrease in proliferative activity after 14 days. Continuous lines of epithelial cells were not obtained. Even in the absence of estrogens, transmission electron microscopy revealed evidence of keratinization of the superficial cells of the multilayer. Scanning electron microscopy of the surface of mature epithelial cells in culture revealed ultrastructural features that closely resembled those present on the surface of exfoliated cells obtained by scraping the vagina in vivo. This in vitro tissue culture model of human vaginal epithelium may provide a simple method of studying factors that influence vaginal epithelium growth, maturation and function.     

Influence of probiotic vaginal lactobacilli on in vitro adhesion of urogenital pathogens to vaginal epithelial cells

AIMS: Lactobacilli, the predominant micro-organisms of the vaginal microbiota, play a major role in the maintenance of a healthy urogenital tract by preventing the colonization of pathogenic bacteria. The aim of the present study was to assess the ability of four vaginal Lactobacillus strains, previously selected for their probiotic features, to block in vitro the adherence of three human urogenital pathogens to vaginal epithelial cells (VEC). METHODS AND RESULTS: Three types of assays were performed in order to determine the inhibitory effect of lactobacilli on adhesion of urogenital pathogens to VEC: blockage by exclusion (lactobacilli and VEC followed by pathogens), competition (lactobacilli, VEC and pathogens together) and displacement (pathogens and VEC followed by the addition of lactobacilli). Bacterial adhesion to VEC was quantified by microscopy (x1000) after Gram's stain. All the strains were able to inhibit by exclusion and competition the adhesion of Staphylococcus aureus to VEC but none was able to decrease the attachment of Escherichia coli by neither of the mechanisms assayed. Only Lactobacillus acidophillus CRL 1259 and Lactobacillus paracasei CRL 1289 inhibited the attachment of Group B streptococci (GBS) to VEC by exclusion and competition respectively. CONCLUSIONS: Lactobacillus of vaginal origin were able to inhibit the attachment of genitouropathogenic Staph. aureus and GBS to the vaginal epithelium. SIGNIFICANCE AND IMPACT OF THE STUDY: The results support the probiotic potential of these Lactobacillus strains as anti-infective agents in the vagina and encourage further studies about their capacity to prevent and manage urogenital tract infections in females.     

Development and evaluation of acid-buffering bioadhesive vaginal tablet for mixed vaginal infections

An acid-buffering bioadhesive vaginal tablet was developed for the treatment of genitourinary tract infections. From the bioadhesion experiment and release studies it was found that polycarbophil and sodium carboxymethylcellulose is a good combination for an acid-buffering bioadhesive vaginal tablet. Sodium monocitrate was used as a buffering agent to provide acidic pH (4.4), which is an attribute of a healthy vagina. The effervescent mixture (citric acid and sodium bicarbonate) along with a superdisintegrant (Ac-Di-sol) was used to enhance the swellability of the bioadhesive tablet. The drugs clotrimazole (antifungal) and metronidazole (antiprotozoal as well as an antibacterial) were used in the formulation along with Lactobacillus acidophilus spores to treat mixed vaginal infections. From the ex vivo retention study it was found that the bioadhesive polymers hold the tablet for more than 24 hours inside the vaginal tube. The hardness of the acid-buffering bioadhesive vaginal tablet was optimized, at 4 to 5 kg hardness the swelling was found to be good and the cumulative release profile of the developed tablet was matched with a marketed conventional tablet (Infa-V). The in vitro spreadability of the swelled tablet was comparable to the marketed gel. In the in vitro antimicrobial study it was found that the acid-buffering bioadhesive tablet produces better antimicrobial action than marketed intravaginal drug delivery systems (Infa-V, Candid-V and Canesten 1).     

乳酸菌阴道胶囊对宫颈癌TP化疗患者炎症及阴道菌群的影响

目的 探讨乳酸菌阴道胶囊对中晚期宫颈癌TP化疗患者血清IL 2、IL 10、TGF β1水平及阴道菌群的影响。 方法 选取2018年10月-2019年8月于本院接受TP化疗的83例中晚期宫颈癌患者为研究对象,以随机数字表法分为两组,观察组43例,对照组40例。两组均接受TP化疗,观察组在对照组基础上接受乳酸菌阴道胶囊治疗。比较两组临床疗效,治疗前后血清IL 2、IL 10、TGF β1水平,阴道菌群分布情况,致病微生物阳性检出率和化疗相关不良反应发生情况。 结果 两组临床疗效等级分布差异显著(P0.05)。 结论 乳酸菌阴道胶囊可提高中晚期宫颈癌TP化疗患者临床疗效,调节患者免疫应答水平,有效改善患者阴道菌群平衡,降低患者阴道致病微生物阳性率,安全性高。     

High incidence of distal vaginal atresia in mice lacking Tyro3 RTK subfamily

Vaginal atresia is a congenital abnormality of the female genitourinary system, and the specific molecular mechanism leading to failure of vaginal development remains to be elucidated. Here, we report that the female mice lacking Tyro3 RTK subfamily (Tyro3, Axl, and Mer) exhibit a high incidence of distal vaginal atresia. The ratios of the vaginal atresia in Tyro3 RTKs mutant female mice are as follows: 2.5% for Mer(-/-) mice, 4.0% for Axl(-/-)Mer(-/-), 3.7% for Mer(-/-)Tyro3(-/-), 16.06% for Tyro(-/-)Axl(-/-)Mer(-/-) mice. We did not find the vaginal atresia in Axl(-/-), Tyro3(-/-), Axl(-/-) Tyro(-/-), and wild-type mice. These observations suggest that Tyro3 RTKs play roles collaboratively in vaginal development, and Mer is more critical, Axl and Tyro3 support the function of Mer. The phenotype of mice with the vaginal atresia was characterized in this study. Tyro3 RTKs mutant mouse could be a useful model to study the mechanism of vaginal atresia formation.     

乳酸菌阴道胶囊干预对孕晚期细菌性阴道病患者阴道菌群及分娩结局的影响

目的 探讨乳酸菌阴道胶囊干预对孕晚期细菌性阴道病（BV）患者阴道菌群及分娩结局的影响。方法 2017年12月‒2019年2月在泰山医学院附属青岛医院接受治疗及分娩的BV患者136例,参照随机数表法分为对照组、研究组各68例。对照组患者接受甲硝唑治疗,研究组患者接受乳酸菌阴道胶囊＋甲硝唑治疗。持续1周后,对比两组患者的疗效、乳酸菌计数情况、用药不良反应发生情况的差异,记录两组患者分娩结局。结果 治疗1周后,两组患者的疗效分级差异有统计学意义（U=7.062、P=0.029）;治疗总有效率的差异无统计学意义（2=2.030、P=0.154）。两组患者的阴道乳酸菌计数分级差异有统计学意义（U=9.179、P=0.047）,研究组患者的乳酸菌计数正常率高于对照组患者（2=6.103、P=0.013）。用药期间,两组患者皮肤瘙痒、下腹疼痛、头晕头痛等不良反应发生率差异无统计学意义（2=0.341、1.007、0.208,P=0.559、0.316、0.649）。两组患者均无一例发生心律失常。研究组患者的胎膜早破、早产、产褥感染、子宫内膜炎发生率低于对照组患者（2=4.087、4.561、3.765、4.121,P=0.043、0.033、0.049、0.042）。两组患者的新生儿感染、新生儿高胆红素血症发生率差异无统计学意义（2=2.030、2.944,P=0.154、0.086）。结论 乳酸菌阴道胶囊干预对优化孕晚期BV患者阴道菌群、改善妊娠结局具有一定积极作用。     

非脱垂子宫阴式全切除136例的临床分析

目的探讨基层医院开展阴式子宫切除术（TVH）的可能性、必要性及其价值。方法对我院妇科2007年8月至2009年1月行阴式、腹式全子宫切除的193例进行研究,比较阴式与腹式两种术式的手术时间、术中出血量、术后胃肠功能恢复时间。结果两种术式的手术时间、术中出血量、术后胃肠功能恢复时间,均有明显差异（P〈0．01）．结论TVH组手术时间短,出血量少,腹部皮肤不留疤痕,术后康复快,住院时间短,医疗费用少,患者满意．     

The probiotic properties and potential of vaginal Lactobacillus spp. isolated from healthy women against some vaginal pathogens

During the last decade, probiotic research has progressed considerably and significant advances have been made in the selection and characterization of specific probiotic strains. The most studied probiotics belong to the genus Lactobacillus. In this study, 80 Lactobacillus spp. isolated from healthy women tolerated low pH and were able to grow in the presence of bile salts. RAPD PCR technique resulted in the identification of 38 different types. These isolates were then evaluated based on adhesion capacity, antibiotic susceptibility and tolerance in simulated gastrointestinal tract. Species-specific PCR and detection of bacteriocin-related genes were also surveyed. Among the isolates, five strains—Lacticaseibacillus rhamnosus NO21, Lacticaseibacillus casei NO1, Lactiplantibacillus plantarum NO4, Lactobacillus acidophilus NO7 and Lactobacillus gasseri NO38—presented acceptable antibiotic susceptibility pattern. Further analysis showed antimicrobial activity of Lacticaseibacillus culture against various bacterial pathogens and real-time PCR showed all five strains were able to prevent the colonization of bacterial pathogens. All five selected strains produced organic acids, hydrogen peroxide and were resistant to the spermicide. In addition, they lacked haemolytic activity with the ability of hydrophobicity, auto-aggregation and co-aggregation with pathogens. These results suggest that the vaginal microbiome could be a good source for the isolation of probiotics and the strains of this study may be considered as good probiotic candidates.     

产复康冲剂治疗药物流产后阴道出血的临床观察

目的：观察产复康冲剂在药物流产后服用,阴道出血时间及出血量的变化,方法与结果：实验组（产复康组）及对照组各120名,结果两组流产率无差异（P＞0．05）。完全流产组二者周内出血停止者实验组多于对照组P＜0．01。三周出血不止者实验组少于对照组,P＜0．01。尽管大于月经量出血的平均天数实验组略长,但点滴出血时间较对照组短P＜0．01。结论：产复康冲剂补气养血,排淤生新,加快恶露排净达到尽快止血目的,是治疗药物流后异常阴道出血的方法之一。     

乳酸菌阴道胶囊辅助治疗对未足月胎膜早破的疗效及对其阴道微生态的影响

目的 观察乳酸菌阴道胶囊辅助治疗对未足月胎膜早破的治疗效果,并探讨其对阴道微生态的影响。 方法 86例未足月胎膜早破患者采用随机数表分为常规组与研究组,各43例。常规组予以常规治疗,研究组予以乳酸菌阴道胶囊联合常规治疗。比较治疗后两组阴道微生态、胎儿宫内感染率和分娩方式、母体和胎儿不良妊娠结局发生情况。 结果 治疗后研究组Chao1指数和Shannon指数均高于常规组(均P结论 乳酸菌阴道胶囊辅助治疗未足月胎膜早破相较于常规治疗可改善阴道微生态,还可降低胎儿宫内感染率、剖宫产率,减少母体和围产儿不良妊娠结局的发生。     

Keratinization of rat vaginal epithelium. III. Effect of estradiol on keratinization

Effect of estradiol-17 beta on rat vaginal epithelial cells (VEC) was studied by transmission electron microscopy. During the normal estrous cycle in adult rats keratinization and exfoliation of VEC was observed only in the estrus phase, though tonofilament bundles were seen distributed in the cytoplasm of the VEC during the proestrus and, to a lesser extent, in the diestrus phase. Electron microscope data on adult ovariectomised, immature and neonatally estradiol-primed rats demonstrated keratinization of the VEC within 24 h of injection. Proliferative activity was observed in the basal cells in primed animals. The basal cells remained cuboidal whereas cells of intermediate and luminal layers became flattened. These cells showed irregular profiles and, therefore, enhanced numbers of cellular junctions (desmosomes) were observed compared to control groups. Many tonofilament bundles were seen in the cytoplasm of cells belonging to intermediate and luminal layers in primed animals.     

The in vitro effect of hydrogen peroxide on vaginal microbial communities

This study presents a series of experiments carried out in order to elucidate the role of H2O2 in antimicrobial activity of lactobacilli. Vaginal swabs were collected from 60 premenopausal women and checked for pH and Nugent score, and Lactobacillus species were cultured, phenotyped and genotyped. The main outcome measures involved: (1) species of vaginal lactobacilli most effective in liberating H2O2, (2) minimal microbicidal concentrations of added H2O2, (3) kinetics of H2O2 liberation in relation to oxygen tension, (4) antimicrobial activity of pure H2O2 versus one produced by selected vaginal lactobacilli and the total activity of their culture supernatants. Results showed that H2O2 was liberated especially by: Lactobacillus delbrueckii, Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus johnsonii and L. gasseri. Hydrogen peroxide reached concentrations from 0.05 to 1.0 mM, which under intensive aeration increased even up to 1.8 mM. Microorganisms related to vaginal pathologies show varied resistance to the action of pure H2O2. Most potent inhibitory activity against bacteria and yeasts was presented by Lactobacillus culture supernate producing H2O2, followed by the nonproducing strain and pure H2O2. To conclude - the antimicrobial activity of lactobacilli is a summation of various inhibitory mechanisms in which H2O2 plays some but not a crucial role, in addition to other substances.     

Correlation between vaginal microbiota and different progression stages of cervical cancer

The process from high-risk human papillomavirus (HR-HPV) infection to cervical cancer is a continuous and long-term process, but the pathogenesis of the whole process is not completely clear. Here, 59 Chinese women were engaged in this study, and divided into five groups: normal healthy group, HR-HPV infections group, low-grade intraepithelial neoplasia (LSIL) group, high-SIL(HSIL) group, and cervical cancer group. With the occurrence of HR-HPV infection and the development of cervical lesions, the diversity of vaginal microbiota species was increased, and the relative abundance of Lactobacillus (L.), the dominant bacteria in maintaining vaginal microecological balance, was decreased gradually. In contrast, the abundance of Actinobacteria in the four disease groups was significantly higher than that in normal group. Furthermore L. iners may be related to the serious progression of cervical cancer. After analyzing the whole process, we found that Gardnerella(G.), Atopobium(A.) and Dialister(D.) have important effects on both persistent HR-HPV infection and the pathogenesis of cervical cancer. In addition, PICRUSt2 and KEGG results showed that the KEGG pathways enriched by the predicted genes of vaginal microbiota in cancer group included metabolic diseases, endocrine system and immune systems when compared with that in normal group. These findings may provide insights into the pathogenesis of cervical cancer, and help to improve the early detection and prevention of cervical precancerous lesions.     

Effects of culture conditions on the growth and auto-aggregation ability of vaginal Lactobacillus johnsonii CRL 1294

AIMS: To evaluate the effects of different physico-chemical factors on the growth and auto-aggregating ability of vaginal Lactobacillus johnsonii CRL 1294. METHODS AND RESULTS: L. johnsonii CRL 1294 was cultivated in different culture media, initial pH and temperature of incubation. The growth parameters were estimated by the Gompertz model, being optimal (higher final biomass and growth rate, and shorter lag phase) at an initial pH of 6.5 and at a temperature of 37 degrees C, both in LAPTg and MRS. The auto-aggregation ability, which was assessed by a model of exponential association, was evidenced in all the growth phases, being higher at pH 5 or 6.5. CONCLUSIONS: The growth of L. johnsonii CRL 1294 was affected in different way by all the physico-chemical factors tested. However, the auto-aggregation ability increased mainly at low initial pH of growth media. SIGNIFICANCE AND IMPACT OF THE STUDY: The auto-aggregation ability under different culture conditions of a vaginal Lactobacillus strain was systematically and statistically evaluated for the first time. The higher cellular aggregation evidenced at low pH could be a fundamental characteristic in the acidic vaginal environment to promote the protective role of lactobacilli.     

Isolation and identification of fungi from vaginal flora in three species of captive Leontopithecus

The ability to reproduce in captivity is an essential component of lion tamarin (Leontopithecus) conservation programs. However, infections such as vaginitis, cervicitis, and endometritis are important diseases that may influence the reproduction of these animals. Therefore, it is important to detect continuous or occasional vaginal microbial populations, and to understand their potential role as an endogenous source of infection [Collins, 1964; Blue, 1983; Pugh et al., 1986]. Vaginal swabs were collected from 25 female tamarins of the three currently available species (L. rosalia, L. chrysopygus, and L. chrysomelas) at the Center of Primatology in Rio de Janeiro, Brazil. The swabs were processed according to standard mycological protocols, and isolates were biochemically characterized. Fungal isolates were recovered from 16 animals (64.0%). The results showed that 70.6% of the isolated microorganisms consisted of yeast, including three species of Candida (mainly C. glabrata). We suggest that this species is a resident member of the normal vaginal flora in Leontopithecus. Filamentous fungi (mainly from Trichosporon, Aspergillus, and Penicilliumgenera) constituted 29.4% of the isolates, and were considered to be transitory contaminants of the genital area. We suggest that colonization of the vaginal environment is related to the endocrine pattern associated with the reproductive status of these animals, but not to parity.     

Radiation treatment for refractory endometriosis: a 38-year-old female presenting with vaginal bleeding

BackgroundEndometriosis is typically managed with a medical or surgical approach, though some patients have medically refractory disease and are poor surgical candidates.Case presentationA 39-year-old woman presented to our facility with uncontrolled bleeding and pain from an endometriotic mass at the vaginal cuff. She had a history of abdominal hysterectomy with bilateral salpingo-oophorectomy, now with medically refractive and inoperable disease due to prior history of vesicovaginal fistula. We prescribed 30 Gy in 10 fractions with 10 MV and 18 MV photons to the target. At follow-up our patient reported a complete resolution of bleeding and pelvic pain.ConclusionRadiation treatment can be an effective treatment for refractory endometriosis.     

Benign glandular cells in posthysterectomy vaginal smears: the incidence is higher than expected

In this study, we have investigated the frequency and clinical significance of glandular cells in posthysterectomy vaginal smears. The slides of vaginal cuff smears of 290 patients were reviewed. The glandular cells were categorized into three groups: (1) squamous metaplastic-like cells; (2) columnar endocervical-like cells; and (3) small round cuboidal cells. Glandular cells were found in 39 (13%) of the 290 vaginal smears. Group 1 type cells were seen in 76% (n = 30), group 2 type cells in 38% (n = 15) and group 3 type cells in 47% (n = 19) of the smears. In 19 (48%) of the smears combination of two or three groups were seen. The presence of glandular cells showed a strong association with inflammation/repair as a background finding in the smears. No correlation could be found between the presence of atrophy and history of chemotherapy or radiotherapy. Apart from these there was a prominent increase in reporting benign glandular cells after the application of the current Bethesda 2001 reporting criteria in our laboratory. As a result our study showed that the finding of glandular cells in posthysterectomy vaginal smears is more frequent than expected and most of them could be related to inflammatory and regenerative processes in the absence of a clearly identified underlying cause.     

Evaluation of characteristics of Pediococcus spp. to be used as a vaginal probiotic

Aims

The aim of our research was to select, identify and characterize an isolate of lactic acid bacteria to be considered as a vaginal probiotic.

Methods and Results

Thirty‐five isolates of Pediococcus spp. showed bacteriocinogenic activity against Listeria monocytogenes and the ability to survive in simulated vaginal fluid (SVF) at pH 4·2. One isolate of Pediococcus spp. was selected and characterized to evaluate its safety before the use as vaginal probiotic. Pediococcus pentosaceus SB83 did not show the presence of virulence factors such as the production of gelatinase, lipase and DNase, haemolytic activity, nor the presence of virulence genes (genes esp, agg, gelE, efaAfm, efaAfs, cylA, cylB and cylM). Pediococcus pentosaceus SB83 was considered sensitive to chloramphenicol, gentamicin, streptomycin, kanamycin, erythromycin and ampicillin. This strain was considered resistant to tetracycline and vancomycin. Pediococcus pentosaceus SB83 was a biofilm producer at different pH values (4·2, 5·5 and 6·5) in SVF and in de Man, Rogosa and Sharpe medium.

Conclusions

The in vitro results provide a basis for the use of P. pentosaceus SB83 as a vaginal probiotic, to prevent colonization by L. monocytogenes in pregnant women.

Significance and Impact of Study

The application of vaginal probiotics could have the potential for preventing vaginal infections and consequently reduce abortion and neonatal infections.     

Novel vaginal drug delivery system: deformable propylene glycol liposomes-in-hydrogel

Deformable propylene glycol-containing liposomes (DPGLs) incorporating metronidazole or clotrimazole were prepared and evaluated as an efficient drug delivery system to improve the treatment of vaginal microbial infections. The liposome formulations were optimized based on sufficient trapping efficiencies for both drugs and membrane elasticity as a prerequisite for successful permeability and therapy. An appropriate viscosity for vaginal administration was achieved by incorporating the liposomes into Carbopol hydrogel. DPGLs were able to penetrate through the hydrogel network more rapidly than conventional liposomes. In vitro studies of drug release from the liposomal hydrogel under conditions simulating human treatment confirmed sustained and diffusion-based drug release. Characterization of the rheological and textural properties of the DPGL-containing liposomal hydrogels demonstrated that the incorporation of DPGLs alone had no significant influence on mechanical properties of hydrogels compared to controls. These results support the great potential of DPGL-in-hydrogel as an efficient delivery system for the controlled and sustained release of antimicrobial drugs in the vagina.