The effect of orally administered probiotic Lactobacillus reuteri-containing tablets in peri-implant mucositis: a double-blind randomized controlled trial.

BACKGROUND AND OBJECTIVES: Probiotics create a biofilm and protect the oral tissues against the action of periodontal pathogenic bacteria. The aim of this study was to evaluate the effects of the oral probiotic Lactobacillus reuteri Prodentis upon the peri-implant health of edentulous patients with dental implants and peri-implant mucositis, establishing comparisons vs implants without peri-implant disease. MATERIAL AND METHODS: A double-blind, placebo-controlled, prospective cross-over study was made. The patients were all edentulous and were divided into two groups, (A) no peri-implant disease, and (B) peri-implant mucositis affecting one or more implants. Patients with peri-implantitis were excluded. The dosage was one tablet every 24 h over 30 d. All patients in both groups initially received the oral probiotic Lactobacillus reuteri Prodentis, followed by placebo. Patients started with probiotic treatment during 30 d, followed by a 6 mo washout period and the administration of placebo for the same period. The following parameters were studied: crevicular fluid volume, modified plaque index, probing depth, modified gingival index, and concentrations of interleukin 1ß, interleukin 6 and interleukin 8. RESULTS: A total of 77 implants were evaluated in 34 patients. Group A involved 22 patients with 54 implants without peri-implant alterations, and group B, 12 patients with mucositis affecting one or more implants (23 implants). After treatment with the probiotic, both the patients with mucositis and the patients without peri-implant disease showed improvements in the clinical parameters, with reductions in cytokine levels. In contrast, no such changes were observed with placebo. CONCLUSIONS: After treatment with the probiotic Lactobacillus reuteri in patients with implants presenting mucositis, the clinical parameters improved, and the cytokine levels decreased - in contraposition to the observations in the placebo group. Probiotic administration may be regarded as a good alternative for both the treatment of peri-implant mucositis and its prevention, as it also improved clinical parameters in the healthy individuals. Further studies involving larger patient series are needed regarding the effects of probiotics upon peri-implant health.

Anti-inflammatory properties of N-acetylcysteine on lipopolysaccharide-activated macrophages.

INTRODUCTION: Innate immune cells play a role in modulating host immune response. Part of the macrophage inflammatory response is the release of an array of inflammatory cytokines, important molecules during the development of innate and adaptative immunity. Several antioxidant agents have been used in the control of the inflammatory response. OBJECTIVE: To evaluate the anti-inflammatory effect of N-acetylcysteine (NAC) on the expression and secretion of inflammatory cytokines and interleukin (IL)-10 in lipopolysaccharide (LPS)-activated THP-1 macrophages under mild oxidative conditions. METHODS: Macrophages were activated by LPS (0.1 and 1 µg/ml) for up to 24 h. The effect of 15 mM NAC was evaluated at 2, 4, 6 and 24 h. mRNA expression of tumor necrosis factor (TNF)-α, IL-1ß, IL-6, IL-8 and IL-10 was assessed by real time PCR. The expression of the corresponding cytokines plus IL-12p70 was analyzed using a bead array for flow cytometry. RESULTS: NAC inhibits the inflammatory cytokines TNFα, IL-1ß and IL-6 in LPS-activated macrophages under mild oxidative conditions. IL-10 mRNA and protein expression are strongly downregulated in NAC-treated cells, which may further modify the inflammatory cytokine profile. CONCLUSION: NAC modulates immune functions during the inflammatory response.

Antioxidant activity of N-acetylcysteine, flavonoids and alpha-tocopherol on endometrial cells in culture.

An appropriate local environment is necessary for successful implantation. Oxidative stress is implicated in the pathogenesis of several pathologies, and may contribute to early pregnancy failure. Antioxidant therapies have been studied in infertility. In this study, we have assessed the antioxidant activity of N-acetylcysteine (NAC), flavonoids (quercetin, catechin) and alpha-tocopherol in an oxidative model of endometrial cells (RL95). Endometrial cells were incubated at several hydrogen peroxide concentrations. Antioxidant effects of NAC (15 mM), quercetin (150 microM), catechin (150 microM) and alpha-tocopherol included in liposomes (1.6 microg) were assessed by measuring cell viability by the MTT assay. Alpha-tocopherol-liposomes taken up by endometrial cells were assessed by HPLC. All liposomes used were able to introduce alpha-tocopherol into cells. The antioxidant effect of NAC and quercetin improved the viability of oxidised cells, and this effect was observed when the oxidant and antioxidant were coincubated. No viability change occurred when the antioxidant was added before or after the oxidant. The antioxidant effect of NAC was better than that of quercetin. When catechin or alpha-tocopherol were used in the same conditions, no antioxidant effect was detected in cells in culture. These results demonstrate that NAC and quercetin are good H2O2 scavengers.

The presence of antibodies to oxidative modified proteins in serum from polycystic ovary syndrome patients.

Polycystic ovary syndrome (PCOS) affects 5-10% of women of reproductive age. Free radicals, as a product of oxidative stress, impair cells and tissue properties related to human fertility. These free radicals, together with the oxidized molecules, may have a cytotoxic or deleterious effects on sperm and oocytes, on early embryo development or on the endometrium. Aldehyde-modified proteins are highly immunogenic and circulating autoantibodies to new epitopes, such as malondialdehyde (MDA), may affect the reproductive system. Autoantibodies or elevated reactive oxygen species (ROS) in serum are often associated with inflammatory response. The purpose of this work is to investigate whether PCOS women show increased levels of oxidized proteins (protein-MDA) and anti-endometrial antibodies (AEA) in their sera, compared with control patients, and to determine whether AEA specificity is related to oxidized protein derivatives. Sera from 31 women [10 patients with PCOS (PCOS group) and 21 women with male factor of infertility (control group)] were chosen from patients attending for infertility. Anti-endometrial antibodies were determined by enzyme-linked immunosorbent assay (ELISA) with an endometrial cell line (RL-95). Antibodies against MDA modified human serum albumin (HSA-MDA) were also determined by ELISA. Oxidized proteins (protein-MDA) in serum were determined by a colorimetric assay. Patients with PCOS have significantly higher levels of AEA and anti-HSA-MDA, as well as oxidized proteins (protein-MDA) in serum than control patients. For the first time, we describe an autoimmune response in PCOS patients, in terms of AEA. The evidence of protein-MDA in the serum of these patients, together with the increased antibody reactivity to MDA-modified proteins (HSA-MDA) in vitro, supports the conclusion that oxidative stress may be one of the important causes for abnormal endometrial environment with poor embryo receptivity in PCOS patients.

Inhibition of in vitro gamete adhesion and in vivo fertility in mice by immunization with a synthetic peptide.

PROBLEM: To evaluate the contraceptive ability of a synthetic peptide in in vitro and in vivo fertility in the mouse. METHOD OF STUDY: A synthetic peptide segment: GELRERAPGQGTNG (SP) was used to immunize female B6CF1 (C57BL/6 x BALB/c) mice. A peptide with an amino acid sequence QQPLSIQQHERG (p2control) was used as control. Anti-SP and anti-p2control antisera were used to evaluate sperm function inhibition in vitro. Fertility of immunized mice was determined by microscopic evaluation of the number and state of preimplantation embryos (8-16 cell stage). RESULTS: In the mouse, anti-SP antisera recognized surface antigens in the acrosome region of mature and capacitated sperm. Anti-SP antisera inhibited in vitro sperm binding to zona pellucida. In vivo, immune response against SP in Freund's adjuvant resulted in a significant increase (P < 0.01) in the number of dead embryos and eggs (a mean of 66%, in contrast with < 25% in control mice). Fertility inhibition in vivo and in vitro was not observed when the p2control peptide was used in the immunizations. CONCLUSIONS: These results would suggest that the SP sequence is involved in gamete adhesion, and an antifertility vaccine against the SP peptide segment could be feasible.

Autoimmune response in women with endometriosis.

PROBLEM: The aim of this study was to investigate the humoral immune response to the female reproductive tissues associated with endometriosis (grades I-III) (n = 52), compared with a group of healthy fertile women (n = 6). METHOD OF STUDY: An ELISA with cultured endometrial cell lines in monolayer was used to determine the presence of anti-endometrial antibodies (AEA). For anti-zona pellucida antibodies (AZPA) assessment a conventional ELISA was employed. The presence of antibodies to human sperm (ASA) was performed by the tray agglutination test (TAT). RESULTS: Endometriosis grade III was associated with AEA in serum in the 45.4% of patients. The presence of AEA in serum is correlated to endometriosis severity. The 8.7% of women with endometriosis showed ASA, and the 10.9% of them were positive for AZPA. Antibodies specific for endometrial cells do not show reaction to any gamete antigen (sperm or oocyte), suggesting that they are not cross reactive. CONCLUSIONS: Severity of endometriosis is correlated with high titers of AEA.

Anti-endometrial autoantibodies in women with a diagnosis of infertility.

PROBLEM: The purpose of this study was to investigate the frequency of anti-endometrial antibodies (AEA) in infertile women. METHOD OF STUDY: Sera from fertile women (n = 6), and from patients with ovulatory dysfunction (n = 11), tubal obstruction (n = 9) and unexplained infertility (n = 5) were investigated for the presence of anti-endometrial membrane antibodies. We used two human endometrial cancer cell lines and human endometrial cells from gynecological biopsies as an antigenic source for analysis. The immunoenzymatic assay (ELISA) was performed with cultured endometrial cells in monolayers. Immunoblot analysis was performed with these two cell lines. RESULTS: A good correlation between the response with each cell line and with human endometrial cells was obtained, indicating that the antigens analyzed were probably similar. Endometrial antibodies were detectable in a high percentage of women with tubal obstruction (77.8 and 66.7%, respectively) and ovulatory dysfunction (54.5 and 45.5%, respectively). Unexplained infertility showed anti-endometrial immunological response (40 and 60%, respectively). Some endometrial antigens in infertile women are the target for autoimmune response. The serum from a patient with tubal obstruction and ovulatory dysfunction showed two antigens by immunoblot, with molecular weights of 97 and 50 kDa. CONCLUSION: The presence of anti-endometrial antibodies, detected by ELISA, is associated with infertility, mainly with ovulatory dysfunction and tubal obstruction. Some endometrial antigens may be involved in these two pathologies.

The erythrocyte density test (EDT) and erythrocyte creatine concentration (ECE) as laboratory indexes of perinatal hypoxia.

To study the influence of gestational age and asphyxia on the behaviour of EDT and ECE a group of cuban control and asphyxiated newborns was investigated. A total number of 203 newborns at the age of 0-3 days of life, all with adequate weight for gestational age, were classified into 4 groups according to their gestational age, the 1 min and 5 min Apgar score and the presence of meconium stained amniotic fluid and/or respiratory distress syndrome immediately after birth. In the group of asphyxiated newborns higher values for both parameters were found when they were compared with the respective control groups (p less than 0.05). These techniques could help the neonatologists to more precisely identify newborns with severe perinatal asphyxia and to improve the prognosis of the outcome.