Fetal Weight Outcomes in C57BL/6J and C57BL/6NCrl Mice after Oral Colonization with Porphyromonas gingivalis.

Porphyromonas gingivalis is considered a keystone pathogen that contributes to the initiation and progression of periodontitis in humans. P. gingivalis has also been detected in human placentas associated with adverse pregnancy outcomes. The spread of P. gingivalis from the oral cavity to the reproductive tract thus represents a potential mechanism whereby periodontitis can lead to adverse pregnancy outcomes. In a murine model of pregnancy and oral infection with P. gingivalis, C57BL/6J mice developed low fetal weight, whereas C57BL/6NCrl mice did not. Although C57BL/6NCrl mice harbor segmented filamentous bacteria that drive a Th17 response, fetal weight was independent of frequency of Th17 or Th1 in either substrain. Low fetal weight was instead correlated with increasing amounts of P. gingivalis DNA in the placentas of the C57BL/6J dams. In contrast, fetal weight in C57BL/6NCrl mice was independent of P. gingivalis in the placenta. Differences in genetics or microbiome that influence the ability of P. gingivalis to colonize the placenta may drive differential fetal weight outcomes between C57BL/6J and C57BL/6NCrl mice and, potentially, between diverse human populations.

Hepatic toxicity caused by PLGA-microspheres containing usnic acid from the lichen C ladonia substellata (AHTI) during pregnancy in Wistar rats.

This study aimed to evaluate the teratogenic and hepatotoxic potential of the usnic acid encapsulated into PLGA-microspheres. In total, 12 female Wistar rats in pregnancy were randomly distributed in the control group (n= 6) that received 1.0 mL of physiological solution and treatment group (n= 6) that received 25 mg/kg of encapsulated usnic acid by oral administration. All females were euthanized at day 20 of pregnancy and their fetuses were removed and analyzed. During the pregnancy was observed a reduction in weight gain. There was no difference in serum transaminases levels analyzed as well as any difference in liver weight in both groups. The histomorphometric analysis of the liver from the treatment group revealed an increase in number of hepatocytes and a decrease in nuclear area of these cells. Moreover, no alteration was observed in cell area of hepatocytes or number of Kupffer cells. The fetuses had an increase in total number of hepatocytes and a reduction in the amount of megakaryocytes. These results show the hepatotoxic potential of usnic acid during pregnancy. However, its toxicity can be minimized by encapsulation in microspheres.

The effect of Porphyromonas gingivalis lipopolysaccharide on pregnancy in the rat.

OBJECTIVE: Periodontitis, mostly associated with Porphyromonas gingivalis, has frequently been related to adverse pregnancy outcomes. We therefore investigated whether lipopolysaccharides of P. gingivalis (Pg-LPS) induced pregnancy complications in the rat. METHODS: Experiment 1: pregnant rats (day 14) received increasing Pg-LPS doses (0.0-50.0 µg kg(-1) bw; n = 2/3 p per dose). Maternal intra-aortic blood pressure, urinary albumin excretion, placental and foetal weight and foetal resorptions were documented. Experiment 2: 10.0 µg kg(-1) bw (which induced the highest blood pressure together with decreased foetal weight in experiment 1) or saline was infused in pregnant and non-pregnant rats (n = 7/9 p per group). Parameters of experiment 1 and numbers of peripheral leucocytes as well as signs of inflammation in the kidney and placenta were evaluated. RESULTS: Pg-LPS infusion in pregnant rats increased maternal systolic blood pressure, reduced placental weight (dose dependently) and decreased foetal weight and induced foetal resorptions. It, however, did not induce proteinuria or a generalised inflammatory response. No effects of Pg-LPS were seen in non-pregnant rats. CONCLUSION: Pg-LPS increased maternal blood pressure, induced placental and foetal growth restriction, and increased foetal resorptions, without inducing proteinuria and inflammation. Pg-LPS may therefore play a role in pregnancy complications induced by periodontitis.

Effects of the histamine H1 antagonist chlorcyclizine on rat fetal palate development.

BACKGROUND: The effects of histamine H1 antagonist chlorcyclizine on rat palate development were characterized following in utero exposure. METHODS: To identify the optimum dose for inducing cleft palate, pregnant rats were administered 30, 60, or 90 mg/kg chlorcyclizine on Gestation Days 11 to 14. Fetal palate gene expression was also assessed after 90 mg/kg chlorcyclizine at 8, 15 and 30 hours post-dose on Gestation Day 14 using microarray and qRT-PCR. RESULTS: Rats in the 60- and 90-mg/kg groups exhibited adverse clinical signs and body weight loss. Rats in the 90-mg/kg group also demonstrated increases in late resorptions and decreases in fetal weight. Effects in the low-dose group were limited to decreases in body weight gain. Fetal assessment on Gestation Day 21 revealed that findings were limited to the 60- and 90-mg/kg groups, and included cleft palate (80% of litters for both groups), high arched palate, small nose, micrognathia, high domed head, digits shortened/absent and small limb. The fetal incidence of cleft palate was higher at 90 mg/kg, thus this dose was selected to assess palate gene expression. The altered genes associated with chlorcyclizine-induced cleft palate included Wnt5a, Bmp2, Bmp4, Fgf10, Fgfr2, Msx1, and Insig1 but the magnitude of the change was relatively small (1.5- to 2-fold). CONCLUSIONS: Expression of several genes involved in palate, limb and digit development was altered in the fetal palate following in utero exposure to chlorcyclizine. The subtle perturbation and interplay of these genes may have profound effects on the dynamics of fetal palate development.

Developmental toxicity assessment of thermoresponsive poly(N-isopropylacrylamide-co-acrylamide) oligomers in CD-1 mice.

BACKGROUND: Although polymers and hydrogels are used successfully in biomedical applications, including implants and drug delivery devices, smaller molecular weight oligomers, such as those investigated here, have not been extensively studied in vivo. Poly(N-isopropylacrylamide-co-acrylamide), or P(NIPAAm-co-AAm), has a unique thermoresponsive behavior and is under investigation as a novel drug delivery system for metastatic cancer treatment. To date, no studies have been published regarding the safety of P(NIPAAm-co-AAm) to the conceptus. METHODS: From gestation days (GD) 6-16, pregnant CD-1 mice were dosed via i.p. injection with aqueous solutions containing 500, 750, or 1,000 mg/kg/d P(NIPAAm-co-AAm). Dams were sacrificed on GD 17 and their litters were examined for abnormalities. RESULTS: P(NIPAAm-co-AAm) caused no statistically significant difference in maternal weight gain or percent resorbed or dead fetuses compared to control values, but fetal weight was significantly decreased in the two highest dosage groups. CONCLUSIONS: At the highest dosages employed, maternal exposure to P(NIPAAm-co-AAm) was associated with decreased fetal weight. However, as the estimated human exposure levels for persons using this system would be some 1,500-fold lower than the lowest dosage administered in this study, the authors feel that this oligomer was not shown to pose a biologically significant risk at relevant human dosages.

Impact of maternal periodontal health on fetus weight in Iraqi pregnant women: a clinical study / Impacto da saúde periodontal materna no peso fetal em gestantes iraquianas: um estudo clínico

Objective: the aim of this study was to investigate the relationship between periodontal health condition and the weight of fetuses in Iraqi pregnant women in order to magnify the importance of periodontal health maintenance during pregnancy. Material and Methods: fetus weight was determined using ultrasound scanning for 222 pregnant women, accordingly they were divided into two groups: group A: normal fetus weight and group B: below normal fetal weight. Their periodontal condition was examined by means of Plaque index (PI), Gingival index (GI), Bleeding on probing (BOP) and Clinical Attachment loss (CAL) using WHO CPITN periodontal probe. WHO charts of normal fetal weight for each week were considered to determine the normality of fetus weight. Results: significant value p=0.00 was obtained when comparing the examined periodontal parameters between groups A and B, mean of periodontal parameters of PI, GI and BOP were higher in group B (1.1964: 1.4541), (1.1877: 1.4925), (0.3553: 1.3748) respectively. Q2 and IQR of PI, BOP and GI in group A were (1.190:0.3), (0.30:0.5), (1.160:0.3) respectively. And (1.460:0.24) (1.50:0.7) (1.460:0.26) in Group B. There were 4 cases of CAL in group A as opposed to 88 cases in group B, Q2 of CAL in group A=0.00, Q2 in group B=1.00. IQR=0.00 in both groups. Non-significant value p=0. 503(p>0.05) was seen when comparing the incidence of low fetal weight between the three trimesters. Conclusion: it is important to maintain a good periodontal condition and oral Hygiene status in pregnant women for healthier fetal weight and healthier pregnancy with less complications. (AU)

Objetivo: o objetivo deste estudo foi investigar a relação entre a condição de saúde periodontal e a peso dos fetos em gestantes iraquianas para ampliar a importância da manutenção da saúde periodontal durante a gravidez. Material e Métodos: o peso do feto foi determinado por ultrassonografia 222 As gestantes, consequentemente, foram divididas em dois grupos: grupo A: peso normal do feto e grupo B: abaixo do peso fetal normal. Sua condição periodontal foi examinada por meio de índice de placa (IP), gengival índice (GI), Sangramento na sondagem (BOP) e Perda de Inserção Clínica (CAL) usando a sonda periodontal CPITN da OMS. Os gráficos da OMS de peso fetal normal para cada semana foram considerados para determinar a normalidade do peso feto. Resultados: obteve-se valor significativo p=0,00 quando comparados os parâmetros periodontais examinados entre nos grupos A e B, as médias dos parâmetros periodontais do IP, GI e BOP foram maiores no grupo B (1,1964: 1,4541), (1,1877: 1,4925), (0,3553: 1,3748), respectivamente. Q2 e IQR do PI, BOP e GI no grupo A foram (1,190:0,3), (0.30:0.5), (1.160:0.3) respectivamente.E (1.460:0.24) (1.50:0.7) (1.460:0.26) no Grupo B. Houve 4 casos de CAL no grupo A em oposição a 88 casos no grupo B, Q2 de CAL no grupo A=0,00, Q2 no grupo B=1,00. IQR=0,00 em ambos os grupos. Valor não significativo p=0. 503(p>0,05) foi observado quando comparada a incidência de baixo nível fetal peso entre os três trimestres. Conclusão:é importante manter uma boa condição periodontal e Estado de higiene bucal em gestantes para maior peso fetal e gravidez mais saudável com menos complicações. (AU)

Mercury concentration in maternal serum, cord blood, and placenta in patients with amalgam dental fillings: effects on fetal biometric measurements.

AIM: We aimed to determine the extent to which mercury is transmitted from the mother to fetus via the umbilical cord in patients with amalgam dental fillings, and its effect on fetal biometric measurements. METHODS: Twenty-eight patients as the study group with amalgam fillings, and 32 of them as the control group were included in this prospective case-control study. The mercury levels were measured in the maternal and cord venous sera, and the placental samples. Two groups were compared in terms of these and the fetal/neonatal biometric measurements. RESULTS: In the study group, the maternal and umbilical cord mercury levels were found to be significantly higher than those from the control group (p = 0.006 and p = 0.010, respectively). These high levels did not affect the fetal biometric measurements. CONCLUSIONS: The presence of high serum mercury levels in pregnant women with amalgam fillings is important, and warrants further long-term studies in order to investigate the fetal neurological effects as well.

Effects of cigarette smoke on the Meckel's cartilage of rat fetus: morphologic, morphometric and stereologic study.

The purpose of this study was to investigate the effects of cigarette smoke on the development of the embryo mandible (Meckel's) cartilage in rat fetuses. When inhaled by female Wistar rats between the 9th and the 12th day of pregnancy, cigarette smoke (5 cigarettes a day) caused intrauterine growth retardation, providing smaller fetuses and placentas. In fetuses from the experimental group, the histopathologic examination revealed a poorly developed Meckel's cartilage with smaller chondroblasts showing a scanty cytoplasm with spherical and paler central nuclei, as well as more abundant cartilage matrix. Morphometric analysis revealed that Meckel's cartilage lacunae were smaller in the fetuses from the experimental group, although not showing any remarkable alteration in shape. The results suggested that inhalation of cigarette smoke by pregnant rats during the organogenic period induced growth retardation and delayed cellular differentiation in rat fetal Meckel's cartilage.

Hepatic toxicity caused by PLGA-microspheres containing usnic acid from the lichen C ladonia substellata (AHTI) during pregnancy in Wistar rats

ABSTRACT This study aimed to evaluate the teratogenic and hepatotoxic potential of the usnic acid encapsulated into PLGA-microspheres. In total, 12 female Wistar rats in pregnancy were randomly distributed in the control group (n= 6) that received 1.0 mL of physiological solution and treatment group (n= 6) that received 25 mg/kg of encapsulated usnic acid by oral administration. All females were euthanized at day 20 of pregnancy and their fetuses were removed and analyzed. During the pregnancy was observed a reduction in weight gain. There was no difference in serum transaminases levels analyzed as well as any difference in liver weight in both groups. The histomorphometric analysis of the liver from the treatment group revealed an increase in number of hepatocytes and a decrease in nuclear area of these cells. Moreover, no alteration was observed in cell area of hepatocytes or number of Kupffer cells. The fetuses had an increase in total number of hepatocytes and a reduction in the amount of megakaryocytes. These results show the hepatotoxic potential of usnic acid during pregnancy. However, its toxicity can be minimized by encapsulation in microspheres.

Therapeutic effects of anticoagulant agents on preeclampsia in a murine model induced by phosphatidylserine/phosphatidylcholine microvesicles.

Preeclampsia (PE) is a placenta-mediated pregnancy complication that results in high maternal and neonatal mortality and morbidity worldwide. Currently, there is no satisfactory pharmacotherapeutic treatment to stop the PE progression. In this study, we investigated the therapeutic effects of anticoagulant agents, including annexin V, heparin, and a fusion protein of annexin V and hirudin (AND), in a murine PE model induced by intravenous injection of phosphatidylserine/phosphatidylcholine (PS/PC) microvesicles. Compared with the control pregnant animals, the pregnant mice injected with PS/PC presented PE-like symptoms, including elevated systolic blood pressure, proteinuria, and reduction of antithrombin III and blood platelets. However, the PE-like symptoms were significantly alleviated after the PS/PC-injected mice were treated with annexin V, AND, or heparin. Furthermore, fibrin deposition in the placentas in the anticoagulant treated mice was remarkably improved, compared with that in the mice injected with PS/PC alone. The data demonstrate that anticoagulants are effective to prevent the occurrence of PE in the murine model and also suggest that hypercoagulation in the placenta is a contributing factor in the pathogenesis of PE.

Effect of repeated stress treatments during the follicular phase and early pregnancy on reproductive performance of gilts.

In pig husbandry, stress is being considered an important cause of impaired reproductive performance. Therefore, an experiment was performed to quantify effects of repeated stressors during the follicular phase and/or during early pregnancy on reproductive performance of gilts. Eighty-one cyclic gilts were assigned to one of four treatments, namely, stress treatment during the follicular phase (n = 20), stress treatment during early pregnancy (n = 20), stress treatment during both phases (n = 21) and no stress treatment (n = 20). All gilts were housed individually, but gilts in the stress treatments had no opportunity for visual or physical contact with other gilts. Further, animals in a stress-treatment were grouped for half an hour at the start of the treatment and during the treatment period nose-sling and an unpredictable feeding scheme were applied regularly. The extent of stress was monitored using heart rate measurements, behavioural observations and saliva cortisol levels during nose-sling fixation. Of the 81 gilts, 93% showed oestrus and were inseminated. Of these, 93% were pregnant at day 35, having 17.9 +/- 0.3 ovulations and 15.6 +/- 0.3 foetuses. These parameters were not affected by treatment. The stress treatment during the follicular phase tended to shorten cycle length (stress: 20.8 +/- 0.20; control: 21.2 +/- 0.17 days, p = 0.07) and weight of foetuses at day 35 (stress: 4.47 +/- 0.08 g; no stress: 4.69 +/- 0.08 g, p = 0.06); stress during early pregnancy did not affect any of the reproduction parameters. Percentage stereotypic behaviour, heart rate and saliva cortisol levels varied greatly between animals and between days, but did not differ between the treatments. No relationships were found between any of the reproductive parameters and any of the stress parameters (heart rate, cortisol, stereotypic behaviour). These results indicate that the repeatedly applied acute stressors did not generate a chronic stress-response and that these stressors during the follicular phase and/or during early pregnancy did not affect reproductive processes. It is not clear how these findings relate to suggested effects of stress(ors) on reproductive performance in pig husbandry.

Insulin-like growth factor and interleukin-1beta levels and subsequent fetal size in response to chronic Porphyromonas gingivalis exposure in the pregnant rabbit.

OBJECTIVE: The purpose of this study was to describe maternal insulin-like growth factor, interleukin-1beta, and fetal size in a rabbit model of Porphyromonas gingivalis exposure. STUDY DESIGN: With the use of a previously described model, 8 New Zealand White rabbits were exposed to either P gingivalis or media during pregnancy and killed at term. Kit and placenta weight were compared between groups. Doe serum insulin-like growth factor system protein and interleukin-1beta levels were compared by analysis of variance for repeated measures; a probability value of <.05 was considered to be significant. RESULTS: No significant differences in kit and placental weights between exposed and unexposed groups were observed. Insulin-like growth factor system proteins increased significantly as pregnancy progressed, but there were no significant differences in insulin-like growth factor system proteins or interleukin-1beta between exposed and unexposed does. CONCLUSION: Chronic P gingivalis exposure does not disrupt insulin-like growth factor system proteins or systemic inflammation and does not impair fetal growth in the pregnant rabbit. Gestational age changes in doe insulin-like growth factor system proteins occur, and the timing of exposure to oral pathogens may influence fetal growth.

Phosphatidylserine/phosphatidylcholine microvesicles can induce preeclampsia-like changes in pregnant mice.

We established a phosphatidylserine (PS)/phosphatidylcholine (PC) microvesicles-induced preeclampsia-like model in mice. PS/PC were prepared by mixing 80% PC and 20% PS, and suspended in 0.05 M Tris-HCl at a concentration of 10 mg/mL. One hundred microliters of PS/PC (n = 6) and saline as a control (n = 10) were injected in tail veins of Institute of Cancer Research (ICR) mice every day from days 5.5 to 16.5 of pregnancy. Systolic blood pressure (SBP) was measured by means of the tail-cuff method. On day 17.5, the mice were anesthetized by diethyl ether and euthanized with the collapse of the circulation by drawing blood from the heart. The animals were dissected and the fetuses and placentas removed. Fetal weight and placental weight were evaluated. Plasma antithrombin activity (AT), thrombin-antithrombin complex (TAT), platelet counts, and proteinuria were measured on day 17.5. Placentas were fixed in 4% paraformaldehyde for histologic studies. Statistical analysis was evaluated by analysis of variance and Welch's t-test. Mice injected with PS/PC showed a significant elevation in SBP (124 versus 101 mm Hg; p < 0.001), a significant increase in TAT levels (23 versus 6.6 mug/L; p < 0.05), a significant decrease in platelet counts (88 versus 102 x 10 (10)/L; p < 0.05), a decrease in AT, an increase in proteinuria, and a significant reduction in fetal weight (1.2 versus 1.3 g; p < 0.0001) and placental weight (0.13 versus 0.15 g; p < 0.001), compared with controls. Placentas of mice injected with PS/PC showed diffuse fibrin depositions in the labyrinth layer. We have demonstrated that the artificial PS/PC vesicles induce intrauterine growth restriction with elevations of SBP. The elevation of plasma TAT and the diffuse fibrin depositions in the placentas indicate enhanced thrombin formation, and the significant elevations of SBP indicate preeclampsia-like changes that can be induced by hypercoagulation in the placenta.

Developmental toxicity study with triethylene glycol given by gavage to CD rats and CD-1 mice.

Triethylene glycol (TEG) is a liquid industrial chemical with a potential for human exposure. The likelihood for developmental toxicity was investigated in two species. Timed-pregnant CD rats and CD-1 mice were dosed daily by gavage with undiluted TEG over gestational days (gd) 5-15 at 0.0 (water control), 1126, 5630 or 11,260 mg kg(-1) day(-1) with rats and 0.0, 563, 5630 or 11,260 mg kg(-1) day(-1) with mice. They were examined daily, and gestational body weights and food and water consumption measured throughout gestation. At necropsy on gd 21 (rats) or gd 18 (mice) dams were examined for body, gravid uterine, liver and kidney weights, and implantation sites. Maternal kidneys were examined histologically. Fetuses were weighed, sex determined, and examined for external, soft tissue and skeletal variations and malformations. Rat dams had reduced body weights, body weight gains, and food consumption, and increased water consumption and relative kidney weights at 11,260 mg kg(-1) day(-1). They also had reduced body weight and increased water consumption at 5630 mg kg(-1) day(-1). Mice had clinical signs and increased relative kidney weight at 11,260 mg kg(-1) day(-1). Renal histology was normal in both species. Neither species had treatment-related effects on corpora lutea or implantations. Fetal body weights were reduced at 11,260 mg kg(-1) day(-1) (both species) and 5630 mg kg(-1) day(-1) (mice). In rat fetuses there was a pattern of delayed ossification in the thoracic region at 11,260 mg kg(-1) day(-1). Mouse fetuses had delayed ossification in the frontal and supraoccipital bones, cervical region, hindlimb proximal phalanges and reduced caudal segments at 11,260 mg kg(-1) day(-1), and in the skull bones at 5630 mg kg(-1) day(-1). These patterns of delayed ossification are consistent with reduced fetal body weights. No biologically significant embryotoxicity or teratogenicity was observed at any dosage in either species. The NOEL for TEG given by gavage over the period of organogenesis was 1126 mg kg(-1) day(-1) in the rat and 5630 mg kg(-1) day(-1) in the mouse for maternal toxicity, and 5630 mg kg(-1) day(-1) (rat) and 563 mg kg(-1) day(-1) (mouse) for developmental toxicology.

Endothelial function and myogenic reactivity in small mesenteric arteries of hyperlipidemic pregnant rats.

Supraphysiological increases in serum triglycerides and cholesterol often occur during pregnancy, but their effects on vascular function are poorly understood. Intraperitoneal injection of the nontoxic surfactant poloxamer 407 (P-407) results in sustained elevation of triglycerides and cholesterol. We asked if P-407-induced hyperlipidemia during late pregnancy adversely affects mesenteric resistance artery vasodilator function. On days 13-15 of pregnancy, rats were given a single intraperitoneal injection of P-407, sterile water vehicle, or non-lipid-altering pluronic F-88 (P-88). Four days postinjection, serum triglycerides, cholesterol, free fatty acids, and the lipid peroxidation product malondialdehyde were significantly increased in P-407-treated rats. Mesenteric arteries from P-407-treated rats displayed significant increases in myogenic reactivity (constrictor responses to step increases in intraluminal pressure). The nitric oxide (NO) blocker N(alpha)-methyl-L-arginine increased the myogenic response in control but not in P-407 arteries, normalizing group differences. Endothelial removal increased myogenic reactivity beyond that of prior NO synthase inhibition in controls and potentiated myogenic reactivity in P-407 arteries such that responses again converged. Relaxation responses to the endothelium-dependent vasodilator methacholine did not differ. We conclude that that P-407-induced hyperlipidemia during pregnancy increases myogenic reactivity due to selective attenuation of an NO-mediated vasodilator component of the myogenic response.

Effects of cigarette smoke on the Meckel's cartilage of rat fetus: morphologic, morphometric and stereologic study

O objetivo deste estudo foi investigar os efeitos da fumaça de cigarros sobre o desenvolvimento da cartilagem mandibular (cartilagem de Meckel) do embrião de rato. Quando inalado por ratas Wistar, entre o 9º e o 12º dia de prenhez, a fumaça de cigarros (5/dia) provocou retardo do crescimento intrauterino, com fetos e placentas menores. Nos fetos do grupo experimental, o exame histopatológico revelou uma cartilagem de Meckel pouco desenvolvida, com condroblastos menores apresentando citoplasma escaso com núcleos mais pálidos, esféricos e centrais, assim como uma matrix cartilaginosa mais abundante. A análise morfométrica revelou que as lacunas da cartilagem de Meckel eram menores nos fetos do grupo experimental, não apresentando alteração significativa de sua forma. Os resultados sugeriram que a fumaça de cigarros inalada pelas ratas prenhes durante o período de organogênese induziu retardo tanto do crescimento quanto da diferenciação celular na cartilagem de Meckel dos fetos.