Huiyi Yan

Increased Fetal Thymocytes Apoptosis Contributes to Prenatal Nicotine Exposure-induced Th1/Th2 Imbalance in Male Offspring Mice

Nicotine, a definite risk factor during pregnancy, is an immunomodulator. This study was designed to investigate the effects of prenatal nicotine exposure (PNE) on the balance of Th1/Th2 in offspring, and further explore the developmental origin mechanisms from the perspective of fetal thymocytes apoptosis. Pregnant Balb/c mice were administered 1.5 mg/kg nicotine subcutaneously twice per day from gestational day (GD) 9 to GD18. Results showed that PNE could cause a Th2 shift in male offspring, manifested as increased ratio of IgG1/IgG2a, IL-4 production in serum, and IL-4/IFN-γ expression ratio in spleen. Increased apoptosis of total thymocytes and CD4SP and reduced cell proportion of CD4SP were found in PNE male offspring on postnatal day (PND) 14 and PND 49. In the fetuses, decreased body weight and organ index of fetal thymus, histological changes in fetal thymus, reduced CD4SP proportion and increased fetal thymocyte apoptosis were observed in nicotine group. The increased mRNA expression of genes involved in Fas-mediated apoptotic pathway and protein expression of Fas were also detected. In conclusion, PNE could cause a Th2 shift in male offspring mediated by reduced CD4+ T cells output, which may result from the increasing apoptosis of total thymocytes and CD4SP.

High-performance therapeutic quercetin-doped adhesive for adhesive–dentin interfaces

Almost half of dental restorations have failed in less than 10 years, and approximately 60% of practice time has been consumed to replace these dental restorations. As such, contemporary dentin adhesives should be modified to treat secondary caries and prevent the degradation of adhesive–dentin interfaces. To achieve this goal, we developed a versatile therapeutic adhesive in the present study by incorporating quercetin, which is a naturally derived plant extract, into a commercial adhesive at three concentrations (100, 500 and 1000 µg/mL). An unmodified adhesive served as a control. The antibacterial ability on Streptococcus mutans biofilm, conversion degree, microtensile bond strength, failure modes, in situ zymography, nanoleakage expression and cytotoxicity of quercetin-doped adhesive were comprehensively evaluated. Results showed that the quercetin-doped adhesive (500 µg/mL) preserved its bonding properties against collagenase ageing and inhibited the growth of S. mutans biofilm. Efficient bonding interface sealing ability, matrix metalloproteinase inhibition and acceptable biocompatibility were also achieved. Thus, a simple, safe and workable strategy was successfully developed to produce therapeutic adhesives for the extension of the service life of adhesive restorations.

α7 nAChR mediated Fas demethylation contributes to prenatal nicotine exposure-induced programmed thymocyte apoptosis in mice

This study aimed to investigate the effects of prenatal nicotine exposure (PNE) on thymocyte apoptosis and postnatal immune impairments in vivo and further explore the epigenetic mechanisms of the pro-apoptotic effect of nicotine in vitro. The results showed that PNE caused immune impairments in offspring on postnatal day 49, manifested as increased IL-4 production and an increased IgG1/IgG2a ratio in serum. Enhanced apoptosis of total and CD4+SP thymocytes was observed both in fetus and in offspring. Further, by exposing thymocytes to 0–100 μM of nicotine in vitro for 48 h, we found that nicotine increased α7 nicotinic acetylcholine receptor (nAChR) expression, activated the Fas apoptotic pathway, and promoted thymocyte apoptosis in concentration-dependent manners. In addition, nicotine could induce Tet methylcytosine dioxygenase (TET) 2 expression and Fas promoter demethylation, which can be abolished by TET2 siRNA transfection. Moreover, the α7 nAChR specific antagonist α-bungarotoxin can abrogate nicotine-induced TET2 increase, and the following Fas demethylation and Fas-mediated apoptosis. In conclusion, our findings showed, for the first time, that α7 nAChR activation could induce TET2-mediated Fas demethylation in thymocytes and results in the upregulation of Fas apoptotic pathway, which provide evidence for elucidating the PNE-induced programmed thymocyte apoptosis.

Effects of Chlorhexidine-Encapsulated Mesoporous Silica Nanoparticles on the Anti-Biofilm and Mechanical Properties of Glass Ionomer Cement

One of the primary causes for the failure of glass ionomer cement (GIC) is secondary caries. To enhance the anti-microbial performance of GIC without affecting its mechanical properties, chlorhexidine (CHX) was encapsulated in expanded-pore mesoporous silica nanoparticles (pMSN) to synthesize CHX@pMSN. CHX@pMSN was added at three mass fractions (1%, 5%, and 10% (w/w)) to GIC powder as the experimental groups. Pure GIC was set as the control group. The mechanical and anti-biofilm properties of GIC from each group were tested. The results demonstrated that CHX was successfully encapsulated on/into pMSN, and the encapsulating efficiency of CHX was 44.62% in CHX@pMSN. The anti-biofilm ability was significantly enhanced in all experimental groups (p < 0.001) compared with that in the control group. CHX was continuously released, and anti-biofilm ability was maintained up to 30 days. In addition, the mechanical properties (compressive strength, surface hardness, elastic modulus, water sorption, and solubility) of 1% (w/w) group were maintained compared with those in the control group (p > 0.05). In conclusion, adding 1% (w/w) CHX@pMSN to GIC led to conspicuous anti-biofilm ability and had no adverse effect on the mechanical properties of this restorative material. This study proposes a new strategy for preventing secondary caries by using CHX@pMSN-modified GIC.

Chlorhexidine-encapsulated mesoporous silica-modified dentin adhesive

OBJECTIVES This work aims to explore the feasibility of chlorhexidine-encapsulated mesoporous silica (CHX@pMSN) as a modifier of a commercial dental adhesive via the evaluation of physicochemical properties and antibacterial capabilities of adhesive-dentin interface. METHODS Therapeutic adhesives were developed in the present study by incorporating CHX@pMSN into a commercial adhesive at four mass fractions (0, 1, 5 and 10 wt.%). The antibacterial capability on Streptococcus mutans (S. mutans) biofilm, conversion degree, adhesive morphology, microtensile bond strength (MTBS) and nanoleakage expression were evaluated comprehensively. RESULTS MTT and CLSM evaluation showed that CHX@pMSN-doped adhesive inhibits S. mutans biofilm growth, while CHX is released from the modified adhesive continuously. The incorporation of CHX@pMSN did not affect immediate bond strength at the concentration of 1% and 5% (P >  0.05). Moreover, these bonds were mainly preserved in 5% CHX@pMSN group after one month of collagenase ageing. Meanwhile, CHX@pMSN-doped adhesive groups exhibited similar nanoleakage distribution compared with the control. CONCLUSION This study showed that the 5% CHX@pMSN-modified adhesive achieved balance amongst unaffected immediate bonding strength, well-preserved bonds against collagenase ageing and effective inhibition of S. mutans biofilm growth. CLINICAL SIGNIFICANCE CHX@pMSN-modified dentin adhesive can potentially extend the service life of adhesive restoration in clinic.

Effects of calcium-containing desensitizers on the bonding stability of an etchand-rinse adhesive against long-term water storage and pH cycling

This study aimed to evaluate the effects of two calcium-containing desensitizing pastes on the bonding stability of an etch-and-rinse (E&R) adhesive to dentine. After dentine hypersensitivity model established, dentine surfaces were assigned one of the following pretreatment: Group 1, no desensitizer; Group 2, CPP-ACP; and Group 3, Novamin. Specimens were then bonded with an E&R adhesive. Beams from each tooth were randomly divided into three subgroups and then subjected to microtensile bond strength (MTBS) test after 24 h; 12 months of water storage; or 15 runs of pH cycling. Failure modes, nanoleakage, and tubule-occluding effectiveness were analyzed. Results showed that CPP-ACP- or Novamin-pretreated specimens mainly preserved the bonding strength after 12 months of water storage, while effective tubule occlusion could be observed. The results suggested that the calcium-containing desensitizers were compatible pretreatment for bonding with E&R adhesives to obtain reliable long-term bonding strength and prevention of post-operative sensitivity.

Maternal Glucocorticoid Elevation and Associated Fetal Thymocyte Apoptosis are Involved in Immune Disorders of Prenatal Caffeine Exposed Offspring Mice

Our previous study showed that prenatal caffeine exposure (PCE) could induce intrauterine growth retardation (IUGR) and glucocorticoid elevation in the fetus. Researchers suggested that IUGR is a risk factor for T helper cell (Th)1/Th2 deviation. However, whether PCE can induce these immune disorders and the underlying mechanisms of that induction remain unknown. This study aimed to observe the effects of PCE on the Th1/Th2 balance in offspring and further explore the developmental origin mechanisms from the perspective of glucocorticoid overexposure-induced thymocyte apoptosis. An IUGR model was established by caffeine administration from gestational day (GD) 9 to GD 18, and the offspring were immunized on postnatal day (PND) 42. The results show that maternal glucocorticoid overexposure increased fetal thymocyte apoptosis by activating both the Fas-mediated and the Bim-regulated apoptotic pathways. After birth, accelerated thymocyte apoptosis and Th1 suppression were also found in the PCE offspring at PND 14 and PND 49. Moreover, the PCE offspring showed immune disorders after immunization, manifesting as increased IgG1/IgG2a ratio and IL-4 production in the serum. In conclusion, PCE could induce fetal overexposure to maternal glucocorticoids and increase thymocyte apoptosis, which could persist into postnatal life and be implicated in Th1 inhibition and further immune disorders.

Prenatal caffeine ingestion increases susceptibility to pulmonary inflammation in adult female rat offspring

This study aimed to investigate the association between prenatal caffeine ingestion (PCI) and risk of postnatal pulmonary inflammation. Pregnant Wistar rats were administered 60mg/kg/d caffeine intragastrically from gestational day (GD) 7 to GD 20. The results showed that PCI obviously increased intrauterine growth retardation rate to 39.2% and suppressed weight growth of the offspring. PCI also enhanced the expression of transforming growth factor β, α-smooth muscle actin, interleukin (IL)-1β, and IL-8 in lungs and caused pulmonary interstitial thickening in the offspring. Further, with lipopolysaccharide stimulation on postnatal day 77, PCI offspring showed more serious inflammatory infiltration, higher injury scores, and higher levels of IL-6 and tumor necrosis factor-α in lungs than those of the control. Our findings showed, for the first time, that PCI is a certainly threat to postnatal pulmonary inflammation. The potential mechanism is that PCI alter the expression of pulmonary interstitial thickening-associated genes in the offspring.

Enhanced thymocyte apoptosis induced by maternal undernutrition in late gestation results in declined mature T cells in rat fetal thymus

This study was designed to observe the effects of maternal food restriction (MFR) on the development of fetal thymus in different gestation periods. Timed pregnant rats were randomized into 3 groups: CN (free access to standard chow throughout gestation), MFR0-21 (50% MFR throughout gestation), MFR0-14 (50% MFR from gestational day (GD) 0 to GD14, early-mid gestation). Results showed that MFR during early-mid period had few impact on the fetal thymus and T cell subpopulations. However, MFR throughout gestation resulted in thymic atrophy, deceased frequency of both CD4+ and CD8+ single positive (SP) T cells and enhanced thymocyte apoptosis in fetus. Our results suggest the fetal thymus is more vulnerable to the adverse intrauterine environments in the late gestation period, and the decreased number of SP T cells could result from the enhanced thymocyte apoptosis.