Yao-Dung Hsieh

Lactobacillus acidophilus attenuates Salmonella-induced intestinal inflammation via TGF-β signaling

BackgroundSalmonella is a common intestinal pathogen that causes acute and chronic inflammatory response. Probiotics reduce inflammatory cytokine production and serve as beneficial commensal microorganisms in the human gastrointestinal tract. TGF-β (transforming growth factor β)/SMAD and NF-κB signaling play important roles in inflammation in intestinal cells. However, the involvement of the signaling in regulating inflammation between Salmonella and probiotics is not fully understood.MethodsL. acidophilus and prebiotic inulin were used to treat human intestinal Caco-2 cells prior to infection with Salmonella. The cells were harvested to examine the cytokines and MIR21 expression with immunoblotting and real-time PCR. NF-κB and SMAD3/4 reporter vectors were transfected into cells to monitor inflammation and TGF-β1 signaling, respectively.ResultsIn this study, we showed that the probiotic L. acidophilus decreased Salmonella-induced NF-κB activation in human intestinal Caco-2 cells. Expression of the inflammatory cytokines, TNF-α and IL-8, in L. acidophilus-pretreated cells was also significantly lower than that in cells infected with Salmonella alone. Moreover, TGF-β1 and MIR21 expression was elevated in cells pretreated with L. acidophilus or synbiotic, a combination of inulin and L. acidophilus, compared to that in untreated cells or cells infected with S. typhimurium alone. By contrast, expression of SMAD7, a target of MIR21, was accordingly reduced in cells treated with L. acidophilus or synbiotics. Consistent with TGF-β1/MIR21 and SMAD7 expression, SMAD3/4 transcriptional activity was significantly higher in the cells treated with L. acidophilus or synbiotics. Furthermore, TGF-β1 antibody antagonized the SMAD3/4 and NF-κB transcriptional activity modulated by L. acidophilus in intestinal cells.ConclusionOur results suggest that the TGF-β1/MIR21 signaling pathway may be involved in the suppressive effects of L. acidophilus on inflammation caused by S. typhimurium in intestinal Caco-2 cells.

The osteoinductive effect of chitosan-collagen composites around pure titanium implant surfaces in rats.

BACKGROUND AND OBJECTIVE The enhancing effects of chitosan on activation of platelets and differentiation of osteoprogenitor cells have been demonstrated in vitro. The purpose of this study was to evaluate the in vivo osteoinductive effect of chitosan-collagen composites around pure titanium implant surfaces. MATERIAL AND METHODS Chitosan-collagen composites containing chitosan of different molecular weights (450 and 750 kDa) were wrapped onto titanium implants and embedded into the subcutaneous area on the back of 15 Sprague-Dawley rats. The control consisted of implants wrapped with plain collagen type I membranes. Implants and surrounding tissues were retrieved 6 wks after surgery and identified by Alizarin red and Alcian blue whole mount staining. The newly formed structures in the test groups were further analyzed by Toluidine blue and Masson-Goldner trichrome staining, and immunohistochemical staining with osteopontin and alkaline phosphotase. The bone formation parameters of the new bone in the two test groups were measured and compared. RESULTS New bone formed ectopically in both chitosan-collagen groups, whereas no bone induction occurred in the negative control group. These newly formed bone-like structures were further confirmed by immunohistochemical staining. Comparison of bone parameters of the newly induced bone revealed no statistically significant differences between the 450 and 750 kDa chitosan-collagen groups. CONCLUSION Our results demonstrated that chitosan-collagen composites might induce in vivo new bone formation around pure titanium implant surfaces. Different molecular weights of chitosan did not show significantly different effects on the osteoinductive potential of the test materials.

Evaluation on the movement of endosseous titanium implants under continuous orthodontic forces: an experimental study in the dog

OBJECTIVES The purpose of this study was to evaluate the movement of pure titanium implants under different continuous forces in the edentulous alveolar ridge. MATERIAL AND METHODS Four pairs of titanium implants were inserted into the right maxillary and mandibular post-extraction edentulous ridge of the experimental dog. Three different levels of continuous force (100, 200, and 500 g) were loaded onto three pairs of adjacent implant abutments using a memory Ni-Ti coil spring for up to 6 months and the remaining two implant abutments as the control group received no force. The positions of implant abutments were observed and the distances between the implants abutment at the top, middle and base levels were measured at the 0th, 2nd, 3rd, 6th and 8th month of the follow-up period. RESULTS There was no significant change in the distances between adjacent abutments loaded with 100 or 200 g continuous forces throughout the entire study period. However, significantly more movement of implant abutments was noted in the 500 g pair after the 3rd month of loading when compared with the 200 or the 100 g pair (both P < 0.001). This change further increased at the 6th month (P < 0.001, 0.01, respectively). Moreover, the difference in the measurements at the top, middle and base level indicated that the two adjacent implants moved in a tipping manner in the 500 g pair after 3 and 6 months of loading. CONCLUSION The osseointegrated implants remained stable and rigid with a pulling force of 100 and 200 g after 6 months of loading. However, when the force reached 500 g, the implants moved in an inward-tipping pattern. The results suggested that endosseous titanium implants might not necessarily be rigid anchorages under all circumstances.

Effect of Methoxychlor on Ca2+ Handling and Viability in OC2 Human Oral Cancer Cells

Abstract:  The effect of the insecticide methoxychlor on the physiology of oral cells is unknown. This study aimed to explore the effect of methoxychlor on cytosolic Ca2+ concentrations ([Ca2+]i) in human oral cancer cells (OC2) by using the Ca2+‐sensitive fluorescent dye fura‐2. Methoxychlor at 5–20 μM increased [Ca2+]i in a concentration‐dependent manner. The signal was reduced by 70% by removing extracellular Ca2+. Methoxychlor‐induced Ca2+ entry was not affected by nifedipine, econazole, SK&F96365 and protein kinase C modulators but was inhibited by the phospholipase A2 inhibitor aristolochic acid. In Ca2+‐free medium, treatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin or 2,5‐di‐tert‐butylhydroquinone (BHQ) inhibited or abolished methoxychlor‐induced [Ca2+]i rise. Incubation with methoxychlor also inhibited thapsigargin‐ or BHQ‐induced [Ca2+]i rise. Inhibition of phospholipase C with U73122 did not alter methoxychlor‐induced [Ca2+]i rise. At 5–20 μM, methoxychlor killed cells in a concentration‐dependent manner. The cytotoxic effect of methoxychlor was not reversed by chelating cytosolic Ca2+ with 1,2‐bis(2‐aminophenoxy)ethane‐N,N,N’,N’‐tetraacetic acid/AM (BAPTA/AM). Annexin V‐FITC data suggest that methoxychlor (10 and 20 μM) evoked apoptosis in a concentration‐dependent manner. Together, in human OC2, methoxychlor induced a [Ca2+]i rise probably by inducing phospholipase C‐independent Ca2+ release from the endoplasmic reticulum and Ca2+ entry via phospholipase A2‐sensitive channels. Methoxychlor induced cell death that may involve apoptosis.

Vimentin is a potential prognostic factor for tongue squamous cell carcinoma among five epithelial–mesenchymal transition-related proteins

We aimed to investigate the association of the expression levels of five epithelial–mesenchymal transition (EMT)-related proteins (Snail, Twist, E-cadherin, N-cadherin, and Vimentin) with tumorigenesis, pathologic parameters and prognosis in tongue squamous cell carcinoma (TSCC) patients by immunohistochemistry of tissue microarray. The expression levels of Snail, E-cadherin, N-cadherin and Vimentin were significantly different between the tumor adjacent normal and tumor tissues. In tumor tissues, lower E-cadherin and higher N-cadherin levels were associated with a higher grade of cell differentiation, advanced stage of disease, and lymph node metastasis. However, higher Vimentin expression was associated with poor cell differentiation and lymph node metastasis. Patients with low E-cadherin expression had poor disease-specific survival (DSS). Conversely, positive N-cadherin and higher Vimentin expression levels were associated with poor DSS and disease-free survival. Notably, our multivariate Cox regression model indicated that high Vimentin expression was an adverse prognostic factor for DSS in TSCC patients, even after the adjustment for cell differentiation, pathological stage, and expression levels of Snail, Twist, E-cadherin, and N-cadherin. Snail, E-cadherin, N-cadherin, and Vimentin were associated with tumorigenesis and pathological outcomes. Among the five EMT-related proteins, Vimentin was a potential prognostic factor for TSCC patients.

Selective cytotoxic effects of low-power laser irradiation on human oral cancer cells

Low‐power laser irradiation (LPLI) is known to regulate cell proliferation and migration in clinical use. Recent studies have shown that LPLI induces cell death in some certain types of cancer cell lines. However, the cytotoxic selectivity of LPLI for cancer cells is not fully understood. The aim of this study was to compare the cytotoxic effects of LPLI in both human oral cancer OC2 cells and normal human gingival fibroblast (HGF) cells.

Propionibacterium acnes in the Pathogenesis and Immunotherapy of Acne Vulgaris

Acne vulgaris, a multi-factorial disease, is one of the most common skin diseases, affecting an estimated 80% of Americans at some point during their lives. The gram-positive and anaerobic Propionibacterium acnes (P. acnes) bacterium has been implicated in acne inflammation and pathogenesis. Therapies for acne vulgaris using antibiotics generally lack bacterial specificity, promote the generation of antibiotic-resistant bacterial strains, and cause adverse effects. Immunotherapy against P. acnes or its antigens (sialidase and CAMP factor) has been demonstrated to be effective in mice, attenuating P. acnes-induced inflammation; thus, this method may be applied to develop a potential vaccine targeting P. acnes for acne vulgaris treatment. This review summarizes reports describing the role of P. acnes in the pathogenesis of acne and various immunotherapy-based approaches targeting P. acnes, suggesting the potential effectiveness of immunotherapy for acne vulgaris as well as P. acnes-associated diseases.

RelA-Mediated BECN1 Expression Is Required for Reactive Oxygen Species-Induced Autophagy in Oral Cancer Cells Exposed to Low-Power Laser Irradiation.

Low-power laser irradiation (LPLI) is a non-invasive and safe method for cancer treatment that alters a variety of physiological processes in the cells. Autophagy can play either a cytoprotective role or a detrimental role in cancer cells exposed to stress. The detailed mechanisms of autophagy and its role on cytotoxicity in oral cancer cells exposed to LPLI remain unclear. In this study, we showed that LPLI at 810 nm with energy density 60 J/cm2 increased the number of microtubule associated protein 1 light chain 3 (MAP1LC3) puncta and increased autophagic flux in oral cancer cells. Moreover, reactive oxygen species (ROS) production was induced, which increased RelA transcriptional activity and beclin 1 (BECN1) expression in oral cancer cells irradiated with LPLI. Furthermore, ROS scavenger or knockdown of RelA diminished LPLI-induced BECN1 expression and MAP1LC3-II conversion. In addition, pharmacological and genetic ablation of autophagy significantly enhanced the effects of LPLI-induced apoptosis in oral cancer cells. These results suggest that autophagy may be a resistant mechanism for LPLI-induced apoptosis in oral cancer cells.

Effects of Salvia miltiorrhiza ethanolic extract on lipopolysaccharide-induced dental alveolar bone resorption in rats

Background/purpose Salvia miltiorrhiza (SM) Bunge (Labiatae/Lamiaceae; common name danshen) is a Chinese medicine that improves blood circulation and inhibits inflammatory response. Thus, it is used for the treatment of cardiac diseases and inflammation. In this study, we aimed to evaluate the effect of an ethanolic extract of SM (SME) on the dental alveolar bone resorption induced by bacterial lipopolysaccharide (LPS) in rats. Materials and methods An ethanolic extract was prepared from roots of SM. The major constituents of this extract were determined by high-performance liquid chromatography. The activity of the extract was evaluated in a rat model in which the dental alveolar bone resorption was induced by injection of bacterial LPS into the palatal gingiva around the maxillary molar teeth. The effect of SME on the bone resorption was studied by histologic and histomorphometric analysis. Results The number of osteoclasts and the percentage of osteoclasts covering the alveolar bone surfaces were significantly increased in the LPS group compared with those in the phosphate-buffered saline (PBS) group. The number and percentage of the osteoclasts on the bony surfaces were significantly reduced in the SME group in comparison with the LPS group, although it was still higher than the numbers observed in the PBS group. Conclusion Because SME reduced bone resorption caused by the injections of bacterial LPS in rats, we suggest that SME might have a protective effect on dental alveolar bone resorption in periodontitis.

Correlation Between Resonance Frequency Analysis and Bone Quality Assessments at Dental Implant Recipient Sites.

PURPOSE To evaluate whether primary implant stability could be used to predict bone quality, the association between the implant stability quotient (ISQ) value and the bone type at the implant site was evaluated. MATERIALS AND METHODS Ninety-five implant sites in 50 patients were included. Bone type (categorized by Lekholm and Zarb) at the implant site was initially assessed using presurgical dental radiography. During the preparation of the implant site, a bone core specimen was carefully obtained. The bone type was assessed by tactile sensation during the drilling operation, according to the Misch criteria. The primary stability of the inserted implant was evaluated by resonance frequency analysis (RFA). The ISQ value was recorded. The bone core specimen was then examined by stereomicroscopy or microcomputed tomography (micro-CT), and the bone type was determined by the surface characteristics of the specimen, based on Lekholm and Zarb classification. Agreement between the bone quality assessed by the four methods (ie, presurgical radiography, tactile sensation, stereomicroscopy, and micro-CT) was tested by Cohens kappa statistics, whereas the association between the ISQ value and the bone type was evaluated by the generalized linear regression model. RESULTS The mean ISQ score was 72.6, and the score was significantly influenced by the maxillary or mandibular arch (P = .001). The bone type at the implant sites varied according to the assessment method. However, a significant influence of the arch was repeatedly noted when using radiography or tactile sensation. Among the four bone-quality assessment methods, a weak agreement existed only between stereomicroscopy and micro-CT, especially in the maxilla (κ = 0.469). A negative association between the ISQ value and the bone type assessed by stereomicroscopy or by micro-CT was significant in the maxilla, but not in the mandible, after adjustments for sex, age, and right/left side (P = .013 and P = .027 for stereomicroscopy and micro-CT, respectively). CONCLUSION The ISQ value was weakly associated with the bone type when assessed by stereomicroscopy or micro-CT in the maxilla. Caution is necessary if RFA is used as a tool to evaluate bone quality at the implant site, especially in the mandible.