Effects of systemic Bifidobacterium longum and Lactobacillus rhamnosus probiotics on the ligature-induced periodontitis in rat.

Background/purpose: Probiotics might be beneficial in preventing periodontitis. Effects of Bifidobacterium and Lactobacillus on periodontitis were examined using the ligature-induced rat model. Materials and methods: Thirty-five male Sprague-Dawley rats were divided into control, ligation, Bifidobacterium longum (BL986), Lactobacillus rhamnosus (LRH09), and combination groups. Periodontitis was induced in maxillary second molars. From the day before ligation, phosphate-buffered saline (for control and ligation groups) or probiotics (2 × 109 CFU/g for probiotic groups) were fed daily. On day 8, gingival mRNA expressions for interleukin (IL)-1ß, IL-6, tissue necrosis factor (TNF)-α, IL-10, and NF-κB were determined via qPCR. Micro-computed tomography (µCT) and histomorphometry were employed to examine periodontal destruction. Results: Compared to the ligation group, mRNA of IL-1ß, TNF-α, IL-6, and NF-κB in probiotic groups were significantly decreased, but IL-10 was increased. Besides, the IL-10 was more significant in the combination group than in single-use group. Through µCT, the cementoenamel junction (CEJ)-to-bone distance and trabecular separation in combination group were less than that in ligation group, although the bone volume fraction and trabecular number/thickness showed an increase in three probiotic groups. Histopathologically, the combination group had significantly smaller gingival inflammatory cell-infiltrated area and CEJ-to-epithelium distance than the ligation group and the group with BL986 or LRH09. Additionally, the CEJ-to-bone distance was significantly smaller in the combination group than in the ligation and BL986 groups. Conclusion: Systemic combination of BL986 and LRH09 had a synergistic effect on enhancing IL-10 and ameliorating the induced experimental periodontitis, although the single-use still presented partially alleviative effects.

Investigating the differences in nutritional status between successfully weaned and unsuccessfully weaned respirator patients.

Long-term respirator users admitted to intensive care units need to be transferred to a respiratory care center (RCC) for weaning. It may cause malnutrition in critical care patients, which may manifest as a reduction in respiratory muscle mass, lower ventilatory capacity, and decreased respiratory tolerance. This study aimed to assess that if the patients' nutritional status were improved, it could help RCC patients to wean from respirators. All participants were recruited from the RCC of a medical foundation in the city and Taipei Tzu Chi Hospital. The indicators include serum albumin level, respirator detachment index, maximum inspiratory pressure (PImax), rapid shallow breathing index, and body composition measurements. We recorded the length of hospital stay, mortality, and RCW (respiratory care ward) referral rate for these participants and analyzed the differences in relevant research indicators between those who were and weren't weaned off. 43 of 62 patients were weaned from respirators, while 19 failed. The resuscitation rate was 54.8%. Patients with respirator weaning had a lower number of RCC admission days (23.1 ± 11.1 days) than respirator-dependent patients (35.6 ± 7.8 days, P < 0.05). The PImax of successfully weaned patients had a greater reduction (- 27.09 ± 9.7 cmH2O) than unsuccessful ones (- 21.4 ± 10.2 cmH2O, P < 0.05). The Acute Physiology and Chronic Health Evaluation II (APACHE II) scores of successfully weaned patients (15.8 ± 5.0) were lower than those who were not (20.4 ± 8.4, P < 0.05). There was no significant difference in serum albumin levels between the two groups. In the successfully weaned patients, the serum albumin concentration was increased from 2.2 ± 0.3 to 2.5 ± 0.4 mg/dL, P < 0.05. Improved nutritional status can help RCC patients to wean from respirators.

Staphylococcus aureus enhances gelatinase activities in monocytic U937 cells and in human gingival fibroblasts.

Background/purpose: Staphylococcus aureus (S. aureus) has been suggested to be an initiative pathogen in peri-implantitis because of the solid affinity to titanium. However, the detail pathogenesis for the peri-implantitis initiation by S. aureus is still lacking. This study aimed to in vitro examine the gelatinases' activities of monocytic U937 cell and human gingival fibroblast after challenges with S. aureus lipoteichoic acid (LTA) and peptidoglycan (PGN). Materials and methods: Releases of gelatinases, including matrix metalloproteinase (MMP)-2 and -9, from cells were measured by zymography. The releases were further examined after being given the S. aureus LTA/PGN. Roles of nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) pathways on the enzyme releases were examined by administrating inhibitors. Results: S. aureus LTA and PGN increased the activities of pro-MMP-9 from U937 cells and pro-MMP-2 and MMP-2 from gingival fibroblasts. By giving the NF-κB inhibitor, the enhanced gelatinase activities in both cells were attenuated. In U937 cells, the enhanced pro-MMP-9 could further be attenuated by MAPK inhibitors, including extracellular signal-regulated kinase 1 and 2 (ERK1/2), P38 MAPK, and c-Jun N-terminal kinase (JNK) inhibitors; however, the attenuation by MAPK inhibitors could not be observed for MMP-2 in gingival fibroblasts. Nevertheless, in gingival fibroblasts, the pro-MMP-2 could be attenuated by JNK inhibitor. Conclusion: S. aureus could enhance gelatinase activities of gingival fibroblasts and U937 cells, via NF-κB. The MAPK pathway was also involved in MMP-9 activity of U937 cells; however, the involvement of MAPK in MMP-2 activity of gingival fibroblasts was questioned.

Cyanidin-3-O-glucoside downregulates ligation-activated endoplasmic reticulum stress and alleviates induced periodontal destruction in rats.

OBJECTIVE: Cyanidin-3-O-glucoside (C3G), a dietary anthocyanin, possesses various biological properties, including alleviating endoplasmic reticulum (ER) stress. This study examined the effect of C3G on periodontitis via ER stress in rats. DESIGN: Periodontitis was induced by placing silk sutures around maxillary second molars. C3G (0, 3, or 9 mg/kg) was fed on the day before ligation (10 rats/group). Further, 10 non-ligation control rats received deionized water. On day 8, gingivae were obtained to determine CCAAT/enhancer-binding protein homologous protein (CHOP), c-Jun N-terminal kinase (JNK), phospho-JNK (p-JNK), and nuclear factor-κB (NF-κB) by immunoblotting. Periodontal destruction was evaluated using micro-computed tomography (µCT) and histology. RESULTS: Gingival expression of CHOP, p-JNK/JNK, and NF-κB significantly increased in ligation rats (0 mg/kg C3G) than that in controls. However, protein expression in ligation groups presented a negative association with C3G concentration. By µCT, the distance of cemento-enamel junction to bone significantly increased in ligation groups; however, distances showed a negative association with C3G concentration. In the region of interest, bone volume and trabecular thickness and number significantly decreased in ligation groups but they were positively associated with C3G concentration. In terms of trabecular separation, opposite results were found. Histologically, infiltrated connective tissue (ICT) and periodontal destructions increased in ligation groups; however, they were negatively associated with C3G concentration. Moreover, ICT area is positively correlated with µCT- and histologically measured destructions and protein expression of CHOP, p-JNK/JNK, or NF-κB. CONCLUSION: C3G promotes favorable modulation of ER stress and alleviates destruction of periodontitis, which may imply a new strategy.

Porphyromonas gingivalis lipopolysaccharide and gingival fibroblast augment MMP-9 expression of monocytic U937 cells through cyclophilin A.

BACKGROUND: Intercellular cross-talking was suggested in matrix metalloproteinase (MMP)-9 expression with unknown mechanisms. Studies showed cyclophilin A (CypA) playing an important role in regulating MMP-9 expression in varied diseases. The aim of the study was to examine the CyPA on the MMP-9 augmentation in monocytic U937 cells after Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS) treatment and human gingival fibroblast (hGF) co-culture. METHODS: In independent culture or co-culture of hGF and U937 cell, quantitative real-time polymerase chain reaction (qPCR) and zymography were selected to examine the mRNA and protein activity of MMP-9, respectively. The CyPA expression was determined by qPCR. RESULTS: LPS could enhance MMP-9 mRNA expression and enzyme activity in U937 cell. However, the enhancements were not observed in hGF. Similarly, LPS enhanced CyPA mRNA in U937, but not in hGF. After co-cultured with hGF, however, MMP-9 and CyPA in U937 increased regardless of the presence/absence of LPS. In U937 cells, the extra-supplied CyPA increased MMP-9 mRNA and enzyme activity, whereas the CyPA inhibitor, cyclosporine A, suppressed the LPS- and co-culture-enhanced MMP-9. Moreover, the inhibitors for MAP kinase, including PD98059 (ERK) and SP600125 (JNK), suppressed the CyPA-enhanced MMP-9 in U937. CONCLUSION: Through the CyPA pathway, the LPS and the hGF could augment the MMP-9 expression in the U937 cells.

Bone formation with functionalized 3D printed poly-ε-caprolactone scaffold with plasma-rich-fibrin implanted in critical-sized calvaria defect of rat.

BACKGROUND/PURPOSE: Space-making is one of the essential factors for bone regeneration in severe bony defect. To test the hypothesis that an appropriately designed scaffold may be beneficial for the bone formation in defect, the new bone formed in the critical-size calvarial defect of rat was examined after implanted with a 3D-printed poly-ɛ-caprolactone (PCL) scaffold, retaining with and without plasma rich fibrin (PRF). MATERIALS AND METHODS: Thirty-two rats were divided into four groups (control, PCL, PRF, and PCL-plus-PRF). A custom-made 3D-printed PCL scaffold, 900 µm in pore size, retaining with and without PRF, was implanted into a critical-sized calvarial defect, 6 mm in diameter. Animals were sacrificed at week-4 or 8 after implantation for assessing the new bone formation by dental radiography, micro-computed tomography (µ-CT), and histology. RESULTS: By radiography and µ-CT, significantly greater mineralization areas/volumes were observed in defects with 3D-printed scaffold groups compared to that without the scaffold in both two-time points. However, no advantage was found by adding PRF. Histology showed that bone tissues grew into the central zone of the critical defect when 3D-printed PCL scaffold was present. In contrast, for the groups without the scaffolds, new bones were formed mostly along defect borders, and the central zones of the defects were collapsed and healed with thin connective tissue. CONCLUSION: Our results suggest that the use of a 900 µm pore size 3D-printed PCL scaffold may have the potential in facilitating the new bone formation.

Recommendations for treating stage I-III periodontitis in the Taiwanese population: A consensus report from the Taiwan Academy of Periodontology.

BACKGROUND/PURPOSE: Based on the fundamental of the S3-level clinical practice guideline (CPG) for treating stage I-III periodontitis developed by the European Federation of Periodontology (EFP), this consensus report aimed to develop treatment recommendations for treating periodontitis in the Taiwanese population. METHODS: The report was constructed by experts from the Taiwan Academy of Periodontology. The following topics were reviewed: (a) the prevalence of periodontitis in Asia and current status of treatment in Taiwan; (b) specific anatomical considerations for treating periodontitis in Asians; (d) educational and preventive interventions and supragingival plaque control; (d) subgingival instrumentation and adjunctive treatment; (e) surgical periodontal therapy; and (f) maintenance and supportive periodontal care. Recommendations were made according to the evidences from the EFP CPG, the published literature and clinical studies in Asians, and the expert opinions. RESULTS: The treatment recommendations for the Taiwanese population were generally in parallel with the EFP CPG, and extra cautions during treatment and maintenance phases were advised due to the anatomical variations, such as shorter root trunk, higher prevalence of supernumerary distolingual root and lingual bony concavity in mandibular posteriors, and thinner anterior labial plate, of the Asian population. CONCLUSION: The EFP CPG could be adopted for treating periodontitis and maintaining periodontal health of the Taiwanese population, and anatomical variations should be cautious when the treatment is delivered.

Fibroblast-enhanced cyclophilin A releasing from U937 cell upregulates MMP-2 in gingival fibroblast.

OBJECTIVE: This in vitro study aimed to evaluate the expression of cyclophilin A (CyPA) in U937 monocytic cells after coculturing with the human gingival fibroblasts (HGFs) and the effect of CyPA on the augmentation of MMP-2 expression in the coculture environment. BACKGROUND: Leukocyte infiltration in gingival connective tissue is one of the major findings in the lesions of inflammatory periodontal diseases. A crosstalk between the resident gingival fibroblasts and the recruited inflammatory cells that promote the expression of matrix metalloproteinases (MMPs) was proposed based on recent findings, whereas the cluster of differentiation 147 (CD147)-CyPA pathway was suggested to be involved with the crosstalk. MATERIAL AND METHODS: CyPA was released into media, in the independent or transwell coculture of HGF and U937 cells, as determined by enzyme-linked immunosorbent assay, whereas intracellular mRNA expressions for CyPA and MMP-2 were examined by quantitative real-time polymerase chain reaction, in the transwell coculture or conditional medium models. Zymography was conducted to analyze the activities of pro-MMP-2/MMP-2 released into the media. RESULTS: (a) A significantly increased CyPA protein level was observed in the transwell coculture media compared with that in the independent culture. (b) The transwell coculture-enhanced mRNA expression for CyPA was noticed in U937 cells but not in HGFs. After adding with HGF-conditioned medium, the mRNA enhancement in U937 cells occurred in a dose-dependent manner. (c) Although the MMP-2 activities significantly increased after transwell coculturing, the MMP-2 mRNA enhancement was observed only in HGFs. (d) Exogenous CyPA could enhance MMP-2 activities in HGFs in a dose-dependent manner. However, the CyPA antagonist reduced the MMP-2 activities in the transwell cocultures. (e) Moreover, the CyPA-enhanced MMP-2 activity in HGF was decreased significantly by the pathway inhibitor for c-Jun amino-terminal kinase (JNK). CONCLUSION: Based on the present findings, we suggest that gingival fibroblasts could enhance the CyPA release from U937 cells, via the JNK pathway, resulting in MMP-2 enhancement in fibroblasts. The finding shed light on a new mechanism of cellular interaction involving MMP-2 and CyPA, in two cells.

Immediate hyperbaric oxygen after tooth extraction ameliorates bisphosphonate-related osteonecrotic lesion in rats.

BACKGROUND: This study aims to assess whether hyperbaric oxygen (HBO) applied immediately after tooth extraction could ameliorate medication-related osteonecrosis of the jaw in rats. METHODS: To evaluate whether osteonecrosis could be successfully induced, healing of extraction maxillary molars was examined in 40 female Sprague Dawley rats received zoledronic acid (7.5 µg/kg) plus dexamethasone (1 mg/kg). Rats were divided into four groups, receiving zero, two, four, or seven injection(s) for 7 days, respectively. Effect of HBO, pressurized to 2.5 atmospheres absolute (ATA) at rate of 0.15 ATA/min with 100% oxygen for 90 minutes, applied immediately after tooth extraction, on the development of osteonecrosis was evaluated. Lesions among groups were compared by size of ulceration, exact area (mm2 ) or relative area (%), and by histology. RESULTS: Unhealed ridge was observed in all nine rats in four and seven injection groups, but none of 10 rats in the control (non-injection) group. Immediate HBO significantly reduced the lesions in rats that received four injections, regardless of the distribution and the total/relative areas of lesions (P <0.01). Histological findings showed the lesions were uncovered epithelium and severe tissue inflammation. CONCLUSION: This is the first in vivo study demonstrating the HBO applied immediately after tooth extraction effectively decreases the development of medication-related osteonecrosis.

Ameliorative effect of hesperidin on ligation-induced periodontitis in rats.

BACKGROUND: This study evaluated the ameliorative effect of hesperidin (HES), an anti-inflammatory flavanone, in rats with ligation (Lig)-induced periodontitis. METHODS: A total of 48 rats were randomly divided into non-ligation group (NL), Lig group, and two ligation-plus-HES groups (L+H). HES was administered immediately after ligature placement at a dose of 75 or 150 mg/kg by intragastric feeding. Destruction of the ligated maxillary second and mandibular first molars were evaluated by dental radiography, microcomputed tomography (micro-CT), and histometry performed after sacrificing the rats on the seventh day. The expression levels of interleukin (IL)-1ß, IL-6, and inducible nitric oxide synthase (iNOS) messenger (m)RNAs in the gingiva were determined by reverse-transcription polymerase chain reaction. The expression of iNOS was examined by immunohistochemistry. RESULTS: The dental radiography and micro-CT findings revealed significantly increased alveolar bone loss in the Lig group, which was significantly prevented by HES. The histometry results revealed less gingival inflammation and connective tissue loss in the L+H groups compared with that in the Lig group. The mRNA expression levels of IL-6, IL-1 ß, and iNOS were significantly increased in the Lig group but were reduced in the L+H groups. The immunostaining results showed that the ligation-induced iNOS expression was also decreased by HES. CONCLUSIONS: Oral administration of HES promotes an ameliorative effect against the ligation-induced alveolar bone loss and effectively inhibits the production of proinflammatory mediators in rats with experimentally induced periodontitis. Therefore, HES may be a good candidate for modulating oral inflammatory diseases.

Accuracy of Implant Placement with a Navigation System, a Laboratory Guide, and Freehand Drilling.

PURPOSE: Computer-aided surgery under navigation system guidance is widely applied in dental implant procedures. However, the accuracy of drilling with such navigation systems has not been comparatively evaluated alongside those of laboratory guide-based and freehand drilling. Therefore, this study aimed to compare the accuracies of these three drilling systems. MATERIALS AND METHODS: A navigation system, a laboratory guide, and freehand drilling were used to drill 150 holes on 30 cast models. Two master models-one each for the maxilla and mandible-were prepared with the idea of placing five implants per cast. After drilling five holes on each cast, postoperative cone beam computed tomography images were acquired to measure the magnitude of errors. RESULTS: The navigation system and laboratory guide were more accurate than freehand placement with respect to total errors at the entry and apex, lateral error at the apex, and angular error. The navigation system was more accurate than the laboratory guide with respect to angular error. Laboratory guide-based drilling was more accurate than freehand drilling in terms of lateral error at entry. CONCLUSION: In comparison with the laboratory guide and freehand placement, the navigation system exhibited lower angular and axial errors. Despite its higher accuracy, the navigation system requires the operator to pay greater attention.

Periodontal status of tooth adjacent to implant with peri-implantitis.

OBJECTIVES: To evaluate the relationship between peri-implantitis and the periodontal health of the adjacent tooth, the periodontal status of the teeth adjacent and contralateral to the implants with and without peri-implantitis. METHODS: Fifty-three subjects with existing dental implants and chronic periodontitis were examined in this cross-sectional study. Seventy implants were categorized into peri-implantitis (n = 42) and healthy/mucositis (n = 28) groups. The periodontal and peri-implant status, including probing depth (PD), clinical attachment level (CAL), and gingival recession (GR) were measured at 6 sites around the implants and the teeth adjacent and contralateral to those implants. In total 560 sites of the 70 teeth/implant sets, the association between the periodontal status at the near and away sites of the teeth (according to implant) and the implant status (without/with peri-implantitis) was examined. RESULTS: A significantly different mean PD (5.01 ±â€¯1.69, 4.42 ±â€¯1.8, 3.55 ±â€¯0.88, and 3.71 ±â€¯1.07 mm, p < 0.001) and CAL (6.02 ±â€¯2.36, 4.89 ±â€¯2.04, 4.35 ±â€¯1.11, and 4.35 ±â€¯1.5 mm, p < 0.001) were noted at the near sites of the teeth adjacent to the implants with peri-implantitis when compared with the away sites of adjacent and contralateral teeth and the near sites of contralateral teeth. With generalized estimating equation (GEE), the presence of peri-implantitis (ß â€¯= 1.041 mm, confidence interval = 0.646-1.435, and p < 0.001; ß â€¯= 0.857 mm, confidence interval = 0.279-1.434, and p < 0.004) and tooth location (ß â€¯= 0.65 mm, confidence interval = 0.4-0.9, and p < 0.001; ß â€¯= 0.682 mm, confidence interval = 0.34-1.024, and p < 0.001) were significantly associated with the values of the PD and CAL of the teeth. Moreover, the factor of examining sites (i.e. near and away sites of the tooth) was significantly associated with CAL (ß = 0.304 mm, confidence interval = 0.019-0.588, and p = 0.036) and GR (ß = 0.136 mm, confidence interval = 0.02-0.252, and p = 0.022). CONCLUSION: The existence of peri-implantitis, the tooth location, and the examining site are significantly associated with the periodontal measurements of the remaining teeth. CLINICAL SIGNIFICANCE: Peri-implant health is related to the periodontal health of the natural teeth close to the dental implant.

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.

Crown morphology of the mandibular first molars with distolingual roots.

BACKGROUND/PURPOSE: Most mandibular first molars have two roots. A major common variation of this tooth is the presence of a distolingual root, which is a common Mongoloid trait in certain populations. The aim of this article was to examine crown morphology in relation to the presence of the distolingual root. MATERIALS AND METHODS: Using dental casts, the crown morphology of 141 mandibular first molars from 71 Taiwanese individuals was analyzed. Periapical radiographs were used to detect distolingual roots. The length and width of the crowns and the crown units (i.e., trigonid and talonid) were measured. Ten intercuspal distances and five cusp angles were examined. RESULTS: The buccolingual dimension of the crown and its ratio to the mesiodistal dimension were significantly increased in molars with a distolingual root, compared to molars without a distolingual root. Mesiodistal crown dimensions were similar; however, the crown unit dimensions were different: molars with a distolingual root had a shorter mesiodistal trigonid dimension but a longer talonid dimension, compared to molars without a distolingual root. The intercuspal distances from the three buccal cusps to the distolingual cusp were significantly longer, however, the distance between the mesiobuccal cusp and mesiolingual cusp was significantly shorter in teeth with a distolingual root than in teeth without a distolingual root. A significantly wider mesiolingual angle and narrower distolingual angle were observed in molars with a distolingual root, compared to molars without a distolingual root. CONCLUSION: The presence of a distolingual root significantly increased the buccolingual dimension of the crown and the location of distolingual cusp is significantly closer to the lingual side.

Gelatinases and extracellular matrix metalloproteinase inducer are associated with cyclosporin-A-induced attenuation of periodontal degradation in rats.

BACKGROUND: The present study aims to examine the inhibitory effect of cyclosporin-A (CsA) on periodontal breakdown and to further explore the correlations of CsA-induced attenuation of periodontal bone loss with the expressions of gelatinases (i.e., matrix metalloproteinase [MMP]-2 and MMP-9) and extracellular matrix metalloproteinase inducer (EMMPRIN). METHODS: Forty Sprague-Dawley rats were randomly divided into four groups: 1) control; 2) CsA; 3) ligature (Lig); and 4) ligature plus CsA (Lig + CsA). The CsA group received 10 mg ⋅ Kg(-1) ⋅ d(-1) CsA for 8 days. The Lig group received silk ligature on selected molars. The Lig + CsA group received silk ligature and CsA treatment. The inhibitory effects of CsA on the ligature-induced periodontal breakdown was examined with microcomputed tomography (micro-CT) and histometric analyses to analyze the amount of attachment loss, crestal bone loss, connective tissue attachment, and the surface area with inflammatory cell infiltration. The effects of CsA on ligature-induced expressions of gelatinases and EMMPRIN in gingival tissues were examined with Western blotting and zymography, respectively. RESULTS: By micro-CT and histology, the Lig + CsA group had significantly more periodontal breakdown than the control and CsA groups but less periodontal breakdown than the Lig group. Consistent results were found for the expressions of gelatinases and EMMPRIN among the groups demonstrating that the Lig + CsA group had significantly less gingival protein expression of gelatinases and EMMPRIN than the Lig group. CONCLUSIONS: CsA inhibited the expressions of gelatinase MMPs and EMMPRIN and partially prevented the periodontal breakdown in ligature-induced experimental periodontitis. The CsA-induced attenuation of periodontal bone loss was strongly correlated positively with the expressions of MMP-2, MMP-9, and EMMPRIN in gingiva.

Cyclosporine A enhances gingival ß-catenin stability via Wnt signaling.

BACKGROUND: Cyclosporine A (CsA) increases ß-catenin messenger RNA (mRNA) and protein expression. The present study demonstrates that Wnt/ß-catenin signaling inhibits ß-catenin degradation in the gingiva. METHODS: Forty 5-week-old male Sprague-Dawley rats were assigned to two study groups after healing from right maxillary molar extractions. The rats in the experimental group were fed 30 mg/kg CsA daily for 4 weeks, whereas the control rats were fed mineral oil. At the end of the study, all rats were sacrificed, and the gingivae were obtained. The gingival morphology after CsA treatment was evaluated by histology, and the genes related to Wnt/ß-catenin signaling were initially screened by microarray. Polymerase chain reaction, Western blotting, and immunohistochemistry were used to examine the mRNA and protein expression of proliferating cell nuclear antigen, cyclin D1, E-cadherin, ß-catenin, Dvl-1, glycogen synthase kinase-3ß, axin-1, and adenomatous polyposis coli (APC). Phosphoserine and ubiquitinylated ß-catenin were detected after immunoprecipitation. RESULTS: In rats treated with CsA, overgrowth of gingivae was observed, and altered expression of genes related to Wnt/ß-catenin signaling was detected by the microarray. The gingival mRNA and protein expression profiles for genes associated with Wnt/ß-catenin signaling further confirmed the effect of CsA: ß-catenin and Dvl-1 expression increased, but APC and axin-1 expression decreased. Western blotting and immunohistochemistry showed decreases in ß-catenin serine phosphorylation (33/37) and ubiquitinylation in the gingivae of CsA-treated rats. CONCLUSION: CsA-enhanced gingival ß-catenin stability may be involved in gene upregulation or ß-catenin degradation via the Wnt/ß-catenin pathway.

Role of Shh and TGF in cyclosporine-enhanced expression of collagen and α-SMA by gingival fibroblast.

OBJECTIVE: Cyclosporine-A (CsA)-induced gingival overgrowth may arise from an alteration in stoma matrix homeostasis. Sonic hedgehog (Shh) plays a key role during embryogenic development and fibrotic progression, and may be involved in CsA-altered gingival matrix homeostasis. METHODS: Using the reverse transcription-polymerase chain reaction and Western blot analysis, we investigated the mRNA and protein expressions of Shh, type 1 collagen (COL1), alpha-smooth muscle actin (α-SMA) and transforming growth factor-beta (TGF-ß) in human gingival fibroblasts after CsA treatments. The effect of Shh on CsA-induced alterations was further evaluated by the extra-supplement or inhibition of Shh or TGF-ß. RESULTS: Cyclosporine-A enhanced COL1, α-SMA, Shh and TGF-ß expressions in human gingival fibroblasts. The exogenous Shh/TGF-ß augmented the expression of COL1 and α-SMA, and the Shh/TGF-ß inhibition suppressed the CsA-enhanced COL1 and α-SMA expressions. Moreover, Shh mRNA and protein expressions increased if extra-supplementing the exogenous TGF-ß, whereas the CsA-upregulated Shh was mitigated by the TGF-ß pathway inhibitor. However, neither exogenous Shh nor the Shh pathway inhibitor alters TGF-ß expression or CsA-up-regulated TGF-ß expression. CONCLUSIONS: Shh, regulated by TGF-ß, mediates CsA-altered gingival matrix homeostasis.

Role of transforming growth factor-beta1 in cyclosporine-induced epithelial-to-mesenchymal transition in gingival epithelium.

BACKGROUND: It has been proposed that cyclosporin A (CsA) may induce epithelial-to-mesenchymal transition (EMT) in gingiva. The aims of the present study are to confirm the notion that EMT occurs in human gingival epithelial (hGE) cells after CsA treatment and to investigate the role of transforming growth factor beta1 (TGF-ß1) on this CsA-induced EMT. METHODS: The effects of CsA, with and without TGF-ß1 inhibitor, on the morphologic changes of primary culture of hGE cells were examined in vitro. The changes of protein and messenger RNA (mRNA) expressions of two EMT markers (E-cadherin and alpha-smooth muscle actin) in the hGE cells after CsA treatment with and without TGF-ß1 inhibitor were evaluated with immunocytochemistry and real-time polymerase chain reaction. RESULTS: The epithelial cells became spindle-like, elongated, and disassociated from neighboring cells and lost their original cobblestone monolayer pattern when CsA was added. However, the epithelial cells stayed in their original cobblestone morphology with treatment of TGF-ß1 inhibitor on top of the CsA treatment. When CsA was given, the protein and mRNA expressions of E-cadherin and α-SMA were significantly altered, and these alterations were significantly reversed with pretreatment of TGF-ß1 inhibitor. CONCLUSIONS: CsA could induce Type 2 EMT in gingiva by changing the morphology of epithelial cells and altering the EMT markers/effectors. The CsA-induced gingival EMT is dependent or at least partially dependent on TGF-ß1.

Association of CCL5 and CCR5 gene polymorphisms with periodontitis in Taiwanese.

BACKGROUND: It has been suggested that genetic factors may predispose individuals to periodontal diseases. The present case-control study aims to test whether the -403 single nucleotide polymorphism of chemokine ligand 5 (CCL5-403) and the 32-bp deletion of CCR5 (CCR5Δ32) polymorphisms are associated with susceptibility to chronic and aggressive periodontitis. METHODS: Taiwanese participants (N = 213) were grouped into control group (CG), generalized aggressive periodontitis (GAgP), or chronic periodontitis (CP) groups. DNA samples were obtained from peripheral blood. CCL5-403, evaluated by polymerase chain reaction-restriction fragment length polymorphism, and CCR5Δ32, evaluated by polymerase chain reaction, were compared among the three groups. RESULTS: There was a significant association between type of periodontitis and having allele A or G in the CCL5-403 polymorphism. GAgP patients were 3.7 times more likely than CP patients and 2.0 times more likely than CG patients to have allele A, instead of allele G, in CCL5-403. GAgP patients were 3.1 times more likely than CG patients to have AG versus GG genotype. GAgP patients were also 5.0 and 19.8 times more likely than CP patients to have AG and AA genotypes, respectively, compared to GG. For the CCR5Δ32 polymorphism, no association was found between the type of periodontitis and having different genotype or allele distributions among GAgP, CP, or CG patients. CONCLUSION: The single nucleotide polymorphism of CCL5-403 G substitution by A may play a role in AgP; however, the CCR5Δ32 polymorphism may not.

Serum hyperglycemia might be not related to fat composition of diet and vegetable composition of diet might improve sugar control in taiwanese diabetic subjects.

OBJECTIVE: This is an Asian study, which was designed to examine the correlations between biochemical data and food composition of diabetic patients in Taiwan. METHODS: One hundred and seventy Taiwanese diabetic patients were enrolled. The correlations between biochemical data and diet composition (from 24-hour recall of intake food) of these patients were explored (Spearman correlation, p < 0.05). Diet components were also correlated with each other to show diet characteristics of diabetic patients in Taiwan. Linear regression was also performed for the significantly correlated groups to estimate possible impacts from diet composition to biochemical data. RESULTS: Postprandial serum glucose level was negatively correlated with fat percentage of diet, intake amount of polyunsaturated fatty acid and fiber diet composition. Hemoglobin A1c was negatively correlated with fat diet, polyunsaturated fatty acid and vegetable diet. Fat composition, calorie percentage accounted by polyunsaturated fatty acid and monounsaturated fatty acid in diet seemed to be negatively correlated with sugar percentage of diet and positively correlated with vegetable and fiber composition of diet. Linear regression showed that intake amount of polyunsaturated fatty acid, calorie percentage accounted by polyunsaturated fatty acid, fat percentage of diet, vegetable composition of diet would predict lower hemoglobin A1c and postprandial blood sugar. Besides, higher percentage of fat diet composition could predict higher percentage of vegetable diet composition in Taiwanese diabetic patients. CONCLUSION: Fat diet might not elevate serum glucose. Vegetable diet and polyunsaturated fatty acid diet composition might be correlated with better sugar control in Taiwanese diabetic patients.