Gingival fibroblast response to cyclosporin A and transforming growth factor β1

This study investigates a potential role for TGFβ₁ in the pathogenesis of cyclosporin A-induced gingival overgrowth (CsA-OG). TGFβ₁ was localized immunohistochemically in the connective tissue of both normal gingiva and CsA-OG. Intense staining for TGFβ₁ was detected at the tips of the dermal papillae of the overgrown gingiva. In addition, fibroblasts derived from healthy gingiva and fibroblasts derived from CsA-OG were cultured both as monolayers or embedded in a 3D-collagen gel. Fibroblast activity was monitored in terms of protein and collagen production in the presence of (i) 1 ng/ml TGFβ₁ (ii) 500 ng/ml CsA, or (iii) 500 ng/ml CsA and 1 ng/ml TGFβ₁. In monolayer culture TGFβ₁ significantly increased protein and collagen production in all cell strains (p<0.05); however, there was no difference in response between fibroblasts from overgrown and healthy tissue. The production of both protein and collagen was significantly lower in the presence of the combination of CsA and TGFβ₁ when compared with the maximal stimulation produced by TGFβ₁ alone. In gel, TGFβ₁ significantly elevated matrix production by all overgrown cell strains (p<0.05) but had little or no effect on the normal cell strains. The combination of CsA and TGFβ₁ in gel cultures reduced protein and collagen production by overgrown cell strains compared with TGFβ¹ alone. It is concluded that the cellular activity of gingival fibroblasts is dependant on culture conditions and that fibroblasts derived from overgrown gingival tissue are more responsive to TGFβ₁ than normal gingival fibroblasts when cultured in type I collagen gel.     

Comparison of physical properties of polycarboxylate-based and conventional root canal sealers.

Several physical properties of nine commercial root canal sealers were evaluated in vitro and were compared with those of two experimental endodontic materials based on polycarboxylate formulations. Flow, setting time, compressive strength, radiopacity, adhesion to root dentin, and solubility were evaluated. The zinc oxideeugenol root canal sealers were typically of low strength and high solubility. These sealers and the Diaket polyvinyl resin sealer showed no adhesion to dentin. The epoxy resin AH26 showed superior properties with respect to strength, flow, radiopacity, and adhesion; solubility of this material was high. The polycarboxylate formulation showed significantly higher values over the commercial sealers in properties of strength, adhesion, and reduced solubility. The tensile adhesive bond strength of the polycarboxylate to root dentin was twice that of AH26. A wide variation in properties of the commercial materials tested showed the empirical nature of these "sealers." Further testing of polycarboxylate endodontic sealers is indicated.     

Effect of tube feeding on hippocampal-dependent memory in SAMP1 mice

This study examined the effect of tube feeding on hippocampal Fos induction and spatial performance in a water maze task in senescenceaccelerated mice (SAMP1). Tube feeding accelerated the age-related decline in spatial memory and decreased Fos induction in the hippocampal CA1 region in aged SAMP1 mice. The results suggest that tube feeding in aged SAMP1 mice reduces input activity in the hippocampus, thereby leading to senile memory deficits.     

Role of gene E2f1 in susceptibility to bacterial adherence of oral streptococci to tooth surfaces in mice

Dental plaque is composed of a biofilm community of microorganisms on teeth that coats the oral cavity, including attaching to the teeth, and provides a protective reservoir for oral microbial pathogens, which are the primary cause of persistent and chronic infectious diseases. Oral streptococci are pioneering organisms that play an important role in biofilm formation on tooth surfaces as well as being primary causative agents of dental caries. The purpose of this study was to clarify the role of the E2f1 gene in susceptibility to dry mouth and bacterial adherence of oral streptococci to tooth surfaces in animal model experiments. A mutation of the E2f1 gene in mice is known to cause enhanced T-lymphocyte proliferation, leading to testicular atrophy, splenomegaly, salivary gland dysplasia, and other systemic and organ-specific autoimmunity. We found a decreased volume of saliva production and protein production rate, along with increased amylase activity, IgA concentration, and mucin 1 concentration in E2F-1(-/-) mice as compared with the control C57BL/6 mice. Further, we quantified the recolonization of oral streptococci in E2F-1(-/-) mice and found that a higher number of some oral streptococci were colonized on the teeth of these mice. In particular, following oral ingestion of 1% sucrose in water, the colonization of Streptococcus mutans increased in comparison with other streptococci. Our results suggest that the E2f1 gene may affect susceptibility for oral biofilm formation by streptococci in humans with dry mouth.     

The supernumerary cheek tooth in tabby/EDA mice-a reminiscence of the premolar in mouse ancestors

OBJECTIVE: A supernumerary cheek tooth occurs mesially to the first molar in tabby/EDA (Ta) mice affected by hypohidrotic ectodermal dysplasia. The supernumerary tooth (S) has been hypothetically homologized to the premolar, which has disappeared during mouse evolution. DESIGN: This hypothesis was tested using available morphological data on the lower cheek teeth in wild type (WT) and Ta mice. RESULTS: The presence of S is accompanied by a reduction in the mesial portion of the M(1) in mutant mice. 3D reconstructions suggest that the S in Ta homo/hemizygous embryos originates from a split off the mesial portion of the first molar (M(1)) cap. In WT embryos, two vestigial tooth primordia are transiently distinct in front of the M(1). The distal vestige has the form of a wide bud and participates during the development of the mesial portion of the M(1). This bud has been homologized with the vestigial primordium of the fourth premolar of mouse ancestors. The premolar disappearance coincided with a mesial lengthening of the M(1) during mouse evolution. The incorporation of the distal premolar vestige into the mesial part of the M(1) in WT embryos can be regarded as a repetition of the premolar disappearance during evolution. CONCLUSION:: Ontogenetic and phylogenetic data support that the S in Ta mice arises due to the segregation of the distal premolar vestige from the molar dentition and thus represents an evolutionary throwback (atavism).     

小鼠Notch1胞外段高抗原区编码基因的克隆与表达

目的 :克隆小鼠Notch1胞外段高抗原区编码基因 ,并进行原核表达及蛋白纯化 ,为制备mNotch1的单克隆抗体做准备。方法 :计算机分析小鼠Notch1全长 ,PCR扩增其胞外段高抗原区编码基因 ,克隆入载体T -vector进行序列测定 ,然后将测定正确的片段连入GST融合表达载体pGEX - 4T - 1,得到pG -NEF ,转化宿主菌DH5α ,经IPTG诱导 ,SDS -PAGE分析 ,以及新生条带表达形式分析以后 ,用GST亲和层析柱纯化。结果 :小鼠Notch1分子胞外段近膜处约 170个氨基酸区域抗原性较高 ,PCR扩增得到大小约 5 0 0bp的特异性片段 ,序列测定结果与文献报导的完全一致 ;原核表达产物大小约Mr 43× 10 3 ,以包涵体形式存在 ,约占总蛋白的2 5 % ,纯化后目的蛋白含量 >95 %。结论 :成功地进行了小鼠Notch1胞外段高抗原区编码基因的基因克隆、原核表达及蛋白纯化。     

ICAM-1-expressing pocket epithelium, LFA-1-expressing T cells in gingival tissue and gingival crevicular fluid as features characterizing inflammatory cell invasion and exudation in adult periodontitis

Activated T lymphocytes constitute a major component of inflammatory cells in the early periodontal lesion, and also appear in the gingival crevicular fluid. In an attempt to clarify the relationship between the ICAM-1 (CD54) expression of pocket epithelium in gingiva and the infiltrating lymphocyte population, we carried out an analysis of CD11a⁺ (LFA-lα), CD25⁺ (IL-2Rα) and CD4⁺ (Th) cells subjacent to ICAM-1-expressing pocket epithelia and CD11a⁺ CD25⁺CD4⁺ cells in gingival crevicular fluid (GCF). GCF was collected by crevicular washing from 16 patients with periodontitis (P group) and 3 subjects with healthy gingiva (H group). Peripheral blood (PB) was collected at the same time. Mononuclear cells were isolated by Ficoll-paque gradient centrifugation from GCF and PB. Monoclonal antibodies (mAb) to CD11a, CD25, and CD4 were used for three-color flow cytometry. Gingival biopsies were obtained from 7 patients in P group and 3 subjects in H group. Serial cryostat sections (6 μm in thickness) were prepared from each biopsy, on which a double staining was performed. The number of CD11a⁺ CD25⁺ CD4⁺ cells and the fluorescence intensity of FITC conjugated anti-CD 11a were significantly higher in GCF than in PB (p≤0.001 to p≤0.01). CD11a⁺ CD25⁺CD4⁺ cells were not detected in GCF in H group. The pocket epithelia expressed CD54 in P group, but not in H group. The number of CD11a⁺, CD25⁺ and CD4⁺cells infiltrating the connective tissue subjacent to the upper, middle and lower parts of the CD54 positive pocket epithelium (n=16) was 141±26, 38±13, 144±29 (cells/0.04 mm²), respectively, whereas in the CD54 negative pocket epithelium, it was (n=5) 9±2, 3±1, 8±3. In P group, the CD11a⁺CD25⁺CD4⁺cell number in GCF correlated with CD25⁺, CD11a⁺cells in the connective tissue subjacent to the CD54⁺pocket epithelium. These results indicate that expression of ICAM-1 in pocket epithelium is relevant to the migration of CD11a, CD25, CD4 positive cells in connective tissue subjacent to the pocket epithelium into the periodontal pocket. Assessing the relationship of our findings and other adhesion molecules would offer important clues to the understanding of T cell migration in affected gingiva.