纯钛表面牙周韧带细胞牙骨质附着蛋白的表达

目的以牙骨质附着蛋白为分化指标，分析牙周韧带细胞在纯钛表面的分化趋势，并探讨诱导矿化对牙周韧带细胞牙骨质附着蛋白表达的影响。方法第三代牙周韧带细胞在商用纯钛表面接种后，分别进行常规培养和矿化液诱导矿化培养，通过免疫荧光原位检测牙骨质附着蛋白的表达，并比较诱导矿化对牙骨质附着蛋白表达的影响。结果牙周韧带细胞在纯钛表面初期呈条纹状生长，常规培养和诱导矿化培养务件下均能表达牙骨质附着蛋白，诱导矿化对牙骨质蛋白的表达无明显影响。结论在常规培养务件和诱导矿化培养条件下，纯钛表面生长的牙周韧带细胞能高度表达牙骨质附着蛋白，提示纯钛表面体外培养的牙周韧带细胞具有形成牙骨质的分化趋势，从而可为种植体周构建牙周韧带结构提供在种植体侧的锚着点。     

A possible mechanism of low molecular weight protein tyrosine phosphatase (LMW-PTP) activity modulation by glutathione action during human osteoblast differentiation

Objective

Low molecular weight protein tyrosine phosphatases (LMW-PTPs) are a family of enzymes strongly involved in the regulation of cell growth and differentiation. Since there is no information concerning the relationship between osteoblastic differentiation and LMW-PTP expression/activity, we investigated its involvement during human osteoblast-like cells (hFOB 1.19) differentiation. It is known that LMW-PTP is regulated by an elegant redox mechanism, so we also observed how the osteoblastic differentiation affected the reduced glutathione levels.

Design

hFOB 1.19 cells were cultured in DMEM/F12 up to 35 days. The osteoblast phenotype acquisition was monitored by measuring alkaline phosphatase activity and mineralized nodule formation by Von Kossa staining. LMW-PTP activity and expression were measured using the p-nitrophenylphosphate as substrate and Western blotting respectively. Crystal violet assay determined the cell number in each experimental point. Glutathione level was determined by both HPLC and DNTB assays.

Results

LMW-PTP modulation was coincident with the osteoblastic differentiation biomarkers, such as alkaline phosphatase activity and presence of nodules of mineralization in vitro. Likewise LMW-PTP, the reduced glutathione-dependent microenvironment was modulated during osteoblastic differentiation. During this process, LMW-PTP expression/activity, as well as alkaline phosphatase and glutathione increased progressively up to the 21st day (p < 0.001) of culturing, decreasing thereafter.

Conclusions

Our results clearly suggest that LMW-PTP expression/activity was rigorously modulated during osteoblastic differentiation, possibly in response to the redox status of the cells, since it seems to depend on suitable levels of reduced glutathione. In this way, we pointed out LMW-PTP as an important signaling molecule in osteoblast biology and bone formation.     

HLA-DR4 and salivary immunoglobulin A reactions to oral streptococci

The aim of this study was to describe and compare salivary immunoglobulin A (IgA) antibody reactions to extracts of strains of three oral streptococci in human leukocyte antigen (HLA)-DR4-positive and -DR4-negative subjects. Whole paraffin-stimulated saliva samples were collected from 27 apparently healthy subjects. Previous HLA typing showed that 20 subjects were DR4 positive and 7 were DR4 negative. HLA-DRB1*04 subtyping was performed among the DR4-positive subjects. Whole-cell antigen extracts from Streptococcus mutans (KPSK 2), Streptococcus sobrinus (OMZ 65) and Streptococcus parasanguis (Nt 62) were separated in SDS-PAGE. The antigens were immunoblotted with diluted saliva (Western blot), scanned and analyzed in a computer system. All immunoblot bands were recorded in DR4-positive and DR4-negative saliva pools, and bands with an optical density >or=0.1 were selected for analysis in individual salivas. The DR4-negative subjects in general had more immunoblot bands and more distinct bands than did the DR4-positive subjects. A higher concentration of total IgA in saliva was correlated with more bands, especially to antigens separated from S. mutans. When the number of bands was calculated per IgA unit, significant differences were observed between DR4-positive and DR4-negative salivas. This was particularly seen for S. mutans and S. parasanguis. As the number of bands was analyzed in relation to DR4 subgroups, DRB1*04, there was a lower salivary IgA activity to S. mutans in the DRB1*0401 and *0404. The variable level of correlation previously demonstrated for S. mutans colonisation and serologically defined DR4 positive subjects might be explained by the heterogeneity in this group, and the relation should be sought on a subgroup level.     

Helicobacter pylori and Campylobacter rectus share a common antigen

AIM: The aim of this study was to investigate the presence of antigens with immunological cross-reactivity in periodontopathogenic bacteria and Helicobacter pylori, the pathogen associated with gastritis and peptic ulcers in human. MATERIALS AND METHODS/RESULTS: Among the putative periodontopathogens tested (Actinobacillus actinomycetemcomitans, Campylobacter rectus, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia and Treponema denticola), cross-reactive bands were only detected in C. rectus by SDS-PAGE/Western immunoblotting analysis using a polyclonal antibody directed to H. pylori cells. One of these cross-reactive antigens, a 64-kDa band antigen, also reacted with a monoclonal antibody directed to the human heat shock protein (HSP) 60. The N-terminal amino acid sequence of this C. rectus protein revealed a high degree of homology with corresponding regions of other HSPs belonging to the HSP60 family, indicating that the 64-kDa antigen was a GroEL protein. The nucleotide sequence of the C. rectus GroEL protein coded for a 547 amino acid protein with a predicted size of 57.8 kDa. Comparison of the alignment of the deduced amino acid sequence of the GroEL protein of C. rectus with that of H. pylori showed a high degree of similarity throughout its length (76.8%). GroEL protein from C. rectus possessed the ability to stimulate production of IL-6 by a confluent monolayer of human gingival epithelial cells and was cytotoxic when used at a high concentration. CONCLUSIONS: This study reveals an immunological relationship between H. pylori and C. rectus, and clearly indicates that one of the shared antigens is a GroEL protein possessing a biological activity that might play a role in the initiation and progression of periodontal disease.     

Enamel matrix derivative alone or in combination with a bioactive glass in wide intrabony defects

This controlled clinical study investigated the clinical and radiographic outcome of wide intrabony periodontal defects treated by enamel matrix derivatives alone or in combination with a bioactive glass over a period of 8 months. Twenty-three chronic periodontitis patients, who received initial therapy and had radiographical interproximal defects with an associated probing depth of 6 mm or more and an intrabony component of at least 4 mm, were included. Each of the patients, contributing at least one intrabony defect, was treated with either enamel matrix derivative alone (group 1, n=10) or the combination (group 2, n=13). In both groups, all clinical and radiographical parameters were improved. Groups 1 and 2 presented a mean pocket reduction of 5.03±0.89 and 5.73±0.80 mm, recession of 0.97±0.24 and 0.56±0.18 mm, relative attachment gain of 4.06±1.06 and 5.17±0.85 mm, and radiographic bone gain of 2.15±0.42 and 2.76±0.69 mm, respectively. An intergroup comparison revealed significant differences for all of the parameters, yielding a more favorable outcome towards the combined approach. Within the limits of the study, both treatments resulted in marked clinical and radiographical improvements, but combined treatment seemed to enhance the results in the treatment of wide intrabony defects.     

煅石膏、骨形成蛋白及羟基磷灰石三元复合人工骨修复骨缺损的实验研究

以煅石膏(PLP）作为颗粒型羟基磷灰石(HA)人工骨粘接成形剂和骨形成蛋白(BMP)的载体，制成三元复合人工骨．分别用HA—bBMP、HA—PLP和单纯HA植入狗下颌骨实验性骨缺损中，采用组织学、定量组织学、免疫组织化学、X线摄片和扫描电镜观察的方法评价该复合人工骨的生物学性能。术后1，2，4，8和16周观察发现，HA—bBMP—PLP复合人工骨具有明显的骨诱导活性．PLP可充当BMP缓慢释放系统载体．增强BMP骨诱导活性，和作为颗粒型HA的粘接成形剂．使复合人工骨具有一定的可塑性和成形性，可达到准确的植入，植入后早期可有效防止HA颗粒移动。本研究证实．HA—bBMP—PLP三元复合人工骨不但可限制植入后HA颗粒的早期移动，更重要的是可以迅速增加新骨形成量．从形态和功能上大大提高了复合人工骨修复骨缺损的质量。     

T‐cell independent production of salivary secretory IgA after hematopoietic stem cell transplantation in children

This study examined the recovery of secretory IgA (S‐IgA) in saliva after hematopoietic stem cell transplantation (HSCT) in 35 children and young people between the ages of 3 and 27 years (mean = 13.6), and compared this recovery with that of serum immunologic constituents. Reference values for human salivary S‐IgA in saliva were obtained from 77 healthy control subjects between the ages of 7 and 25 years (mean = 11.4). In the 35 patients, a nadir of secretory IgA concentrations in saliva (S‐IgA) was observed between the 3rd and the 4th month, and a return to normal values 1 year after HSCT. Serum IgA concentrations reached their nadir in the 6th month, and normalized in the 18 months after HSCT. The recovery of T‐helper cells (CD4⁺/3⁺) was also delayed to beyond 18 months. We found a significant correlation between the reconstitution pattern of S‐IgA and that of T‐helper lymphocytes, but no correlation was found between the post‐transplant evolutions of S‐IgA and serum IgA, or between S‐IgA and T‐helper cells. The recovery of S‐IgA was more rapid than that of serum IgA and appeared to be T‐helper cell independent.     

骨形成蛋白家族与牵引成骨术

牵引成骨术(DO)中新骨的形成具有胚胎骨的发生和正常骨折愈合的某些特征,骨形成蛋白家族(BMPs)在其中扮演了重要角色。本文就BMPs与DO的生物学联系、作用调节机制、外源性BMPs的应用等方面作一综述。     

The effect of periodontal treatment on glycemic control in patients with type 2 diabetes mellitus

BACKGROUND, AIMS: This study was designed to explore the effect of periodontal therapy on glycemic control in persons with type 2 diabetes mellitus (DM). METHODS: 36 patients with type 2 DM (treatment group) received therapy for adult periodontitis during an 18-month period. A 36-person control group was randomly selected from the same population of persons with type 2 DM who did not receive periodontal treatment. RESULTS: These groups were well matched for most of the parameters investigated. During the nine-month observation period, there was a 6.7% improvement in glycemic control in the control group when compared to a 17.1% improvement in the treatment group, a statistically significant difference. Several parameters that could confound or moderate this glycemic control were explored. These included the treatment of non-dental infections, weight and medication changes. No moderating effect was associated with any of these variables. However, there were too few subjects in the study to have the statistical power necessary to assess these possible moderators of glycemic control. CONCLUSIONS: We interpret the data in the study to suggest that periodontal therapy was associated with improved glycemic control in persons with type 2 DM.     

Use of a composite pedicled muscle flap and rhBMP-7 for mandibular reconstruction

The aim of this study was to investigate the feasibility of reconstructing critical size continuity osteoperiosteal defects of the mandible using a composite of recombinant BMP-7 contained in a bovine type-1 collagen carrier wrapped in a pedicled sterno-occipitalis muscle flap. At 3 months following surgery, bridging of the surgical defect was noted in three subjects (60%). Histologically, the induced bone regenerate showed maturation from woven to lamellar bone. Islands of cartilage were distributed throughout the defect. Replacement ossification of the degenerated muscle was a common feature in all specimens. Microradiography showed a gradual increase in the calcification of mineralized tissue from the margin to the centre of the newly generated bone. This research represents a proof of the concept that bone can be satisfactorily formed within a muscular scaffolding at the site of the created defect in a one-stage procedure.