切胶免疫制备腮腺液高丰度蛋白多克隆抗体

目的 通过切胶免疫制备人腮腺液高丰度蛋白多克隆抗体,为下一步腮腺液高丰度蛋白单克隆抗体制备提供抗原解决方案.方法 将腮腺液经超滤法浓缩蛋白后,进行SDS-PAGE分析,切取分子量约为50-65kDa的高丰度蛋白条带,研磨后注射新西兰大耳白兔诱导产生免疫应答并制备兔抗人腮腺液高丰度蛋白多克隆抗体,并应用蛋白免疫印记(Western blot)等方法进行抗体鉴定.结果 成功制备和鉴定了人腮腺液高丰度蛋白多克隆抗体.结论 成功制备和鉴定了人腮腺液高丰度蛋白多克隆抗体,为下一步腮腺液高丰度蛋白单克隆抗体制备及唾液蛋白质组学研究奠定基础.     

Periodontal tissue engineering and regeneration: Consensus Report of the Sixth European Workshop on Periodontology

Aim: To review the scientific preclinical background and clinical studies of current methods of periodontal regeneration in the treatment of infrabony defects and soft tissue deficiencies
Method: Five commissioned review papers including two systematic reviews were scrutinized by a group of experts in order to derive consensus conclusions, clinical relevance/implications and to propose future research requirements.
Results: The following five papers were assessed:
  1. Biological mediators and periodontal regeneration: a review of enamel matrix proteins at the cellular and molecular levels.
  2. Regeneration of periodontal tissues: combination of barrier membranes and grafting materials – Biological foundation and preclinical evidence.
  3. Clinical outcomes with bioactive agents alone or in combination with grafting or GTR
  4. Treatment of gingival recession with coronally advanced flap procedures. A systematic review.
  5. Soft tissue management at implant sites     

釉基质蛋白体外诱导大鼠外胚间充质细胞骨桥素和骨钙素mRNA的表达

目的：了解体外培养环境中，大鼠外胚间充质细胞在两种不同形式釉基质蛋白(粗提型EMPs及纯化型EMD)诱导条件下，成骨分化标志骨桥素及骨钙素mRNA的表达水平。方法：酶消化法原代培养SD大鼠颌突外胚间充质细胞，在含釉基质蛋白(EMPs或EMD或EMPs EMD)的培养液中进行体外诱导培养，提取总RNA，采用RT-PCR法，以β-actin cDNA为内参照，检测骨桥素及骨钙素mRNA的表达水平。结果：1)各诱导实验组细胞均出现骨桥素mRNA的表达；2)仅EMPs EMD组出现骨钙素mRNA的表达。结论：外胚间充质细胞经釉基质蛋白特别是粗提型EMPs及纯化型EMD体外复合和诱导培养，可向成骨或成牙骨质方向分化，出现骨桥素及骨钙素mRNA的表达。     

Separation of the outer membrane and identification of major outer membrane proteins from Porphyromonas gingivalis

The outer membrane of Porphyromonas gingivalis, an oral strict anaerobe, was isolated by sucrose density gradient centrifugation. The outer membrane obtained by the differential detergent extraction method, previously reported, showed an essentially similar protein pattern on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), confirming that the latter method is suitable for the study of outer membrane proteins in this organism. N-terminal amino acid sequence analysis revealed that major outer membrane proteins in this organism included Arg-gingipain, Lys-gingipain, RagA (a TonB-linked receptor), and putative porins that were homologous to Escherichia coli OmpA.     

Antibacterial effect of ozone on cariogenic bacterial species

Objective

The aim was to evaluate the antibacterial effect of ozone on cariogenic bacterial species with and without the presence of saliva and a possible effect on the salivary proteins.

Methods

Suspensions of Actinomyces naeslundii (ACTCC 12104^T), Lactobacilli casei (N CTC 151) and Streptococcus mutans (NCTC 10449), in salt buffer or in saliva, were exposed to ozone gas delivered by the ozone generator Healozone™ 2130C. Aliquots of the suspensions were taken after 10, 30 and 60 s ozone exposures and cultivated on agar plates. Initial number of bacteria per ml was 8.0 × 10⁷ (SD 2.2 × 10⁷) (A. naeslundii), 1.0 × 10⁸ (SD 3.1 × 10⁶) (L. casei) and 1.0 × 10⁸ (SD 7.0 × 10⁵) (S. mutans), respectively. The proteins were separated by SDS electrophoresis and visualized by silver staining.

Results

In salt buffer 92%, 73% and 64% of the initial numbers of A. naeslundii, S. mutans and L. casei, respectively, were killed already after 10 s ozone exposure, while approximately 99.9% of the bacteria were dead after a 60 s exposure. After 10 and 30 s, but not after 60 s exposure to ozone, S. mutans and L. casei were less efficiently killed in saliva compared to the salt buffer. Various saliva proteins were degraded by ozone after a 60 s exposure.

Conclusions

The cariogenic species S. mutans, L. casei and A. naeslundii were almost eliminated following 60 s of ozone treatment. This killing was reduced in the presence of saliva although increasing the ozone application time to 60 s overcame these reductants in saliva. Detection of altered salivary proteins indicates that saliva components constitute additional targets for ozone.     

体内诱导法建立的Tca8113细胞耐卡铂细胞株的耐药性表达研究

目的探讨通过体内诱导法得到的耐药细胞株的耐药性表达情况。方法以人舌癌Tca8113细胞系为研究对象,实验组采用荷瘤耐药动物的肿瘤细胞培养获得,对照组为体外连续培养诱导其产生耐药性。用四唑蓝法检测细胞耐药指数,标记生物素链亲和素法免疫细胞化学检测耐药相关蛋白P糖蛋白、谷胱甘肽S转移酶π、多药耐药蛋白和拓扑异构酶Ⅱ的表达,逆转录PCR检测多药耐药基因、多药耐药蛋白、拓扑异构酶Ⅱ和谷胱甘肽S转移酶π的表达。结果免疫细胞化学和逆转录PCR显示2组细胞多药耐药蛋白、谷胱甘肽S转移酶π、拓扑异构酶Ⅱ等表达均升高,但体内诱导较体外诱导可以获得更高、更稳定的耐药性。诱导后的Tca8113细胞对氨甲喋呤、卡铂、平阳霉素、长春新碱耐药性明显升高,而对5-氟尿嘧啶耐药性增加不明显。结论体内诱导得到的细胞株耐药性明显高于体外诱导株,肿瘤细胞的耐药现象受多种基因的调节。体内诱导Tca8113卡铂耐药细胞株Tca8113/CBP-vo可以作为肿瘤耐药研究的平台。     

A clinical study evaluating the treatment of supra-alveolar-type defects with access flap surgery with and without an enamel matrix protein derivative: a pilot study

Aim: There is evidence that regenerative treatment of intra-bony and mandibular class II furcation defects with access flap and an application of an enamel matrix protein derivative (EMD) can result in a clinical benefit compared with access flap alone. The aim of this pilot study was to check if the results of access flap surgery in suprabony defects are improved by additional application of EMD.
Material and Methods: Thirty-nine adult subjects with supra-alveolar-type defects were randomly assigned to a test ( n =25) and a control group ( n =14). Seventy teeth were treated with EMD; 28 teeth were treated by access flap. Probing depth (PD), clinical attachment level and bleeding on probing were evaluated at baseline and after 12 months.
Results: PD of the operated teeth was improved in both groups ( p <0.001 to p =0.041) but always better in the test group. The attachment gain was 2.72±1.80 mm at sites with an initial PD 7 mm in the test group and 0.78±0.62 mm in the control group ( p =0.004). In the test group the mean attachment gain was 0.97±0.92 mm ( p <0.001); the mean reduction of PD was 1.55±0.90 mm ( p <0.001).
Conclusions: The data suggest a significant clinical benefit of supplementary application of EMD during surgical treatment of periodontitis of supra-alveolar pockets, especially in deeper pockets.     

Five-year results following treatment of intrabony defects with enamel matrix proteins and guided tissue regeneration

BACKGROUND: Treatment with enamel matrix proteins (EMD) or guided tissue regeneration (GTR) has been shown to enhance periodontal regeneration. However, until now there are limited data on the long-term results following these treatment modalities. Aim: The aim of the present clinical study was to present the 5-year results following treatment of intrabony defects with EMD, GTR, combination of EMD and GTR, and open flap debridement (OFD). MATERIAL AND METHODS: Forty-two patients, each of whom displayed one intrabony defect of a probing depth of at least 6 mm, were randomly treated with one of the four treatment modalities. The following parameters were evaluated prior to surgery, at 1 year and at 5 years after: plaque index, gingival index, bleeding on probing, probing pocket depth (PPD), gingival recession, and clinical attachment level (CAL). No statistically significant differences in any of the parameters were observed at baseline between the four groups. RESULTS: The sites treated with EMD demonstrated a mean CAL gain of 3.4+/-1.1 mm (p<0.001) and of 2.9+/-1.6 mm (p<0.001) at 1 and 5 years, respectively. The sites treated with GTR showed a mean CAL gain of 3.2+/-0.8 (p<0.001) at 1 year and of 2.7+/-0.9 mm (p<0.001) at 5 years. The mean CAL gain at sites treated with EMD+GTR was 3.0+/-1.0 mm (p<0.001) and 2.6+/-0.7 mm (p<0.001) at 1 and 5 years, respectively. The sites treated with OFD demonstrated a mean CAL gain of 1.6+/-1.0 mm (p<0.001) at 1 year and 1.3+/-1.2 mm (p<0.001) at 5 years. At 1 year, the only statistically significant difference between the four different treatments was found in terms of PPD reduction and CAL gain between EMD and OFD (p<0.05). However, at 5 years there were no statistically significant differences in any of the investigated parameters between the four different treatments. CONCLUSION: Within the limits of the present study, it may be concluded that the short-term clinical results following treatment with EMD, GTR, EMD+GTR, and OFD can be maintained over a period of 5 years.     

Adsorption of human plasma proteins to modified titanium surfaces

OBJECTIVES: The aim of this study was to examine the effect of modified titanium (Ti) surfaces on the initial events of plasma proteins adsorption. MATERIALS AND METHODS: 'Ti disks' with three types of surface modifications were compared: machined, acid-etched and acid-etched and blasted. Physical and chemical characterizations of the surfaces were performed via scanning electron microscopy (SEM), atomic force microscopy (AFM) used for analysis of surface topography, characterization of the titanium oxide (TiO2) layer was carried out by X-ray photoelectron spectroscopy (XPS) and characterization of surface energy by the determination of contact angles. Evaluation of plasma proteins' adsorption to the treated Ti surfaces was performed by mass spectrometry, confocal laser scanning microscopy and XPS. Quantitative proteins' assessment was carried out by enzyme-linked immunosorbent assay. RESULTS: SEM images revealed major differences in the topography of the examined surfaces. Acid-etched and blasted Ti surfaces were found to have higher roughness values and a thicker TiO2 layer as compared with acid-etched and machined surfaces. Moreover, acid-etched and blasted surfaces showed high surface area differentiation, pointing to a high increase in the three-dimensional (3D) surface area over the 2D surface area compared with the other surfaces. Adsorption of plasma proteins to the acid-etched and blasted Ti surfaces was both qualitatively and quantitatively more intense compared with the machined and acid-etched surfaces. This was shown for each examined protein, total proteins and by the removal degree of the protein coat. CONCLUSIONS: The preferential adsorption of plasma proteins to the acid-etched and blasted Ti surfaces may be explained by its topographical characteristics and by the increase of the 3D surface area of this modified surface.     

热休克蛋白60、70在口腔扁平苔藓中表达的研究

目的　探讨热休克蛋白 (heatshockprotein ,HSP) 60和HSP70在口腔扁平苔藓病变中的作用。方法　对 62例口腔扁平苔藓、1 0例正常口腔粘膜、2 1例慢性盘状红斑狼疮、1 0例粘膜良性淋巴组织增生病 ,46例白斑进行免疫组织化学SP法染色 ,分析HSP60、HSP70在口腔扁平苔藓中的表达 ;对 1 2例口腔扁平苔藓和 5例正常粘膜进行逆转录PCR实验 ,观察HSP60、HSP70mRNA的变化。结果　HSP60在口腔扁平苔藓病损区的表达较其他各组显著增强 ,差异有显著性 (P <0 0 0 1 )。HSP70在糜烂型口腔扁平苔藓病损区的表达下调。RT PCR结果显示 ,HSP60、HSP70mRNA表达增强。结论　HSP60及HSP70在口腔扁平苔藓的发病中起重要作用