Effect of biofilm formation on virulence factor secretion via the general secretory pathway in Streptococcus mutans

Objectives

To investigate the role of SecA in protein secretion, and to evaluate the effect of biofilm formation on protein secretion in Streptococcus mutans.

Design

S. mutans strains UA159 and GS-5 were used in this study. Cells grown in biofilm and planktonic conditions were observed using immunogold electron microscopy. The mRNA levels of ftf, gtfB, gtfC, gtfD, Pac and secA were analysed in different growth conditions using real-time quantitative polymerase chain reaction. The levels of wall proteins and whole-cell protein extracts were examined using Western blot analysis.

Results

A microdomain colocalised with SecA and virulence factors such as Pac (AgI/II) and glucosyltransferase (GTF) was observed. The mRNA level of secA was upregulated in the biofilm condition. The level of protein expression of SecA and wall protein levels of GTF, fructosyltransferase (FTF) and Pac (AgI/II) in the biofilm condition were significantly higher than in the planktonic condition.

Conclusions

These data suggest that S. mutans utilises the Sec pathway to secrete virulence factor proteins such as Pac (AgI/II), GTF and FTF, and protein secretion occurred at a distinct microdomain. The level of SecA, the key factor in the Sec pathway, was influenced significantly by biofilm formation in S. mutans.     

Type II collagen and aggrecan mRNA expression by in situ hybridization in rabbit temporomandibular joint posterior attachment following disc displacement

Pathological changes and mRNA expression were studied in the posterior attachment of 40 adult Japanese white rabbits. The right temporomandibular joints of 28 rabbits were subjected to surgical disc displacement. Joints were studied by histochemistry and in situ hybridization. The collagen in the posterior attachment became dense, especially near the posterior band of the disc. Chondrocytes were found not only in the disc but also in the posterior attachment. Sometimes cartilage formation was seen. Type II collagen mRNA expression was first detected in the posterior attachment 4 days postoperatively and became progressively stronger with time. Aggrecan expression in the posterior attachment decreased at first, then increased gradually. It was concluded that, in the temporomandibular joint, chondrocytes appear in the posterior attachment as a result of biomechanical stimuli and the attachment becomes fibrocartilaginous following disc displacement.     

德莱尔面具结合螺旋开大器矫治早期骨性Ⅲ类错(牙合)的临床应用

目的探讨德莱尔面具结合螺旋开大器矫治早期骨性前牙反(牙合)的疗效,并分析颌骨、牙齿及软组织的改变.方法选择上颌骨发育不足导致的早期骨性前牙反(牙合)16例,采用德莱尔面具结合螺旋开大器进行前方牵引.牵引前后均摄头颅侧位定位片,并对牵引前后X线头颅侧位片进行测量分析及临床表现比较.结果 A点前移,上前牙唇向倾斜;下颌骨生长的方向、形态改变,上下颌骨矢状向关系趋于正常,凹面型明显改善.结论德莱尔面具结合螺旋开大器是矫治早期骨性前牙反(牙合)的有效方法.     

Coptidis rhizoma inhibits growth and proteases of oral bacteria

OBJECTIVE: The aim of this study was to investigate the antibacterial effect of Coptidis Rhizoma (CR), a traditional medicinal plant, on oral bacteria. MATERIALS AND METHODS: CR extract was prepared by boiling CR in water for 2 h. Alkaloids contained in CR extract were assayed by high performance liquid chromatography (HPLC). Antibacterial activity of CR extract was estimated from the lowest concentration that did not permit bacterial growth (minimum inhibitory concentration, MIC) and the concentrations that inhibited 50% of bacterial proteolytic activity (IC50). RESULTS: CR extract inhibited the growth of Actinomyces naeslundii, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Actinobacillus actinomycetemcomitans at MIC of 0.031-0.25 mg ml(-1), whereas it had less inhibitory effect (MIC: 0.5-2 mg ml(-1)) on the growth of Streptococcus and Lactobacillus. The major active component of CR extract was berberine (Ber), an alkaloid, and its inhibiting specificity to bacterial growth was similar to that of CR extract. CR extract and Ber were bacteriostatic at the MICs against most of the bacteria, and bacteriocidal at the concentrations higher than the MICs. Ber inhibited the activities of collagenase from P. gingivalis and A. actinomycetemcomitans. CONCLUSION: CR extract and Ber had an inhibitory effect on periodontopathogenic bacteria. These results suggest the possibility of their clinical application for the treatment of periodontal diseases.     

混旋聚乳酸-纳米羟基磷灰石复合板固定下颌骨骨折的实验研究

目的 探讨混旋聚乳酸—纳米羟基磷灰石(poly DL lactide—nano—hydroxyapatite，PDLLA—nano HA)复合板(简称复合板)作颌骨骨折内固定的可行性。方法 在兔两侧下颁骨体部造成实验骨折，分别采用复合板和纯PDLLA板固定。在不同时期行大体观察、组织学检查，测量骨痂横截面积。结果 各时期骨折固定良好；术后第2、3周，各有1只动物的PDLLA板侧出现炎性包块，形成瘘道；复合板侧均正常愈合。术后2、3、4周时复合板侧骨痂形成量与对侧差异有显著性。组织学检查发现，骨折愈合早期复合板侧成骨细胞和成纤维细胞及其分泌的细胞基质更为丰富。结论 复合板在体内降解速度适中，对骨折愈合过程无不良影响，比纯PDLIA板更适用于颌骨骨折。     

颞颌关节镜治疗不同类型内错乱疗效的研究

为评价颞颌关节镜治疗不同类型内错乱的疗效，人进行８１例次关节手术。内错乱分期：Ⅱ期（早中期）２５例，Ⅲ期（中期）１８例，Ⅳ期（中晚期）２４例，Ⅴ期（晚期）１４例，中位随访期３．２年（０．５－７年），总有效率７４％（６０／８１）。内错乱分期与有效率的关系：Ⅱ期２４／２５，Ⅲ期１６／１８，Ⅳ期１３／２４，Ⅴ期７／１４。经统计学处理：Ⅱ、Ⅲ期之间及Ⅳ、Ⅴ期之间无显著性差异（Ｐ〉０．０５）；Ⅱ、Ⅲ期与Ⅳ、     

Immunohistochemical study of hard tissue formation in the rat pulp cavity after tooth replantation

While mineralized tissue is formed in the pulp cavity after tooth replantation or transplantation, little is known of this hard tissue formation. Therefore, we conducted histological and immunohistochemical evaluations of hard tissue formed in the pulp of rat maxillary molars after tooth replantation. At 5 days after replantation, degenerated odontoblasts were lining the pulp cavity. At 14 days, dentin- or bone-like tissue was present in the pulp cavity. Immunoreactivity for osteopontin (OPN) and bone sialoprotein (BSP) was strong in the bone-like tissue, but weak in the dentin-like tissue. Conversely, dentin sialoprotein (DSP) was localized in the dentin-like tissue, but not in the bone-like tissue. Cells positive for BMP4, Smad4, Runx2, and Osterix were found around the blood vessels of the root apex at 5 days. At 14 days, these cells were also localized around the bone-like tissue. Cells expressing alpha-smooth muscle actin (SMA) were seen around the newly formed bone-like tissue, whereas no such cells were found around the newly formed dentin-like tissue. In an experiment involving the transplantation of a green fluorescent protein (GFP)-transgenic rat tooth into a wild-type rat tooth socket, GFP-positive cells were detected on the surface of the bone-like tissue and over all dentin-like tissue. These results indicate that the original pulp cells had the ability to differentiate into osteoblast-like cells as well as into odontoblast-like cells.     

Education of dentists in China

China is geographically located in the east of Asia and its population exceeds 1.3 billion. An understanding of dental education in China is thus of interest. However, as there is little published information on this topic, this paper provides information about China regarding its dental history, dental school system including curriculum and dental licensure. High school graduates take a nationwide entrance examination to apply for dental school, of which there are more than 50 in China. A five year dental education leads to the BDS degree. Dental school graduates must then pass the nationwide licensure examination to practise dentistry. Currently, there are not adequate numbers of dentists to provide the necessary oral health care for people living outside metropolitan areas.     

Modulation of the antibody response by Porphyromonas gingivalis and Fusobacterium nucleatum in a mouse model

Successive immunization of mice with Fusobacterium nucleatum and Porphyromonas gingivalis has been shown to modulate the specific serum IgG responses to these organisms. The aim of this study was to investigate these antibody responses further by examining the IgG subclasses induced as well as the opsonizing properties of the specific antibodies. Serum samples from BALB/c mice immunized with F. nucleatum (gp1-F), P. gingivalis (gp2-P), P. gingivalis followed by F. nucleatum (gp3-PF) F. nucleatum followed by P. gingivalis (gp4-FP) or saline alone (gp5-S) were examined for specific IgG1 (Th2) and IgG2a (Th1) antibody levels using an ELISA and the opsonizing properties measured using a neutrophil chemiluminescence assay. While IgG1 and IgG2a subclasses were induced in all immunized groups, there was a tendency towards an IgG1 response in mice immunized with P. gingivalis alone, while immunization with F. nucleatum followed by P. gingivalis induced significantly higher anti-P. gingivalis IgG2a levels than IgG1. The maximum light output due to neutrophil phagocytosis of P. gingivalis occurred at 10 min using nonopsonized bacteria. Chemiluminescence was reduced using serum-opsonized P. gingivalis and, in particular, sera from P. gingivalis-immunized mice (gp2-P), with maximum responses occurring at 40 min. In contrast, phagocytosis of immune serum-opsonized F. nucleatum demonstrated peak light output at 10 min, while that of F. nucleatum opsonized with sera from saline injected mice (gp5-S) and control nonopsonized bacteria showed peak responses at 40 min. The lowest phagocytic response occurred using gp4-FP serum-opsonized F. nucleatum. In conclusion, the results of the present study have demonstrated a systemic Th1/Th2 response in mice immunized with P. gingivalis and/or F. nucleatum with a trend towards a Th2 response in P. gingivalis-immunized mice and a significantly increased anti-P. gingivalis IgG2a (Th1) response in mice immunized with F. nucleatum prior to P. gingivalis. Further, the inhibition of neutrophil phagocytosis of immune serum-opsonized P. gingivalis was modulated by the presence of anti-F. nucleatum antibodies, while anti-P. gingivalis antibodies induced an inhibitory effect on the phagocytic response to F. nucleatum.