山羊颞下颌关节盘细胞透射电镜观察

目的 用透射电镜的方法对两只1月龄健康山羊四只颞下颌关节盘的细胞及胶原结构进行了观察.方法 切取山羊双侧颞下颌关节盘,经组织学处理后行透射电镜观察细胞和胶原超微结构特征.结果 颞下颌关节盘由不均分布的细胞和胶原纤维组成,细胞有成纤维细胞样细胞和软骨细胞样细胞两种,其中成纤维细胞样细胞较软骨细胞样细胞占优势.成纤维细胞样细胞表现有细长的突起,核大,梭形或不规则形,细胞器较少.软骨细胞样细胞胞核呈圆形或椭圆形,周围少有电子透射区,细胞膜不明显,胞突少见.胶原纤维由平行排列的有周期性横纹特征的胶原原纤维组成.结论 颞下颌关节盘细胞有成纤维细胞样和软骨细胞样细胞两种,前者占优势,胶原原纤维表现有周期性横纹特征.这为颞下颌关节盘组织工程研究中细胞来源及表型分析奠定了一定基础.     

Low-energy laser stimulates tooth movement velocity via expression of RANK and RANKL

OBJECTIVE: Recent studies have demonstrated that low-energy laser irradiation stimulates bone formation in vitro and in vivo. However, very little is known about the effects of laser irradiation on osteoclastogenesis. The receptor activator of the nuclear factor-kB (RANK) / RANK ligand (RANKL) / osteoprotegerin (OPG) system is essential and sufficient for osteoclastogenesis. The present study was designed to examine the effects of low-energy laser irradiation on expressions of RANK, RANKL, and OPG during experimental tooth movement. DESIGN: To induce experimental tooth movement in rats, 10 g of orthodontic force was applied to the molars. Next, a Ga-Al-As diode laser was used to irradiate the area around the moved tooth and the amount of tooth movement was measured for 7 days. Immunohistochemical staining with RANK, RANKL, and OPG was performed. Real time PCR was also performed to elucidate the expression of RANK in irradiated rat osteoclast precursor cells in vitro. RESULTS: In the irradiation group, the amount of tooth movement was significantly greater than in the non-irradiation group by the end of the experimental period. Cells that showed positive immunoreactions to the primary antibodies of RANKL and RANK were significantly increased in the irradiation group on day 2 and 3, compared with the non-irradiation group. In contrast, the expression of OPG was not changed. Further, RANK expression in osteoclast precursor cells was detected at an early stage (day 2 and 3) in the irradiation group. CONCLUSION: These findings suggest that low-energy laser irradiation stimulates the velocity of tooth movement via induction of RANK and RANKL.     

Prostaglandin E2 and interleukin-1 levels in smokeless tobacco-induced oral mucosal lesions

Inflammatory mediators released as a result of smokeless tobacco (ST)-induced irritation may play a role in the development of oral mucosal lesions at habitual tobacco placement sites in ST users. The present study examined levels of interleukin-1 (IL-1) and prostaglandin E2 (PGE2) in ST-induced mucosal lesions and compared these to mediator levels in clinically normal mucosa. Soft tissue biopsies were obtained from white mucosal lesions at habitual placement sites and normal alveolar mucosal tissue at non-placement sites in 18 ST users. Fifteen non-tobacco using subjects also provided normal alveolar mucosal biopsies. IL-1 and PGE2 were recovered from the specimens, and mediator levels were determined by enzyme immunoassay. Prostaglandin E2 levels (pg/mg) were lower in both regions in the ST subjects, but values did not vary significantly between the regions with 2.77±0.72 and 2.86±0.99 at placement and non-placement sites, respectively, in ST users and 7.31±3.84 in non-tobacco users. Both IL-1α and IL-lβ (pg/mg) were significantly (p < 0.0I) elevated in ST lesions (IL-lã=25.56±4.00; IL-1β=7.76±1.68) compared to either non-placement sites in ST users (IL-lα=14.64±2.65; IL-lβ=1.63±0.72) or non-tobacco users (IL-lα=12.84±2.60; IL-lβ=2.04±0.75). In view of IL-l's role in keratinocyte proliferation and its inflammatory effects, this cytokine may contribute to mucosal and gingival alterations observed in ST users.     

Molecular mediators of Porphyromonas gingivalis‐induced T‐cell apoptosis

Porphyromonas gingivalis produces virulence factors which can modify the molecular and cellular components of the host immune response. In the present work we investigated the role of specific virulence factors from P. gingivalis in the induction of apoptosis in Jurkat T cells. P. gingivalis culture supernatants mimicked the effect of butyric acid on T‐cell apoptosis and this effect was associated with an increase in histone H4 acetylation. A role for proteases was excluded in experiments which demonstrated that neither protease inhibitors nor use of P. gingivalis mutants defective in protease synthesis had any effect on the stimulation of T‐cell apoptosis in this system.     

Modulation of osteogenic potential by recombinant human bone morphogenic protein-2 in human periodontal ligament cells: effect of serum, culture medium, and osteoinductive medium

BACKGROUND AND OBJECTIVE: Bone morphogenic proteins are known, in animal models, to promote many developmental processes, including osteogenesis. Clinical trials are currently underway to evaluate the potential of bone morphogenic proteins to promote bone and periodontal regeneration in humans. The aim of this study was to establish an optimal cell culture condition for using to study the biological effects of recombinant human bone morphogenic protein-2 on periodontal ligament cells. MATERIAL AND METHODS: The roles of serum concentration, types of culture medium (alpha-modified essential medium or Dulbecco's modified Eagle's medium), the presence of osteoinductive medium (including dexamethasone, ascorbic acid and beta-glycerophosphate), and timing of addition of the osteoinductive medium and recombinant human bone morphogenic protein-2, on the expression of alkaline phosphatase were investigated in cultured periodontal ligament cells. Cytochemical stainings and biological assay of alkaline phosphatase were also demonstrated. RESULTS: Our results suggested that an increased concentration of serum might mask the effect of recombinant human bone morphogenic protein-2 on the expression of alkaline phosphatase in periodontal ligament cells. alpha-Modified essential medium was found to induce a stronger cytochemical staining of the alkaline phosphatase than Dulbecco's modified Eagle's medium under similar culture conditions. Pre-incubation of cells with osteoinductive medium before the addition of various concentrations of recombinant human bone morphogenic protein-2 enhanced greater alkaline phosphatase expression than the simultaneous presence of both osteoinductive medium and recombinant human bone morphogenic protein-2. CONCLUSION: The findings of this study suggest that the effect of recombinant human bone morphogenic protein-2 on periodontal ligament cells could be efficiently investigated after the proper selection of culture variables and temporal sequence of adding bioactive factors. The optimal culture condition identified in this study might be useful in further studies to elucidate the regulatory mechanism of periodontal ligament cells in periodontal regeneration after stimulation with recombinant human bone morphogenic protein-2.     

Effect of soaking in Hank's balanced salt solution or milk on PDL cell viability of dry stored human teeth

Abstract— This study was designed to evaluate the effect of soaking in either Hank's balanced salt solution (HBSS) or milk on peri⁻odontal ligament (PDL) cell viability in avulsed teeth. Dry storage times of 30, 60, and 90 min were evaluated. PDL cell viability was determined after removal of the cells from the root surfaces of extracted teeth using a modification of the procedure described by Nakashima (Arch Oral Biol 1991;36:655–63). After trypsinization and subsequent treatment in collagenase, the cells were stained with trypan blue, and viable and non-viable cells were counted using a hemocytometer and converted to percentages for statistical comparison. The results of this study demonstrated no significant difference in the number of viable cells with or without soaking in HBSS or milk at any of the dry storage times. In addition, there was no significant difference in PDL cell viability between the 30-and the 60-min dry periods. Although the soaking procedure had no obvious negative consequence, no simcant improvement in PDL cell viability by the addition of this step was demonstrated under the conditions of this study.     

Expression of receptors for basic fibroblast growth factor on human periodontal ligament cells

Basic fibroblast growth factor (FGF-2; bFGF) is a major mitogen for connective tissue cells, and participates in the healing process. It has already been reported that FGF-2 could be applicable to enhance periodontal regeneration. In the present study, we examined FGF receptor (FGFR) expression on human periodontal ligament (PDL) cells. The binding of [¹²⁵I]-labeled FGF-2 to human PDL cells was studied by radioreceptor assay. The binding of [¹²⁵I]-FGF-2 to PDL cells reached a plateau after 2.5 h incubation at 4°C and was inhibited by the addition of unlabeled FGF-2 and acidic FGF (FGF-1; aFGF), but not insulin-like growth factor-I, platelet-derived growth factor and transforming growth factor-β₁. Scatchard analysis revealed the presence of approximately 1.0 × 10⁵ FGF-2 binding sites per cell with an apparent Kd of 1.2 × 10-¹⁰ M. Interestingly, the binding of [¹²⁵I]-FGF-2 on PDL cells reached its maximum at d 6 of the culture and then gradually decreased. Scatchard analysis also demonstrated that the number of FGFRs on a PDL cell was altered during the course of the culture, while the affinity between FGF-2 and its receptor was not. The responsiveness of PDL cells to FGF-2, which was monitored by the inhibitory effect on alkaline phosphatase activity, was reduced in proportion to the decrease in the number of FGFRs on the PDL cells. The present study suggests that PDL cells alter the responsiveness to FGF-2 during the course of the culture by changing the density of its receptor, and that the density of FGFR expression might be a marker of the cytodiflerentiation of PDL cells into mineralized tissue forming cells.     

Differential expression of salivary S100A7 in oral submucous fibrosis

Aim

To investigate the expression of salivary S100A7 levels among patients with oral submucous fibrosis (OSF) and healthy controls.

解整合素样-金属蛋白酶8和12在颌骨巨细胞病变组织中表达的研究

目的 检测解整合素样-金属蛋白酶(ADAM)8和12在颌骨巨细胞病变中的表达,探讨它们在颌骨巨细胞病变细胞发生中的作用。方法 采用免疫组织化学的方法检测40例颌骨中心性巨细胞病变,10例颌骨周围性巨细胞病变,9例巨颌症,6例颌骨动脉瘤性骨囊肿石蜡组织中的ADAM8、ADAM12的基因表达。结果 颌骨中心性巨细胞病变、颌骨周围性巨细胞肉芽肿、巨颌症、颌骨动脉瘤性骨囊肿中的几乎所有多核巨细胞和部分圆形单核基质细胞均为ADAM8、ADAM12阳性;颌骨中心性和周围性巨细胞病变中部分短梭形的细胞ADAM12阳性。结论 颌骨巨细胞病变中的多核破骨样细胞可能由圆形单核基质细胞融合而成,而ADAM8、ADAM12参与了此融合过程,同时,ADAM12可能还参与了病变中梭形单核基质细胞的成熟过程。