TGF—β1对人牙髓细胞内钙影响的激光共聚焦显微镜动态观察

目的：探讨ＴＧＦ－β１信号转导过程与人牙髓细胞内钙［（Ｃａ＾２＋）ｉ］的关系。方法：体外培养人牙髓细胞经钙荧光指示剂Ｆｌｕｏ－３负载后,用激光扫描共聚焦显微镜测定ＴＧＦ－β１影响前后的人牙髓细胞内钙随时间变化情况。结果：ＴＧＦ－β（０．１ｎｇ／Ｌ）使人牙髓细胞内钙先升高后降低,最后维持在此加药前稍高的静息钙水平上。结论：通过ＴＧＦ－β１可引起牙髓细胞内Ｃａ＾２＋水平的显著变化,证明Ｃａ＾２＋参与人     

氢氧化钙对人牙髓细胞内游离钙的影响

目的 检测Ca(OH)2对体外培养的人牙髓细胞内Ca^2 的影响。方法 在RPMI1640培养液中加入Ca(OH)2、CaC12和NaHCO4,培养第6代人牙髓细胞,用Flou-3荧光探针负载后,再次用Ca(OH)2刺激,激光扫描共聚焦显微镜检测细胞内Ca^2 。结果 含Ca(OH)2培养液培养的牙髓细胞,再用Ca(OH）2刺激时胞内Ca^2 升高;其他条件培养的细胞,Ca(OH2)刺激时Ca^2 无变化。结论 Ca(OH)2可导致人牙髓细胞内Ca^2 升高。     

TGF-β_1对人牙髓细胞内钙影响的激光共聚焦显微镜动态观察

目的 :探讨 TGF- β1 信号转导过程与人牙髓细胞内钙〔( Ca2 ) i〕的关系。方法 :体外培养人牙髓细胞经钙荧光指示剂 Fluo- 3负载后 ,用激光扫描共聚焦显微镜测定 TGF- β1 影响前后的人牙髓细胞内钙随时间变化情况。结果 :TGF- β1 ( 0 .1ng/ L)使人牙髓细胞内钙先升高后降低 ,最后维持在比加药前稍高的静息钙水平上。结论 :通过 TGF-β可引起牙髓细胞内 Ca2 水平的显著变化 ,证明 Ca2 参与了人牙髓细胞 TGF-β1 信号转导过程并参与将信号转入核内     

羟基磷灰石,氢氧化钙对鼠牙髓细胞碱性磷酶活性的影响

检测羟基磷灰石和氢氧化钙颗粒对鼠原代牙髓细胞碱性磷酸酶活性的影响。方法：采用０．５％胰酶－０．１％ＥＤＴＡ消化法获取ＳＤ鼠原代牙髓细胞。细胞接种后，分别加入氢氧化钙和羟基磷灰石颗粒，在７ｄ或１４ｄ分别测定鼠原代牙髓细胞的碱性磷酸酶活性。     

流体切应力增加骨髓来源破骨细胞内游离钙离子浓度的研究

目的研究流体切应力对培养于载玻片上的破骨细胞内Ca2+浓度的影响。方法应用激光扫描共聚焦显微镜和Fluo- 3/AM荧光探针标记技术,测定加力前后破骨细胞内游离Ca2+的荧光强度,获得的图像在计算机上用图像分析软件进行比较分析。结果在37 ℃、Fluo- 3/AM终浓度为10 μmol/L的条件下,破骨细胞可获得良好的标记效果,与未加力的对照组相比,流体切应力使破骨细胞内Ca2+荧光信号的平均强度值显著增加。结论体外培养于载玻片上的破骨细胞内Ca2+浓度对流体切应力敏感,Ca2+可介导流体切应力对破骨细胞功能的调节。     

纳米羟基磷灰石对牙釉质再矿化影响的实验研究

目的：通过体外实验了解纳米羟基磷灰石对釉质早期龋再矿化的影响。方法：制备牙釉质早期龋样本，分别使用再矿化液和纳米羟基磷灰石液处理，应用扫描电镜观察脱矿釉质表面经处理后的形态学改变，采用激光共聚焦扫描电镜观察荧光染料进入脱矿釉质内的量，应用釉质块荧光面积、总荧光量和病损平均荧光量等3个参数对再矿化效果进行量化，统计采用方差分析并作Dunnett T检验。结果：脱矿釉质块经过再矿化液和纳米羟基磷灰石液处理后，表面形态发生改变；激光共聚焦扫描电镜观察各参数均有显著降低，表明矿化液和纳米羟基磷灰石可促进脱矿釉质再矿化，且纳米羟基磷灰石显著优于再矿化液的作用。结论：纳米羟基磷灰石可促进脱矿釉质再矿化。     

机械牵张应变对人牙周膜细胞内游离钙离子浓度的影响

目的:观察机械牵张应变作用下,人牙周膜细胞(HPDLCs)内游离Ca2 浓度的变化。方法:体外培养HPDLCs,利用动态机械应变细胞加载装置,对1%、10%和20%应变组的细胞分别进行30、60和120min的动态牵张加载,通过流式细胞仪和激光共聚焦显微镜检测细胞内游离Ca2 浓度的变化,并应用SPSS13.0软件包对数据进行方差分析。结果:当HPDLCs加载30min时,各组检测到的胞内游离Ca2 浓度与对照组无显著差异(P>0.05);60min时,各个应变组的胞内Ca2 浓度明显升高,与对照组相比均有非常显著的差异(P<0.01);加载时间延长到120min时,各组Ca2 浓度又降至对照组水平(P>0.05)。结论:机械牵张应变可通过钙离子信号系统影响HPDLCs的生物学活性。     

Miconazole对破骨细胞在羟基磷灰石涂层吸收的影响

目的:研究钙离子通道抑制剂miconazole对破骨细胞对羟基磷灰石涂层的吸收的影响。方法:在羟基磷灰石涂层表面诱导破骨细胞,破骨细胞形成后,分别加入不同浓度的miconazole细胞培养基,培养48h后,扫描电镜观察羟基磷灰石表层的吸收陷窝。结果:miconazole可抑制破骨细胞的羟基磷灰石涂层的吸收陷窝面积,且吸收陷窝面积随着miconazole浓度的增加而增加。结论:miconazole可抑制破骨细胞对羟基磷灰石涂层的吸收。     

不同比例的聚乳酸-聚乙醇酸共聚物与羟基磷灰石复合支架对人牙髓干细胞增殖及矿化能力影响研究

目的　探讨不同比例的聚乳酸-聚乙醇酸共聚物与羟基磷灰石（PLGA/HA）复合支架的降解速率及其对人牙髓干细胞（human dental pulp stem cells,hDPSCs）增殖及矿化能力的影响。方法    采用溶液浇筑/颗粒沥析技术构建含质量分数分别为10%、20%及30% HA的PLGA/HA复合支架的3个实验组,单纯PLGA支架作为对照组。采用降解实验检测支架的降解速率。将hDPSCs接种于不同比例的PLGA/HA复合支架进行培养,扫描电镜观察细胞形态,应用MTT法检测细胞增殖能力。hDPSCs接种于各组支架培养72 h后,将复合支架植入裸鼠体内,分别于饲养1个月及3个月后处死裸鼠取出支架,应用HE染色观察组织学形态,应用牙本质基质蛋白1（DMP1）免疫组化染色观察细胞的矿化程度。结果    10%HA组和20%HA组具有较适宜的降解速率。各组均具有较高的促hDPSCs增殖能力,促进效果随HA含量的增加而加强。HE染色和免疫组化染色结果显示,HA可促进hDPSCs在体内的成牙本质向分化,分化程度与HA含量成正比,20% HA组及30% HA组矿化程度均较佳。结论   含20%HA的PLGA/HA复合支架具有较佳的降解速率及促细胞增殖和矿化能力,是比较理想的细胞支架材料。     

Three-dimensional topographic and metrologic evaluation of dental implants by confocal laser scanning microscopy

Background: Surface topography of dental implants has changed during the past few years; however, the last systematic study on this topic is dated 1993. Purpose: The aim of this study was to correlate dental implants by surface analysis. Materials and Methods: A microtopographic analysis of 35 dental implants was performed using confocal laser scanning microscopy. Roughness value (S_a) and developed surface area (ratio, S_dr) were calculated. Implants were grouped according to their surface treatment: “minimally rough” with no further surface treatment (n= 2); ablative structured using etching or blasting (n= 17); titanium plasma spray coated (TPS; n= 9); coated with hydroxyapatite (HA; n ‐ 7). Results: Most implants (n= 16) showed S_a values between 3.0 and 5.0 um. The developed surface area has a mean value of 13.5 and an SD of 6.52. Minimally rough implant surfaces show the lowest S_a values (mean 0.5 um). Implants with ablative surface treatment have mean S_a values of 3.1 m. Both groups with additive surface treatment (TPS and HA) present similar roughness values with a mean of 6.0 um and 5.8 um, respectively. Ratio Sdr ranges from 3.1 for the minimally rough implants to 11.4 for the ablative treated implants and 14.3 for TPS‐coated and 18.4 for HA‐coated implants. There is a significant difference between the roughness and ratio values of the different groups. The topographic images show a typical surface according to the underlying surface treatment. Conclusions: We can confirm the “classic” grouping of dental implants by type of surface treatment into the groups minimally rough, ablative, TPS coated, and HA coated as these treatments lead to different ascending S_as; however, the additional value of the ratio S_dr including both spatial and amplitude aspects of the surface could not be confirmed in this study. Functional parameters describing the topographic differences are still lacking.     

Architecture of intact natural human plaque biofilms studied by confocal laser scanning microscopy

Determination of the structure of human plaque will be of great benefit in the prediction of its formation and also the effects of treatment. However, a problem lies in the harvesting of undisturbed intact plaque samples from human volunteers and the viewing of the biofilms in their natural state. In this study, we used an in situ device for the in vivo generation of intact dental plaque biofilms on natural tooth surfaces in human subjects. Two devices were placed in the mouths of each of eight healthy volunteers and left to generate biofilm for 4 days. Immediately upon removal from the mouth, the intact, undisturbed biofilms were imaged by the non-invasive technique of confocal microscopy in both reflected light and fluorescence mode. Depth measurements indicated that the plaque formed in the devices was thicker round the edges at the enamel/nylon junction (range = 75-220 microm) than in the center of the devices (range = 35-215 microm). The reflected-light confocal images showed a heterogeneous structure in all of the plaque biofilms examined; channels and voids were clearly visible. This is in contrast to images generated previously by electron microscopy, suggesting a more compact structure. Staining of the biofilms with fluorescein in conjunction with fluorescence imaging suggested that the voids were fluid-filled. This more open architecture is consistent with recent models of biofilm structure from other habitats and has important implications for the delivery of therapeutics to desired targets within the plaque.     

氢氧化钙激动人牙髓细胞内钙释放及其通道的研究

目的：检测Ｃａ（ＯＨ）２激动体外培养的人牙髓细胞内Ｃａ＾２＋的释放和钙释放通道。方法：在ＲＰＭＩ１６４０培养液中分别加入Ｃａ（ＯＨ）２、Ｃａ（ＯＨ）２和肝素、Ｃａ（ＯＨ）２和普鲁卡因,培养第６代人牙髓细胞,用Ｆｌｕｏ－３荧光探针负载后,再次用Ｃａ（ＯＨ）２作用于牙髓细胞,用激光共聚焦显微镜检测胞内Ｃａ＾２＋。结果：含Ｃａ（ＯＨ）２、Ｃａ（ＯＨ）２和普鲁卡因的培养液培养的细胞内Ｃａ＾２＋浓度升高;含     

激光扫描共焦显微镜活体组织诊断技术的应用进展

激光扫描共焦显微镜作为一种新的无创诊断技术,能对人体体表组织细胞在活体进行观察,相对于常规组织学检查,它不需要标本切取、固定、包埋、切片、染色等繁琐的过程,具有很大的优势。它能在活体组织观察细胞形态和组织结构,甚至在使用增强对照的试剂后,能在分子水平观察细胞的活动。口腔黏膜、体表皮肤由于其特殊的位置及结构,使激光扫描共焦显微镜在这些方面得到广泛的应用。作者对激光扫描共焦显微镜在口腔黏膜、皮肤病变的活体组织诊断以及外科手术中的应用进行综述。     

龋病病因相关因素的激光扫描共聚焦显微镜研究

牙菌斑生物膜作为细菌生长的微环境,在龋病形成和发展中起重要作用;口腔环境中相关因素如口腔黏膜中钙离子含量等与龋病也有一定关系。利用新兴的激光扫描共聚焦显微镜对菌斑生物膜及其他相关因素进行研究,取得了一系列新的进展,本文就此领域的相关问题作一综述。     

Inter-odontoblastic fibres in human dentine observed by scanning electron microscopy

Crowns of healthy premolars with closed apical foramina were fixed, resin-impregnated and freeze fractured. Fragments were critical-point dried and examined by scanning electron microscopy. Radial inter-odontoblastic fibres with a diameter of 0.1-0.7 micron were observed. Most of these radial fibres originated from the odontoblast cell body, but some of them arose from the pulp. Both of them entered the predentine at the rim of dentinal tubules and their branches formed the fibrous network of predentine. The odontoblast processes had only a few short side branches at the level of predentine. Some fibrils were observed in the mineralized intertubular dentine, as well.     

Scanning electron microscopy of human dental pulp

Premolar teeth, extracted from 13- and 14-yr-old patients requiring orthodontic treatment, were cleaved longitudinally to expose the entire pulp chamber. Most of the pulp was dissected from the chamber, exposing parts of the odontoblast layer and leaving only a thin portion of pulp tissue covering the dentine. Following fixation and dehydration, the fractured teeth were prepared for scanning electron microscopy (SEM) either by vacuum or critical-point drying.Marked disruption of the odontoblast layer was observed in vaccum-dried preparations. Groups of odontoblasts were scattered over the predentine, the odontoblastic processes displaying either marked stretching or complete separation from the cell body. Critical-point dried specimens showed minimal disturbance of the odontoblast layer and adjacent pulp tissue. Odontoblasts, fibroblasts and collagen fibrils of the pulp and predentine were well preserved by critical point drying, permitting detailed examination by SEM.The pulp dissection and critical-point drying techniques used in this investigation constitute a satisfactory procedure for studying by SEM the morphology and interrelationships of connective tissue elements of the pulp, odontoblasts and predentine.