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     

关于唾液分泌机制的研究（Ⅱ）

目的：研究生理状态下细胞内Ｃａ＾２＋池的分布以及ＩＰ３引起Ｃａ＾２＋释入的量,以进一步了解唾液分泌机制。方法：利用全细胞膜片钳技术对分离的大鼠颌下腺腺泡细胞进行膜电流测量。结果：乙酰胆碱引起定量Ｃａ＾２＋释放;光分解Ｃａｇｅｄ ＩＰ３引起定量Ｃａ＾２＋释放;电流振动是细胞内Ｃａ＾２＋浓度振动性变的结果。结论：ＩＰ３敏感的细胞内Ｃａ＾２＋池其释放量依赖于ＩＰ３浓度,主要分布在ＣＬ＾－通道存在的细胞膜     

炎症状态下人牙髓神经纤维内Ca^2＋浓度的变化

利用细胞内Ｃａ２＋浓度测定技术，直接测定了牙髓神经纤维内Ｃａ２＋浓度，结果发现：无论是临床炎症性痛过敏牙髓，或是炎症介质组织胺、５－羟色胺处理的牙髓，其神经纤维内Ｃａ２＋浓度均比正常牙髓显著升高。而当降低细胞外液Ｃａ２＋浓度时，虽经炎症介质处理，神经纤维内Ｃａ２＋浓度与正常组无显著差异。说明牙髓神经纤维内Ｃａ２＋升高可能与其炎性痛过敏的发生有关。增加的Ｃａ２＋可能来源于细胞外液的Ｃａ２＋跨膜通道内流     

硬组织替代材料对成骨细胞内游离钙及钙调素水平的影响

目的;研究羟基聚磷酸钙钠（ＨＰＡ）、羟基磷灰石（ＨＡ）、生物玻璃陶瓷（ＢＧＣ）和纯钛（Ｔｉ）４种硬组织替代材料对成骨细胞的细胞内游离钙（〖Ｃａ＾２＋〗ｉ）浓度、钙调素（ＣａＭ）含量以及细胞膜ＡＴＰ酶活性的影响,探讨这４处材料的生物相容性。方法：选用ＳＤ乳鼠颅顶骨皮骨细胞进行体外培养,把ＨＰＡ、ＨＡ、ＢＧＣ和Ｔｉ粉分别制成材料浸提液,将培养成活的成骨细胞接种于材料浸提液中培养,３ｄ后测定成骨细胞内（     

正常及炎性牙髓中T,B淋巴细胞的微波免疫组化研究

本研究旨在探讨Ｔ、Ｂ细胞与牙髓炎发生、发展及转归间的内在联系。采用微波免疫组化技术，对人正常及炎性牙髓中Ｔ、Ｂ细胞进行分类识别和定量分析．结果表明：正常牙髓中Ｔ细胞散在分布于基质中央区，ＣＤ８＾＋细胞居多（ＣＤ４＾＋／ＣＤ８＾＋＝０．４８），未检测出Ｂ细胞。炎性牙髓中Ｔ、Ｂ细胞增多，且主要集中在炎症中心。可复性牙髓炎时ＣＤ４＾＋／ＣＤ８＾＋＝０．５８，出现Ｂ细胞（ＣＤ３＾＋／ＣＤ２０＾＋＝１１．３     

SOD与SOD复合Ca(OH)_2切髓术的组织学疗效对比

目的：观察超氧化物岐化酶（ＳＯＤ）、ＳＯＤ复合氢氧化钙〔Ｃａ（ＯＨ）２〕（ＣＨ）作为盖髓剂的组织学疗效。方法：将ＳＯＤ、ＳＯＤ复合Ｃａ（ＯＨ）２用于猴磨牙切髓术，观察组织学效果。结果：术后６个月，均可诱导牙本质桥形成，但组织学效果不同，ＳＯＤ组以骨样牙本质为主；ＳＯＤ复合Ｃａ（ＯＨ）２组以典型规则的管样牙本质为主；对照组Ｃａ（ＯＨ）２以不规则管样牙本质和骨样牙本质为主。结论：ＳＯＤ复合Ｃａ（ＯＨ）２在诱导牙本质桥形成的组织学效果方面优于单纯使用ＳＯＤ。     

SOD与SOD复合Ca（OH）2切髓术的组织学疗效相比

观察超氧化物歧岐化酶、ＳＯＤ复 氢氧化钙「Ｃａ（ＯＨ）２」（ＣＨ）作为盖髓剂的组织学疗效。方法：将ＳＯＤ、ＳＯＤ复合Ｃａ（ＯＨ）２用于猴磨牙切髓术、观察组织学效果。结论：ＳＯＤ复合Ｃａ（ＯＨ）２在诱导牙本质桥形成的组织学效果方面优于单纯使用ＳＯＤ。     

年轻恒牙冠折后Ca(OH)_2护髓的疗效观察

目的：观察年轻恒牙釉—牙本质折断后,使用Ｃａ（ＯＨ）２制剂护髓的疗效及影响疗效的因素。方法：对１９８８～１９９５年间因釉—牙本质折断而在北医大口腔医院儿科就诊的５２例（６４个牙）患儿的病历资料进行了临床回顾性分析。其中４８例５９个牙使用Ｃａ（ＯＨ）２制剂进行间接盖髓术护髓。结果：使用Ｃａ（ＯＨ）２间接盖髓,其成功率为７８％;外伤牙的受伤程度及外伤后的就诊时间影响其疗效。结论：年轻恒牙釉—牙本质折断后,使用Ｃａ（ＯＨ）２做护髓剂成功率高     

细胞外Ca^2＋浓度对培养腮腺腺细胞分化和功能的影响

为研究高于生理浓度的细胞外Ｃａ２＋对腮腺腺细胞分化和分泌功能的影响，以１０倍和２０倍生理浓度的Ｃａ２＋作用于培养中的腮腺腺细胞，应用免疫组织化学技术，观察细胞外Ｃａ２＋对细胞连接蛋白及与分化有关的蛋白表达的影响。结果显示，细胞粘接分子（ＣＡＭ）、缝隙连接蛋白（Ｃｘ３２）随细胞外Ｃａ２＋升高而表达升高；ｃｊｕｎ蛋白随Ｃａ２＋浓度增高表达更广泛；而表皮生长因子受体（ＥＧＦγ），ｃｆｏｓ及腮腺特异性分泌蛋白α淀粉酶、富含脯氨酸蛋白（ＰＲＰ）表达无明显变化。研究结果提示，细胞外Ｃａ２＋在一定程度上可促进细胞分化，但对细胞分泌功能无明显影响。     

儿童冠折80例未露髓牙的处理

作者选择儿童牙科８０份病历完整的酃露髓冠折病例，对其不同的临床处理方法进行对比分析，试图优选 处理方法提高医疗质量。常用的处理方法有三种：Ｃａ（ＯＨ）２护髓后即刻金属带环固位；Ｃａ（ＯＨ）护髓后玻璃离子水门汀固位；Ｃａ（ＯＨ）２护髓后光敏树脂固位修复。     

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。     

维生素D3刺激髁突软骨细胞内钙释放及机械压力对其影响的研究

目的　探讨维生素D3(1,25(OH)2D3)刺激体外培养的兔髁突软骨细胞内钙离子的释放通道及机械压力对其影响。方法　消化法培养新西兰白兔髁突软骨细胞,分别进行肝素(20 g/L)、普鲁卡因(1 g/L)钙通道阻断处理, 及用自行设计制作的可控液压细胞加载装置分别进行90 kPa 60 min和90 kPa 360 min的加压处理,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10-8mol/L1,25(OH)2D3100μL刺激前后髁突软骨细胞内钙随时间变化情况,同时设有对照组。结果　1,25(OH)2D3刺激后对照组细胞内荧光强度随时间明显升高;普鲁卡因处理组变化与其相似,而肝素处理组在刺激前后胞内荧光强度无明显变化;90 kPa加压60 min处理组在接受1,25(OH)2D3刺激后胞内荧光强度也随时间明显升高,且其升高幅度显著高于对照组,而90 kPa加压360 min处理组在刺激后胞内荧光强度虽也有升高,但与对照组无显著差异,且在记录末期出现下降趋势。结论　1,25(OH)2D3能刺激髁突软骨细胞内三磷酸肌醇受体(IP3R)钙释放通道开放,使细胞内钙离子水平显著升高;一定的机械压力预调可改变该细胞内IP3通道对刺激的敏感性。     

人牙乳头细胞分化过程中胞内Ca~(2 )浓度变化和1,25(OH)_2D_3对Ca~(2 )影响

目的 :探讨人牙乳头细胞不同分化阶段和 1,2 5 (OH) 2 D3 作用下细胞内Ca2 的变化。方法 :体外培养人牙乳头细胞 ,利用激光共聚焦显微镜观察细胞内Ca2 浓度的变化。结果 :具有某些成牙本质细胞表型的细胞 (2 1d)胞内Ca2 浓度比无分化表型的细胞 (14d)高约 2倍 ;1,2 5 (OH) 2 D3 使 2 1d组细胞胞内Ca2 明显升高 ,而 14d组则无明显变化。结论 :人牙乳头细胞在分化过程中胞内Ca2 浓度上调 ;1,2 5 (OH) 2 D3 能提高牙乳头细胞静息内Ca2 水平以达新的Ca2 稳态 ,促进牙乳头细胞分化     

The effect of extracellular calcium ion on gene expression of bone-related proteins in human pulp cells

Calcium hydroxide is often used for induction of reparative dentin formation in endodontic treatment. However, little is known about the mechanism by which calcium hydroxide works. The calcium ion (Ca2+) is an important regulator of cell functions. In this study, we examined the effect of extracellular Ca2+ on gene expression of bone-related proteins in human cultured pulp cells in serum-free conditions. A Ca2+ level elevated by 0.7 mM induced an increase in mRNA expression of osteopontin and bone morphogenetic protein (BMP)-2. However, mRNA levels of BMP-4 and alkaline phosphatase decreased under the elevated Ca2+ culture condition. The same concentration of additional magnesium ions had little effect on expressions of the examined bone-related protein mRNAs. These findings suggest that Ca2+ in Ca(OH)2 specifically modulates osteopontin and BMP-2 levels during calcification in pulp.     

牙髓干细胞分化过程中L型钙离子通道羧基末端的表达

目的:探讨牙髓干细胞(DPSCs)分化过程中L型钙离子通道羧基末端的表达。方法:利用酶消化法体外分离、培养大鼠牙髓干细胞;吉姆萨染色法检测大鼠牙髓干细胞的克隆形成能力;神经诱导体系下诱导牙髓干细胞向神经样细胞分化,免疫荧光染色检测细胞分化后胶质纤维酸蛋白(glial fibrillary acidic pro-tein,GFAP)的表达和细胞分化前后L型钙离子通道Cav 1.2及羧基末端的表达。结果:牙髓干细胞的克隆形成能力为每1 000个细胞形成2～17个克隆;免疫荧光染色检测诱导后细胞GFAP表达阳性;免疫荧光染色检测显示:牙髓干细胞分化前L型钙离子通道Cav 1.2羧基末端表达于细胞膜上,细胞分化后羧基末端同时表达于细胞膜上和细胞核中。结论:L型钙离子通道Cav 1.2羧基末端在牙髓干细胞分化过程中发生核转位,羧基末端可能在牙髓干细胞的分化过程中发挥着一定的作用。     

Histological observations of hard tissue barrier formation in amputated dental pulp capped with α-tricalcium phosphate containing calcium hydroxide

Abstract This study was performed to evaluate the pulpal response to α-tricalcium phosphate (αTCP) containing calcium hydroxide (Ca(OH)₂). The dental pulps of monkeys were amputated and dressed with four agents: αTCP, αTCP containing 1% Ca(OH)₂, αTCP containing 5% Ca(OH)₂, and Ca(OH)₂ being used as a control. The pulpal responses were histologically evaluated after 4 and 8 weeks. The pulp tissue treated with αTCP proliferated above the level of the original wound surface, and a thin layer of hard tissue barrier was formed directly against the capping agent. The barrier demonstrated atubular matrix lined with flattened or cuboidal cells, but occasionally appeared irregular in form. Ca(OH)₂ dressing resulted in destruction of pulp tissue, with a thick hard tissue barrier being formed below the level of the exposure site. The barrier consisted coronally of osteodentin and pulpally of tubular dentin lined with odontoblast-like cells. By contrast, 1% Ca(OH)₂ added to αTCP produced a slight proliferation of pulp tissue. An atubular matrix barrier, pulpally lined with cuboidal cells, formed above the exposure site. It was later followed by the formation of tubular matrix lined with columnar cells. Teeth treated with 5% Ca(OH)₂, showed a thin necrotic layer and a thick barrier formation. The barrier was composed of tubular dentin-like tissue lined with odontoblast-like cells. It would appear that αTCP containing a small amount of Ca(OH)₂ may be clinically useful as a capping agent, as it induced consistent hard tissue formation, without excessive destruction of underlying pulp tissue.     

机械压力及细胞内钙释放通道阻滞对髁突软骨细胞骨架影响的研究

目的：探讨髁突软骨细胞骨架随力学刺激的变化过程及其是否受到细胞内钙及其IP3通道的影响。方法：消化法培养2周新西兰白兔的髁突软骨细胞,用自行设计制作的可控液压细胞加载装置分别在90kPa压强下加载60min、360min、20g／L肝素阻断处理后再分别进行90kPa压强下加载60min、360min,考马斯亮蓝染色方法观察各实验组细胞骨架形态的变化特征。结果：对照组MCC细胞骨架系统显示为围绕胞核呈丝网状紧密有序排列,弥散性颗粒较均匀地分布其中;90kPa加压60min后MCC细胞骨架增多且排列致密;加压时间延长至360min时,MCC细胞骨架表达较60min时减弱,且多呈网状排列结构;IP3通道阻断后,再经90kPa加压60min,MCC细胞骨架收缩聚集于胞膜周边,呈现稀疏、不连续的排列状态;加压时间延长至360min时,MCC细胞骨架有序的网状结构重新出现,有些细胞出现特征性“鸟喙”样改变。结论：MCC细胞骨架系统参与了髁突软骨细胞力学信号转导过程,适宜的压力可使骨架表达增强、排列致密,在此机械力导致MCC细胞骨架的重排过程中,细胞内钙及其IP3通道起一定的调节作用。     

Comparative evaluation of diffused calcium and hydroxyl ion release from three different Indirect pulp capping agents in permanent teeth – An in vitro study

BackgroundIndirect pulp capping therapy has gained increased popularity in paediatric dentistry since it is less invasive, and is of low cost. The aim of the present study was to evaluate and to compare the diffusion of calcium (Ca2+) and hydroxyl (OH–) ions through coronal dentin into pulp after indirect pulp capping in vitro using TheraCal LC, ProRoot MTA and Calcimol LC.Materials and methodsTotal of 60 human caries-free maxillary first premolars were selected for the study. Samples were divided into 4 groups with 15 in each group: Group 1 TheraCal LC; Group 2 ProRoot MTA; Group 3 Calcimol LC; Group 4 Control Group. Indirect pulpcapping on the coronal RDT (remaining dentine thickness) system was performed using pulp-capping materials, such as TheraCal LC, ProRoot MTA and Calcimol LC, on the respective samples. The control group was completely filled with composite. Ca2+ ions (ppm) and OH– ions (pH) were analysed in deionized water using a multimeter connected to a calcium probe (calcium ion electrode) and pH metre connected to a temperature-compensated pH probe after 3 h, 24 h, 7 days, 14 days, 28 days and 60 days.ResultsCalcium release was significantly higher (P < 0.05) in the TheraCal LC group than in the other groups. Slightly alkaline pH values were observed in all the groups except for the control.ConclusionTheraCal is a new light-curable pulp capping material that initially releases high Ca2+ ions and creates an environmental pH close to physiological pH after 60 days.     

The comparison of a dentin adhesive with calcium hydroxide as a pulp-capping agent on the exposed pulps of human and sheep teeth.

OBJECTIVE: This study evaluated the biocompatibility of a one-step dentin bonding agent (Prime&Bond 2.1) in pulp capping compared with calcium hydroxide [Ca(OH)2]. METHOD AND MATERIALS: Thirty sheep teeth and 20 intact human premolars were used. After cavity preparation, pulp exposure was achieved with a bur (#390). Adhesive pulp capping was performed in 25 teeth (15 sheep and 10 human). In the control group (12 sheep and 10 human teeth), pulps were capped with Ca(OH)2 and all of the cavities in both groups were sealed with resin composite. Three of the sheep teeth were used as intact controls. Teeth were extracted 7 or 90 days following treatment and prepared for histological examination and bacterial detection. RESULTS: At 7 days, severe inflammatory responses underlying the bonding agent and in the coronal pulp were observed with soft tissue disorganization in both human and sheep teeth capped with Prime&Bond 2.1. All of the teeth capped with Ca(OH)2 exhibited mild inflammatory reactions limited with the perforation area. After 90 days with the bonding agent, in 3 of 9 sheep teeth, chronic inflammatory reactions were significant, while slight pulpal reactions were observed in the others and dentin bridge formation in all of the sheep teeth was found. However, in human pulps, persistent, unresolved inflammation with the lack of dentin bridge formation was observed. In the Ca(OH)2 group, pulp repair with dentin bridging was found in all of the teeth, both sheep and human. No correlation was found between the presence of inflammation and bacterial staining using Spearman rank correlation test (P > .05). CONCLUSION: Prime&Bond 2.1 facilitates enhanced pulp healing and bridge formation in sheep teeth, but in human teeth it was not as successful as Ca(OH)2 as a pulp capping agent.