牙骨质基质提取物附着于牙根表面的实验研究

目的：观察牙骨质基质提取物是否能良好的附着在牙根表面上。方法：应用同位素标记原理用１３１Ｉ标记法标记牙骨质基质提取物，将牙片放置于含已标记的牙骨质基质提取物的培养液中培养。再将牙片取出，干燥后放置在Ｘ线感光胶片上，然后对感光照片作灰度值分析。结果：实验组的灰度值平均为１０８，而对照组为６４，实验组感光照片灰度值显著高于对照组。结论：牙骨基质提取物能较好的附着到牙根表面上，这就为牙骨质基质提取物作为牙根表面生物材料的更深入的研究奠定了一定的基础。     

牙骨质基质胍提取物促牙龈成纤维细胞、成骨细胞粘附研究

目的 :鉴定牙骨质基质胍提取物内牙骨质蛋白的生物活性。方法 :制取牙骨质基质胍提取物 ,检测人牙龈成纤维细胞、MC3T3 E1成骨细胞两种细胞在含牙骨质基质提取物浓度分别为 2 .5、5、10、2 0μg/ml的DMEM培养液中孵化 1h的贴壁率并以加牛血清蛋白 1mg/ml为空白对照组。检测两种细胞在含牙骨质基质提取物浓度 10 μg/ml的DMEM培养液中孵化 30、60、90、12 0min贴壁率。 结果 :不同浓度的牙骨质基质提取物均可促进牙龈成纤维细胞和成骨细胞的粘附 ,并呈浓度依赖性 ,尤以 10 μg/ml的牙骨质基质提取物的浓度为较佳浓度。细胞的贴壁率随牙骨质基质提取物作用时间的延长逐步升高 ,并以 90min为较理想的作用时间。结论 :人牙骨质基质胍提取物含有可促进牙龈成纤维细胞和成骨细胞粘附的牙骨质活性蛋白     

不同钛表面对血管内皮细胞早期粘附与增殖活性的影响

目的:观察血管内皮细胞在两种纯钛表面(光滑表面和粗糙表面)上的粘附情况.方法:医用纯钛片预备成光滑表面和粗糙表面两组,将猪血管内皮细胞接种于两组钛片表面,分别培养24、48和72 h后,通过吖啶橙染色和MTT活性测定,观察细胞在两种不同表面上的粘附和增殖活性.结果:荧光显微镜显示两组钛片上的内皮细胞均随着培养时间延长而增多,24 h时粗糙表面钛片上粘附的内皮细胞明显多于光滑表面组,而且细胞形态伸展好、成梭形.MTT结果显示粗糙表面组24 h和48 h的0D490值显著高于光滑表面组,而在培养72 h时,两组间没有统计学差异.结论:纯钛经双酸蚀处理形成的粗糙表面较其光滑表面更有利于血管内皮细胞的早期粘附,并促进其增殖.     

五倍子水提取物抑制LPS对牙周膜细胞附着牙骨质片表面的影响

目的观察五倍子水提取物对LPS抑制PDLC在牙骨质片表面附着的影响。方法采用细胞培养技术,细胞计数法及扫描电镜进行观察。结果加入五倍子水提取物溶液后,对LPS抑制PDLC在牙骨质片表面的附着有明显拮抗作用,该作用随五倍子水提取物浓度增加而增加,到10μg/mL时,达到峰值。另外,培养液中先加入不同浓度的五倍子水提取物溶液,培养24h后再加入LPS时(A组),与LPS和五倍子水提取物溶液同时加入组(B组)相比,除对LPS抑制PDLC在牙骨质片表面附着有同样拮抗趋势外,当五倍子水提取物浓度为10μg/mL、20μg/mL时,A组的细胞附着数明显高于B组。扫描电镜显示,A组与B组牙骨质片表面细胞数量较多,细胞形态丰满,细胞胞突伸展明显,并相互交织成栅栏状、网状结构,部分细胞呈叠层生长,其中A组效果更明显。结论五倍子水提取物对LPS抑制PDLC在牙骨质片表面附着有拮抗和保护作用。     

钛表面贻贝蛋白包被对人真皮成纤维细胞粘附和增殖的影响

目的:观察人真皮成纤维细胞在两种贻贝足丝蛋白提取物Usun-afix和BD CELL-TAKTM试剂包被钛表面的粘附和增殖状况,研究贻贝蛋白包被法是否能够用于经皮种植体表面处理。方法:分别制备光滑钛片和两种贻贝蛋白包被的钛片样本,选取P3代人真皮成纤维细胞接种到3种培养基底上:①不做处理的光滑钛表面(对照组);②Usun-afix包被的钛表面;③BD CELL-TAKTM包被的钛表面。MTT法检测1、3、5 d细胞在3种钛表面的增殖数量;利用DiI-AcLDL荧光染色结合MTT法检测15、30 min和1、2、4 h时间点细胞的粘附状况。结果:培养1、3、5 d,成纤维细胞在3种表面活性均较好,3组之间无明显统计学差异(P>0.05);DiI-AcLDL荧光染色显示,在培养15、30 min和1 h各时间点,Usun-afix和BD CELL-TAKTM包被钛表面粘附细胞数目明显多于光滑钛表面,在1、2、4 h各时间点,两包被组与对照组相比有更好的细胞伸展形态;不同时间点MTT检测显示,两包被组在接种15、30 min和1、2 h各时间点的吸光度值均明显高于对照组(P<0.05),在4 h时,虽两包被组OD值比对照组略高,但无统计学差异(P>0.05);两包被组在各个时间点的促粘附效果相似,差异均无统计学意义(P>0.05)。结论:成纤维细胞接种早期(<4 h),可更容易粘附到两种贻贝足丝蛋白提取物Usun-afix和BD CELL-TAKTM试剂包被的钛表面,而贻贝蛋白对成纤维细胞的增殖能力没有明显的促进效果。     

两种钛表面成骨细胞附着静态与动态观察

目的体外构建钛表面成骨细胞生长模型,对细胞生长情况进行动静态观察。方法将光滑钛片（光滑钛片组）及多孔海绵状钛（海绵钛组）处理为酸碱表面和钙磷沉积表面,接种成骨细胞,12h、48h实时观察,48h后钛表面细胞固定,扫描电镜观察。结果光滑钛片组可见细胞充分延展附着,其中钙磷表面组细胞附着更多,可见细胞下钙磷沉积。海绵钛组可动态观察成骨细胞在海绵钛内立体生长,在钛珠问形成细胞桥。结论本研究构建形成钛表面成骨细胞生长体外模型,可进行钛表面细胞生长的实时观察与分析。     

牙骨质基质提取物对两种细胞在牙根表面的细胞生物学研究

目的：观察牙骨质基质提取物能否促进牙周膜成纤维细胞、牙龈成纤维细胞向牙根表面移行、附着和趋向。方法：用细胞培养法和图像分析法分析细胞的附着和趋向。结果：发现牙骨质基质提取物能明显提高牙龈成纤维细胞和牙周膜成纤维细胞在牙根表面的附着，而且，随着培养时间的延长，牙骨质基质提取物促牙龈成纤维细胞和牙周膜成纤维细胞附着的功能更加显著。结论：牙骨质基质提取物可较好地促进牙龈成纤维细胞和牙周膜成纤维细胞在未脱矿的牙根表面上的附着、移行和趋向     

牙骨质基质提取物促进牙周结缔组织细胞在牙根表面上增殖的实验研究

目的：证实牙骨质基质提取物可促进牙周结缔组织细胞在牙根表面上增殖。方法：用细胞培养法和增殖细胞记数法。结果：加入牙骨质基质提取物的实验组无论是牙龈成纤维细胞还是牙周膜细胞的增殖能力均有显著的提高。结论：牙骨质基质提取物可促进牙龈成纤维细胞和牙周膜成纤维细胞在牙根表面上的增殖，牙骨质基质提取物对牙龈成纤维细胞的促有丝分裂作用强于对牙周膜成纤维细胞的作用     

钛种植体表面粗糙度对细菌黏附的影响

目的：研究钛种植体的不同表面粗糙度对变形链球菌及血链球菌黏附的影响.方法：用光电3-D表面测量系统测定两种纯钛片机械切割表面和大颗粒喷砂酸蚀表面的表面粗糙度.将钛片与变形链球菌和血链球菌共同培养,培养时间分别为4h,1d和5d.通过菌落形成单位（CFU）计数法及结晶紫染色法,比较不同时间点两种细菌在两种粗糙度钛片上的黏附量.结果：机械切割表面和大颗粒喷砂酸蚀表面钛片的表面粗糙度Ra值分别为1.25 μm和4.25 μm.CFU计数显示,在不同的培养时间点,两组钛片上的变形链球菌及血链球菌活菌黏附数量相近,差异无统计学意义（P＞0.05）.结晶紫染色显示,在不同的培养时间点,大颗粒喷砂酸蚀组钛片上血链球菌的细菌黏附总量均多于机械切割组钛片,差异有统计学意义（P ＜0.05）.在培养早期（4h）,大颗粒喷砂酸蚀组钛片上变形链球菌的细菌黏附总量大于机械切割组钛片;但在培养后期（1 d,5 d）两组钛片上变形链球菌的细菌黏附总量相近,差异无统计学意义（P＞0.05）.结论：钛片不同表面粗糙度对变形链球菌和血链球菌的活菌黏附数量无影响,但粗糙表面上黏附的细菌及基质总量大于中度粗糙表面.对于变形链球菌而言,粗糙度对细菌及基质黏附总量的影响随着生物膜的成熟而消失.     

四种钛种植体表面在模拟体液中诱导磷灰石沉积的研究

目的：检测四种钛植入体表面在细胞培养液中诱导生成磷灰石的能力及形貌特征。方法：直径15mm钝钛片打磨至600目，进行以下处理：（1）单纯光滑钛片；（2）钛片粗化处理，采用喷砂＋酸蚀法；（3）光滑钛片＋碱热处理；（4）粗化钛片＋碱热处理。以上四种表面之钛片分别浸入DMEM＋10％FCS细胞培养液中，分别于第6、12、18、24天取出一组钛片，扫描电镜观察表面形貌，能谱分析仪分析表面元素组成比例。对24d时钛片用X射线衍射仪分析表面组分。结果：在各时间点的光滑表面及单纯粗化表面均未见磷灰石形成，而在经碱热处理的实验组中，第6天时在碱热处理形成的网孔中可见磷灰石的形核的形成。粗化表面＋碱热处理组形核为纳米级的针状结晶物，随时间增长，形核逐渐长大成密集球状物；光滑＋碱热处理组表面为颗粒状形核。粗化表面＋碱热处理组较光滑钛片＋碱热处理组Ca、P沉积速度快，对第24天时经碱热处理试样XRD分析表明，表面沉积物主要为羟基磷灰石。结论：在含血清的细胞培养液中浸泡后，经碱热处理的钛表面显示了明显的诱导磷灰石沉积能力。表面沉积物成分为羟基磷灰石，粗化＋碱热处理表面沉积的磷灰石为纳米级针状结构且沉积速度较快。     

Surface properties of titanium and zirconia dental implant materials and their effect on bacterial adhesion

Objectives

Zirconia ceramic material has been widely used in implant dentistry. In this in vitro study the physiochemical properties of titanium and zirconia materials were investigated and the affinity of different bacteria to different materials was compared.

Methods

Disc samples with different surface states were used: polished partially stabilized zirconia (PZ), titanium blasted with zirconia (TBZ), titanium blasted with zirconia then acid etched (TBZA), and polished titanium (PT) as a control. Surface topography was examined using scanning electron microscopy and profilometry. Contact angle, surface free energy (SFE), surface microhardness and chemical composition were determined.Disc samples were separately incubated with Streptococcus mitis and Prevotella nigrescens, either with or without pre-coating with human saliva, for 6 h and the surface area covered by bacteria was calculated from fluorescence microscope images.

Results

PZ and TBZ exhibited lower surface free energy and lesser surface wettability than PT. Also, PZ and TBZ surfaces showed lower percentage of bacterial adhesion compared with control PT surface.

Conclusions

The zirconia material and titanium blasted with zirconia surface (TBZ surface) showed superior effect to titanium material in reducing the adhesion of the experimented bacteria especially after coating with saliva pellicle. Modifying titanium with zirconia lead to have the same surface properties of pure zirconia material in reducing bacterial adhesion.SFE appears to be the most important factors that determine initial bacterial adhesion to smooth surface.     

Influence of titanium surface charge on fibroblast adhesion

Background: Although dental implants have a high success rate, failure owing to the absence of adhesion between the gingival connective tissue and the implant surface is still being reported. Purpose: This study was designed to evaluate the effect of a titanium surface charge on fibroblast adhesion. Material and Methods: An electrical chamber was custom‐made to generate negative and positive surface charges on commercially pure titanium cylinders with a potential difference of 4.5 V. Twenty‐seven titanium cylinders were divided into three experimental groups. In each group, cell attachment to a positively charged titanium cylinder, a negatively charged titanium cylinder, and a titanium cylinder (control) was studied at three time intervals of 15, 30, and 60 minutes. NCTC clone 929 fibroblasts were used in these experiments. The effect of the potential difference in the pH of Dulbecco's Modified Eagle Medium (DMEM) was also evaluated using two new specimens at time intervals of 15, 30, 60, and 80 minutes. Results: The fibroblast cell attachment was more statistically significant to the positively charged titanium cylinder than the negatively charged titanium cylinder (p =.002) and the control (p=.000), whereas the cell adhesion difference between the control and the negatively charged titanium cylinder was not statistically significant (p=.808). The range of pH difference of the DMEM in the negative and positive parts of the electrical chamber was 0.46 and 0.30, respectively. Conclusion: Within the limitations of this in vitro study, the positive surface charge of the titanium cylinder results in significantly favorable cell adhesion.     

不同管径钛纳米管对成纤维细胞增殖、伸展和胶原分泌功能的影响

目的:研究不同管径Ti-TiO2纳米管对成纤维细胞增殖、伸展和胶原分泌功能的影响.方法:以1、5、10、20 V电压阳极氧化制备不同管径Ti-TiO2纳米管试件,试件表面培养成纤维细胞,MTT法检测细胞增殖,扫描电镜观察形态,天狼星红苦味酸染色检测细胞胶原分泌功能.结果:1、5、10、20 V制备的纳米管径依次为15、30、50和100 nm.细胞在抛光表面的增殖均高于纳米管表面;第5天时,100 nm纳米管表面细胞的增殖显著高于其他(P＜0.01).第3天时,抛光表面细胞呈典型长梭型,纳米管表面细胞伪足明显.100 nm纳米管表面细胞的试件胶原分泌功能最大(P＜0.05).结论:100 nm纳米管对细胞增殖的抑制作用较小并能促进细胞的胶原纤维分泌功能.     

自酸蚀牙本质粘接剂对人牙髓成纤维细胞的毒性作用

目的比较和评价自酸蚀粘接剂XenoⅢ（XO）和Adper Prompt（AP）以及全酸蚀粘接剂Single bond2（SB）三者的细胞毒性大小。方法将3种牙本质粘接剂XO、AP和SB涂布于直径为5.0 mm、厚度为0.5 mm的牙本质圆片的两面,置于DMEM培养液中获得材料的浸提液,然后将培养液稀释成100.0%、50.0%、25.0%和12.5%四种体积分数。选用组织块法体外原代培养人牙髓成纤维细胞,并将不同体积分数的材料浸提液与第5代人牙髓成纤维细胞共同培养,通过MTT法评价材料24、72、120 h的细胞毒性。结果牙本质粘接系统XO、AP和SB在体外对人牙髓成纤维细胞均有一定程度的细胞毒性,两种自酸蚀粘接剂XO和AP的细胞毒性明显低于全酸蚀粘接剂SB,其差异有统计学意义（P<0.05）。结论自酸蚀牙本质粘接剂XO和AP的细胞毒性小于全酸蚀牙本质粘接剂SB。     

纯钛铸件化学抛光的实验研究

目的:探讨纯钛铸件的化学抛光效果。方法:3组板状铸件(15mm×15 mm×1.4 mm)分别给予以下3种处理:A组铸件喷砂后不做其它处理;B组铸件喷砂后常规手工研磨;C组铸件喷砂后化学抛光。测试B、C两组铸件的减重率(-wt％),各处理组铸件的表面粗糙度(Ra值)和表面氧化膜厚度。结果:肉眼观察经化学抛光的纯钛铸件表面呈镜面状。化学抛光组的减重率高于手工研磨组,Ra值小于手工研磨组(P<0.05)。其表面存在约8μm厚的氧化膜,且均匀一致。结论:纯钛铸件化学抛光效果良好,具有临床推广价值。     

In vitro biocompatibility of a new titanium-29niobium-13tantalum-4.6zirconium alloy with osteoblast-like MG63 cells

BACKGROUND: Titanium-29niobium-13tantalum-4.6zirconium (TiNb) has recently been developed as a new implant material. TiNb is composed of non-toxic elements and has a lower modulus of elasticity than the other titanium alloys. However, its biocompatibility has not been adequately characterized. The aim of this study was to evaluate the biocompatibility of TiNb using an osteoblast-titanium co-culture system. METHODS: MG63 cells were cultured on three kinds of titanium disks: TiNb, pure titanium (pTi), and titanium-6aluminum-4vanadium (TiAl), prepared with two different surfaces, a polished and acid-etched surface and a machined-grooved surface. The surface topography and roughness were evaluated by scanning electron microscopy (SEM). After 48 hours culture, the number of proliferating cells and prostaglandin E2 (PGE2) production in the culture supernatant were determined. RESULTS: There was no significant difference in surface roughness among the three titanium disks with a polished and acid-etched surface. After 48 hours of culture, the number of cells was significantly reduced on pTi and TiAl compared to TiNb and the control. PGE2 production was significantly higher on pTi than on TiAl, TiNb, and the control. We further examined the effect of surface roughness on PGE2 production using machine-grooved titanium disks. While pTi and TiAl stimulated the production of PGE2 depending on surface roughness, roughened TiNb did not affect PGE2 production. CONCLUSIONS: These results suggest that TiNb may exhibit favorable biocompatibility because it has an efficient surface topography for cell proliferation, and the level of PGE2 production does not depend on surface roughness. We conclude that TiNb may be useful as an implant material.     

SEM evaluation of the in-vitro effects of an air-abrasive system on various implant surfaces.

This investigation was conducted to determine the effects of an air-abrasive polishing system on various implant surfaces. Four each of the following types of implants were obtained from the manufacturer: Stryker DB (a titanium alloy implant), Denar Steri-Oss (a pure titanium implant), IMZ (pure titanium polished collar/plasma-sprayed body), and IMZ transmucosal implant extensions (highly polished pure titanium). Four samples of each type of implant material were placed in a group to be treated with an air-abrasive polishing system for 0.5, 1, 5, or 10 seconds, for a total of 16 samples. Scanning electron photomicrographs taken of each sample before and after treatment were analyzed by three examiners who were blind to the treatment conditions. They determined if the abrasion on the treated surface was greater than, equal to, or less than the pretreatment control. No perceptible difference was noted between the pretreatment and posttreatment photomicrographs regarding the surface integrity of the implant material surfaces.     

Microbial community composition on modified dental implant surfaces: an in vivo study

Purpose: The aim of the present in vivo study was to examine alterations of the microbial community structure in biofilms on different dental implant surfaces over the time. Materials and Methods: Zirconium nitride-coated glass (ZrN-glass) and ZrN-coated polished titanium (ZrN-Ti) disks were used as substrates and polished titanium (Ti-pol) was used as a control. The specimens were mounted on removable intraoral splints in one adult. After 24 hours and 14 days of intraoral exposure, the microbial biofilms were analyzed by generating 16S rRNA gene clone libraries. Results: ZrN coating of a Ti surface altered the microbial composition early on (24 hours), with a tendency to augment Lactobacillus-related phylotypes later. Long-term exposure (14 days) of dental implant surfaces to microbes resulted in a significantly different composition of the biofilm on all three tested surfaces. Conclusions: This preliminary study showed that a ZrN-Ti disk surface harbors a significantly different microbial composition from a polished Ti surface. Further improvement of ZrN physical vapor deposition coatings might help to influence the adhesion of bacteria that are less pathogenic, thereby reducing the risk of peri-implantitis.     

Influence of a Nanoporous Zirconia Implant Surface of on Cell Viability of Human Osteoblasts

Purpose : The dense nonretentive surface of zirconia implants was modified into a nanoporous surface using selective infiltration etching surface treatment. The aim of this study was to investigate the influence of such a nanoporous modified zirconia surface on the attachment of human osteoblasts. Materials and Methods : Human osteoblasts were cultured for 21 days on (i) selective infiltration etched zirconia (nanoporous surface), (ii) polished zirconia, (iii) polished titanium, or (iv) airborne particle abraded acid etched (SLA) titanium disks. After the culture period the following parameters were assessed: number of cells, the morphology of the cells, the attachment of the cells, alkaline phosphatase activity, and the level of total protein (α= 0.05). Results : Statistical analysis revealed a significantly higher cell count on the third (F = 17.4, p < 0.001) and eighth day (F = 163, p < 0.001) for nanoporous zirconia and SLA titanium surfaces compared to polished specimens. The number of cells (nanoporous zirconia 160 ± 20/mm², SLA titanium 133 ± 15/mm²) and cell size (nanoporous zirconia 50.7 ± 3 μm, SLA titanium 42.5 ± 4 μm) were significantly higher than polished specimens. Nanoporous zirconia specimens demonstrated comparable alkaline phosphatase activity (0.0036 ± 0.0035 ng/μl) and intracellular protein content (72.7 ± 0.9 ng/μl) compared to other tested groups. Scanning electron microscopy revealed that cells attached on the polished surface using finger‐like processes, whereas on the nanoporous surface, finger‐like processes were not observed, as the cell membrane appeared to be in close proximity to the underlying surface. Conclusion : The findings of this study suggest that a nanoporous zirconia surface favors cell growth and attachment compared to a polished surface. It was proposed that a nanoporous zirconia surface may improve clinical performance of zirconia implants.     

Comparison of castability and surface roughness of commercially pure titanium and cobalt-chromium denture frameworks.

STATEMENT OF PROBLEM: Titanium is a biocompatible material, but it is not widely used in clinical dentistry for conventional removable denture frameworks. Little research exists on its applicability. PURPOSE: This study compared the casting accuracy and roughness of titanium and cobalt-chromium denture frameworks. MATERIAL AND METHODS: Twenty Kennedy Class II, Division 1 removable partial denture frameworks were fabricated with commercially pure titanium (n = 10) and a cobalt-chromium alloy (n = 10). The casting accuracy of each framework was determined by visual, radiographic, and microscopic methods. The roughness of each polished framework surface was analyzed with atomic force microscopy. RESULTS: The clinical fit, porosities, and microporosities of both types of metal frameworks were qualitatively similar. The surface roughness of polished pure titanium and cobalt-chromium frameworks was 104.43 +/- 69.24 nm and 133.91 +/- 40.92 nm, respectively. This difference was not statistically significant (P >.05). CONCLUSION: The clinical fit, porosity, and surface roughness of the titanium and cobalt-chromium frameworks fabricated for this study were comparable.