碱性成纤维细胞生长因子对成骨细胞粘附的调控作用

目的探讨碱性成纤维细胞生长因子(bFGF)对成骨细胞在钛片上粘附的调控作用。方法体外培养大鼠成骨细胞,利用存活细胞检测(MTT)法以及扫描电镜(SEM)观察bFGF对成骨细胞在钛片上粘附情况的影响。结果在0~27 ng/ml浓度之间,bFGF对促进成骨细胞在钛片上的粘附作用具有剂量依赖效应,27 ng/ml bFGF能促进成骨细胞在粘附初期的形态的改变以及在材料上的铺展。在高浓度的环境下,bFGF对促进成骨细胞在钛片上的粘附作用有一定的抑制作用。结论一定浓度的bFGF能促进成骨细胞在钛片上的粘附。     

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？…

目的：证实牙骨质基质提取物可是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认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。     

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。     

碱性成纤维细胞生长因子对成骨细胞和牙周膜成纤维细胞迁移、增殖的影响

目的　明确碱性成纤维细胞生长因子(bFGF)对体外成骨细胞和牙周膜成纤维细胞迁移、增殖的影响,以 探讨在牙种植体组织界面局部应用bFGF诱导类牙周膜形成的可行性。方法　同一只SD大鼠来源的成骨细胞和 牙周膜成纤维细胞经传代培养至第4代,建立体外创伤模型,分别在普通培养基和含bFGF的培养基中培养,观察 细胞迁移情况,四唑盐比色实验(MTT)测定细胞的增殖速度。结果　普通培养基中,成骨细胞迁移速度快于成纤维 细胞。加bFGF培养基中牙周膜成纤维细胞迁移速度明显快于其他各组,同时MTT结果显示加入bFGF能明显促进 两种细胞的增殖。结论　bFGF能明显促进牙周膜成纤维细胞的增殖、移行。     

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细胞在牙根表面附着的研究

目的 明确附着的牙根表面的牙骨质基质提取物具有促进牙周细胞附着的方法。方法 采集健康的牙龋组织和牙周膜,体外培养牙龈成纤维细胞和牙周膜成纤维细胞。从因正畸需拔除的健康牙体表面获得牙骨质基质提取物,分别观察不同作用时间,不同浓度的牙骨质基质提取物对牙龈成纤维细胞和牙周膜成纤维细胞在牙体表面附着的影响,结果 牙龈成纤维细胞,牙周膜成纤维细胞均对牙骨质基质提取物有浓度和时间依赖性,最佳作用时间为２小时,     

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     

牙骨质基质提取物吸附于光滑钛表面上的实验研究

目的:探讨牙骨质提取物能否良好地吸附在光滑钛表面上.方法:以含131I标记的牙骨质基质胍提取物的DMEM培养液为实验组,以131I处理的DMEM培养液为对照组,将金属钛片放置在以上两组溶液中孵化1h,钛片干燥后放置X线感光胶片上曝光约3h,分析感光照片的灰度值.结果:牙骨质基质提取物处理组的图象灰度面积和灰度值均显著高于仅用碘化物处理的空白对照组(P＜0.01).结论:牙骨质基质提取物能较好地吸附在光滑的钛表面上.     

重组人成纤维细胞生长因子21调节成牙骨质细胞矿化作用及其机制研究

目的 探讨重组人成纤维细胞生长因子21 (rhFGF21)对成牙骨质细胞增殖和矿化的作用及其机制。方法 采用苏木精-伊红染色、免疫组织化学染色和免疫荧光法检测成纤维细胞生长因子21 (FGF21)在大鼠牙周组织和成牙骨质细胞OCCM-30中的表达和分布;采用CCK-8法检测经不同浓度rhFGF21处理OCCM-30的增殖情况;采用碱性磷酸酶染色和茜素红染色分别检测OCCM-30矿化诱导3、7 d后的矿化状态;通过实时定量聚合酶链反应（PCR）及蛋白免疫印迹法（Western blot）检测成骨相关基因Runx2和Osterix的转录及蛋白表达情况;通过PCR阵列分析评估OCCM-30内转化生长因子β (TGFβ)/骨形态发生蛋白（BMP）信号通路基因的表达变化。结果 FGF21在大鼠牙周组织和OCCM-30中存在表达;rhFGF21对OCCM-30的增殖能力无明显影响,但50 ng/mL rhFGF21能促进OCCM-30矿化能力增强（P<0.001）;实时定量PCR结果显示Runx2和Osterix的转录水平在50 ng/mL rhFGF21矿化诱导3 d时上升,5 d时下降;...     

成骨细胞和牙周膜成纤维细胞交互作用的初探

目的　了解牙周组织再生中两种重要的细胞(成骨细胞和牙周膜成纤维细胞)之间的相互调控作用,以探讨牙周膜形成和维持的相关因素及其对牙种植体周围结构的影响机制。方法　采用原代培养法获取同一SD大鼠的成骨细胞和牙周膜成纤维细胞,将两种细胞分别接种于培养板上,再置于细胞培养池中以建立体外共同培养模型,分别测试两种细胞的碱性磷酸酶(ALP)及Ⅰ型胶原表达。结果　两种细胞在共同培养后,其ALP活性都发生显著变化,牙周膜成纤维细胞ALP表达明显提高,而成骨细胞ALP则降低;Ⅰ型胶原未发生显著变化。结论　当两种细胞在体外共同培养时,牙周膜成纤维细胞能抑制成骨细胞的成骨作用,而成骨细胞则能诱导牙周膜成纤维细胞向成骨样细胞分化。     

烟草浸提液对大鼠成骨细胞在钛板上附着和增殖的影响

目的:观察烟草浸提液(smokeless tobacco extract,ST)对大鼠成骨细胞(rat osteoblasts,ROBs)在钛板上附着和增殖的影响。方法:采用酶解-组织块法进行ROBs的原代培养,观察细胞生长状态;用碱性磷酸酶(ALP)组织化学染色法、矿化结节茜素红染色法对成骨细胞进行鉴定。不同浓度的ST作用于附着在钛板上的ROBs,观测细胞的铺展面积,MTT法检测钛板表面细胞的附着、增殖情况。采用SPSS 13.0软件包对数据进行双因素方差分析。结果:ALP染色显示,ROBs细胞内有黑色颗粒,矿化结节茜素红染色为红色,证明该细胞为成骨细胞。随着ST浓度的升高,ROBs在钛板上的铺展面积逐渐变小;增殖呈下降趋势,ST实验组与阴性对照组之间存在显著差异(P<0.05)。各实验组内前3 d存在显著差异(P<0.05),但3.2～50g/L组第4～7天间的差异无统计学意义(P>0.05)。结论:ST可影响ROBs在钛板上附着和增殖,并随ST浓度的增加而增强,提示吸烟不利于口腔种植的骨整合。     

Protein production by human gingival fibroblasts is enhanced by guanidine EDTA extracts of cementum

Nonconfluent cultures of human gingival fibroblasts were exposed to both guanidine and guanidine EDTA extracts of cementum for 48 hours. To compare the effects of cementum extracts on fibroblasts with other mineralized tissue extracts, cells were also exposed to guanidine and guanidine EDTA extracts of dentin and alveolar bone. The cells were radioactively labeled during the last 24 h. Total protein production was measured via the incorporation of radioactive proline. Collagen production was estimated by digestion of the radioactive protein mixture with bacterial collagenase. All guanidine EDTA extracts elicited statistically significant increases in total protein production compared to controls. At 50 μg/ml of extract, the increases in protein production were 340%, 143% and 338% for bone, cementum and dentin, respectively. Similar results were obtained for collagen production. In contrast, the guanidine extracts had no effect on either protein or collagen production by human gingival fibroblasts. These data indicate that the functions of gingival fibroblasts can be altered by proteins from associated mineralized tissues. Identifying such proteins and understanding their biological functions will enhance our knowledge of the mechanisms that regulate connective tissue regeneration.     

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目的    探讨烟草提取液（ST）和尼古丁液对人牙龈成纤维细胞（human gingival fibroblast,HGF）的影响。方法    2009年6—12月于昆明医学院口腔实验室,用含不同浓度的ST和尼古丁液的培养液体外培养HGF 5 d,应用噻唑蓝（MTT）快速比色法,检测细胞生长增殖情况。结果    与对照组相比,ST质量浓度从5 g／L开始时,抑制HGF的增殖,差异有统计学意义（P < 0.05）,随着浓度的加大,抑制作用更明显,呈浓度依赖性。同样,高质量浓度的尼古丁液从0.01 g／L开始对细胞增殖有抑制作用,并呈浓度依赖性,差异有统计学意义（P < 0.05）。结论    高浓度的ST和尼古丁均能抑制HGF的增殖,低浓度时则对细胞的增殖无影响。烟草对口腔黏膜上皮细胞的增殖呈现抑制作用,并随着浓度的加大,抑制作用愈发明显。     

Effect of various root surface treatments on the attachment and growth of human gingival fibroblasts: histologic and scanning electron microscopic evaluation

Human teeth extracted because of advanced periodontal disease were obtained. The portions of the roots which had been exposed in periodontal pockets were either untreated or were treated with root planing or citric acid, or root planing followed by citric acid. Human gingival fibroblasts were then added to the roots so treated and were allowed to incubate for 72 h. The ability of cells to attach to and grow onto these roots was assessed by means of gross evaluation of staining intensity and by histologic and scanning electron microscopic observation. The results of multiple experiments in each root-treatment category indicated that only roots which had been planed, whether or not citric acid demineralization was used, promoted cell attachment and growth. In addition, there were no discernible morphologic differences in the cells which were plated onto roots which were root planed only, compared to those which were root planed and citric-acid treated. In both situations too, the cells displayed morphology typical of human gingival fibroblasts in culture.     

Interleukin-6 production by human gingival fibroblasts

The ability of human gingival fibroblasts to synthesize interleukin-6 (IL-6) was studied using in vitro and immunohistochemical techniques. Culture supernatants of human gingival fibroblasts contained significant quantities of IL-6 activity which could be stimulated by fetal calf serum, recombinant interleukin-1 beta and lipopolysaccharide. The activity in the supernatants was specifically attributed to IL-6 since up to 97% of the activity could be inhibited by an anti-IL-6 antibody. Immunohistochemical studies on low-density human gingival fibroblast cultures indicated that the cells were associated with material reactive to the anti-IL-6 antibody. This localization was seen on the cell surface and in the cytoplasm of the cells. Immunoreactivity towards IL-6 was also noted in sections of human gingivae. Moderate staining was seen in the connective tissues and lower portions of the gingival epithelium, while intense staining was seen at foci of inflammation. The identification of IL-6 with human gingival tissues and cells implicates this lymphokine in the molecular events associated with the inflammatory periodontal diseases.     

Effects of polyhexamethylene guanidine phosphate on human gingival fibroblasts

Objective: Polyhexamethylene guanidine phosphate (PHMG-P) was compared to chlorhexidine (CHX) in order to determine potential cytotoxic and immune-modulatory effects on human gingival fibroblasts.

Materials and methods: Cytotoxic effects of PHMG-P and CHX on human gingival fibroblasts were assessed using cell viability assay at various time points and concentrations. The effects of PHMG-P and CHX on the secretion of prostaglandin (PG) E₂, interleukin (IL)-6, IL-8 and matrix metalloproteinase (MMP)-1 by non-stimulated or IL-1β stimulated fibroblasts were evaluated by enzyme-linked immunosorbent assays.

Results: PHMG-P concentration 0.00009% led to the total loss of fibroblast viability within 24?h, whereas inhibition of fibroblast viability by CHX occurred at significantly higher concentrations of 0.0009% (p?p?2, IL-6, IL-8 and MMP-1. Treatment of IL-1β stimulated fibroblasts in combination with PHMG-P or CHX at concentrations of 0.000045 or 0.0.00009% resulted in significantly decreased PGE₂, IL-6, IL-8 and MMP-1 levels. PHMG-P or CHX alone did not affect the baseline secretion of PGE₂, IL-6, IL-8 or MMP-1 by gingival fibroblasts.

Conclusions: Cytotoxic effects on gingival fibroblasts were triggered by both PHMG-P and CHX at concentrations below those used in clinical practice. The tested antiseptics did not cause inflammation and reduced IL-1β-induced secretion of inflammatory mediators and collagenase by gingival fibroblasts, which suggests anti-inflammatory properties.     

富血小板血浆和米诺环素对牙周膜成纤维细胞附着增殖的影响

目的：研究富血小板血浆（platelet-richplasma,PRP）和米诺环素对牙周膜成纤维细胞（periodontalliga-ment fibroblast,PDLFs）在病变牙骨质上附着增殖的影响。方法采用组织块法培养原代PDLFs,二步密度梯度离心法制备PRP,取因重度牙周病拔除的患牙,制取牙根片40片,随机分为4组。 A组单独使用PRP;B组单独使用米诺环素;C组联合使用PRP和米诺环素;D组为对照组。采用四唑化合物—吩嗪硫酸甲酯比色法检测病变牙根片上的细胞活性,采用SPSS l1．5统计软件分析结果。结果附着实验A、B、C、D组的光密度值分别为0．4420±0．0165、0．4983±0．0176、0．5443±0．0094和0．3857±0．0047。增殖实验A、B、C、D组的光密度值分别为0．4767±0．0049、0．6533±0．0022、0．6947±0．0011和0．4023±0．0092。 PRP与米诺环素单独作用均显著促进PDLFs在牙骨质上的附着和增殖,而且两者具有协同作用。结论 PRP及米诺环素均对PDLFs在病变牙骨质上的附着和增殖有促进作用,且两者有协同作用。     

Expression of fibrillins and tropoelastin by human gingival and periodontal ligament fibroblasts in vitro

The elastic system fibers consist of three different types, oxytalan, elaunin and elastic fibers, which differ in the relative content of microfibrils and elastin. In periodontal tissues, oxytalan fibers are known to be distributed in the periodontal ligament and gingiva, while elaunin and elastic fibers are present only in the gingiva. We examined the in vitro synthesis of microfibrils and elastin by human gingival fibroblasts (HGF) and periodontal ligament fibroblasts (HPLF). The two kinds of HGF and HPLF were cultured in MEM containing 10% newborn calf serum for 30 days. Since fibrillin-1 and fibrillin-2 are the major components of microfibrils involved in elastogenesis, we investigated the synthesis of fibrillins and tropoelastin in the conditioned medium of HGF and HPLF. Western blot analysis revealed fibrillin-1 and fibrillin-2 to occur in the HGF and HPLF culture medium, HGF exhibiting a higher level of synthesis than HPLF. Tropoelastin, on the other hand, was detected only in the medium of HGF after day 24. In addition, analysis of RNA extracted from HGF and HPLF on day 30 showed that only HGF expressed mRNA encoding tropoelastin. Immunohistochemically, accumulation of tropoelastin in the perinuclear area was found only in HGF. These results show that HGF expressed microfibrils and elastin, while HPLF expressed only microfibrils for the experimental period, and suggest a biochemical basis for the different distribution of elastic system fibers of the gingiva and periodontal ligament in vivo.     

护牙素与多乐氟对放射线照射后牙骨质显微硬度的影响

目的 探讨护牙素与多乐氟对放射线照射后牙骨质显微硬度值的影响.方法 收集因正畸减数拔除的前磨牙40颗,随机分为4组,其中3组在20℃下对牙根表面开窗区进行放射线照射：直接照射组直接采用放射线照射开窗区;护牙素组每次照射前先在开窗区涂上护牙素,再进行放射线照射,照射结束后去离子水冲洗表面;多乐氟组首次照射前在开窗区按照操作步骤涂多乐氟,再次照射前无需再涂;另设空白对照组,只在相同环境中保存,开窗区表面不做任何处理.照射前后分别测量各组的牙骨质表面显微硬度值.结果 照射前直接照射组、护牙素组、多乐氟组、空白对照组的显微硬度值分别为86.8±6.88、90.9±9.34、84.5±7.87、91.2±9.81,各组牙骨质表面显微硬度值差异无统计学意义（P＞0.05）.照射后,直接照射组显微硬度值（30.7±5.62）明显低于护牙素组（113.4±10.15）、多乐氟组（135.9±8.52）、空白对照组（91.2±9.81） （P ＜0.05）,护牙素组显微硬度值明显高于空白对照组（P＜0.05）,而多乐氟组显微硬度值明显高于空白对照组与护牙素组（P＜0.05）.结论 应用护牙素或多乐氟能够有效预防放射线所致的牙骨质脱矿,并能促进其再矿化,但多乐氟的防龋效果优于护牙素.