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。     

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。     

人牙骨质基质提取物的提取及蛋白组分分析

目的：探讨制取牙骨质某质提取的可行性并分析提取物的蛋白组分，方法：收集因正畸而拔除的牙周末感染的恒牙，用钝性刮匙去除牙根表面的结缔组织，在解剖显微镜下从牙本质表面刮下牙髓质，分别制取牙髓质基质醋酸和胍2种取物。采用SDS－聚苯烯酰胺凝电泳法分析2种提取物的蛋白组分。结果：牙骨质基质醋酸提取物主要含有Mr55000和Mr68000 2种蛋白，而牙骨质基质胍提取物则含Mr62000、Mr60000、Mr55000及Mr41000 4种蛋白。结论：所采用的蛋白质提取方法是可行的。     

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     

牛成牙骨质细胞在体内形成牙骨质样组织的初步研究

目的：培养成牙骨质细胞（cementoblasts,CBs)，观察以CBs自身作为组织工程支架在裸鼠内形成牙骨质的可行性。方法：在矿化培养液中培养新生牛的成牙骨质细胞，使其形成多层CBs作为载体（CBs-made carrier)与CBs混合，植入裸鼠皮下。以CBs与胶原膜载体（collagen carrier) 复合体作为对照。7周后取材并做HE、von Kossa、茜素红及牙骨质附着蛋白单克隆抗体免疫组化染色。结果：CBs在体内可形成牙骨质样矿化组织，中心矿化程度较高，四周有尚未钙化的类牙骨质及CBs包绕，并有牙骨质细胞埋入矿化组织。以CBs自身作为载体所形成的牙骨质较以胶原膜作为载体所形成的牙骨质样组织多。结论：本组分离培养的CBs在体内可形成牙骨质样矿化组织；以多层细胞自身作为载体是一种较理想的生物支架材料。     

牙骨质相关细胞的研究进展

到目前为止，人们对牙骨质的了解仍较少。人牙骨质再生被认为是牙周病暴露的根面上牙周组织再生的关键。本文回顾了学者们对牙骨质及其相关细胞的研究，阐述了形成牙骨质前体细胞的位置以及成牙骨质细胞的起源。     

人牙龈成纤维细胞和牙周膜成纤维细胞的培养和生物学特征

目的：探讨采用不同方法建立体外培养人牙龈成纤维细胞和人牙周膜成纤维细胞并对其生物学特性作初步探讨。方法：分别采用消化培养法和组织块培养人的牙龈和牙周膜成纤维细胞。通过倒置显微镜活体观察，以及光镜、透射电镜、免疫组化和生长曲线等方法对其生物学特性作初步观察。结果：消化培养法成功率低于组织块培养法。光镜下和透射电镜下观察两种细胞在形态和结构上相似。免疫组化染色证实此两种细胞均来源于中胚层的纤维母细胞。生长曲线表明牙龈成纤维细胞的倍增时间比牙周膜成纤维细胞稍短，而牙周膜成纤维细胞的增殖活性比牙龈成纤维细胞高。结论：组织块培养法适用于此二种细胞的培养。细胞生物学特征方面有许多相似形，提示这两种细胞来于同一个细胞群。     

硝苯地平和机械力对人牙周膜成纤维细胞基质金属蛋白酶的表达影响

目的研究机械静态拉伸作用下,硝苯地平对人牙周膜成纤维细胞（HPDLF）基质金属蛋白酶（MMP）-10和MMP-13的表达影响。方法按弹力形变不同将细胞随机分为4组,分别为0%、8%、12%、16%形变组,每组再按照硝苯地平浓度不同分为4个亚组,分别为0、10、30、50 μm亚组。硝苯地平孵育1 h后,给予细胞12 h静态拉伸,免疫组化染色检测细胞胞浆中MMP-10和MMP-13的表达。结果弹性形变为0%时,各亚组中MMP-10和MMP-13的表达无明显变化,与对照组相比差异无统计学意义（P>0.05）。当硝苯地平浓度为0 μm时,8%、12%、16%形变组胞浆内MMP-10和MMP-13均呈高表达,且随形变的增大,表达呈上升趋势（P<0.001）;当加入硝苯地平后,随硝苯地平浓度的增加,MMP-10和MMP-13表达均呈下降趋势（P<0.001）。结论硝苯地平对机械力诱导的MMP-10和MMP-13表达有抑制作用,提示钙离子通道参与机械力对HPDLF中MMP-10和MMP-13的诱导表达。     

人牙周膜成纤维细胞体外培养的研究进展

人牙周膜成纤维细胞是牙周细胞生物学、药理学、毒理学和牙周组织工程学研究的基础,对口腔各个领域意义重大。由于受到牙周组织的解剖结构和取材数量的限制,牙周膜成纤维细胞的体外培养较为困难,因此本文就人牙周膜成纤维细胞的体外培养方法和应用等研究进展作一综述。     

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.     

釉基质蛋白对人牙周膜细胞粘附、伸展的影响

目的：了解釉基质蛋白对人牙周膜细胞黏附，伸展的影响。方法：改良组织块法培养人牙周膜细胞，固相结合分析方法观察釉基质蛋白对人牙周膜细胞黏附，伸展的影响。结果：实验组PDLC黏附率与对照组无差别，实验组PDLC伸展率高于对照组。结论：EMPs对PDLC黏附无影响，但可促进其伸展。     

Glucose modulates growth of gingival fibroblasts and periodontal ligament cells: correlation with expression of basic fibroblast growth factor

Diabetes mellitus is a systemic disease with profound effects on oral health and periodontal wound healing. Uncontrolled diabetes adversely affects surgical wound healing and is often associated with abnormal proliferation of fibroblasts, excessive angiogenesis and poor bone regeneration. Human gingival fibroblasts and periodontal ligament cells from both diabetics and non-diabetics were evaluated for growth responses following culture in 20 mM glucose, a concentration compatible with blood glucose levels in uncontrolled diabetics. Gingival fibroblasts derived from 9 non-diabetic patients and 3 insulin-dependent diabetics either proliferated or showed little change of growth in elevated glucose. Enhanced proliferation was observed following 1 wk of culture in glucose. Growth of periodontal ligament cells from 5 non-diabetic patients was inhibited by 20 mM glucose. Fibroblasts that were markedly growth stimulated were probed for expression of basic fibroblast growth factor (bFGF) using a reverse-transcribed polymerase chain reaction (RT-PCR). Results indicate that fibroblasts exhibiting the greatest increase in growth in response to high glucose also exhibited increased expression of bFGF. No changes were observed in mRNA expression for platelet-derived growth factor-AA, platelet-derived growth factor-BB, insulin-like growth factor and transforming growth factor-β₁. Mitogenic effects induced by the cytosol of fibroblasts exhibiting increases of growth in 20 mM glucose were abrogated by neutralizing antibodies to bFGF. In addition, some periodontal ligament cells that were growth inhibited by high glucose had reduced expression of bFGF. These data suggest that bFGF may play a role in the abnormal wound healing associated with periodontal surgery of uncontrolled diabetics.     

Lectins inhibit periodontal ligament fibroblast attachment, spreading and migration on laminin substrates

The ways in which carbohydrate signals from the extracellular matrix influence the responses of periodontal ligament (PDL) fibroblasts are essentially unknown. The purpose of this study, using video digital image analysis, was to examine the effects of lectins on the attachment, spreading and migrational responses of rat periodontal ligament fibroblasts on the highly glycosylated glycoprotein laminin (LN) in vitro. PDL fibroblasts were isolated from rat molar teeth and grown in culture. Bound LN and control substrates (bovine serum albumin and untreated plastic) were incubated with solutions of either wheatgerm agglutinin (WGA), specific for N-acetylglucosamine, or concanavalin A (ConA), specific for mannose, in 96-well plates. Unbound lectin was rinsed away and 10.0 x 10(3) cells were seeded per well and allowed to attach for 1.5 h. Incubation of LN substrates with WGA, prior to the addition of any cells, inhibited PDL fibroblast binding more than 5-fold. ConA, however, had no effect on cell binding but inhibited mean individual cell spreading nearly 3-fold under similar assay conditions. The effects could be prevented by adding each lectin's respective specific sugar. The lectins had no effects on the control substrates. In a 24-h cell migration assay WGA and ConA both significantly inhibited migration of PDL fibroblasts. It is likely that WGA inhibited cell attachment and cell migration, by binding to oligosaccharides and blocking access to adjacent polypeptide cell recognition sequences on LN. The results from the ConA experiments, where binding was allowed but spreading was severely inhibited, suggest a possible informational role for the carbohydrates present on LN.     

两种生长因子对人牙周膜成纤维细胞在钛金属表面附着和生长的影响

目的：研究两种生长因子对牙周膜成纤维细胞在然金属表面附着和生长的影响。方法：将纯钛、钛75试件入在12孔培养板内，取生长良好的第五代人牙周膜成纤维细胞（PDLF）接种在试件表面，分别在接种后4h、12h、24h、72h进行贴壁细胞地数。结果：接种后4h、12h、24h、72h、bFGF组纯钛、钛75表面细胞附着数与空白对照组的差异均有显著性（P＜0．05），rhBMP－2组纯钛、然75表面细胞附着数在初期（24h）与空白对组无显著性差异（P＞0．05）。72h时与空白对照组差异有显著性（P＜0．05），表明bFGF促进细胞附着和生长作用显著，而rhBMP－2促进细胞生长作用较促附着作用明显。结果：PDLF在钛金属表面的附着和生长可被生长因子所增强，但不同的生长因子对细胞附着和生长的生物学效应不尽相同。     

体外培养碱性成纤维细胞因子对人牙周膜成纤维细胞生物学特性的影响

目的：探讨体外培养过程中,碱性成纤维细胞生长因子（bFGF）对人牙周膜成纤维细胞（hPDLFs）生物学特性的影响。方法：体外分离培养人牙周膜成纤维细胞并鉴定,加入不同浓度bFGF（1ng／ml、10ng／ml、50ng／ml、100ng／ml）培养,MTT方法检测细胞增殖情况;并对细胞进行矿化诱导,检测细胞的碱性磷酸酶活性。结果：hPDLFs呈星形或长梭形,免疫组化波丝蛋白阳性,角蛋白阴性,证实该细胞来源可靠。bFGF在一定浓度范围内,与细胞增殖成正比,而在本实验培养条件下（10％FBS）bFGF最大效应浓度为10ng／ml。矿化诱导条件下,碱性磷酸酶活性明显增加,在联合应用bFGF的情况下,100ng／ml组碱性磷酸酶活性明显高于其他组。结论：不同浓度bFGF对人牙周膜成纤维细胞生物学行为的影响不同,在一定浓度条件下,低浓度bFGF促进人牙周膜成纤维细胞的增殖,而高浓度bFGF作用于人牙周膜成纤维细胞可能更易于分化。