氟化钠及续断组分对成骨细胞增殖的影响

本研究利用［３Ｈ］－ＴｄＲ掺入法观察不同浓度的氟化钠及中药续断组分对体外培养的正常成人成骨细胞的影响。结果显示，氟化钠能直接影响成骨细胞的增殖，且具有双向调节效应，在一定浓度范围内（１．２５～１０μｍｏｌ／Ｌ）能促进细胞增殖，但其有效浓度范围相对狭窄，浓度过高（＞２０μｍｏｌ／Ｌ）反而抑制成骨细胞的生长。中药续断的两种组分对成骨细胞的增殖均具有促进作用，但有各自不同的有效浓度范围。本研究提示氟化钠对成骨细胞具有直接的作用。中药续断对成骨细胞也有促增殖作用，理论上有望从促骨形成方面用于骨质疏松的预防及治疗，同时也为中药的筛选提供了一条新路。     

补肾中药HU-ECS对培养成骨细胞增殖、分化及矿化功能的影响

目的：为了解补肾中药HU-ECS对体外培养成骨细胞的作用。方法：应用MTT法、对硝基苯磷酸盐法及茜素红染色方法观察HU-ECS对体外培养民骨细胞的增殖、ALP表达及矿化结节形成的影响。结果：HU-ECS具有刺激骨细胞增殖作用（10^-6g/ml组,P＜0.01-0.001）,提高ALP活性及矿化结节形成的数量。结论：HU-ECS具有刺激体外培养成骨细胞增殖及分化成熟的功能。     

骨疏康对成骨细胞刺激作用的研究

目的 通过体外成骨细胞培养,观察补肾中药骨疏康对成骨细胞的直接刺激作用。方法 采用第２代成骨细胞加入浓度为５μｇ／ｍｌ、１０μｇ／ｍｌ、２０μｇ／ｍｌ的骨疏康药液,测定ＡＫＰ、ＢＧＰ、ＰＤＹ的含量并进行细胞计数。结果 加药后细胞数随药物浓度增加而增加,ＡＫＰ和ＢＧＰ水平也相应提高。２０μｇ／ｍｌ组的ＬＡＰ和ＢＧＰ水平较５μｇ／ｍｌ组和１０μｇ／ｍｌ组增加明显,有显差异Ｐ〈０．０５。ＰＹＤ含量随药     

氟化钠对大鼠破骨样细胞及其凋亡的影响

目的探讨氟化钠对大鼠破骨样细胞及其凋亡的影响,进而推测地方性氟病的发病和氟化物治疗骨质疏松的机制.方法以5mg/L NaF和15mg/L NaF饲喂正常和去卵巢(OVX)大鼠6个月,观察了体内和培养基中的NaF对大鼠破骨样细胞(OLC)形成和凋亡的影响.结果氟摄入2个月时,15mg/L NaF明显抑制正常和OVX大鼠OLC形成;第5个月时,5mg/L NaF和15mg/L NaF均抑制正常和OVX大鼠的OLC形成.氟摄入2周时,发现NaF促进OVX大鼠OLC凋亡;1个月时,NaF促进正常和OVX大鼠OLC凋亡,随时间的延长,其促进作用逐渐增强.以上作用均表现为15mg/LNaF的作用明显大于5mg/L NaF.体外NaF抑制OLC形成并促进OLC凋亡,其作用程度为15mg/LNaF＞lOmg/L NaF＞5mg/L NaF.结论体内NaF抑制正常和OVX大鼠OLC的形成并促进正常和OVX大鼠OLC凋亡,其作用是时间和剂量依赖性的.培养基中NaF抑制OLC形成并促进正常和OVX大鼠OLC的凋亡,其作用是剂量依赖性的.     

内皮素受体拮抗剂对前列腺癌细胞PC3和成骨细胞相互作用的影响

目的 在细胞共培养微环境下,观察内皮素(ET)受体拮抗剂阻断内皮素轴对前列腺癌细胞PC3和成骨细胞SaOS_2相互作用的影响.方法 利用细胞体外共培养系统,比较前列腺癌细胞PC3和成骨细胞SaOS_2在共培养微环境下和单独培养环境下细胞增殖的差异.共培养微环境下,分别以ET_A受体(ET_AR)拮抗剂BQ123和ET_B受体(ET_BR)拮抗剂BQ788阻断相应的ET受体,观察其对前列腺癌细胞和成骨细胞增殖的影响.同时,通过酶联免疫吸附试验(ELISA)法测定各组培养液中的ET-I浓度,通过逆转录.聚合酶链反应(RT-PCR)测定PC3细胞、SaOS_2:细胞ET-I及其受体的mRNA表达,比较它们的表达差异.结果 与单独培养比较,共培养微环境下的PC3细胞和SaOS_2细胞的增殖数量均显著升高(P均＜0.05).ET_AR拮抗剂BQ123能阻断共培养微环境对SeOS:细胞的生长促进作用,部分阻断共培养微环境对PC3细胞的生长促进作用,而ETBR拮抗剂BQ788则无显著的干预作用.EUSA结果显示共培养组培养液中ET-1浓度[(14.26±1.06)βg/L/10~5个细胞]显著高于单独培养组[(6.58±0.74)pc/L/105个细胞,P＜0.05].RT-PCR结果显示PC3细胞表达ET-1和ET_AR,SaOS_2细胞只表达ET_AR;与单独培养比较,共培养微环境下PC3细胞ET-l的表达显著升高(P＜0.05),而PC3细胞和SaOS2细胞ET_AR的表达差异均无统计学意义(P＞0.05).结论 ET-1及ET_AR构成的内皮素轴是共培养微环境中前列腺癌细胞PC3和成骨细胞之间相互作用促进增殖的重要链接因子,而ETAR拮抗剂能有效地抑制两者间的相互作用,抑制PC3细胞增殖.     

流体剪切力对成骨细胞影响的研究进展

骨是一个富含多孔的组织,被组织间液持续灌流,由于受血压和活体外机械装载的驱使,组织间液经过骨组织中的腔隙、小管网和骨内膜表面的骨内层产生明显的流体剪切力（fluid shear stress,FSS）。大量研究表明,成骨细胞是其中重要的感受与效应细胞,可通过力敏感离子通道、G蛋白与酪氨酸激酶、整合素受体与细胞骨架等多种途径感受体内外力学刺激,并将力学刺激信号转化为细胞生物化学信号,介导力相关敏感基因表达,合成各种酶类等活性物质,激活信号网络级联反应,     

番茄红素对氧化应激后成骨细胞的影响

目的研究具有强抗氧化作用的番茄红素对氧化应激后成骨细胞的影响。方法制造氧化应激状态成骨细胞模型。将细胞随机分为4组:高、中、低浓度番茄红素组及对照组。然后用流式细胞术检测各组成骨细胞生长指数,用酶标仪检测各组成骨细胞内碱性磷酸酶的活性。在各组细胞中另加入矿化结节诱导液,观察各组成骨细胞矿化能力。结果各实验组成骨细胞较对照组生长指数高,碱性磷酸酶表达增加,形成的矿化结节多。结论番茄红素可以增加氧化应激状态成骨细胞的增殖、碱性磷酸酶的表达及矿化功能。     

阿伦磷酸钠对人成骨细胞增殖的影响

目的 :改良成人成骨细胞消化培养方法 ,研究阿伦磷酸钠对成人成骨细胞增殖的影响。方法 :手术中获取病人的髂骨块 ,酶消化法得到成骨细胞 ,传 4代后以 1× 10 5/ml的浓度种于 2块 96孔板中。MTT方法检测阿伦酸钠对成骨细胞增殖的影响。结果 :高于 10 4M浓度的阿伦磷酸钠明显抑制成骨细胞的增殖 ,浓度低于 10 5M时 ,成骨细胞的增殖不受影响。结论 :高浓度的阿伦磷酸钠抑制人成骨细胞的增殖     

神经肽对人成骨细胞生物学影响机理的研究

目的：研究神经肽类物质对成骨细胞的钙离子通道及胞间通讯的影响。探讨其对细胞生物学特征影响的机制。方法：借助CLSM及Fluo-3/AMt CFDA荧光染色,观察0．1ng/ml的SP、NPY对成骨细胞内Ca^2 浓度及胞间通讯的影响。结果：借助于CLSM观察到SP、NPY及BMP均使成骨细胞的胞内Ca^2 浓度增加,并可加强细胞间的通讯联系。结论：神经肽引起胞间通讯的加强可使成骨细胞对各种因子刺激的反应更趋一致性,可提高成骨的质量。     

尼莫地平对体外培养成骨细胞的影响

目的 观察尼莫地平对体外培养成骨细胞的作用.方法 在新生SD大鼠头颅骨第2继代成骨细胞(OB2)培养液中分别加入不同浓度(10-1～10 9g/L)尼莫地平,分别观察OB2的增殖功能(用波长570nm处OD值表示),分化功能(用碱性磷酸酶ALP活性表示)和矿化功能(用矿化结节数量/视野表示).结果 增殖功能OD值为0.12±0.01～0.41±0.04;ALP活性为0.09±0.01U/mg蛋白质;矿化结节数量/视野为1.3±0.9个.结论 与对照组比较,尼莫地平各浓度对OB2的增殖、分化和矿化功能作用各异.当尼莫地平浓度为10 6g/L时,对OB2的增殖和分化功能具有刺激作用,对OB2的矿化功能具有显著的抑制作用;当浓度升高(10-2g/L)时,对OB2的增殖功能具有明显的抑制作用;当浓度降低(10 8g/L)时,对OB2的增殖功能失去作用.     

Radiopacity and fatigue characterization of a novel acrylic bone cement with sodium fluoride

Acrylic bone cement must provide good radiographic visibility and good long-term mechanical resistance in joint replacements. A new formulation of cement with 6% barium sulfate and 6% sodium fluoride was developed (Fluoride Bone Cement). Barium sulfate is a necessary addition to allow radiographic visibility although it reduces the mechanical strength of the material. Sodium fluoride promotes bone formation. However, its effect on the mechanical behavior is currently unknown while its influence on radiopacity can only be roughly estimated. The aim of this investigation was to establish if the new formulation would be suitable for clinical trials. In this respect, a mechanical (fatigue test) and radiographic (optical density measurements on x-ray films) characterization was performed on a typical commercially available cement with barium sulfate added and on the Fluoride Bone Cement. It was demonstrated that the fluoride cement has a (marginally) superior fatigue strength and comparable radiopacity to commercial radiopaque cements.     

Effect of controlled local release of sodium fluoride on trabecular bone.

Systemic sodium fluoride has been used in the treatment of osteoporosis. Recent studies have shown that it has a positive risk/benefit ratio for use in increasing spinal trabecular bone density. However, thinning of the cortices of the long bones with a resulting increase in fracture incidence has been observed. This study was designed to determine the response of bone to sodium fluoride released from a biodegradable polymer matrix, a technique which could potentially deliver it locally to a site of need in the skeleton which has a positive response to fluoride. In one group of mature New Zealand white rabbits, cylindrical poly(D,L-lactic acid) (PLA) implants, with or without impregnated sodium fluoride, were implanted into the contralateral femoral trochanters and tibial metaphyses. In a second group, similar implants were placed in adjacent vertebrae. Four weeks postimplantation, the femora, tibiae, and vertebrae were removed, sectioned, cleaned of all but mineralized tissue, and the surfaces of the sections stained. The stained surfaces were imaged and analyzed for morphometric properties of the trabeculae. Comparing contralateral vertebrae, those exposed to sodium fluoride had significantly thickened trabeculae, with decreased spacing between them and a greater bone fraction. A similar increase in trabecular width was found in the subchondral bone of the proximal tibiae exposed to local release fluoride. Femoral sections showed no difference, possibly due to the lack of extensive trabecular bone in the region chosen for study.     

The effects of sodium fluoride on bone breaking strength

The therapeutic use of sodium fluoride has been recommended in a variety of osteopenic bone diseases. The recommendations are based mainly on the known osteosclerotic effects of sodium fluoride and little information is available as to its effect on bone strength. The influence of various concentrations of sodium fluoride on bone strength in growing rats on high and low calcium diets was studied. The administration of sodium fluoride increased bone diameter, indicating stimulation of periosteal bone formation, but bone strength was reduced or not affected by fluoride ingestion.

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Zusammenfassung Die therapeutische Verwendung von Natriumfluorid ist für eine Anzahl von Knochenmangel-Krankheiten empfohlen worden. Die Empfehlungen basieren hauptsächlich auf den bekannten osteosklerotischen Wirkungen von Natriumfluorid; über dessen Effekt auf die Knochenstärke ist wenig bekannt. In dieser Arbeit wurde der Einfluß verschiedener Konzentrationen von Natriumfluorid auf die Knochenstärken von wachsenden Ratten mit hoher und niederer Calciumeinnahme untersucht. Die Verabreichung von Natriumfluorid erhöhte den Knochendurchmesser, was auf eine Stimulierung der Periostbildung hinwies, die Knochenstärke wurde jedoch durch Fluorideinnahme herabgesetzt oder nicht beeinflußt.

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Résumé L'utilisation thérapeutique de fluorure de sodium a été recommandée dans le traitement de diverses maladies osseuses ostéopéniques. Cet emploi est basé principalement sur les effects ostéosclérotiques bien connus du fluorure de sodium, mais on connait mal son effet sur la force de résistance osseuse. L'influence de diverses concentrations de fluorure de sodium sur la force de résistance osseuse de jeunes rats soumis à des régimes riches et pauvres en calcium a été étudiée. L'administration de fluorure de sodium augmente le diamètre osseux, indiquant une stimulation de la formation de l'os périosté, mais la force de résistance osseuse est réduite ou non modifiée par l'ingestion de fluor.

     

Influence of food and antacid administration on fluoride bioavailability from enteric-coated sodium fluoride tablets

The relative bioavailability of enteric-coated sodium fluoride (NaF) tablets (10 mg F⁻) has been assessed following administration with a standard calcium-rich breakfast or calcium-poor lunch, and 2 h before or simultaneously with antacid administration (2.4 g aluminum-magnesium hydroxide), versus intake on an empty stomach. Twelve volunteers were studied 3 times according to an open, three-way crossover design over a 24 h period at weekly intervals. Meals were found to decrease the peak serum concentration of NaF from 122 μg/L during fasting (after baseline subtraction) to 71 and 88 μg/L with breakfast and lunch respectively, and to slow its absorption rate with Tmax increasing from 3.3 to 7.3 and 11.2 hours, without altering its bioavailability. Antacid impaired the bioavailability of NaF by 80% when administered simultaneously, with AUC decreasing from 987 to 155 μg.h/ L, but had no significant effect when taken 2 h before NaF. In conclusion, the enteric-coated NaF tablets used in this study can be administered with food or after a 2-hour delay following antacid administration, but should not be taken simultaneously with antacid.     

Effects of low-dose long-term sodium fluoride preventive treatment on rat bone mass and biomechanical properties

Effects of fluoride on bone strength and cortical bone mass remain controversial. We compared 9-month, low-dose sodium fluoride (NaF) treatment with estrogen replacement therapy. Female Wistar rats 4.5 months old were divided into baseline, sham-operated (sham), sham-treated with NaF at 0.5 mg NaF/kg/day in drinking water, and ovariectomy (OVX), OVX treated with NaF and with estrogen. Bone mass was measured by dual X-ray absorptiometry (DXA)in vitro. Dimensions of the first lumbar vertebral body (L1) were determined by radiogrammetry. The right femur was processed undecalcified to obtain a midshaft cross-section to determine cross-sectional moments of inertia (CSMIs). L1 compressive test and left femoral torsional test were performed. OVX induced significant bone loss in L1 and femoral midshaft. Bone mass was increased to a greater extent in NaF-treated rats than in rats receiving estrogen replacement therapy. Femoral CSMIs in OVX rars, both L1 sizes and femoral CSMIs in NaF-treated rats, were significantly increased. Estrogen treatment had the least dimension expansion. OVX significantly decreased L1 compressive variables. There was no statistical difference in compressive parameters between NaF-treated groups and controls. OVX significantly increased femoral torsional strength but NaF treatment did not. Bone fluoride content was significantly increased after treatment with NaF. No significant difference in bone mineralization degree (ash and calcium) was found between treated and control rats. The discrepancy that an increase in bone mass and geometric properties in both trabecular and cortical bones by low-dose, long-term NaF treatment did not increase vertebral strength nor proportionally improve femoral strength indicated that the bone intrinsic biomechanical properties could be changed by NaF treatment.     

格列美脲对大鼠下颌骨成骨细胞PI3K／Akt通路的影响

目的研究格列美脲（glimepiride）对大鼠下颌骨成骨细胞PI3K／Akt通路的影响。方法分离培养原代下颌骨成骨细胞,进行成骨细胞鉴定,将成骨细胞以1×10^5接种于25cm^2培养瓶中,细胞融合后加入10μmol／L格列美脲分别干预0、15、30、60、120min。提取蛋白,取等量蛋白,采用凝胶电泳,分别加入兔抗大鼠PI3KP85、Akt／P-Akt抗体、IRS-1／P-IRS-1抗体和G肌动蛋白,37℃孵育4h,洗膜后,经辣根过氧化物酶标记的羊抗兔IgG孵育50min,化学发光法显示抗原抗体复合物,X线片显影并定影,所有杂交信号在Labworks4．5成像分析仪系统测定条带密度。蛋白目的条带水平用与β肌动蛋白的比值表示。以上实验重复3次。结果在格列美脲诱导下,PIKP85、Akt／P—Akt、IRS-1／P—IRS-1表达显著提高。结论格列美脲在成骨细胞中也能激活PI3K／Akt通路。     

Relationship between bone fluoride content and histological evidence of calcification defects in osteoporotic women treated long term with sodium fluoride

Fluoride treatment is used to increase bone formation and cancellous bone mass in patients suffering from postmenopausal osteoporosis with vertebral fractures. Patients submitted to similar therapeutic protocols have shown various histological responses to the treatment, some developing calcification defects and others not. In fact, the bone histological response to fluoride salts depends on the cumulative uptake of fluoride by bone. To clarify the relationship between the presence of calcification defects (identified by the presence of mottled bone and linear formation defects) and the bone fluoride content, a retrospective study was performed on 29 women with type 1 osteoporosis and treated for several months (11–24) with sodium fluoride (50 mg/day), calcium and vitamin D. Bone fluoride content always significantly increased after treatment, but it was significantly higher in patients showing calcification defects than in those having no defects. These differences between the two groups of patients were not due to differences in clinical details (no significant differences concerning age, duration of treatment, total amount of fluoride ingested, renal function) or in their bone remodelling activity. Thus, it may be hypothesized that the high bone fluoride uptake is due to different individual responses from one patient to another concerning the bioavailability of the same dose of fluoride. This is difficult to predict, except by testing the individual bioavailability of the compound to be used in each patient before starting long-term treatment.     

乳铁蛋白对大鼠成骨细胞增殖与分化的影响

目的观察乳铁蛋白(Lf)对大鼠成骨细胞增殖分化的影响,初步探讨Lf对成骨细胞的作用机制。方法用不同浓度的Lf干预成骨细胞,在不同时间分别测定细胞数目的增殖、细胞内碱性磷酸酶活性,并用半定量RT-PCR法检测成骨细胞内RANKL/OPG mRNA基因的表达。结果Lf呈时间和浓度依赖性地促进大鼠成骨细胞增殖及碱性磷酸酶活性,每组实验在不同时间的差异均有显著的统计学意义(P<0.05),其中400μg/ml组在72h时细胞增殖数目达到最大。在碱性磷酸酶活性方面,400μg/ml组在72h时则有所下降。不同浓度Lf、不同作用时间均能促使成骨细胞中OPG mRNA表达增加,同时抑制RANKL mRNA表达。结论Lf能促进成骨细胞的增殖分化,并且能够调节成骨细胞RANKL/ OPG mRNA表达,从而抑制破骨细胞介导的骨吸收。     

氟化钠对人黄韧带细胞碱性磷酸酶活性及骨钙素合成的影响

目的 探讨氟化钠(NaF)对体外培养条件下人黄韧带细胞碱性磷酸酶(alkaline phosphatase,ALP)活性及骨钙素(bone gla protein,BGP)合成的影响.方法 依据手术标本来源不同,分为正常黄韧带细胞(NLF)组(取自急性外伤性胸腰椎骨折截瘫患者,7例)、退变黄韧带细胞(DLF)组(取自退变性腰椎管狭窄症患者,9例)及骨化黄韧带细胞(OLF)组(取自胸椎黄韧带骨化患者,8例).采用组织块贴壁法进行体外细胞培养,共获得24组传代细胞.取第五代细胞加入不同浓度的NaF,观察细胞形态变化,并测定NaF对各组人黄韧带细胞的ALP活性和BGP合成的影响.结果 不同来源的人黄韧带细胞均可在体外增殖并传代,DLF与OLF组人黄韧带细胞呈多形性表现,并可形成钙结节.体外培养条件下,高浓度(1.0mmol/L)NaF可导致人黄韧带细胞发生中毒反应;低浓度(0.01～0.125 mmol/L)NaF可促进DLF组黄韧带细胞增殖、钙结节形成,ALP活性上调,BGP合成明显增加,而对OLF与NLF组人黄韧带细胞则无明显效应.结论 体外培养条件下,低浓度(0.01～0.125 mmol/L)NaF可促进人退变黄韧带细胞的ALP活性增高及BGP合成增加,提示低浓度NaF可能促进人退变黄韧带细胞向成骨方向分化.