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1.
背景:骨形态发生蛋白被证实有成骨诱导作用,然而关于骨形态发生蛋白7诱导骨膜细胞成骨的超微结构研究尚未见报道。目的:观察骨膜细胞经成骨因子骨形态发生蛋白7诱导后的生物活性及超微结构。方法:实验取材于成人胫骨。分离出原代骨膜细胞后行常规体外培养,实验分为实验组和对照组。实验组加入骨形态发生蛋白7和细胞培养辅助剂,对照组仅仅加入了成骨细胞培养辅助剂。每组分别在第5,10,15天设3个时间点,每个时间点设3个样本,分别行钙结节的染色及采用相差显微镜观察大体形态结构,在透射电镜下观察超微形态结构。结果与结论:实验组和对照组形成的成骨细胞增殖良好,形态一致。细胞早期呈梭形,立体感强,饱满透明,分裂期呈短柱状或立方形,电镜下见成骨细胞内有大量的粗面内质网和高尔基复合体;后期细胞由长梭形逐渐变成宽梭和不规则形,后期射透电镜下可见细胞内有大量基质小泡,有膜包被,胞质内上有碱性磷酸酶及钙结合蛋白,泡内有钙盐结晶,成骨细胞的基底部和侧面出现突起与邻近的骨细胞突起相连。由骨形态发生蛋白7诱导的成骨细胞在体外增殖更快,细胞分裂及成骨速度更快。结果提示骨形态发生蛋白7在体外培养中具有明显增强成骨细胞增殖的能力。  相似文献   

2.
目的研究黄芪甲苷促进成骨细胞增殖分化的作用,并初步探讨作用机制。方法将成骨细胞MC3T3-E1分为对照组和实验组,对照组正常培养成骨细胞,实验组采用不同浓度黄芪甲苷干预成骨细胞,并孵育不同时间,MTT法观察对细胞增殖的影响,检测碱性磷酸酶(ALP)活性观察对细胞分化的作用,Western blot检测成骨细胞Toll样受体-4(TLR-4)蛋白表达。结果与对照组比较,随着黄芪甲苷浓度的增加,成骨细胞活性或ALP活性逐渐上升,并趋于稳定。延长黄芪甲苷作用时间,低浓度组细胞活性显著升高,而各组细胞ALP活性均显著上升。与对照组比较,实验组细胞中TLR-4表达显著下降。结论黄芪甲苷能够促进成骨细胞增殖和分化,其作用机制可能与TLR-4通路有关。  相似文献   

3.
目的 研究水飞蓟宾-磷脂酰胆碱复合物(Silybin-phosphatidylcholine Compound,SPC)诱导肝癌HepG2细胞凋亡作用及其机制.方法 不同浓度SPC处理肝癌HepG2细胞.通过MTT法、细胞形态学观察、流式细胞仪和激光共聚焦显微镜观察SPC时HepG2细胞的生长抑制作用及其细胞凋亡的影响.结果 SPC可抑制HepG2细胞的增殖.其GI50为46.8μg/ml;50μg/ml的SPC作用HepG2细胞24h后,细胞出现早期调亡的形态;75μg/ml、100μg/ml的SPC对HepG2细胞凋亡率分别为(17.22±3.45)%和(25.50±5.72)%;50μg/ml、75μg/ml、100μg/ml的SPC作用于HepG2细胞24h后,细胞内的Ca2+浓度显著升高(P<0.05,P<0.01).结论 SPC能够诱导肝癌细胞系HepG2细胞凋亡,其机制与SPC升高细胞内Ca2+浓度有关.  相似文献   

4.
新型改性聚乳酸与成骨细胞相容性的研究   总被引:5,自引:0,他引:5  
探讨了成骨细胞与新型生物可降解材料乙二胺改性聚乳酸(EMPLA)的细胞相容性。在PLA、EMPLA及玻璃(对照组)上培养成骨细胞,采用细胞形态学观察法和细胞增殖法,在相差显微镜下观察细胞在上生长情况;分别于1、2、4、6d用MTT法记数,绘制生长曲线图。实验结果表明EMPLA组的成骨细胞比PLA组和对照组的形态好,增殖快,说明EMPLA比PLA表现出更好的细胞相容性,EMPLA在生物医学,特别是在组织工程领域存在着广泛的应用性。  相似文献   

5.
目的评价新型有序纳米介孔生物活性玻璃(80SMBG)的浸提浓度对成骨细胞的影响。方法采用大鼠成骨细胞体外培养,按细胞增殖度法,取0.25%、0.5%、1%、2%、4%共五种浓度的80SMBG浸提培养液进行检测,分别于3、4、5天用倒置显微镜观察细胞生长情况;用四唑盐(MTT)比色法计数计算细胞相对增殖度,用6级毒性分类法评级;用偶氮偶联法(PNPP)染色,分析碱性磷酸酶活性的变化。结果 80SMBG浸提液在高浓度时轻度抑制细胞增殖,促进细胞分化;低浓度促进细胞增殖,轻度抑制细胞分化。结论 80SMBG的体外生物活性较高,对体液微环境的影响轻微,细胞相容性好,作为骨组织修复替代材料或骨组织工程支架材料具有较高的研究和应用价值。  相似文献   

6.
羊胎盘免疫调节因子E花环活性评价的优化   总被引:1,自引:1,他引:0  
目的改进E花环实验方法,运用改进的实验方法评价羊胎盘免疫调节因子(GPIF)生物活性效果。方法2次将分离的T细胞(猪、山羊胸腺及其外周血和人外周血T细胞)45℃30min灭活,分别加入不同稀释度的GPIF和神经氨酸酶处理的绵羊血红细胞(SRBC),以观察在抑制(病理)条件下,GPIF对T细胞生物活性的影响。结果猪、山羊胸腺及其外周血和人外周血T细胞的E花环实验筛选显示猪胸腺效果最好,其阳性对照组与阴性对照组比差异显著(P<0.05),实验组与阴性对照组比差异极其显著(P<0.01),实验组在0.5mg/mL质量浓度与阴性对照组差异极其显著(P<0.01);使神经氨酸酶处理后的SRBCE花环结环率提高了7%左右。结论经过改进的E花环评价GPIF生物活性具有方法简便、可操作性强、重复性好的特点;实验室研制的GPIF对增强T细胞活性具有剂量效应。  相似文献   

7.
曾彬  林国生  蔡军 《解剖学报》2006,37(6):715-719
目的添加转化生长因子_1β(TGF_β1)以及与内脏内胚层样END_2细胞共培养,定向诱导胚胎干细胞(ESCs)分化,探索联合使用化学诱导法与共培养法对ESCs的心肌细胞定向诱导分化作用。方法将ESCs悬浮培养形成2~3 d类胚体(EBs),再向培养液内添加TGF_β1,或(和)将2~3 d EBs与END_2细胞或END_2细胞条件培养液共培养。自然分化为对照组。免疫荧光技术检测心肌细胞特异性肌动蛋白(α_actin)及肌钙蛋白T(TnT)的表达,透射电镜观察分化心肌细胞的超微结构。结果向培养液添加TGF_β1或将2~3 d EBs与END_2细胞或END_2细胞条件培养液共培养,各自有(43±2.08)%(P<0.01),(69±3.61)%(P<0.01),(65±3.06)%(P<0.01)的EBs出现自发节律性收缩,均表达心肌细胞特异性蛋白α_Actin和TnT,观察到心肌样超微结构。自然分化组发生自发节律性收缩的EBs只有(12±1.53)%,尤其是联合使用两种诱导方法,自发性收缩的EBs高达(91±1.52)%(P<0.01),收缩区域较单一诱导组大,且细胞形态较单一。结论联合使用化学诱导和共培养2种诱导法对ESCs的心肌细胞定向分化有协同作用。  相似文献   

8.
目的评价新型有序纳米介孔生物活性玻璃(80SMBG)对成骨细胞的生物相容性。方法以Novabone为对照,用大鼠成骨细胞与80SMBG浸提培养液、Novabone浸提培养液和正常的空白培养液培养,分别于3、4、5d用四唑盐(MTT)比色法计数计算细胞相对增殖度,用6级毒性分类法评级,同时用偶氮偶联法(PNPP)染色,分析碱性磷酸酶活性的变化。结果80SMBG毒性评级为0~1级,对成骨细胞分化无明显的抑制作用。结论80SMBG的体外生物活性较高,对体液微环境的影响轻微,细胞相容性好,作为骨组织修复替代材料或骨组织工程支架材料具有较高的研究和应用价值。  相似文献   

9.
目的 检测在脂多糖(lipopolysaccharide,LPS)作用下,A549细胞中SP-B、NFкB、RARα表达情况,探讨感染因素(脂多糖)对SP-B表达及其有关调节因素(RARα)表达的影响,并观察在此过程中是否有NFкB表达的改变.方法 培养的A549细胞分为对照组和实验组,实验组分别用不同浓度的脂多糖处理24 h.免疫细胞化学法检测SP-B和RARα在各组A549细胞中的表达情况,免疫荧光共聚焦法检测NFкB在各组A549细胞中的表达变化.结果 各实验组SP-B的表达均较对照组减弱(P<0.01),且呈药物浓度依赖性(P<0.05).各实验组胞核内RARα表达均较对照组减弱(P<0.01),且呈明显的药物浓度依赖性(P<0.01).而各实验组胞质RARα表达均较对照组增强(P<0.01),也呈药物浓度依赖性(P<0.01).与对照组相比,实验组中NFкB部分转位至A549细胞胞核中表达.结论 脂多糖作用后,A549细胞中SP-B的表达减少,且NFκB和RARα在细胞内的分布都发生了变化,表明在呼吸窘迫综合征的发病中,炎症过程可能起到重要作用,NFκB和RARα可能在此阶段中都参与了SP-B表达调控.  相似文献   

10.
目的 研究新型复合材料3-羟基丁酸-co-3-羟基戊酸共聚物-溶胶-凝胶生物活性玻璃(PHBV-SGBG)对骨髓基质细胞的细胞相容性,并建立动物骨缺损模型,探讨PHBV-SGBG的体内成骨效能.方法 采用MTT法和直接接触法检测复合材料PHBV-SGBG的细胞相容性.制备犬胫骨骨缺损模型,实验组在骨缺损区植入PHBV-SGBG材料,对照组不作处理,分别于术后2、4、8、12周取材,扫描电镜和X线能谱分析观察该材料的体内成骨效能.结果 复合材料PHBV-SGBG的细胞毒性分级在0~1之间,骨髓基质细胞可紧密附着于材料表面,黏附、生长、增殖,并有突起向材料的连通微孔内伸展.实验组术后2周已有明显新骨生成,术后4周时Ca/P元素比值(1.086±0.034),对照组为(0.793±0.053),术后12周时骨缺损区充填新生的成熟骨组织,扫描电镜和能谱分析显示实验组Ca峰值逐渐升高:12周时Ca/P元素比值(1.603±0.067),优于对照组(11456±0.036)(P<0.05).结论 复合材料PHBV-SGBG无细胞毒性,具有很好的骨传导性和骨再生能力,有望成为新型的骨组织工程材料.  相似文献   

11.
12.
Yang S  Kim HM 《Biomaterials》2012,33(10):2902-2915
The proliferation of anchorage-dependent cells of mesenchymal origin requires the attachment of the cells to substrates. Thus, cells that are poorly attached to substrates exhibit retarded cell cycle progression or apoptotic death. A major disadvantage of most polymers used in tissue engineering is their hydrophobicity; hydrophobic surfaces do not allow cells to attach firmly and, therefore, do not allow normal proliferation rates. In this study, we investigated the molecular mechanism underlying the reduced proliferation rate of cells that are poorly attached to substrates. There was an inverse relationship between the activity of the small GTPase RhoA (RhoA) and the cell proliferation rate. RhoA activity correlated inversely with the strength of cell adhesion to the substrates. The high RhoA activity in the cells poorly attached to substrates caused an increase in the activity of Rho-associated kinase (ROCK), a well-known effector of RhoA that upregulated the activity of phosphatase and tensin homolog (PTEN). The resulting activated PTEN downregulated Akt activity, which is essential for cell proliferation. Thus, the cells that were poorly attached to substrates showed low levels of cell proliferation because the RhoA-ROCK-PTEN pathway was hyperactive. In addition, RhoA activity seemed to be related to focal adhesion kinase (FAK) activity. Weak FAK activity in these poorly attached cells failed to downregulate the high RhoA activity that restrained cell proliferation. Interestingly, reducing the expression of any component of the RhoA-ROCK-PTEN pathway rescued the proliferation rate without physico-chemical surface modifications. Based on these results, we suggest that the RhoA-ROCK-PTEN pathway acts as a molecular switch to control cell proliferation and determine anchorage dependence. In cells that are poorly attached to substrates, its inhibition is sufficient to restore cell proliferation without the need for physico-chemical modification of the material surface.  相似文献   

13.
介绍了生物力学领域中细胞生长、增殖与分泌方面的实验研究进展.着重介绍了基底加载实验中不同环境对细胞分裂与增殖的作用;力学载荷对细胞黏附及生长的影响;力学应变对体外培养成骨细胞增殖与分泌的影响;流体环境对细胞生长增殖的作用.  相似文献   

14.
可对细胞施加双向应变的体外培养系统   总被引:4,自引:1,他引:3  
为考察体外培养细胞在动应变作用下的响应,参考Winston等人的方法,我们制作了一套细胞体外培养系统,培养皿用有机玻璃作成,培养皿底为硅橡胶膜,下面有一封闭压力腔,通过对硅橡胶管和挤压轮间隙的调整,压力腔内的液压使硅橡胶膜产生设计应变,其应变范围可从0~4%调整,改变步进电机的转动方式,加载频率可从0.1~5Hz变化。  相似文献   

15.
16.
E-cadherin表达缺失在白血病细胞行为中的作用   总被引:4,自引:3,他引:1       下载免费PDF全文
目的:确定上皮-钙黏蛋白(E-cadherin)表达缺失在白血病细胞行为中的作用,最终阐明E-cadherin介导的骨髓微环境与造血细胞相互作用改变在白血病发生中的作用。 方法:细胞黏附实验测定 HL-60细胞经5-氮杂脱氧胞苷(5-Aza-CdR)诱导E-cadherin表达后细胞黏附能力的改变,MTT法测定细胞增殖能力,细胞迁移实验确定细胞的迁移能力。结果:HL-60细胞经5-Aza-CdR诱导E-cadherin表达后,其对E-cadherin-Fc的黏附能力增加,细胞在E-cadherin-Fc上的增殖能力下降,细胞黏附导致细胞迁移能力下降。结论:白血病细胞中由于E-cadherin表达丧失,引起细胞黏附降低,增殖能力增强,细胞迁移活跃,表明E-cadherin表达缺失与白血病细胞行为的获得有关。  相似文献   

17.
目的:探讨蛇六谷提取物(TuAKe)抑制人慢性髓系白血病K562细胞增殖和诱导分化的作用机制。方法:不同浓度的TuAKe处理K562细胞,MTT比色法和半固体集落形成实验检测K562细胞的增殖能力;瑞氏-姬姆萨染色观察细胞形态;流式细胞术检测分化相关抗原CD11b、CD14和CD42b的阳性表达率;Western blot法检测细胞周期蛋白依赖性激酶2(CDK2)、细胞周期蛋白E1(cyclin E1)和红系分化核转录因子GATA-1的蛋白表达水平。结果:TuAKe能够抑制K562细胞增殖,改变细胞形态,抑制K562细胞进入S期,并阻滞在G_2/M期,提高分化相关抗原CD11b、CD14和CD42b阳性表达率,上调GATA-1蛋白表达,同时下调CDK2和cyclin E1蛋白表达(P0.05)。结论:TuAKe可能通过调控细胞周期抑制K562细胞增殖,同时诱导K562细胞多向分化。  相似文献   

18.
The development of clinical therapeutics that interfere with the migration of leukocytes has revolutionized the treatment of multiple sclerosis and holds great promise for the treatment of a wide range of inflammatory diseases. As the molecules essential for the multi‐step adhesion cascade that mediates cellular migration have been elucidated, the number of potential targets available to modulate leukocyte trafficking has increased exponentially. In this Viewpoint, we briefly review our current understanding of these mole‐cular targets and how these targets vary by tissue and leukocyte subset with emphasis on T cells. We then describe the two currently approved therapeutics that target cell migration, natalizumab and fingolimod, and discuss how an improved understanding of their function could pave the way for the development of safer and more efficacious therapies for inflammatory and autoimmune diseases.  相似文献   

19.
We report on a simple method for self loading and culture of mammalian cells in microfluidic multi-chambers for high throughput screening. The device was obtained by using one layer soft lithography with polydimethylsiloxane (PDMS) and thermal bonding on a glass slide. Self loading of cell suspension could be possible after degassing of the PDMS device for 30 min. Both cell loading efficiency and cell proliferation behaviors have been analyzed with triangle chambers of different sizes, all connected to the main flow channels with small entrances. We found that the number of cells loaded into the micro-chamber increased with the side length of the triangle, showing well size dependence and that self loading at a single cell level was possible for small chambers. For large chambers, the cell area density after loading and proliferation is however quite heterogeneous. For demonstration, HeLa cell growth behavior has been followed for 11 days until the total area of the largest chambers was fully filled.  相似文献   

20.
Summary In this study, electron microscopy was used to study cell clustering in the postnatal mouse striatum. From the date of birth (PO) through postnatal day 7 (P7), groupings of eight to ten striatal neurons were delimited easily in low magnification electron micrographs. Often, within individual groupings, adjacent neurons were separated only by a thin, 10 nm gap, and formed cell pairs or cell triads. Coincident with marked expansion of the striatal neuropil in the second postnatal week, striatal neurons formed more dispersed cell clusters consisting only occasionally of cell pairs or triads.Single, pyknotic neuronal nuclei were seen in clusters of normal neurons exhibiting different stages of maturation but were absent from clusters consisting only of well-differentiated neurons. The neuropil surrounding cell clusters with pyknotic neurons or that adjacent to neighboring cell clusters often contained degenerating dendrites and axon terminals. Whereas this naturally occurring neuronal cell death was present in the tissue throughout the first postnatal week, only degenerating dendritic and axonal profiles were seen in the P15 striatum. This latter fact suggests that the occurrence of pyknotic neuronal somata does not account entirely for the more localized degeneration of other neuronal profiles and raises the possibility that other degenerative processes may be occurring simultaneously in the tissue.Preliminary reports of portions of this work were presented at the NINCDS Huntington's Disease Symposium, San Diego, Ca., November 16–18, 1978, and at the Ninety-first annual meeting of the American Association of Anatomists, Vancouver, British Columbia, April 3–6, 1978  相似文献   

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