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1.
An in vivo study was carried out on uncoated and hydroxyapatite (HA)-coated nanostructured Ti13Nb11Zr alloy in comparison with high-grade Ti6Al4V, to investigate the effect of the different surfaces on osteointegration rate. A highly effective method to obtain a fast biomimetic deposition of a thin layer of nanocrystalline HA was applied to coat both substrates. Cylindrical pins were implanted in rabbit cortical bone and evaluated at 4 and 12 weeks by histomorphometry and microhardness tests. The results confirmed the ability of the slightly supersaturated Ca/P solution to induce a fast deposition of nanocrystalline HA on Ti alloys' surfaces. HA-coated Ti13Nb11Zr had the highest osteointegration rate at 4 and 12 weeks. Both HA-coated surfaces showed an affinity index significantly higher than those of native surfaces at 4 weeks (Ti13Nb11Zr+HA: 37%; Ti6Al4V+HA: 26%). Microhardness test showed a significantly higher bone mineralization index of HA-coated Ti13Nb11Zr in comparison with that of HA-coated Ti6Al4V surface. The study suggests that the HA coating on both alloys enhances bone response around implants and that there is a synergic effect of Ti-Nb-Zr alloy with the HA coating on bone remodeling and maturation.  相似文献   

2.
Long-term stability of arthroplasty prosthesis depends on the integration between the bone tissue and the implanted biomaterials, which requires the contribution of osteoblastic precursors and their continuous differentiation into the osteoblastic phenotype. Classically, these interactions are tested in vitro using mesenchymal stem cells (MSCs) isolated and ex vivo expanded from bone marrow aspirates. Human adipose tissue-derived stromal cells (AMSCs) may be a more convenient source of MSCs, according to their abundance and accessibility, but no data are available on their in vitro interactions with hard biomaterials. The aim of this work is to compare the osteogenic potential of human AMSCs and bone marrow-derived MSCs (BMMSCs) and to evaluate their response to Ti6Al4V alloy in terms of adhesion, proliferation and differentiation features, using the human osteosarcoma cell line SaOS-2 for comparison. The overall results showed that AMSCs have the same ability to produce bone matrix as BMMSCs and that Ti6Al4V surfaces exhibit an osteoinductive action on AMSCs, promoting their differentiation into functional osteoblasts and increasing bone formation. In conclusion, adipose tissue is a promising autologous source of osteoblastic cells with important clinical implications for bone tissue engineering.  相似文献   

3.
背景:以往研究表明骨髓间充质干细胞的获取来源主要是股骨或髂骨,但下颌骨来源骨髓间充质干细胞的研究至今少有报道。 目的:观察兔下颌骨来源的骨髓间充质干细胞体外分离、培养及其生物学特性。 方法:体外分离培养兔下颌骨骨髓间充质干细胞,取第2或3代检测。MTT法检测细胞增殖,克隆形成试验检测细胞的克隆形成率变化,并对其进行成骨和成脂肪诱导分化。 结果与结论:MTT法检测细胞增殖,细胞生长曲线显示下颌骨骨髓间充质干细胞经历潜伏期、对数生长期和平台期。细胞克隆形成实验显示,细胞呈成纤维集落形成单位外观。茜素红、油红O染色结果显示,成骨诱导、成脂诱导后形成钙结节及脂滴。结果证实,兔下颌骨分离出的骨髓间充质干细胞有强大的自我复制及增殖能力,具有多向诱导分化的潜能。  相似文献   

4.
背景:国内外对骨髓间充质干细胞的体外成骨诱导分化研究手段、测定指标均不够全面。 目的:建立并完善一整套人骨髓间充质干细胞的分离培养及鉴定方法,探讨其体外成骨分化能力。 方法:采用密度梯度离心法分离培养人骨髓间充质干细胞,流式细胞仪鉴定细胞表面表型。传至第3代时更换成骨诱导培养基进行成骨分化诱导。 结果与结论:人骨髓间充质干细胞生长旺盛,传代后增殖旺盛,第3代骨髓间充质干细胞表面表型CD44、CD73、CD90表达阳性,CD34表达阴性。诱导后的成骨细胞碱性磷酸酶活性增加,Gomori、Von kossa、茜素红染色均阳性。RT-PCR检测诱导后细胞有Ⅰ型胶原、碱性磷酸酶、骨钙素、骨唾液酸蛋白、骨桥蛋白及骨连接蛋白基因的表达,证明了人骨髓间充质干细胞成功向成骨方向分化。表明实验建立了一整套稳定、成熟的骨髓间充质干细胞分离、培养、扩增方案。  相似文献   

5.
背景:前期实验发现丝素蛋白、壳聚糖以适当的比例混合,可以互相弥补各自的不足,表现出良好的理化性质和生物学特性。 目的:观察骨髓间充质干细胞在丝素蛋白/壳聚糖混合三维支架材料上的生长情况。 方法:将诱导后的兔骨髓间充质干细胞接种在丝素蛋白/壳聚糖支架材料上,检测细胞黏附率,倒置显微镜及扫描电镜观察细胞生长情况。 结果与结论:细胞黏附率随时间的延长而增加。倒置显微镜观察显示,丝素蛋白/壳聚糖支架上的细胞看不清,随着时间的延长,支架周围细胞增多,且有细胞伸入支架内;扫面电镜观察显示,细胞生长活跃、增殖分裂正常,细胞周围见颗粒状、丝状基质物质,细胞的微丝与支架材料黏附紧密;细胞不仅可以在材料表面贴附生长,并伸入材料之中。说明丝素蛋白/壳聚糖混合支架材料具有良好的细胞生物相容性。  相似文献   

6.
背景:拥有超弹性、形状记忆效应及良好抗疲劳性能的镍钛合金成为封堵器支架核心部分的最佳选择,但镍钛合金含50%的镍元素,其生物相容性仍有争议。 目的:通过体外浸泡实验和电化学实验测试用于治疗先天性心脏病的封堵器镍钛合金多弧离子镀改性前后的抗腐蚀性能。 方法:利用多弧离子镀在镍钛合金表面形成具有纳米梯度的TiN/Ti多层薄膜,通过镍钛合金在PBS中静态浸泡1个月,检测镍离子浓度;采用开路电位-时间曲线和极化曲线观察镀膜前后镍钛合金抗腐蚀性能。 结果与结论:在PBS静态浸泡中镍钛合金在1个月时释放量达到顶峰,而经过多弧离子镀改性后的镍钛合金没有镍离子的释放。开路电位-时间曲线和极化曲线表明改性后的镍钛合金的抗腐蚀性得到了很大提高。  相似文献   

7.
背景:与骨髓、外周血、脂肪及胚胎等来源的间充质干细胞相比,脐带和胎盘组织来源的间充质干细胞具有来源广泛、易于获得、不引起供者不适以及不存在道德伦理争议等优势。 目的:探讨胎盘间充质干细胞体外分离、培养、传代的方法及其分化潜能,为胎盘间充质干细胞的临床应用提供基础研究依据。 方法:应用计算机检索2000年1月至2011年12月PubMed数据库、中国期刊全文数据库、万方数据库与胎盘间充质干细胞有关的文章,中文检索词为“胎盘间充质干细胞,体外,诱导分化”,英文检索词为“Placenta-derived mesenchymal stem cells,Induction and differentiation in vitro”。共检索到文献98篇,最终纳入符合标准的33篇文献。 结果与结论:目前国内胎盘间充质干细胞还处于基础研究阶段,国外胎盘间充质干细胞在实验研究方面已经取得了一定的进展,而且也在临床应用研究方面有了一定的突破。胎盘间充质干细胞具有来源广泛,取材方便,不引起供者不适以及不涉及伦理道德问题,与捐献骨髓或采集动员外周血相比,供者无痛苦,感染机会少, 扩增能力强等,在临床治疗方面有待于进一步深入研究。  相似文献   

8.
In vitro secreting profile of human mesenchymal stem cells   总被引:2,自引:0,他引:2  
In addition to their multilineage potential, mesenchymal stem cells (MSCs) also have a wide range of functionality. Not only can MSCs reconstruct a tissue, but they also have the ability to control or cure other cells and can reconstruct a coordinating function. The opportunity to control other cells depends on MSCs being able to secrete factors like cytokines and chemokines. Therefore, we focused on asking, Which factors can be secreted by human MSCs? To answer this question, we analyzed the secreting profile of in vitro-expanded MSCs by using cytokine arrays. The media concentrations of 44 of the 120 analyzed cytokines were significantly increased by MSCs. Conversely, concentrations of 40 cytokines given with the sera were significantly decreased. The data presented here provide an overview about a large range of factors that were secreted by MSCs under cell culture conditions. These data indicate that MSCs demonstrate all previously described functions in cellular interactions without an external stimulus. The MSCs secreted angiogenic, immunosuppressive, anti-apoptotic, and proliferation-stimulating factors.  相似文献   

9.
背景:目前常用的成脂刺激剂主要由胰岛素、地塞米松、IBMX和吲哚美辛组成,该体系涉及处理因素多、诱导周期长且对细胞毒性大,不利于脂肪形成抑制效应的研究。 目的:建立快速诱导大鼠骨髓间充质干细胞成脂分化的培养体系。 方法:大鼠全骨髓细胞原代培养,胰酶消化传代,富集形态均质的间充质干细胞,利用过氧化物酶体增殖物活化受体γ激动剂噻唑烷二酮类药物罗格列酮和吡咯列酮体外单独诱导大鼠骨髓间充质干细胞成脂分化,设立无糖、低糖和高糖DMEM三种基础培养体系,以经典的成脂刺激剂作为阳性对照,在诱导的不同时间点对分化细胞进行形态学观察和油红O染色。 结果与结论:与经典成脂刺激剂相比,罗格列酮或吡咯列酮单独均能诱导大鼠骨髓间充质干细胞向脂肪细胞分化,吡咯列酮诱导48 h和罗格列酮诱导72 h均可观察到富含脂滴或脂泡以及油红O染色的阳性细胞,吡咯列酮与罗格列酮的最佳诱导浓度分别为0.125 mmol/L和10 μmol/L,而高糖环境利于大鼠骨髓间充质干细胞成脂分化。说明基于高糖培养环境的以吡咯列酮或罗格列酮做为成脂刺激剂的培养体系可快速诱导大鼠骨髓间充质干细胞向脂肪细胞分化。  相似文献   

10.
目的探讨一种安全、有效并易于推广的临床研究用间充质干细胞的产业化制备策略。方法常规方法分离获得原代脐带间充质干细胞,P1~P3代以含小牛血清培养基进行稳定传代后,以无血清培养基对P4~P6代细胞继续培养和扩增,并以含血清培养基作为对照。倒置显微镜每天定时观察细胞生长状态,CCK-8法比较P4代和P6代细胞在两种培养基中的增殖能力,并对P4~P6代细胞的细胞裂解液和细胞培养上清中的牛血清白蛋白(BSA)残留量进行检测。结果对分离获得的原代脐带间充质干细胞进行传代培养,在含血清培养基中,P1~P3代细胞增殖能力稳定,以4×104/ml密度接种,3.5~4d可达到90%融合;细胞进入P6后,在无血清培养基培养条件下增殖能力与含血清培养基相近;BSA残留量检测结果显示,无血清培养基组中细胞的培养上清和细胞裂解液中残留的BSA量低于10ng/106细胞,满足《中国生物制品规程》对相关细胞治疗产品中BSA残留量的标准要求。结论含血清培养与无血清培养相结合的程序性培养扩增体系,可为临床提供数量充足、质量可控的间充质干细胞。  相似文献   

11.
背景:羊膜来源间充质干细胞植入机体不同类型组织后是否可以分化为相应组织靶细胞呢? 目的:检测血管内皮细胞生长因子在体外诱导人羊膜间充质干细胞分化为血管内皮细胞的可行性。 方法:分离培养羊膜间充质干细胞,鉴定其表面抗原表达,用含体积分数2%胎牛血清以及50 μg/L血管内皮生长因子的条件培养基诱导,诱导后细胞通过内皮细胞标志物血管内皮生长因子受体2以及v-WF染色鉴定。 结果与结论:羊膜间充质干细胞表面抗原CD29、CD44、CD105阳性,CD34、CD45、CD106以及HLA-DR阴性。诱导后细胞形态明显改变,内皮细胞标志物血管内皮细胞生长因子受体2以及v-WF染色结果阳性。提示羊膜间充质干细胞在体外具有分化为血管内皮细胞的能力。  相似文献   

12.
Mobilization of stem cells and their differentiation into cardiomyocytes are known to have protective effects after myocardial infarction. The integrity of transplanted mesenchymal stem cells for cardiac regeneration is dependent on cell–cell or cell–matrix interaction, which is adversely affected by reactive oxygen species in an ischemic environment. Treatment with erythropoietin was shown to protect human adipose tissue derived mesenchymal stem cells in an ischemic injury in vitro model. The analyses indicated that expression of erythropoietin receptors played a pivotal role in erythropoietin mediated cell survival. In this study, the anti-apoptotic effect of erythropoietin on stem cells was analyzed in apoptosis-induced human mesenchymal stem cells. Apoptosis was induced in cultured adult human adipose tissue derived mesenchymal stem cells by hydrogen peroxide. A group of cultured cells was also treated with recombinant human erythropoietin in a concentration of 50 ng mL−1. The degree of apoptosis was analyzed by flow-cytometry and immunohistochemical staining for Caspase 3. The average percentages of apoptotic cells were significantly higher in H2O2-induced stem cells than in cells co-cultured with erythropoietin (63.03 ± 4.96% vs 29 ± 3.41%, p < 0.01). We conclude that preconditioning with erythropoietin suppresses apoptosis of mesenchymal stem cells and enhances their survival.  相似文献   

13.
背景:骨髓间充质干细胞作为良好的种子细胞,将其复合于生物支架治疗骨缺损取得了良好的进展,是当今研究的一大热点。 目的:观察大鼠骨髓间充质干细胞体外诱导向成骨样细胞分化的效果。 方法:采用贴壁筛选法分离培养SD大鼠骨髓间充质干细胞,取生长状态良好的第3代骨髓间充质干细胞,分为2组,对照组细胞仅用DMEM/F12培养基培养;实验组以含成骨诱导剂的DMEM/F12培养基培养。 结果与结论:全骨髓贴壁法培养的原代骨髓间充质干细胞呈梭形或多角形贴壁生长;经成骨诱导剂诱导后骨髓间充质干细胞呈圆形或卵圆形贴壁生长,碱性磷酸酶活性明显强于对照组(P < 0.01);茜素红染色出现阳性的钙化结节;Western blotting检测Ⅰ型胶原蛋白表达较对照组有明显增加(P < 0.01);骨钙素ELISA定量分析较对照组明显升高(P < 0.01)。提示全骨髓贴壁法培养骨髓间充质干细胞方法简单、实用,所培养的骨髓间充质干细胞在成骨诱导后表现了成骨细胞的形态学和生物学特性。  相似文献   

14.
Tissue engineering represents a promising method for generating chondrogenic grafts for reconstructive surgery. In cultured chondrocytes, the dedifferentiation of cells seems unavoidable for multiplication. Stem cells, however, displaying unlimited self-renewal and the capacity to differentiate towards chondrocytes, might be usable after further characterization. As the interactions between the extracellular matrix and the cellular compartment can alter the cellular behaviour, we investigated the expression of integrins using microarray analysis during chondrogenic differentiation of human mesenchymal stem cells (MSC) in comparison with de-differentiating human chondrocytes (HC) harvested during septoplasty. During chondrogenic differentiation of MSC, the fibronectin-receptor (Integrin beta1alpha5), fibronectin and the GPIIb/IIIa-receptor were downregulated. The components of the vitronectin-receptor (Integrin alphavbeta3) and CD47 were constantly expressed and ILK was downregulated. Vitronectin and osteopontin were not expressed by the cells. In HC, Integrin beta1alpha5 in conjunction with the ligand fibronectin were upregulated during dedifferentiation, Integrin alphavbeta3 as well as the GBIIb/IIIa-receptor were activated on day 21 but neither vitronectin nor osteopontin were expressed by the cells. The integrins, beta2, beta4, beta6, beta8 and alpha2, alpha4, alpha6, alpha7, alpha11, were not expressed at any time. ILK, CD47, and ICAP were activated with ongoing dedifferentiation. In conclusion, a candidate for signal-transmission is the fibronectin receptor (integrin alpha5beta1) in conjunction with its ligand fibronectin. Other receptors, e.g. for vitronectin and osteopontin (alphavbeta3), or their ligands do not seem to be involved in signal transmission for dedifferentiation. The GPIIb/IIIa-receptor might assist the process of dedifferentiation. Intracellularly, ILK, ICAP1 and CD47 might be involved in the transduction of integrin-dependent signals.  相似文献   

15.
背景:目前神经干细胞多由动物获得,不适合人类临床移植治疗。 目的:探索体外环境下人胚胎纹状体来源神经干细胞的培养方法,同时观察其生物学特性。 方法:取经水囊引产的孕8-16周人胚胎纹状体,体外用无血清DMEM培养基进行培养,待细胞形成神经球后进行传代,并应用含体积分数10%胎牛血清的DMEM/ F12培养液进行诱导分化。 结果与结论:体外培养的人胚胎纹状体来源神经干细胞生长迅速,表达神经干细胞标志物nestin。克隆形成实验显示细胞克隆形成率为6.0%-7.0%;BrdU掺入实验显示细胞增殖率为37.9%。免疫荧光染色显示经诱导分化的细胞表达神经元标志物Ⅲ型β微管蛋白、星形胶质细胞标志物胶质纤维酸性蛋白及神经干细胞标志物nestin,但不表达少突胶质细胞标志物髓鞘碱性蛋白。可见人胚胎纹状体来源神经干细胞在体外无血清条件下可保持其生物学特点,具有自我更新能力,经胎牛血清诱导后可向神经元及星形胶质细胞分化。  相似文献   

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文题释义:角膜上皮干细胞:属于单能干细胞,具有细胞周期长、低分化状态、增殖潜力大、不对称分裂等特点,定位于角膜缘基底细胞层,又称之为角膜缘干细胞,对角膜上皮细胞更新及维持角膜透明起着重要作用。角膜缘干细胞的体外培养方法:主要包括酶消化培养法和组织块培养法。酶消化培养法是利用DispaseⅡ酶破坏角膜缘上皮细胞与基底膜之间的半桥粒连接,然后剥取角膜缘上皮层,再使用胰酶将其消化为单个细胞进行培养。组织块培养法没有经过酶的双重消化,将剖取的角膜缘组织块进行贴壁,细胞游离出组织块进行贴壁生长,需要一个漫长的过程。  摘要背景:角膜上皮干细胞定位于角膜缘,又称之为角膜缘干细胞,临床上由于眼表严重热烧伤、化学性烧伤、慢性炎症等原因引起的角膜缘干细胞缺乏或功能障碍治疗较为棘手。目前利用组织工程技术体外培养角膜上皮干细胞并进行临床移植成为新型有效的治疗方向。目的:探讨在无血清培养条件下采用改良组织块培养法培养人角膜上皮干细胞的可行性。方法:人角膜缘组织来自河南省眼库,植片直径小于8 mm角膜移植术后的供者剩余眼球材料,手术显微镜下剖取角膜缘上皮层外2/3区域,采用2种方法培养人角膜上皮干细胞,常规组织块培养组是将组织块上皮面向上贴壁,加入K-SFM培养液后置于 37 ℃、体积分数为5%CO2细胞培养箱中培养;改良组织块培养组是先将组织块浸泡于K-SFM培养液中,置于细胞培养箱中孵育12 h,然后组织块上皮面向下贴壁培养。组织块周边有细胞游离出贴壁生长记作“培养第1天”,每日相差显微镜下观察细胞生长变化。利用免疫荧光染色技术检测改良组织块培养第5,10,14天时原代细胞中p63及K3的表达。结果与结论:①改良组织块培养组出膜时间明显短于常规组织块培养组(P < 0.05),出膜率明显高于常规组织块培养组(P < 0.05);②改良组织块培养组细胞生长状态良好,培养第10天可见小体积细胞较多,聚集成灶状分布;培养第14天可见细胞克隆灶,克隆灶内细胞体积较小,形态均一;③培养第5天,K3表达量较多,p63表达量较少;培养第10天,K3和p63表达量均增多;培养第14天,K3表达量未见明显增多,p63表达量明显增多;④在无血清培养条件下,改良组织块培养法能显著促进角膜上皮干细胞的游离,提高体外培养细胞数量,为人角膜缘上皮组织片的构建提供种子细胞。ORCID: 0000-0001-8370-174X(许中中) 中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程  相似文献   

18.
目的 探讨兔骨髓间充质干细胞(rBMSCs)与纳米羟基磷灰石(nano-HA)支架材料的相容性,进一步验证nano-HA材料作为骨组织工程支架材料的可行性。 方法 将rBMSCs与nano-HA支架材料在体外复合培养,通过倒置显微镜、扫描电镜观察细胞与材料的复合情况,用MTT法、碱性磷酸酶活性检测法检测材料对细胞增殖、分化的影响。 结果 rBMSCs可以在nano-HA支架材料表面及孔隙中良好的黏附、迁移、增殖和分化,复合培养5 d后nano-HA支架材料对rBMSCs的增殖分化表现出一定的促进作用。 结论 rBMSCs与nano-HA支架材料具有良好的生物相容性, nano-HA支架材料可以作为rBMSCs良好的载体。  相似文献   

19.
目的 研究密度梯度离心法结合贴壁法分离大鼠骨髓间充质干细胞(BMSCs)及不同首次换液时间对细胞增殖的影响,探索一种简单易行、扩增效率高的方法。方法 3月龄雄性SD大鼠提取原代BMSCs,密度梯度离心法结合贴壁法分离培养,并应用3组不同首次半量换液时间(24h,48h,72h)进行培养传代,测定生长曲线和细胞的存活率;免疫细胞化学染色分析细胞表面抗原CD44和CD166的表达。结果 分离的BMSCs48h后细胞完全贴壁生长,呈形态均一的纺锤形单核细胞状,第3代细胞培养1w后呈长梭型,呈平行排列生长。48h半量换液组细胞增殖活性及生存率最高,其次为24h和72h组;表面标志物CD44和CD166均呈阳性。结论 联合应用密度梯度离心法及贴壁筛选法可成功分离BMSCs,且效率高、纯度高,48h首次半量换液更适合BMSCs的生长繁殖。  相似文献   

20.
背景:目前,对骨髓间充质干细胞的分离、纯化和扩增还没有统一的、标准化的方法。CM-DiI作为荧光标记物稳定、可靠、标记率高、标记简便。 目的:建立SD大鼠骨髓间充质干细胞体外分离培养及标记的方法。 方法:取2只体质量50-100 g雄性SD大鼠,无菌条件下采集双侧股骨、胫骨骨髓,用全骨髓贴壁分离法和密度梯度离心法培养出原代骨髓间充质干细胞,通过及时、反复传代对细胞进行扩增纯化,在体外用荧光活性染料CM-DiI标记第3代骨髓间充质干细胞后作为供体细胞来源。 结果与结论:用全骨髓贴壁分离法和密度梯度离心法两种方法均能成功体外分离培养骨髓间充质干细胞,经流式细胞仪分析,培养出的细胞CD34阳性率为17.5%,CD44阳性率为97.9%、CD90阳性率为91%,与骨髓间充质干细胞表面抗原一致。但培养出的细胞数量全骨髓贴壁分离法明显多于密度梯度离心法,两种方法培养骨髓间充质干细胞的细胞活力和增殖能力无明显差异。CM-DiI能够成功荧光标记骨髓间充质干细胞,CM-DiI作为荧光标记物稳定、可靠、标记率高、标记简便。中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接:  相似文献   

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