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1.
目的体外培养人牙囊细胞,探讨牙囊细胞是否具成骨、成牙骨质细胞表型特征,是否可作为牙骨质再生的种子细胞。方法酶消化联合组织块法获得人牙囊细胞,免疫酶细胞化学染色技术检测其Ⅰ型胶原、Ⅲ型胶原、骨桥蛋白、骨粘连蛋白的表达,RT-PCR技术检测细胞的骨涎蛋白、骨钙素、碱性磷酸酶的表达。矿化诱导并连续培养牙囊细胞,行Vonkossa染色检测体外矿化形成情况。结果免疫组化结果:牙囊细胞中Ⅰ型胶原、Ⅲ型胶原、骨桥蛋白、骨粘连蛋白呈不同程度的阳性表达;RT-PCR结果显示骨涎蛋白、骨钙素、碱性磷酸酶表达。经矿化诱导后的牙囊细胞在20d时可形成矿化结节,vonKossa染色阳性。结论牙囊细胞具有成骨细胞、成牙骨质细胞的部分特性,体外培养的人牙囊细胞具有分泌合成矿化组织的能力,可试将其作为牙周组织再生的种子细胞。  相似文献   

2.
目的体外培养人牙囊细胞,探讨牙囊细胞是否具成骨、成牙骨质细胞表型特征,是否呵作为牙骨质再生的种子细胞。方法酶消化联合组织块法获得人牙囊细胞,免疫酶细胞化学染色技术检测其Ⅰ型胶原、Ⅲ型胶原、骨桥蛋白、骨粘连蛋白的表达.RT-PCR技术检测细胞的骨涎蛋白、骨钙素、碱性磷酸酶的表达。矿化诱导并连续培养牙囊细胞,行Von kossa染色检测体外矿化形成情况。结果免疫组化结果:牙囊细胞中Ⅰ型胶原、Ⅲ型胶原、骨桥蛋白、骨粘连蛋白呈不同程度的阳性表达:RT-PCR结果显示骨涎蛋白、骨钙素、碱性磷酸酶表达。经矿化诱导后的牙囊细胞在20d时可形成矿化结节,von Kossa染色阳性。结论牙囊细胞具有成骨细胞、成牙骨质细胞的部分特性,体外培养的人牙囊细胞具有分泌合成矿化组织的能力,可试将其作为牙周组织再生的种子细胞。  相似文献   

3.
目的研究富血小板纤维蛋白在诱导骨髓间质干细胞向成骨细胞转化的作用。方法取7天新西兰大白兔股骨,分离骨髓间质干细胞进行体外培养,在各自的培养条件下分为三组:实验组(不同剂量的PRF组)、空白对照组(MSCS组)和阳性对照组(BMP-2组)。三组细胞的比较采用MTT测定细胞增值率、PNPP法测定碱性磷酸酶的表达、免疫组化标记Ⅰ型胶原蛋白的表达、检测细胞为成骨细胞。结果细胞形态学观察,MSCS分化后,细胞形态从长梭形变成三角形,多角形,立方形;MTT测定细胞增殖显示,随着PRF膜剂量的增加,细胞增殖数量增加,PRF剂量为4,5mL时,与BMP-2组差别不大;ALP活性结果显示,MSCS分化后,ALP活性远高于MSCS细胞。两者差异具有显著统计学意义(t=24.608,p=0.000);免疫组化标记Ⅰ型胶原结果显示,PRF组分化后的MSCSⅠ型胶原表达显著。结论富血小板纤维蛋白可以诱导骨髓间质干细胞分化为成骨细胞,分化的类成骨细胞具有成骨细胞的特性,可以作为自体材料应用于口腔种植学领域里骨缺的修复。  相似文献   

4.
培养的人骨髓间充质干细胞向成骨细胞、脂肪细胞分化   总被引:5,自引:2,他引:3  
目的探讨体外培养的人骨髓间充质干细胞(MSCs)向成骨细胞、脂肪细胞分化的方法,以及钙结节形成、Ⅰ型胶原表达等情况. 方法分离人骨髓间充质干细胞,经过原代培养和传代培养,分别加入成骨诱导、脂肪诱导培养体系,经过倒置显微镜观测与HE染色了解诱导后细胞形态变化,并通过von Kossa染色、Ⅰ型胶原免疫组化染色、苏丹黑染色与油红O法染色鉴定细胞的性质.另取2代细胞体外培养,加入成骨诱导分化试剂,以碱性磷酸酶(ALP)为检测指标计数细胞阳性率,比较成骨诱导1、2、3、4周时细胞碱性磷酸酶的变化.结果体外培养的MSCs在合适的条件下能够向成骨细胞、脂肪细胞分化.细胞经成骨诱导2~3周时细胞碱性磷酸酶阳性率最高,达到85%.结论 MSCs体外培养在一定条件下向成骨细胞、软骨细胞与脂肪细胞分化,体外培养时形成钙结节,并表达Ⅰ型胶原.培养2~3周后细胞经成骨诱导分化细胞ALP阳性率较高.  相似文献   

5.
目的 以单层贴壁方法,高浓度维甲酸处理小鼠胚胎干细胞,以期建立一种高效成骨诱导的新体系. 方法将差速贴壁后的小鼠胚胎干细胞以1.5×104个/cm2的密度接种,成骨培养液+10 μmol/ml维甲酸诱导,4 d后更换为成骨诱导液,直至28 d,同时通过细胞的形态学观察、RT-PCR、茜素红染色对其成骨分化进行鉴定.结果 随着分化时间的延长,胚胎干细胞逐渐分化为梭形细胞,且呈漩涡状排列,至分化15 d左右产生结节样团块,21 d时结节样物质较普遍.RT-PCR检测表明成骨诱导14 d时,表达CD73+和I型胶原,说明细胞继续向成骨分化,且已进入了基质分泌阶段.21 d时,表达Ⅰ型胶原和骨钙素,表明已产生了成熟的成骨细胞,并进入了矿化后期.同时成骨诱导28 d时,茜素红染色显示细胞已产生大量钙结节.结论 以高浓度维甲酸,在单层贴壁状态下对小鼠胚胎干细胞进行诱导,可获得大量成熟、具备矿质化特性的成骨细胞,为成骨细胞的体外诱导分化提供了新的途径,同时该体系也成为体外探讨成骨细胞表型决定机制的一个重要模型.  相似文献   

6.
Cui L  Yin S  Deng CL  Yang GH  Chen FG  Liu W  Liu DL  Cao YL 《中华医学杂志》2004,84(15):1304-1309
目的应用软骨形态发生蛋白(CDMP)生长因子,体外诱导真皮成纤维细胞向成软骨细胞表型分化,探讨其作为组织工程化软骨种子细胞的可行性.方法取包皮环切术后丢弃真皮组织共3例,成纤维细胞体外培养扩增至第二代,以1×104/cm2密度接种,12 h后加入细胞诱导液(10%胎牛血清F-12培养液,CDMP1终浓度为100 ng/ml)单层培养,未加的成纤维细胞培养用CDMP1为对照.诱导7 d后观察细胞形态变化,Image Plus图像分析软件计算细胞长宽比例.激光共聚焦显微镜分别观察诱导前后细胞内Ⅰ、Ⅱ、Ⅲ型胶原表达及分布,Western印迹检测诱导前后细胞Ⅱ型胶原蛋白表达.反转录聚合酶链式反应(RT-PCR)检测诱导后成软骨相关的Sox9、聚集蛋白聚糖与Ⅱ、Ⅸ型胶原mRNA表达;同时检测与成骨相关的X型胶原、碱性磷酸酶(AKP)mRNA表达.培养第二代细胞以2×107个细胞/ml细胞密度进行离心管法培养,诱导14 d后行阿辛蓝与Ⅱ型胶原免疫组化染色.结果诱导后可见单层培养的成纤维细胞形态由长梭形向软骨细胞样多角形转变,Image Plus图像分析软件计算细胞诱导后的长宽比例为(1.40±0.15),较诱导前(7.40±1.30)差异有显著意义(P<0.05);与正常软骨细胞(1.29±0.24)差异无显著意义.免疫荧光激光共聚焦观察发现诱导后细胞内出现Ⅱ型胶原表达,Ⅰ、Ⅲ型胶原表达呈阳性.RT-PCR检测成软骨相关的Sox9,聚集蛋白聚糖,Ⅱ、Ⅸ型胶原mRNA表达阳性;X型胶原、AKP未检测到mRNA阳性表达.Western印迹检测细胞诱导后Ⅱ型胶原蛋白表达阳性.离心管培养诱导7 d后阿辛蓝染色示糖胺聚糖(GAG)均匀分布于基质中,免疫组化染色示基质中Ⅱ型胶原表达阳性.结论第二代成人真皮成纤维细胞在CDMP1生长因子诱导下可向成软骨细胞表型分化,并能分泌软骨细胞特异性基质,有望成为软骨组织工程新的种子细胞来源.  相似文献   

7.
目的:探讨不同接种密度对人脐静脉间充质干细胞(hUV-MSCs)原代培养的影响,从而确定原代培养hUV-MSCs的适宜条件。方法:分离人脐静脉内皮及内皮下层细胞,按不同接种密度分组进行原代培养,记录原代培养时间,然后进行传代和成骨诱导培养,并检测细胞钙结节的表达情况。结果:hUV-MSCs原代分离培养消化30min,种植细胞密度为5×105~1×106/cm2的方法较合理。经化学药物(如地塞米松、β-甘油磷酸、维生素C)成骨诱导后,可形成钙结节。结论:合理hUV-MSCs原代培养方法,可以快速获得最大量的自体种子细胞,从而为组织工程化骨的体外构建创造条件。  相似文献   

8.
目的应用软骨形态发生蛋白-1(CDMPI)生长因子,体外诱导真皮成纤维细胞向成软骨细胞表型分化,探讨其作为组织工程化软骨种子细胞的可行性.方法取成人真皮成纤维细胞,体外扩增至第2代,以加入CDMPl生长因子(100ng/mL)的含10%胎牛血清的F-12培养液诱导,每3 d换液1次,进行单层培养,对照组为不加CDMPl培养的成纤维细胞.7 d后,免疫荧光检测Ⅱ型胶原分泌,RT-PCR检测Aggrecan、Ⅱ型胶原mRNA表达.结果诱导后成纤维细胞细胞形态由梭形向软骨细胞样多角形、多边形转变.Ⅱ型胶原免疫荧光检测表达阳性,RT-PCR检测Aggrecan、Ⅱ型胶原mRNA表达阳性.结论成纤维细胞在CDMPI生长因子诱导下能向成软骨细胞表型分化,并能分泌软骨细胞特异性基质,有可能成为软骨组织工程新的种子细胞来源.  相似文献   

9.
目的研究体外使用音猬因子(sonic hedgehog,SHH)体外诱导人牙髓干细胞(dental pulp stemcells,DPSCs)分化为成骨细胞的可行性,以优化DPSCs向成骨细胞分化的诱导条件。方法从因正畸或阻生拔除的第一前磨牙或第三磨牙中进行牙髓的提取,通过酶消化法和过滤法获得单细胞悬液,之后再采用有限稀释法对分离出来的原代DPSCs进行培养,克隆化培养,对基质细胞抗原(stromal cell antigen-1,STRO-1)的表达进行检测。MTT法检测SHH因子对细胞增殖能力的影响,通过RT-PCR实验进行成骨向特异的基因I型胶原(type-I collagen,COL-I)表达的检检,利用透射电镜进行诱导前后的细胞超微结构的观察。结果克隆来源细胞的STRO-1表达阳性。人牙髓干细胞经SHH因子体外诱导表现出成骨细胞特性,碱性磷酸酶和VonKossa染色结果均为阳性,RT-PCR检测成骨分化相关基因I型胶原(COL-I)仅诱导组有阳性表达。结论 SHH可以在体外有效诱导人DPSCs转化为成骨细胞。  相似文献   

10.
目的:掌握体外分离、培养扩增胎盘间充质干细胞的方法,探讨其成骨细胞定向诱导分化的能力。方法:用IV型胶原酶消化人胎盘组织,获得单个核细胞,接种于10%新生牛血清(FBS)低糖培养基中(DMEM),贴壁后常规换液、传代,流式细胞仪检测其表面标志,用β 甘油磷酸钠,维生素C,地塞米松联合诱导,使胎盘贴壁细胞分化为成骨细胞,诱导后10?d检测碱性磷酸酶(ALP)活性,并行茜素红染色。结果:培养7~14?d后有少量贴壁细胞生长,逐渐呈成纤维细胞状,表达CD29、CD44和CD105,不表达CD34、CD45、CD19;在成骨诱导10?d后细胞ALP活性明显增强,茜素红染色可见钙盐沉积。结论:胎盘组织可能是新的间充质干细胞来源。  相似文献   

11.
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.  相似文献   

12.
Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.  相似文献   

13.
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.  相似文献   

14.
Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.  相似文献   

15.
Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.  相似文献   

16.
A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care(~'4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.  相似文献   

17.
Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.  相似文献   

18.
Journal of Nanjing Medical University (English Edition) JNMU, sponsored by Nanjing Medical University, was established in 1987. It is a bimonthly comprehensive English medical journal published locally and abroad.Since 2007, Journal of Nanjing Medical University (English Edition )was granted Elsevier the full publishing and distribution rights worldwide for the Electronic Edition, excluding the People's Republic of China.  相似文献   

19.
目的:评价使用安心颗粒对急诊经皮冠状动脉介入术(PPCI)术后生活质量的影响.方法:将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组(术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次)和对照组(仅接受基础药物治疗).所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表(MIDAS)、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果:入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异(P>0.05);出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低(P<0.05);组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低(P<0.05).两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血(P>0.05).两组发生轻度血小板减少症的患者数比较无差异(P>0.05).结论:PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.  相似文献   

20.
Objective To measure Derpl and Blot5 allergen levels in asthmatics' homes in Hongkong.Methods Seventy houses were enrolled for a mite indoor environment study. Dust samples were obtained from two sites of each patients' house: bed and floor. Derpl and Blot5 levels were quantified by a two-site monoclonal antibody-based ELISA technique.Results The levels of Derpl allergens found in bed (geometric mean (GM) 3.43 μg/g of dust; 95%CI, 1.89-4.96 μg/g)and on the floor (GM 1.12 μg/g of dust; 95%CI, 0.71-1.53 μg/g) indicated significant differences (P=0.005). However, the levels of Blot5 allergens found in bed (GM 19.00 μg/g of dust; 95%CI, 0.89-38.90 μg/g) and on the floor (GM 6.14 μg/g of dust; 95%CI, 0.40-11.90 μg/g) showed no statistically significant difference. In addition, in regards to the exposure index for Derpl and Blot5 allergens found in bed and on the floor, 17.6% in bed and 8.6% on the floor had levels of Blot5 ≥ 10 μg/g of dust, higher than those obtained for Derp1 (7.2% and 0% in bed and on the floor respectively, P< 0.05); higher percentages in bed and on the floor (25.0% and 35.7%) were observed for levels of Blot5 =0 μg/g of dust as compared with Derpl in bed and on the floor (4.3% and 14.5% respectively, P< 0.05).Conclusions Derpl and Blot5 are the major allergens found in this regional study, Blot5 is a more potent allergen in Hongkong, probably reflecting the high level of exposure to Blomia tropicalis (Bt). Bt and Dermatophagoides pteronyssinus (Dp) allergens should be included for precise diagnosis and effective immuno-therapeutic treatment of mite allergy in Hongkong.  相似文献   

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