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71.
Objective To investigate the potential application of human transforming growth factor-beta-1 (hTGF-β1) gene mediated by type 2 recombinant adeno-associated virus (rAAV2) vector inducing chondrogenic differentiation of canine mesenchymal stem cells (MSCs) in vitro. Methods Canine MSCs from bone marrow were isolated and cultured in vitro by density gradient centrifngation and adherence screening methods. The morphology of MSCs was observed by inverted phase contrast microscope and Giemsa stain. Flow eytometry was used to detect surface antigens of MSCs, The third generation of MSCs were transfected by rAAV2-hTGF-β1 with or without MOI of 1 ×105 v.g./cell or 5×105 v.g./cell. The expression of hTGF-β1 was detected by Western blot after 10 days, and TGF-β1 synthesis was determined by ELISA at 3, 6 and 9 day, respectively. After 2 weeks of culturing, mRNA expressions of type Ⅱ collagen and aggrecan were determined by RT-PCR and the collagen Ⅱ protein was detected by immunocytochemistry. Results The MSCs appeared to be morphologically spindle-shaped and showed active capability of proliferation both in primary and passage generations. Flow cytometry analysis indicated that MSCs were universally positive for CD29, CD44 and CD105, but negative for CD34 and CD45. TGF-β1 expression can be observed by Western blot after 10 days in two transfection groups, MOI of 5 × 105 group and MOI of 1× 105 group. With the extension of time, the contents of hTGF-β1 increased in the two groups detected by ELISA, while there was a significant difference between them two (P < 0.01). After 2 weeks of transfection of MSCs by rAAV2-hTGF-β1, the expression of collagen Ⅱ and Aggreacan mRNAs were positive. It also showed positive of collagen Ⅱ detected by immunocytochemistry. Conclusion Canine MSCs show chondrogenesis differentiation after induction by Type 2 rAAV mediated transfer of TGF-β1 gene. The process is a potential application for cartilage tissue engineering. 相似文献
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TGF-β1缓释载体体外构建组织工程软骨 总被引:3,自引:14,他引:3
目的制备负载TGF—β1壳聚糖缓释微球的三维多孔壳聚糖支架,检测TGF—β1缓释载体对软骨细胞功能的影响.方法乳化交联法制备TGF—β1壳聚糖缓释微球并将其与壳聚糖支架复合,检测其体外降解并通过ELISA法检测微球的载药、释药性能。将软骨细胞置于复合载体中立体培养,通过苏木素伊红染色、Ⅱ型胶原免疫组化染色、扫描电镜观察复合载体对细胞的增殖、功能的影响。结果缓释微球的TGF—β1包封率达90.1%,并具有良好的药物缓释性能,软骨细胞在复合载体中增殖良好,并能够保持其表型及Ⅱ型胶原分泌功能。结论负载TGF—β1壳聚糖缓释微球的壳聚糖支架作为软骨细胞的载体在组织工程软骨的构建及软骨损伤的修复中有良好的应用前景。 相似文献
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目的 探讨尼莫地平联合认知功能训练对急性脑脑梗死认知功能障碍患者的影响.方法 选择我院2007-11~2009-11急性脑梗死认知功能障碍患者81例,将以上患者随机分为2组,观察组和对照组.对照组患者给予常规神经内科治疗,同时给予物理治疗、作业治疗等.在对照组治疗基础上,观察组给予认知功能训练和尼莫地平治疗.2组患者均治疗3个月.患者治疗前和治疗3个月后,采用神经行为认知状态测试(NCSE)量表、画钟试验(CDT)、欧洲神经功能缺损评分(ESS)、Fugl-Meyer(FMA)量表、Barthel Index对患者进行评分.结果 观察组治疗3个月后NCSE总评分和画钟试验(CDT)评分分别与对照组治疗3个月后比较,差异有统计学意义(P<0.05);观察组治疗3个月后ESS评分、FAM评分和日常生活活动能力评分分别与对照组治疗3个月后比较,差异有统计学意义(P<0.05).结论 尼莫地平联合认知功能训练有助于改善急性脑脑梗死后认知功能障碍,有利于提高患者的肢体运动功能和日常生活活动能力,临床效果显著,值得借鉴. 相似文献
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目的 研究猪去抗原半月板在体外的降解规律,以进一步调整其合适降解速度。方法选用4~6月龄健康小型猪5只,取其双膝关节半月板。一部分采用环氧化物进行去抗原处理,制备成去抗原半月板;另一部分不作任何处理,为新鲜半月板。将两种半月板冻干处理后,置于含胶原酶、木瓜蛋白酶、透明质酸酶和胰酶的PBS液中,于37C酶解3、7、15、30d。另取两种半月板在PBS液37℃下水解30d。采用失重法、紫外光谱分析及扫描电镜测定其降解特性,并测定两种半月板的亲水性。结果两种半月板于4种酶中的酶解效果显著,其中以胰酶最明显,但去抗原半月板酶解程度低于新鲜半月板。新鲜半月板及去抗原半月板水解率分别为0.76%、0.81%。两种半月板在4种酶的降解液中紫外吸收值范围大致相同,为200~300nm。扫描电镜观察,去抗原半月板的空间结构改变不及新鲜半月板明显。去抗原半月板的亲水性较新鲜半月板稍低:吸水率为157%、188%;保水率为152%、167%。结论去抗原半月板降解较新鲜半月板慢,为调整去抗原半月板在体内降解速度提供依据。 相似文献
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Objective To investigate the potential application of human transforming growth factor-beta-1 (hTGF-β1) gene mediated by type 2 recombinant adeno-associated virus (rAAV2) vector inducing chondrogenic differentiation of canine mesenchymal stem cells (MSCs) in vitro. Methods Canine MSCs from bone marrow were isolated and cultured in vitro by density gradient centrifngation and adherence screening methods. The morphology of MSCs was observed by inverted phase contrast microscope and Giemsa stain. Flow eytometry was used to detect surface antigens of MSCs, The third generation of MSCs were transfected by rAAV2-hTGF-β1 with or without MOI of 1 ×105 v.g./cell or 5×105 v.g./cell. The expression of hTGF-β1 was detected by Western blot after 10 days, and TGF-β1 synthesis was determined by ELISA at 3, 6 and 9 day, respectively. After 2 weeks of culturing, mRNA expressions of type Ⅱ collagen and aggrecan were determined by RT-PCR and the collagen Ⅱ protein was detected by immunocytochemistry. Results The MSCs appeared to be morphologically spindle-shaped and showed active capability of proliferation both in primary and passage generations. Flow cytometry analysis indicated that MSCs were universally positive for CD29, CD44 and CD105, but negative for CD34 and CD45. TGF-β1 expression can be observed by Western blot after 10 days in two transfection groups, MOI of 5 × 105 group and MOI of 1× 105 group. With the extension of time, the contents of hTGF-β1 increased in the two groups detected by ELISA, while there was a significant difference between them two (P < 0.01). After 2 weeks of transfection of MSCs by rAAV2-hTGF-β1, the expression of collagen Ⅱ and Aggreacan mRNAs were positive. It also showed positive of collagen Ⅱ detected by immunocytochemistry. Conclusion Canine MSCs show chondrogenesis differentiation after induction by Type 2 rAAV mediated transfer of TGF-β1 gene. The process is a potential application for cartilage tissue engineering. 相似文献
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【目的】 通过Meta分析,探讨保守和手术两种方法治疗急性髌骨脱位的临床疗效有无差异。【方法】 在PubMed、Cochrane图书馆、MEDLINE、EMBase等数据库(均截止至2014年2月)中检索比较保守和手术治疗急性髌骨脱位的随机对照研究,使用RevMan5.2 软件进行Meta 分析。临床指标为髌骨脱位复发率、髌骨半脱位的复发率、Kujala评分、再次手术率以及膝关节不稳等情况。【结果】 纳入8篇随机对照研究(400例),Meta分析结果显示:对于急性髌骨脱位患者,两种治疗方法在半脱位复发率(RR= 0.73,95%CI:0.46-1.15)、Kujala评分(WMD=5.56,95%CI:-3.54-14.67)、再次手术率(RR = 1.09,95%CI:0.72-1.65)、膝关节不稳症状发生率(RR= 0.57,95%CI:0.25-1.30)等方面差异均无统计学意义(P > 0.05),手术治疗后脱位复发率(RR = 0.65,95%CI:0.48-0.88)低于保守治疗,且2年随访期内,手术治疗Kujala评分(WMD=13.94,95%CI:5.36-22.52)方面优于保守治疗(P< 0.05)。【结论】 对于急性髌骨脱位,手术治疗后脱位复发率较保守治疗低,且膝关节短期功能恢复及稳定性优于保守治疗,但是保守和手术两种治疗方法在膝关节Kujala评分、半脱位复发率、再次手术率以及膝关节不稳症状发生率方面并无统计学差异。 相似文献
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背景:目前缺乏与人类月骨缺血性坏死病理过程类似的Kienböck病动物模型。目的:采用医用TH胶栓塞法建立一种新型月骨缺血坏死兔模型,并探讨造模方法的合理性。方法:选择健康成年新西兰兔30只,雌雄不限。采用自身对照方法随机分为实验侧和对照侧,实验侧采用月骨中心钻孔,注射医用TH胶0.2 mL,反复注药3次;对照侧月骨中心钻孔注射同等量的生理盐水。于实验4,8,12周行X射线、大体观察、Micro-CT 骨计量学及组织病理学检测。结果与结论:大体标本、X射线及组织学检测结果可见实验侧造模后8周月骨出现缺血坏死表现,12周月骨缺血坏死更为典型。Micro-CT 骨小梁微观参数中,实验侧骨小梁密度参数骨体积分数及骨小梁数量均明显低于对照侧(P < 0.05);骨小梁空间参数明显增高,其中实验侧骨小梁离度及结构模型指数明显高于对照侧。提示注射医用TH胶栓塞后8周实验侧出现月骨缺血坏死表现,注药后12周可建立月骨缺血坏死兔模型,可以实现在周围组织无明显损伤情况下出现月骨内血运受损、骨小梁骨折、空骨陷窝等与人类月骨坏死类似的改变。
中国组织工程研究杂志出版内容重点:肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接: 相似文献
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目的 探讨采用壳聚糖与 型胶原复合制作新型组织工程软骨三维多孔支架的方法,并对其理化性能进行检测。 方法 将精制88%脱乙酰度壳聚糖溶于0 .2 mol/ L 醋酸溶液制成2 %溶液,高纯度猪 型胶原溶于0 .5 m ol/ L醋酸溶液制成1%溶液,两者按4∶1(重量比)充分搅拌混合,采用冷冻干燥法制成壳聚糖与 型胶原复合支架。采用碳化二亚胺/ N-羟基琥珀酰亚胺对支架进行交联,力学测定比较支架交联前后的强度变化,扫描电镜观察其超微结构,于2、4、6和8周经溶菌酶体外降解实验测定其体外降解性。 结果 制备的复合支架成形良好,交联后其力学强度明显增加。扫描电镜显示壳聚糖与 型胶原成分在支架内分布均匀,支架内孔洞相互连通似海绵状,孔径10 0~2 5 0 μm。各时间段复合支架体外降解较单纯壳聚糖支架快。 结论 壳聚糖与 型胶原复合成功地制作成了三维多孔复合支架。其理化性能及体外降解测定,可以作为支架载体,应用于组织工程软骨的构建。 相似文献
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壳聚糖与Ⅱ型胶原复合制作组织工程软骨支架及其性能研究 总被引:10,自引:1,他引:10
目的探讨采用壳聚糖与Ⅱ型胶原复合制作新型组织工程软骨三维多孔支架的方法,并对其理化性能进行检测. 方法将精制88%脱乙酰度壳聚糖溶于0.2 mol/L醋酸溶液制成2%溶液,高纯度猪Ⅱ型胶原溶于0.5 mol/L醋酸溶液制成1%溶液,两者按4∶1(重量比)充分搅拌混合,采用冷冻干燥法制成壳聚糖与Ⅱ型胶原复合支架.采用碳化二亚胺/ N-羟基琥珀酰亚胺对支架进行交联,力学测定比较支架交联前后的强度变化,扫描电镜观察其超微结构,于2、4、6和8周经溶菌酶体外降解实验测定其体外降解性. 结果制备的复合支架成形良好,交联后其力学强度明显增加.扫描电镜显示壳聚糖与Ⅱ型胶原成分在支架内分布均匀,支架内孔洞相互连通似海绵状,孔径100~250 μm.各时间段复合支架体外降解较单纯壳聚糖支架快. 结论壳聚糖与Ⅱ型胶原复合成功地制作成了三维多孔复合支架.其理化性能及体外降解测定,可以作为支架载体,应用于组织工程软骨的构建. 相似文献