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Injured tendons have limited repair ability after full‐thickness lesions. Tendon regeneration properties and adverse reactions were assessed ex vivo in an experimental animal model using a new collagen I membrane. The multilamellar membrane obtained from purified equine Achilles tendon is characterized by oriented collagen I fibers and has been shown to sustain cell growth and orientation in vitro. The central third of the patellar tendon (PT) of 10 New Zealand White rabbits was sectioned and grafted with the collagen membrane; the contralateral PT was cut longitudinally (sham‐operated controls). Animals were euthanized 1 or 6 months after surgery, and tendons were subjected to histological and Synchrotron Radiation‐based Computed Microtomography (SRµCT) examination and 3D structure analysis. Histological and SRµCT findings showed satisfactory graft integration with native tendon. Histological examination also showed ongoing angiogenesis. Adverse side‐effects (inflammation, rejection, calcification) were not observed. The multilamellar collagen I membrane can be considered as an effective tool for tendon defect repair and tendon augmentation. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31: 738–745, 2013  相似文献   
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背景:去细胞肌肉生物支架联合人脐带问充质干细胞移植将是治疗脊髓损伤的一项重要措施。但两者是否具有良好的相容性,人脐带间充质干细胞能否在去细胞肌肉生物支架中长期存活并均匀分布,尚未得到证实。目的:观察大鼠去细胞肌肉生物支架与人脐带间充质干细胞的相容性。方法:改良化学法制备大鼠去细胞肌肉生物支架,将第3代人脐带间充质干细胞Hoecliest33342荧光标记后分为3组进行实验,细胞+支架组、细胞+支架大鼠体内组和单纯细胞组。分别应用苏木精-伊红、Masson染色方法观察去细胞肌肉生物支架的组织形态,以荧光倒置相差显微镜和扫描电镜观察人脐带间充质干细胞的吸附和生长情况。结果与结论:人脐带间充质干细胞与去细胞肌肉生物支架充分附着,生长增殖活跃,细胞在支架内分布均匀。细胞+支架体内组与细胞+支架组相比在移植后1—7d人脐带间充质干细胞数量差异无显著性意义(P〉0.05),在移植14d细胞+支架体内组人脐带间充质干细胞数量大于细胞+支架组(P〈0.05)。提示去细胞肌肉生物支架与人脐带间充质干细胞有较好的相容性,体内环境更有利于细胞增殖和两者融合。  相似文献   
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目的 通过制备去细胞化肝脏生物支架,研究自主神经递质对再细胞化过程的影响.方法 使用全器官灌注法制备C57小鼠去细胞化肝脏生物支架,行HE和Massion染色鉴定并检测DNA残留量;RT-PCR和免疫荧光化学检测再细胞化肝脏中种子细胞(L02细胞株)肾上腺素能受体及亚型表达情况;检测平面以及生物支架内异丙肾上腺素对种子细胞增殖的作用,以及对再细胞化肝脏白蛋白分泌和尿素合成水平的影响.结果 成功制备去细胞化肝脏生物支架,L02细胞株表达肾上腺素能受体,并且β2受体亚型表达相对较高;异丙肾上腺素显著促进平面及再细胞化肝脏内L02细胞株增殖,并增强再细胞化肝脏分泌白蛋白和尿素.结论 异丙肾上腺素可促进去细胞化肝脏生物支架再细胞化进程.  相似文献   
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Articular cartilage injuries experienced at an early age can lead to the development of osteoarthritis later in life. In situ three‐dimensional (3D) printing is an exciting and innovative biofabrication technology that enables the surgeon to deliver tissue‐engineering techniques at the time and location of need. We have created a hand‐held 3D printing device (biopen) that allows the simultaneous coaxial extrusion of bioscaffold and cultured cells directly into the cartilage defect in vivo in a single‐session surgery. This pilot study assessed the ability of the biopen to repair a full‐thickness chondral defect and the early outcomes in cartilage regeneration, and compared these results with other treatments in a large animal model. A standardized critical‐sized full‐thickness chondral defect was created in the weight‐bearing surface of the lateral and medial condyles of both femurs of six sheep. Each defect was treated with one of the following treatments: (i) hand‐held in situ 3D printed bioscaffold using the biopen (HH group), (ii) preconstructed bench‐based printed bioscaffolds (BB group), (iii) microfractures (MF group) or (iv) untreated (control, C group). At 8 weeks after surgery, macroscopic, microscopic and biomechanical tests were performed. Surgical 3D bioprinting was performed in all animals without any intra‐ or postoperative complication. The HH biopen allowed early cartilage regeneration. The results of this study show that real‐time, in vivo bioprinting with cells and scaffold is a feasible means of delivering a regenerative medicine strategy in a large animal model to regenerate articular cartilage.  相似文献   
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文题释义:骨髓间充质干细胞复合海螵蛸支架:以中药海螵蛸为载体支架,骨髓间充质干细胞为种子细胞,二者共同培养构建复合生物支架,经过各项生物安全性测评后应用于骨缺损的治疗。制作过程为:选用第3代骨髓间充质干细胞,经0.25%胰酶消化制成细胞悬液,将细胞浓度调整为5×108 L-1,用移液枪将骨髓间充质干细胞悬液缓慢逐滴滴加于24孔板中的海螵蛸上,200 μL/孔,分2次缓慢接种,尽量不使细胞从材料上溢出,置于培养箱中培养4 h,待细胞充分黏附于海螵蛸支架材料后,缓慢逐滴加入培养液700 μL,将支架与细胞复合培养8 d,两三天换液1次。细胞毒性评估:是采用受试物与细胞共同培养的方法,对样品进行毒理学风险评估。生物材料应用于体内后,可能与体内细胞的细胞膜、细胞器、蛋白质合成、DNA合成等相互作用,产生细胞毒性。根据ISO10993-5标准,在复合支架应用于临床治疗前,需通过复合材料浸提液试验以测定其对细胞活性、功能、遗传的影响。  摘要背景:作为骨组织工程的修复材料,应具有良好的生物相容性及降解吸收性,许多学者在此方面也做了较深入的研究,但中药复合细胞生物支架的研究较少。目的:参照《医疗器械生物学评价标准》,对兔骨髓间充质干细胞-海螵蛸生物复合支架进行体外细胞毒性检测,评估复合支架的毒性等级,为临床应用生物支架提供实验依据。方法:依照ISO标准按“材料面积∶浸提介质体积=3-6 cm2∶1 mL”制作骨髓间充质干细胞-海螵蛸生物复合支架材料浸提液。制备L-929细胞悬液,将细胞浓度调整至1×107 L-1进行接种培养,设置阳性对照组(含苯酚的DMEM培养液)、实验组(材料浸提液)、阴性对照组(新鲜DMEM培养液)。培养24,48,72 h采用MTT法检测L-929细胞吸光度值,计算各组细胞的相对增殖率,评估复合支架毒性等级。结果与结论:实验组、阴性对照组、阳性对照组吸光度值在不同时间点不完全相同(P=0.000 < 0.01),各时间点内比较,实验组与阴性对照组吸光度值均明显高于阳性对照组(P < 0.01);兔骨髓间充质干细胞-海螵蛸生物支架细胞毒性为1级。结果说明兔骨髓间充质干细胞-海螵蛸生物支架无明显毒性作用,符合生物材料应用要求。  ORCID: 0000-0002-0003-6228(彭雅) 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   
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Esophageal reconstruction with ECM and muscle tissue in a dog model   总被引:5,自引:0,他引:5  
An in vivo study was conducted to determine if an extracellular matrix (ECM) scaffold co-localized with autologous muscle tissue could achieve constructive remodeling of esophageal tissue without stricture. ECM derived from the porcine urinary bladder was processed, decellularized, configured into a tube shape, and terminally sterilized for use as a bioscaffold for esophageal reconstruction in a dog model. Twenty-two dogs were divided into four groups, three groups of five and one group of seven. Groups 1 and 2 were repaired with either ECM alone or muscle tissue alone, respectively. Groups 3 and 4 were repaired with ECM plus either a partial (30%) covering with muscle tissue or a complete (100%) covering with muscle tissue, respectively. Animals in groups 1 and 2 were sacrificed within approximately 3 weeks because of the formation of intractable esophageal stricture. Four of five dogs in group 3 and six of seven dogs in group 4 were survived for 26 days to 230 days and showed constructive remodeling of esophageal tissue with the formation of well organized esophageal tissue layers, minimal stricture, esophageal motility, and a normal clinical outcome. Mechanical testing of a subset of the remodeled esophageal tissue from animals in groups 3 and 4 showed progressive remodeling from a relatively stiff, non-compliant ECM tube structure toward a tissue with near normal biomechanical properties. We conclude that ECM bioscaffolds plus autologous muscle tissue, but not ECM scaffolds or muscle tissue alone, can facilitate the in situ reconstitution of structurally and functionally acceptable esophageal tissue.  相似文献   
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单纯去细胞化组织材料具有较多缺点,有必要通过交联剂来改善材料的各种性能。新型交联剂京尼平细胞毒性低,能与去细胞化材料生成环状结构,从而大大改善去细胞化生物材料的性能,有利于材料的进一步应用。本文从京尼平的交联特性以及交联产物性能等方面作一综述,为后期的实验提供一定的理论基础。  相似文献   
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目的考察经门静脉移植后的胰岛细胞团在肝脏血管中停留的位置及栓塞部位细胞团的相关情况。方法取成年SD大鼠,从门静脉在体灌注去污剂十二烷基硫酸钠溶液使肝脏去细胞化,再输注经二苯基硫卡巴腙染色后的大鼠胰岛,在体视显微镜和倒置显微镜下观察去细胞后的肝脏内部的血管和胰岛在肝脏内的栓塞情况。结果大鼠肝脏在体去细胞后,可得到完全透明、清晰的肝脏支架,肝内血管分支丰富,经灌注(800±250)IEQ胰岛细胞团后,胰岛主要栓塞在大鼠肝脏边缘血管末梢,有时一个血管末梢栓塞有多个细胞团。结论本方法模拟大鼠门静脉内胰岛移植,呈现了胰岛在肝脏内血管的驻留情况,有利于进一步研究改进胰岛在肝内移植后的存活和功能发挥。  相似文献   
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目的:观测原代小鼠胰岛在小鼠肝脏脱细胞支架上的生长及功能,探讨组织工程学治疗糖尿病的新方法?方法:运用灌注法制备结构完整的全肝脏脱细胞支架并检测,运用胶原酶P灌注消化法获取小鼠胰岛细胞并行结构和功能完整性分析;将分离纯化的原代小鼠胰岛经门静脉灌入全肝脏脱细胞支架,在三维培养体系培养后进行相关检测?结果:肝脏脱细胞支架检测未见细胞结构,胶原结构保存完整?支架内剩余DNA定量检测结果为(38 ± 11)ng/mg dsDNA,生物相容性良好?双硫腙(dithizone,DTZ)染色胰岛呈现特异性猩红色,葡萄糖刺激实验提示在高糖组中培养的原代小鼠胰岛胰岛素分泌功能明显高于低糖组,差异具有统计学意义(P < 0.01);将胰岛移植入肝脏脱细胞支架培养5 d后胰岛定植,且胰岛素基因表达显示脱细胞支架三维培养中胰岛素表达水平与平面培养相比,差异具有统计学意义(P < 0.01)?结论:肝脏脱细胞支架三维培养体系中的胰岛细胞比传统平面培养具有更佳的细胞活力及更有效的胰岛素分泌功能?  相似文献   
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