脂肪干细胞复合PLGA体外诱导成软骨的实验研究

目的 探讨脂肪干细胞接种于PLGA支架复合培养体外诱导成软骨的能力.方法 收集脂肪干细胞,调整细胞悬液密度为4.0×1010>/L,接种在PLGA支架材料上进行复合,在特定培养基条件下,对脂肪干细胞/PLGA进行成软骨诱导,于诱导3周后应用扫描电镜观察,藩红O/固绿染色和Ⅱ型胶原免疫组织化学检测成软骨表型;逆转录-聚合酶链反应(RT-PCR)检测前Ⅱ型胶原蛋白、aggrecan和Sox9的mRNA基因表达情况.结果 脂肪干细胞接种子PLGA材料后,复合体表面逐渐光滑润泽,质地逐步增韧,体外诱导后细胞/PLGA支架复合体基本保持原状;3周时取材行扫描电镜观察,可见细胞在材料表面附着生长,基质分泌明显,连串成片;石蜡切片藩红O/固绿染色可见诱导组细胞胞外基质分泌旺盛,和支架材料连接成片,呈强阳性红染;胞外基质Ⅱ型胶原免疫组织化学着色阳性;RT-PCR检测前Ⅱ型胶原蛋白、aggrecan和Sox9的mRNA基因均阳性表达.结论 脂肪干细胞能够和PLGA复合培养,在特定诱导培养基条件下可以向软骨方向分化,形成类软骨样组织.     

自体富血小板血浆联合骨髓基质干细胞复合改建脱细胞真皮基质修复兔关节软骨的可行性研究

目的:研究自体富血小板血浆（PRP）联合骨髓基质干细胞（BMSCs）复合改建脱细胞真皮基质（ADM）修复兔关节软骨的修复效果及临床价值。方法:选择60只股骨髁关节损伤的青紫蓝兔作为研究对象,随机分为研究组、对照组和空白组,各20只。首先制备PRP,再分离BMSCs,并进行体外培养,接种于ADM支架上。研究组培养基添加10% PRP,其他条件同对照组,空白组兔不做任何处理。将BMSCs-ADM复合物植入兔关节缺损处,在1、2、3个月末对构建的软骨组织进行免疫力学、组织学、生物力学检测,比较PRP-BMSCs-ADM联合修复关节缺损的效果。结果:随着培养时间延长,BMSCs的细胞数目显著增多,呈现S形生长曲线,第3天后进入对数期,第9天后进入平台期;从第3天开始,细胞数目OD值研究组均明显高于对照组（P<0.01）;随着时间的延长,各组标本软骨样细胞数目增多,软骨陷窝逐渐成熟,软骨表面逐渐光滑,IRCS宏观评分和组织学评分均呈现增长趋势（P<0.01）,标本压力弹性模量有上升趋势（P<0.05）;同时期研究组标本评分与压力弹性模量均显著高于对照组和空白组（P<0.01）;3个月后,研究组软骨缺损修复状况明显优于对照组和空白组。结论:自体PRP联合BMSCs复合改建ADM修复兔关节软骨的修复效果良好,具有可行性。     

间充质干细胞治疗骨关节炎的临床研究进展

骨关节炎(OA)是一种进展性疾病,表现为软骨缺损、滑膜炎症、软骨下骨硬化及骨赘形成,关节表面软骨再生能力弱,无法实现自我修复.骨关节炎患者的主要临床表现为疼痛和关节活动受限,肢体功能和生活质量受影响.理想的治疗方案应缓解疼痛、促进软骨修复,以延缓或阻止病程进展.间充质干细胞(MSCs)在治疗骨关节炎方面具备以上优势且无...     

兔脱细胞软骨基质颗粒复合SD大鼠脂肪干细胞对软骨内成骨的促进作用

目的:探讨兔脱细胞软骨基质颗粒(ACM)的生物相容性,阐明其对SD大鼠脂肪干细胞(ADSCs)软骨内成骨的促进作用。方法:提取原代SD大鼠ADSCs进行三向分化实验,流式细胞术检测细胞干性。采集3月龄新西兰白兔耳软骨,采用研磨浸泡法制备ACM备用。实验分为ACM组和ADSCs+ACM共培养组,在扫描电镜下观察ACM和ACM上ADSCs的超微形态表现,采用钙黄绿素/碘化丙啶(Calcein-AM/PI)荧光染色法观察ADSCs+ACM共培养组中ADSCs的生长情况。将ADSCs分为对照组和实验组(ADSCs与ACM共培养),采用CCK-8法检测2组细胞增殖活性,实时荧光定量PCR (RT-qPCR)法检测成软骨诱导7和14 d时2组细胞中Ⅱ型胶原(COL-Ⅱ)、Ⅹ型胶原(COL-Ⅹ)、SOX转录因子9 (SOX-9)、血管内皮生长因子(VEGF)、碱性磷酸酶(ALP)和Runt相关转录因子2 (RUNX-2) mRNA表达水平。结果:成功制备兔ACM,提取的SD大鼠ADSCs可以进行三向分化且表达干细胞特异性表面标志物。ADSCs可以在ACM上黏附生长。与对照组比较,实验组ADSCs增殖...     

耳内镜下嵌入蝶形软骨鼓膜成形术治疗鼓膜前缘穿孔效果及对患者听力恢复的影响

目的 研究耳内镜下嵌入蝶形软骨鼓膜成形术治疗鼓膜前缘穿孔效果及对患者听力恢复的影响.方法 回顾性分析2017年11月至2019年11月宣城市人民医院进行鼓膜前缘穿孔治疗的100例患者资料,依据患者的治疗方式不同,将其分为观察组与对照组,每组各50例.观察组行耳内镜下嵌入蝶形软骨鼓膜成形术,对照组行显微镜下颞肌筋膜修补术...     

经鼻填充鼻中隔软骨修复眶内侧壁骨折

目的：探讨应用鼻内镜填充鼻中隔软骨,修复眶内侧壁骨折,经鼻做眼部手术的可能性。方法：选取眶内侧壁骨折患者11例（11眼）,术前检查均有不同程度的眼球内陷、复视、视力减退症状,行眼眶CT诊有眶内容物疝入筛窦并伴积液,其中6例内直肌肿胀。手术开始在内镜直视指引下,经鼻腔开放筛窦,摘除筛房,暴露骨折的纸板,将疝入物回纳眶内,鼻中隔软骨覆盖骨折区。结果：患者手术后全部治愈。双眼突出度相差≤1mm,平均0.11mm。视力较术前不改变或者略有改善。鼻内镜检查见筛窦术腔上皮化,填充的鼻中隔软骨无移动,无感染及排斥现象。结论：经鼻填充鼻中隔软骨修复眶内侧壁骨折方法简便,成功率高。自体鼻中隔软骨无排斥反应。     

Auricular skin-cartilage sandwich graft technique for full-thickness eyelid reconstruction

Full-thickness deficiency of eyelid tissues can result in coloboma or retraction or both. Here we report our initial experience on the use of auricular skin-cartilage sandwich graft technique for full-thickness eyelid deformities. Five patients (4–32 years) underwent the procedure. Patients with full-thickness eyelid deformity were included. Three patients were operated for large-sized coloboma and two for eyelid retraction. One patient had congenital, and four patients had acquired etiology. The following parameters were specifically assessed: correction of deformity, ocular surface problems, graft status, and epithelization of skin-cartilage graft. All the patients had a good correction of eyelid position, except one patient who had severe eyelid retraction (8 mm) at presentation. None of our patients had corneal erosion/defect, persistent ocular surface redness, or graft loss. The auricular skin-cartilage sandwich graft technique produces optimal results with no graft loss. Advancement of orbicularis muscle in between the auricular skin and cartilage grafts (sandwich technique) is an imperative step that leads to the survival of both grafts.     

自体鼻中隔软骨联合耳廓软骨的鼻中隔延伸物移植术在鼻尖整形中的应用

目的:探讨运用鼻中隔软骨联合耳廓软骨构建鼻中隔延伸物复合体重置鼻尖位置和重塑鼻尖形态的可行性。方法:2018年9月至2019年10月,以中国医学科学院整形外科医院17例低鼻患者为研究对象,17例患者中男2例,女15例;年龄19~39岁,平均27岁。17例患者均为初次鼻整形病例。术中在全身麻醉下行鼻整形术。切取鼻中隔软骨...     

三维支架中骨髓间充质干细胞及软骨细胞和脂肪源性成熟基质细胞的成软骨效应

背景：软骨修复的关键是种子细胞在三维支架中的定向分化,但细胞放入三维支架中很难有透明软骨表现。 目的：考察3种种子骨髓间充质干细胞,软骨细胞和脂肪源性成熟基质细胞在支架中的成软骨特性。 方法：抽取羊骨髓,胰酶消化后获得原代骨髓间充质干细胞细胞、软骨细胞和脂肪源性成熟基质细胞,单层培养扩增。取P1骨髓间充质干细胞,P3软骨细胞和P3脂肪源性成熟基质细胞种植入不同比例(10%,20%,50%,80%,100%)的胶原/透明质酸支架中, 在无血清培养液中三维培养。2周后,用SO染色和免疫组织化学染色分析,观察硫酸软骨素和Ⅱ型胶原合成情况。 结果与结论：种子细胞在三维支架中特定条件下有成软骨效应,骨髓间充质干细胞在含20%透明质酸的胶原/透明质酸支架扩增少于3代有成软骨效应,软骨细胞传代数更高仍然有成软骨效应,但脂肪源性成熟基质细胞成软骨效应能力较弱。结果提示,在同样条件下骨髓间充质干细胞、软骨细胞较脂肪源性成熟基质细胞有更好的成软骨性能。 关键词：骨髓间充质干细胞;软骨细胞;组织工程;软骨;脂肪干细胞 doi:10.3969/j.issn.1673-8225.2010.16.004     

Magnetic resonance imaging of anterior cruciate ligament tears: reevaluation of quantitative parameters and imaging findings including a simplified method for measuring the anterior cruciate ligament angle

We evaluated the diagnostic utility of magnetic resonance imaging (MRI) for predicting anterior cruciate ligament (ACL) tears using both quantitative parameters and nonquantitative imaging findings. MRI examinations were retrospectively evaluated in a group of patients with arthroscopically confirmed complete ACL tear and in a control group with arthroscopically confirmed intact ACL. We evaluated multiple MRI features to compare their sensitivity and specificity for detecting ACL tears. Particular emphasis is put on the evaluation of three different quantitative parameters, including a simplified method for measuring the ACL angle. With a threshold value of 45° the ACL angle reached a sensitivity and specificity of 100% for detecting ACL tears. With a threshold value of 0° the Blumensaat angle had a sensitivity of 90% and a specificity of 98%. Finally, a threshold value of 115° gave the posterior cruciate ligament angle a sensitivity of 70% and a specificity of 82%. Discontinuity was found to be the most useful of the ACL abnormalities. Of the secondary findings anterior tibial displacement was the best predictor of ligamentous injury. However, ACL abnormalities and secondary findings, alone or combined, failed to surpass the diagnostic value of the ACL angle for predicting ACL tears. Quantitative parameters are thus good predictors of ACL tears and may increase the overall sensitivity and specificity of MRI. The ACL angle may be confidently measured in a single MRI section and can be considered to be the most reliable quantitative parameter for detecting ACL tears.