不同持续时间的周期性压力对构建组织工程软骨的影响

Objective To determine the best duration for exerting the cyclic pressure under which the tissue-engineered cartilage is constructed. Methods Free chondrocytes isolated from rabbit articular cartilage were seeded into polylactic acid-co-glycolic acid(PLGA) scaffolds after expansion in vitro, and ran-domized into 4 groups. In Groups 1 to 3, chondrocytes were cultured under daily cyclic pressure (0 ～ 200 kPa, 0.1Hz) for 4 hours, 8 hours, 12 hours respectively; Group 4 was a control in which no pressure was exerted. In each group, after 2 weeks of culture, the tissue engineered cartilages were observed in vitro and assessed by his-tological staining of liE. Next, the content of DNA and the secretion of type Ⅱ collagen and GAG in cartilages were detected quantitatively. Results Under the daily cyclic pressure (0 ～200 kPa, 0.1 Hz), the scaf-fold-chondrocytes complex in the group of 8 hours got the largest volume, smooth, lucidus, and elastic surface, the most queuing chondrocytes, and the highest content of type Ⅱ collagen and GAG (P < 0.01). Conclusions Since chondrecytes are baro-senstive, the metabolism of chondrocytes can be affected by the time of cyclic pressure. Under the effect of 0 ～200 kPa, 0.1Hz, the daily cyclic pressure of 8 hours may be optimal for chondrocytes to multiply and synthesize extracellular matrixes such as type Ⅱ collagen and GAG.     

G蛋白耦联受体激酶结合蛋白1通过调节细胞外调节激酶1/2的活性促进成骨细胞迁移

目的探索G蛋白耦联受体激酶结合蛋白1(GITI)在成骨细胞迁移中的作用,并分析其机理。方法通过Western blot方法检测GIT1蛋白在鼠的成骨细胞内的表达；用免疫荧光染色方法确定：在血小板衍生生长因子(PDGF)不刺激和刺激的条件下,GIT1和细胞外调节激酶1/2(ERK1/2)在成骨细胞内的位置；用共同免疫沉淀的方法测定GIT1和ERK1/2相互结合,并且用免疫荧光双染的方法确定这两种蛋白相互结合的位置；用包含GIT1-RNA发夹结构的腺病毒感染成骨细胞后,用免疫荧光染色方法确定磷酸化ERK1/2(pERK1/2)在成骨细胞内的位置,用划痕愈合法检测在PDGF刺激下的迁移能力。结果在成骨细胞内,PDGF刺激导致了GIT1和ERK1/2的相互结合,并且这种结合发生在成骨细胞的局部粘附内。包含GIT1-RNA发夹结构的腺病毒明显抑制了pERK1/2招募至成骨细胞局部粘附内以及PDGF所刺激的成骨细胞的迁移。结论在PDGF刺激下,GIT1招募pERK1/2至成骨细胞的局部粘附内,从而促进成骨细胞的迁移。     

不同持续时间的周期性压力对构建组织工程软骨的影响

Objective To determine the best duration for exerting the cyclic pressure under which the tissue-engineered cartilage is constructed. Methods Free chondrocytes isolated from rabbit articular cartilage were seeded into polylactic acid-co-glycolic acid(PLGA) scaffolds after expansion in vitro, and ran-domized into 4 groups. In Groups 1 to 3, chondrocytes were cultured under daily cyclic pressure (0 ～ 200 kPa, 0.1Hz) for 4 hours, 8 hours, 12 hours respectively; Group 4 was a control in which no pressure was exerted. In each group, after 2 weeks of culture, the tissue engineered cartilages were observed in vitro and assessed by his-tological staining of liE. Next, the content of DNA and the secretion of type Ⅱ collagen and GAG in cartilages were detected quantitatively. Results Under the daily cyclic pressure (0 ～200 kPa, 0.1 Hz), the scaf-fold-chondrocytes complex in the group of 8 hours got the largest volume, smooth, lucidus, and elastic surface, the most queuing chondrocytes, and the highest content of type Ⅱ collagen and GAG (P < 0.01). Conclusions Since chondrecytes are baro-senstive, the metabolism of chondrocytes can be affected by the time of cyclic pressure. Under the effect of 0 ～200 kPa, 0.1Hz, the daily cyclic pressure of 8 hours may be optimal for chondrocytes to multiply and synthesize extracellular matrixes such as type Ⅱ collagen and GAG.     

不同持续时间的周期性压力对构建组织工程软骨的影响

Objective To determine the best duration for exerting the cyclic pressure under which the tissue-engineered cartilage is constructed. Methods Free chondrocytes isolated from rabbit articular cartilage were seeded into polylactic acid-co-glycolic acid(PLGA) scaffolds after expansion in vitro, and ran-domized into 4 groups. In Groups 1 to 3, chondrocytes were cultured under daily cyclic pressure (0 ～ 200 kPa, 0.1Hz) for 4 hours, 8 hours, 12 hours respectively; Group 4 was a control in which no pressure was exerted. In each group, after 2 weeks of culture, the tissue engineered cartilages were observed in vitro and assessed by his-tological staining of liE. Next, the content of DNA and the secretion of type Ⅱ collagen and GAG in cartilages were detected quantitatively. Results Under the daily cyclic pressure (0 ～200 kPa, 0.1 Hz), the scaf-fold-chondrocytes complex in the group of 8 hours got the largest volume, smooth, lucidus, and elastic surface, the most queuing chondrocytes, and the highest content of type Ⅱ collagen and GAG (P < 0.01). Conclusions Since chondrecytes are baro-senstive, the metabolism of chondrocytes can be affected by the time of cyclic pressure. Under the effect of 0 ～200 kPa, 0.1Hz, the daily cyclic pressure of 8 hours may be optimal for chondrocytes to multiply and synthesize extracellular matrixes such as type Ⅱ collagen and GAG.     

髋关节直接外侧切口在关节置换中的应用

应用髋关节直接外侧切口进行髋关节假体置换１１３例，其中全髋关节置换６４例，双极人工股骨头置换４９例。平均随访时间５．７个月，未见重要神经、血管损伤及早期脱位等并脱位等并发症。作通过尸解发现臀中肌后１／３纤维及其肌腱增厚，其纤维方向与股骨干纵轴一致，并位于股骨轴线后方。该切口最适合在关节置换中应用。     

帕米磷酸钠和降钙素对人工关节磨损微粒引起单核细胞分泌溶骨性因子的影响

目的　探讨帕米磷酸钠和降钙素防治人工关节松动的可能性。方法　采集人体外周血 ,分离单核细胞 ,分组培养。实验分 4组 ,第 1组 :仅单核细胞 ,为对照组 ;第 2组 :单核细胞及微粒 ,为微粒组 ;第 3组 :单核细胞、微粒及帕米磷酸钠 (阿可达 ) ,为阿可达组 ;第 4组 :单核细胞、微粒及降钙素 (密钙息 ) ,为密钙息组。培养 48h后 ,检测细胞上清液中肿瘤坏死因子 (TNF) α、白细胞介素 (IL) 1和IL 6的含量。结果　微粒组的细胞上清液中溶骨性因子含量明显高于对照组 (P <0 .0 1) ;而阿可达组和密钙息组的溶骨性因子含量明显低于微粒组 (P <0 .0 1)。结论　阿可达和密钙息能有效地抑制微粒刺激单核巨噬细胞分泌溶骨性因子 ,从而间接抑制破骨细胞的活性。     

交叉韧带重建手术中可吸收挤压螺钉的应用

目的探讨交叉韧带重建术中可吸收挤压螺钉的使用方法和疗效。方法总结53例交叉韧带重建病例使用可吸收挤压螺钉的情况，在术中、术后并发症、术后康复、膝关节功能状况等方面进行回顾性分析。结果三例出现韧带切割现象。两例股骨侧螺钉拧入后导引针无法拔出。一例挤压螺钉断裂。术后Lysholm评分平均92.4±4．1。结论交叉韧带重建术中使用可吸收挤压螺钉固定的方法固定牢固，术后恢复快，利于早期康复。     

药物对微粒刺激单核细胞溶骨性因子分泌的影响

目的建立人工关节无菌性松动的体外模型，探讨药物防治人工关节松动的可能性。方法(1)体外模型：人体外周血单核细胞分别与骨水泥、聚乙烯和钛合金微粒混合培养，测定上清中溶骨性因子。(2)实验分组：采集志愿献血外周血，分离单核细胞。分骨水泥、聚乙烯和钛合金三组。每组再分4亚组，I亚组：单核细胞；H亚组：单核细胞十微粒；Ⅲ亚组：单核细胞十微粒十帕米膦酸钠；IV亚组：单核细胞十微粒十降钙素。培养48小时，检测上清TNF-α、IL-l和IL-6的含量。结果Ⅱ亚组上清中溶骨因子含量高于I、Ⅲ、IV(p＜0．01)；I、Ⅲ、IV组间差异无统计学意义(P＞0．05)。结论二磷酸盐类药物和降钙素能够抑制微粒对单核巨噬细胞的激活作用，并能直接抑制破骨细胞，因此有望成为防治人工关节无菌性松动的有效药物。     

人工膝关节置换术后夹闭引流管4h对术后出血的影响

目的：探讨人工膝关节置换术（TKA）后引流管夹闭4h和不夹闭对患者术后出血量的影响。方法TKA患者60例均分为A、B两组。A组术后夹闭引流管4h后开放,B组术后不夹闭引流管。比较两组术后引流量、住院时间及术后并发症的发生率。结果A组术后24‐h引流量较B组少[（512.4±253.5）mlvs．（873.4±301.5）ml]（P＜0．05）;A组术后接受输血治疗患者的比例低于B组（46.7％vs．90.0％）（P＜0．05）。在未输血的患者中,A组术后血红蛋白的下降值低于B组[（15.5±7.2）g／Lvs．（21.8±10.3）g／L]（P＜0．05）。两组患者术后发生切口红肿、发热及康复情况差异无统计学意义。结论TKA术后夹闭引流管4h能有效减少患者术后出血量,不增加患者术后感染及功能恢复的风险。     

金属对金属人工全髋关节置换术后早期血清金属离子变化及对免疫功能的影响

目的 探讨金属对金属人工全髋关节置换术后早期血清金属离子的变化及其对免疫功能的影响.方法 将2008年1月至2010年12月期间在南京医科大学第一附属医院接受Durom大直径金属对金属人工全髋关节置换的32例(32髋)患者作为研究对象(Durom组),在同期接受金属对超高分子聚乙烯(Trilogy)人工全髋关节置换的患者中,根据患者年龄、性别、体重指数等选取与Durom组相匹配的32例作为Trilogy组,相同方法选取32位健康志愿者作为对照组.末次随访时,采用ICP-MS法检测患者血清钴、铬离子浓度,并通过检测外周血中血清C3、C4、IgA、IgG、IgM含量评估体液免疫的变化,检测T细胞亚群数量及Th细胞分泌功能评估细胞免疫功能的改变.结果 Durom组和Trilogy组所有患者均完成随访.Durom组平均随访时间为24.9个月(15～34个月),Trilogy组平均随访时间为25.53个月(15～48个月).末次随访时Durom组血清钴离子和铬离子浓度明显升高,分别为Trilogy组的4.33倍和1.95倍,Trilogy组和对照组血清钴离子和铬离子浓度无差异.Durom组与Trilogy组、对照组相比,血清C3、C4、IgA、IgG、IgM无明显差异.Durom组T细胞总数、CD4+和CD8+细胞数明显低于Trilogy组和对照组.对三组血清中的IFN-γ和IL-4进行检测,Durom组的IFN-γ明显高于Trilogy组和对照组,Trilogy组和对照组的IFN-γ无差异;三组的IL-4含量无明显差异.结论 金属对金属人工全髋关节置换术后早期患者血清钴离子、铬离子浓度明显升高,对细胞免疫功能有一定程度的影响.