应用APOFIX治疗齿状突骨折并寰枢椎脱位围术期及康复护理

枢椎齿突骨折是累及寰枢椎区域稳定性的严重损伤,尤其是合并脊髓损伤的患者,其发生率约占脊椎损伤患者的10%.因其解剖结构特殊,此类骨折不愈合的发生率较高,常可导致急性或延迟性颈部脊髓压迫并危及生命[1].2002年1月以来,我科采用椎板夹内固定系统(APOFIX)治疗齿状突骨折并寰枢椎脱位40例,在为齿状突骨折患者提供了坚强的内固定的基础上配合一系列行之有效的护理.术后效果好,恢复快,现将护理体会报告如下.     

人股骨头坏死微观结构及成骨、破骨细胞活性的区域性分布特征

 目的 比较人股骨头坏死标本不同区域的骨微观结构及成、破骨细胞活性。方法 收集2011年3月至2013年5月行全髋关节置换的非创伤性股骨头坏死患者术后的股骨头标本10例（Ficat Ⅳ期）,男6例,女4例;年龄40～57岁,平均47.7岁。Micro-CT扫描后,根据影像学识别骨质密度不同,将每个标本分为软骨下骨区、坏死区、硬化区、健康区,通过病理学检测、纳米压痕、实时荧光定量PCR、免疫组化染色等方法对不同区域的骨微观结构、微观力学性能及成骨、破骨细胞活性进行比较。结果 Micro-CT结果显示,股骨头坏死标本软骨下骨区及坏死区的骨小梁连续性破坏;硬化区的骨小梁数目增多,间隙变窄;正常区域骨小梁结构完整,厚度分布均匀。软骨下骨区、坏死区、硬化区和健康区骨小梁的弹性模量分别为（13.808±4.22） GPa、（13.999±3.816） GPa、（17.266±3.533） GPa和（11.927±1.743） GPa;硬度分别为（0.425±0.173） GPa、（0.331±0.173） GPa、（0.661±0.208） GPa和（0.423±0.088） GPa。抗酒石酸酸性磷酸酶（Trap）染色结果显示,软骨下骨区和坏死区可见Trap染色阳性细胞,硬化区及健康区未见Trap染色阳性细胞。免疫组化染色结果显示,骨形成相关因子Runx2和BMP2在硬化区及健康区表达高于其他区域;骨吸收相关因子RANK和RANKL在软骨下骨区及坏死区表达高于其他区域。结论 股骨头坏死塌陷过程中,骨微观结构发生明显改变,而坏死区骨小梁微观力学强度较健康区无显著降低。股骨头坏死标本中软骨下骨区及坏死区破骨细胞活性增强,硬化区成骨细胞活性增强。     

多媒体数字化系统在细胞生物技术研究中的应用

目的 在细胞生物学研究中，利用先进的多媒体技术对传统的图像资料采集方法进行改革，提高资料采集、储存和保管的质量。方法 实验于2002-02／2004-05在解放军总医院全军骨科研究所完成。利用计算机、数码照相机、扫描仪、刻录仪、计算机图像采集系统、彩色激光打印机及各种软件收集和再现各方面的信息资料，按研究目的制作不同数据库及多媒体文件，建立影像资料库，简化资料的处理过程。结果 2002-02起建立细胞生物学多媒体数字化系统，截至2004-05，累计采集图像16万张，相关资料容量60G。结论 此套系统效率高，成本低，技术易掌握，资料分类、保存、查找容易，便于医学科研工作。     

人脱细胞软骨细胞外基质对异种软骨种子细胞的影响

目的观察人脱细胞软骨细胞外基质(hACAM)对异种兔软骨种子细胞增殖和表型的影响。方法将差速梯度离心法制作的人脱细胞软骨细胞外基质,配制成0.5%的浆料,分别铺于6孔细胞培养板和96孔细胞培养板,形成1 mm厚的薄膜,以此培养板为实验组。空白对照组培养板仅使用单纯培养液培养。在培养板上培养兔关节软骨细胞。通过hochest33258染色验证人软骨细胞外基质脱细胞完全与否;分别在第5、15天两个时间点,通过倒置显微镜、甲苯胺蓝染色观察两种培养方法的兔软骨细胞的生长形态、细胞表型和增殖情况,用CCK-8细胞增殖实验比较两种培养方法在第1、3、7、10天的增殖情况。结果通过hochest33258染色发现差速梯度离心的人软骨细胞外基质脱细胞完全。通过倒置显微镜观察第5和15天的实验组细胞增殖情况优于空白对照组,甲苯胺蓝染色的结果也证明这个观点;CCK-8细胞增殖试验显示第7天hACAM组在细胞增殖方面明显地优于单纯培养液的对照组(P=0.0298),hACAM组的软骨细胞与空白组的软骨细胞在第10天的增殖情况相比没有统计学差异。结论 hACAM免疫原性低,无细胞毒性,能够很好地促进异种软骨细胞的增殖。     

低浓度唑来膦酸对成骨细胞和破骨细胞影响的体外实验

目的：观察低浓度(10-6 mol/L)唑来膦酸(zoledronate acid,ZA)对体外大鼠破骨细胞及成骨细胞的影响。方法体外分别培养大鼠来源的成骨细胞和破骨细胞,将两种细胞各分为两组：空白对照组及低浓度(10-6 mol/L)ZA组。应用抗酒石酸酸性磷酸酶染色、图像分析计算骨吸收陷窝面积,检测破骨细胞形态及骨吸收情况。碱性磷酸酶(alkaline phosphatase, ALP)染色、四甲基偶氮唑盐比色法了解成骨细胞的形态及增殖情况。结果培养1周后破骨细胞具有典型的形态特征,并在骨片上形成了吸收陷窝;ZA组与对照组相比,破骨细胞数量及生成吸收陷窝的数目和面积减少,差异有统计学意义(P＜0.05)。成骨细胞有典型的梭形、ALP染色阳性特征,培养至第7天ZA组成骨细胞光吸收值(3.37±0.11)高于对照组(2.87±0.12),差异有统计学意义(P＜0.05)。结论低浓度(10-6 mol/L)的ZA能够抑制破骨细胞的增殖和活性,促进成骨细胞的增殖,选择恰当给药方式和剂量能够在抑制破骨的同时促进成骨。     

生物蛋白胶作为骨髓间充质干细胞移植载体的体外相容性研究

[目的]构建骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)-生物蛋白胶复合体评价其生物相容性,探讨其作为种子细胞载体的可行性。[方法]本实验分为两组,实验组5×107骨髓间充质干细胞(BMSCs)与生物蛋白胶相混制备MSC-生物蛋白胶复合体,空白组骨髓间充质干细胞(BMSCs)。通过Dead/live染色观察两组的细胞形态及21 d内细胞的存活情况,同时分别在1、3、7、14 d各取上清进行Elisa检测并计算其分泌释放NGF和BDNF的浓度。[结果]实验组培养3 d后光镜下观察呈多个突起,细胞形态良好,空白组呈菱形和多角形;Dead/live染色结果提示体外培养1 d,两组细胞的存活率都在90%,比较无差异,无统计学意义(P﹥0.05),7、14、21 d两组细胞的存活率有差异,具统计学意义(P﹤0.05),存活率在60%以上;Elisa检测结果显示两组中NGF在1 d时有差异,具统计学意义(P﹤0.05),3、7、14 d时NGF和BDNF的含量比较无统计学意义(P﹥0.05),结果显示生物蛋白胶对MSC分泌的细胞因子NGF和BDNF的释放没有明显影响。[结论]生物蛋白胶是一种理想的可用于临床的细胞载体,具有良好的生物相容性,BMSCs-生物蛋白胶复合体能持续稳定的释放神经生长因子BDNF和NGF。     

化学去细胞异体神经研究进展

周围神经损伤是引起肢体残疾的重要原因。寻找有效替代自体神经的移植物是国内外研究热点。前期研究显示CEANA具有进一步研究应用的可行性。本综述将从CEANA基础研究和临床研究这两方面对该领域进行简要概述。     

生物反应器中的力学刺激促进组织工程软骨再生

BACKGROUND: Cartilage tissue engineering has been widely used to achieve cartilage regeneration in vitro and repair cartilage defects. Tissue-engineered cartilage mainly consists of chondrocytes, cartilage scaffold and in vitro environment. OBJECTIVE: To mimic the environment of articular cartilage development in vivo, in order to increase the bionic features of tissue-engineered cartilage scaffold and effectiveness of cartilage repair. METHODS: Knee joint chondrocytes were isolated from New Zealand white rabbits, 2 months old, and expanded in vitro. The chondrocytes at passage 2 were seeded onto a scaffold of articular cartilage extracellular matrix in the concentration of 1×106/L to prepare cell-scaffold composites. Cell-scaffold composites were cultivated in an Instron bioreactor with mechanical compression (1 Hz, 3 hours per day, 10% compression) as experimental group for 7, 14, 24, 28 days or cultured statically for 1 day as control group. RESULTS AND CONCLUSION: Morphological observations demonstrated that the thickness, elastic modulus and maximum load of the composite in the experimental group were significantly higher than those in the control group, which were positively related to time (P < 0.05). Histological staining showed the proliferation of chondrocytes, formation of cartilage lacuna and synthesis of proteoglycan in the experimental group through hematoxylin-eosin staining and safranin-O staining, which were increased gradually with mechanical stimulation time. These results were consistent with the findings of proteoglycan kit. Real-time quantitative PCR revealed that mRNA expressions of collagen type I and collagen type II were significantly higher in the experimental group than the control group (P < 0.05). The experimental group showed the highest mRNA expression of collagen type I and collagen type II at 21 and 28 days of mechanical stimulation, respectively (P < 0.05). With the mechanical stimulation of bioreactor, the cell-scaffold composite can produce more extracellular matrix, such as collagen and proteoglycan, strengthen the mechanical properties to be more coincident with the in vivo environment of cartilage development, and increase the bionic features. With the progress of tissue engineering, the clinical bioregeneration of damaged cartilage will be achieved.   中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

AZ31镁合金材料植入物在兔股骨髁内的降解：Micro-CT评价

BACKGROUND: Magnesium can be degraded voluntarily in vivo, so a second surgery is avoided. However, its alloys have not been widely used in the clinical orthopedics because there is a lack of accurate and reliable methods to assess its degradation in vivo. OBJECTIVE: To explore the degradation of micro-arc-oxidized AZ31 magnesium alloy in the femoral condyle of rabbits based on micro-CT images and relative data. METHODS: Forty micro-arc-oxidized AZ31 magnesium alloys were implanted into the right femoral condyle of 40 New Zealand rabbits. Then 10 right femoral condyles were removed at 5, 10, 15 and 20 weeks after surgery, respectively, to quantitatively analyze and evaluate the degradation of AZ31 magnesium alloys by micro-CT images and relative data. RESULTS AND CONCLUSION: The surface of AZ31 alloys was corroded progressively with dark color and distorted appearance at 5-20 weeks post implantation. Micro-CT images showed that in the first 5 weeks, the degradation was inactive, and at the 10th week, it turned active; at the 15th week, the corrosion pits were obviously increased in number, and the corrosion area and corrosion speed were enlarged and fastened, respectively. Up to the 20th week, the alloy surfaces were full of corrosion pits besides roughness and discontinuity. Relevant data analysis showed that the volume fraction of magnesium alloy was 98.6%, 97.1% and 86.4% at the 5th, 10th and 20th weeks after implantation, respectively, and it had a significant decrease from the 10th to 15th week and from the 15th to 20th week (P < 0.05). Within 15-20 weeks, the volume fraction of magnesium alloy was decreased by 6.5% that was the maximum volume reduction per unit cycle. With the progress of corrosion, the surface continuously became rough and vague, and its surface area was enlarged; the ratio of surface area to volume continuously increased, and there was a significant difference at 15 and 20 weeks (P < 0.05). Because of the increasing number of corrosion pits, the cross-sectional radius decreased, which was reflected by the trabecular thickness decreasing from 1.00 to 0.87 mm. From the view of the slope of curve, the trabecular thickness decreased most rapidly at 10-15 weeks. The mineral density of magnesium alloy continuously decreased from 649.302 to 356.445 mg/cm3 during the whole experiment period (P < 0.05). In addition, the micro-CT image density decreased from 679.710 to 644.947 mg/cm3, but there was no significant difference. To conclude, the degradation speed is peaked at 10-20 weeks after implantation, and the content of magnesium alloys decrease with degradation, but the magnesium density has no significant change. 中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

组织工程化人工神经的研究与进展

自体神经移植是治疗周围神经缺损的标准方法,但会造成供区神经损伤.随着生物工程技术的发展,目前已应用组织工程技术构建了人工神经导管,为修复长段周围神经缺损提供了新的方法和思路.