Structure of rat tail tendon collagen examined by atomic force microscope

The Atomic Force Microscope (AFM) was used to inspect collagen fibrils deposited on mica sheets at different fibrillogenesis times. Collagen was obtained from rat tail tendon fibers. Various fibril forms were observed, together with the characteristic periodic intra-fibril structure (D-bands). The fibril thickness, width, D-band periodicity and depth were measured and the statistical distribution of these parameters at 1, 2, 5, 10 and 15 days of in vitro fibril formation time was calculated. The fibrils showed an increasing size with time, but the band interval measure remained stable. The band depth, after an initial increase, exhibited a relative steadiness. The results indicate that AFM offers, at low resolution, images qualitatively similar to those obtained with electron microscopy, but with less manipulation of the sample. A quantitative evaluation of collagen structural features in the nanometer scale is made possible by AFM.     

心肌胞外间质的比较解剖学研究

借助S-570扫描电镜观察了五种脊椎动物(鱼、青蛙、蛇、鸡、鼠)的心肌胞外间质胶原的形态结构和分布。结果表明，心肌胞外间胶原的结构与分布随着几种动物进化阶段呈现出由简单到复杂、由低级向高级发展的趋势。本为进一步从心脏的超微结构揭示心脏与运动的关系提供了一定的理论基础。     

不同应变水平拉伸对成骨细胞生理功能的影响

采用四点弯曲加载装置对大鼠颅盖骨中分离出的成骨细胞施加周期性拉伸刺激，研究成骨细胞对不同应变水平的拉伸刺激的生理响应。结果表明，500微应变的周期性拉伸刺激促进了成骨细胞的增殖，细胞的^3H－脯氨酸掺入量增加，碱性磷酸酶（ALP）活性和向胞外分泌钙均升高；而1000微应变的周期性拉伸抑制了成骨细胞的增殖、^3H－脯氨酸掺入量、碱性磷酸酶活力和向胞外分泌钙都降低，说明成骨细胞能够分辨不同变水平的力学刺激，500微应变的周期性拉伸刺激有利于细胞的生长和分化，而1000微应变的周期性拉伸对细胞的生理活性有抑制作用。而且细胞在增殖、基质合成和分化、矿化上表现出一致的变化趋势。     

木瓜提取液对大鼠骨组织中ColⅠ和Cath L表达的影响

将50只SD雌性大鼠随机分为5组,连续给木瓜提取液12周后,测定大鼠Ⅰ型胶原(ColⅠ)、组织蛋白酶L(Cath L)、Ⅰ型前胶原氨基端前肽(PINP)、Ⅰ型胶原交联羧基末端肽(ICTP)并行股骨并进行形态学分析.结果表明:木瓜提取液组与雌激素组能显著增加大鼠的骨小梁的数量、降低Cath L的表达,木瓜高剂量组与雌激素组无明显差异(P0.05),这一作用在高剂量组更加明显.木瓜提取液能够降低破骨细胞Cath L的表达来抑制骨基质中ColⅠ的分解,从而起到抗骨质疏松的疗效和骨保护作用.     

Ⅳ型胶原在胃癌中的表达与临床对比分析

为提高胃癌的诊断率,探讨胃液、血清、胃组织中Ⅳ型胶原在胃癌诊断中的意义。121例临床病例被分为3组,即胃癌组50例、胃癌前病变组41例和浅表性胃炎组30例。采用放射免疫法和免疫组化法测定Ⅳ型胶原在各组中的表达。胃癌组胃液中Ⅳ型胶原含量明显高于其余两组(P0.05),胃癌组Ⅳ型胶原的连续性表达明显低于其余两组(P0.01),胃癌组血清Ⅳ型胶原含量高于癌前病变组(P0.05)。Ⅳ型胶原的测定对胃癌的辅助诊断有重要的临床意义。     

1,25双羟维生素D3对人增生性瘢痕成纤维细胞增殖和胶原合成的影响

探讨1,25双羟维生素D3对人增生性瘢痕成纤维细胞增殖和胶原合成的影响.以体外培养的人瘢痕成纤维细胞为实验模型,将1,25双羟维生素D31nmol/L、0.1nmol/L、0.01nmol/L加入细胞培养液中进行干预并与乙醇对照组比较,用乳酸脱氢酶实验检测药物的细胞毒性,并通过MTT实验和羟脯氨酸测定实验分别检测药物对成纤维细胞增殖活性和胶原合成的影响.0.1nmol/L、1nmol/L的1,25双羟维生素D3组对成纤维细胞的生长有抑制作用,与对照组比较差异有显著性(P＜0.01),0.01nmol/L、0.1nmol/L、1nmol/L的1,25双羟维生素D3对瘢痕成纤维细胞胶原合成具有抑制作用(P＜0.01),并呈剂量依赖关系.1,25双羟维生素D3在体外对瘢痕成纤维细胞细胞增殖和胶原合成具有抑制作用.     

银杏叶提取物对大鼠真皮FB增殖及胶原蛋白分泌的影响

原代培养大鼠真皮成纤维细胞（Fibroblast,FB）传代培养后,以不同浓度的银杏叶提取物（EGb761,20、50、100mg／L）对其生长进行干预,四甲基偶氮唑盐（MTT）法检测药物干预后24、48、72h时FB细胞活力变化,碱水解法测定药物干预后3、6d时培养液中羟脯氨酸含量变化,以分别反映EGb761对大鼠真皮FB增殖及胶原蛋白分泌的影响。研究表明EGB761在一定范围内能促进大鼠真皮FB增殖及及胶原蛋白分泌,且呈一定的剂量、时间相关性。     

两种不同基质对体外培养成骨细胞促贴壁生长的影响

目的 比较多聚赖氨酸(PoLy lysine PLL)与鼠尾胶原对成骨细胞体外培养生长的影响。方法 分别采用 PLL和鼠尾胶原铺制培养皿,然后接种成骨细胞。于24 h、48 h、72 h镜下观察细胞形态及占培养皿面积的百分比。 结果 24 h PLL组,细胞贴壁呈均匀分布状,贴壁面积比为(30.6±12.9)％;鼠尾胶原组细胞贴壁呈团状,贴壁面 积比为(42.5±15.7)％,两者比较差异显著(P ＜0.05);48 h PLL组,细胞呈均匀分布生长,贴壁面积比为(43.8± 14.5)％;鼠尾胶原组,细胞团相互连接,生长面积比为(61.3±17.1)％,差异极显著(P ＜0.01);72 h PLL组细胞相 互连接生长面积比为(56.3±15.4)％;鼠尾胶原组,细胞连成片, 生长面积比为(94.4±14.1)％,两者比较差异极 显著(P＜0.01)。结论 对体外培养成骨细胞鼠尾胶原促贴壁生长比PLL更为适宜。     

龟皮胶原蛋白的提取及结构表征(Ⅰ)

采用酶法从乌龟皮中提取胶原蛋白,用聚丙烯酰胺凝胶电泳(SDS-PAGE)方法检测龟皮胶原蛋白多肽产品分子量的分布,并将龟皮胶原蛋白和Ⅰ型胶原蛋白样品进行液相色谱分析(HPLC),比较其氨基酸成分和含量.SDS-PAGE电泳结果分析表明龟皮胶原为Ⅰ型胶原蛋白,分子量约为288ku.龟皮胶原蛋白中,甘氨酸的含量最高,占总量的24.69%;其次是谷氨酸、精氨酸和丙氨酸,分别为9.95%、8.08%和9.31%;天冬氨酸、赖氨酸、丝氨酸、苏氨酸、亮氨酸、苯丙氨酸、异亮氨酸、缬氨酸、酪氨酸等含量在5.58%～1.28%之间;组氨酸和蛋氨酸的含量最低,仅占0.79%和0.09%.由于龟皮胶原蛋白的含量高于Ⅰ型胶原蛋白,且与Ⅰ型胶原蛋白的氨基酸含量差异显著,因此,龟皮胶原蛋白可以作为一种重要的皮胶原进行开发和利用.     

Spatially defined oxygen gradients and vascular endothelial growth factor expression in an engineered 3D cell model

Tissue hypoxia results in rapid angiogenesis in vivo, triggered by angiogenic proteins, including vascular endothelial growth factor (VEGF). Current views of tissue viability are founded on whether ‘deeper-lying’ cells receive sufficient nutrients and oxygen for normal activity and ultimately survival. For intact tissues, levels of such essential nutrients are governed by micro-vascular perfusion. However, there have been few effective quantitatively defined 3D models, which enable testing of the interplay or interdependence of matrix and cell density, and path diffusion on oxygen consumption in vitro. As a result, concepts on cell vulnerability to low oxygen levels, together with the nature of cellular responses are ill defined. The present study has adapted a novel, optical fibre-based system for in situ, real-time oxygen monitoring within three-dimensionally-spiralled cellular collagen constructs, which were then unfurled to enable quantitative, spatial measurements of VEGF production in different parts of the same construct exposed to different oxygen levels. A VEGF response was elicited by cells exposed to low oxygen levels (20 mmHg), primarily within the construct core. Received 3 August 2007; received after revision 24 October 2007; accepted 29 October 2007 An erratum to this article is available at .