内皮细胞衬里人工血管抗血小板沉积的研究

使人工血管体外形成内皮衬里并评价其抗血小板沉积特性。方法 将培养扩增的在人大隐静脉内皮细胞铺植于贫血小板血浆和纤维连接素预衬的聚四氟乙烯人工血管，并体外培养９天，再用含＾１２５Ｉ标记血小板的全血以１００ｍｌ／ｍｉｎ的流速灌注６０分钟。人工血管内壁形态以扫描电镜观察，血小板沉积量以ｒ计数测定。     

内皮细胞衬里人工血管静脉旁路移植10例

为进一步证实内皮细胞衬里人工血管的临床效果,报告临床应用１０例的长期随访。方法,本组１０例布加综合征病人,行腔房转流术６例,肠房转流术２例,肠颈转流术和肠腔转 流术各１例。应用人工血管内径１４ｍｍ,长８－３０ｃｍ,其中针织涤纶２根,编织涤纶７根,带外支持环ＰＴＦＥ１根。     

基因工程内皮细胞种植人工血管的研究进展

自体和非自体的生物血管不能满足临床血管外科的需要。使用理化方法或单纯种植内皮细胞的方法处理人工血管因腔内血栓形成而效果小理想。为了增加人工血管的抗血栓活性，利用基因工程的方法对内皮细胞进行改造，可使其获得稳定表达抗凝血因子的能力。用这种基因工程改造的内皮细胞种植于人工血管，有可能获得较理想的效果。而最终要获得理想的血管移植物，尚有许多需要解决的问题。     

小口径人工血管的研究进展

为提高小口径人工血管的远期通畅率，一方面需要寻找顺应性更好的材料；另一方面则是在现有人工血管内壁种植内皮细胞。这包括单期种植法，二期种植法以及自体静脉碎片快速种植法。用于种植的内皮细胞可来源于自体静脉、人脐静脉和皮下脂肪微血管。通过在人工血管内壁上衬附可吸收的细胞外黏附蛋白、细胞生长刺激因子，利用生物素与抗生物素蛋白的特异结合能力，或改变植入的内皮细胞的带电性，以及在体外对植入的内皮细胞进行流体切应力锻炼等方法，可以提高内皮细胞的黏附力。植入基因工程改造过的内皮细胞，也是提高小口径人工血管通畅率的一种新尝试。     

人工血管内皮化的研究进展

人工血管内皮化的研究进展潘明新综述杨继震何红兵审校作者单位：５１０２８２第一军医大学珠江医院普外科血管病研究室众所周知，人工血管置换病变的静脉和口径小于６ｍｍ的动脉，通畅率十分低下。这与人工血管的内表面难以自发形成抗血栓形成的内皮细胞衬里有关。理论上...     

微血管内皮细胞与人工血管内皮化（综述）

人工血管的应用日趋广泛，如何实现其内皮化是应用的关健。人工血管内皮化的效果决定于三个因素：种植的细胞数、细胞贴壁率、贴壁细胞抗切应力能力。为此，人们进行了内皮细胞获取、培养增殖、微血管内皮细胞获取的研究，尝试了从低密度种植、高密度种植、铺植到离体衬里的一系列内皮化方法，确实了人工血管内皮化的研究的方向：寻求可靠、丰富、方便的细胞来源，建立完善的离体衬里内皮化方法。     

微血管内皮细胞与人工血管内皮化

人工血管的应用日趋广泛,如何实现其内皮化是应用的关键.人工血管内皮化的效果决定于三个因素:种植的细胞数、细胞贴壁率、贴壁细胞抗切应力能力.为此,人们进行了内皮细胞获取、培养增殖、微血管内皮细胞获取的研究,尝试了从低密度种植,高密度种植、铺植(Sodding)到离体衬里(Lining)的一系列内皮化方法,确立了人工血管内皮化的研究的方向:寻求可靠、丰富、方便的细胞来源,建立完善的离体衬里内皮化方法.     

人大隐静脉内皮细胞种植人工血管的实验研究

目的　探讨人自体静脉内皮细胞移植到人工血管上的可行性。方法　将人大隐静脉内皮细胞在体外扩增培养１３ ．０８ ±１ ．２４ 天, 将扩增培养的内皮细胞衬里用于纤维蛋白胶预衬的膨体聚四氟乙烯（ｅｘｐａｎｄｅｄ ｐｏｌｙｔｅｔｒａｆｌｕｏｒｅｔｈｙｌｅｎｅ ｐａｔｃｈ 〔ＧｏｒｅＴｅｘ〕Ｒ, ｅＰＴＦＥ） 人工血管继续培养９ 天。结果　所培养的细胞为二倍体细胞,纯度为９９ ％ 。原代及传代细胞培养上清液中６酮前列环素（６ｋｅｔｏＰＧＦ１α） 和Ｖｏｎ Ｗｉｌｌｅｂｒａｎｄ 因子（ｖ ＷＦ） 含量差异无显著性意义（ Ｐ＞ ０ ．０５） 。细胞种植后第９ 天人工血管腔面见一层均匀的基质,其表面有一层连续的内皮细胞单层,内皮细胞排列紧密,呈梭形,形态饱满。结论　人自体大隐静脉内皮细胞可有效地移植到人工血管,为人自体细胞内皮化人工血管应用于临床奠定了理论基础。     

体外扩增恒河猴自体内皮细胞人工血管移植研究（二）

目的：为自体内皮细胞移植的临床应用奠定理论基础。方法：将１５只恒河猴随机分为两组。实验组１０例，用衬里有自体静脉内皮细胞的人工血管置换髂总动脉；对照组５例，用未衬里内皮细胞塞外，其余８条人工血管术后四周均保持通畅，内膜下组织碍度为８０±１２μｍ；对照组５条人工血管全部阻塞。结论自体内皮移植可有效地南昌市人工血管的通畅率。     

ePTFE人工血管材料内衬自体骨髓内皮细胞的实验研究

目的:探讨自体骨髓内皮细胞衬里对ePTFE人工血管材料通畅性的影响。方法:24只犬随机分入实验组(12只)和对照组(12只)。实验组取犬自体骨髓,提取单核细胞种植ePTFE人工血管,行左髂动脉人工血管置换术;对照组采用普通ePTFE人工血管。分别于术后1月、3月行彩色多普勒超声观察其通畅率,并行组织染色及扫描电镜观察,比较人工血管表面内皮化情况。结果:术后1月时实验组人工血管通畅率及内膜厚度同对照组均无明显差别;但术后3月时实验组通畅率高于对照组,内膜厚度明显小于对照组(P0.05)。结论:自体骨髓内皮细胞衬里技术可明显提高ePTFE人工血管的短期通畅率,但其对远期通畅率的影响尚有待进一步观察。     

Growth Regulation of Endothelial Cells by Tissue Fragments: Analysis on Mechanism of Rapid Endothelialization of Tissue-Seeded Vascular Prosthesis in a Three-Dimensional Culture System

Abstract: Rapid endothelialization of the inner surface was reported in an autologous tissue-seeded vascular prosthesis. We applied a three-dimensional in vitro culture system to elucidate the precise mechanism of rapid endothelial coverage of a tissue-seeded vascular prosthesis. Human venous, omental, adipose and striated muscle tissue fragments were harvested from surgical specimens. They were embedded in collagen gel, and 2.0 times 10⁵ bovine aortic endothelial cells (BAECs) were seeded on the gel. The number of BAECs was counted on Days 2 and 7. Growth rate of BAECs was facilitated on the collagen gel with omental and striated muscle tissue fragments (p < 0.05). Factor VIII-negative spindle cells migrated around tissue fragments, especially around the omental and striated muscle tissue fragments. Rapid endothelialization of a tissue-seeded vascular prosthesis may result from facilitation of EC proliferation by viable tissue fragments and migrated cells. These results confirm tissue fragments regulate EC growth, and are useful as bioengineering tools     

血管内皮生长因子165基因重组腺病毒载体的构建及转染内皮祖细胞的实验研究

目的：利用细菌内同源重组法构建含血管内皮生长因子（VEGF）165基因的重组腺病毒，并观察其在内皮祖细胞（EPCs）中的表达，为进一步研究其在慢性肾脏病中的作用奠定基础。方法：用限制性内切酶XbaⅠ＋HindⅢ从质粒载体中切出VEGFl65基因片段，与KpnⅠ＋HindⅢ双酶切的pAdTrack-CMV形成转移质粒pAdTrack-CMV-VEGFl65，PmeI酶切线性化后与腺病毒基因组质粒pAdEasy-1共转化大肠杆菌BJ5183，由人胚肾293细胞包装成Ad-VEGFl65，PCR和westem blot法鉴定其表达。贴壁法体外培养获得大鼠骨髓来源的EPCs，Ad-VEGFl65基因体外转染EPCs，检测转染后目的基因的表达情况。结果：利用CSCI，法由pAdTrack-VEGFl65和pAdEasy-1共转化BJ5183感受态茵，可获得阳性重组体细菌克隆。PCR检测表明重组腺病毒已含有目的基因，病毒滴度1．4×10^10pfu／ml。Ad-VEGFl65转染培养第6天的EPCs后24h开始，培养上清中VEGF蛋白表达较对照组和转染前增加。结论：成功制备的重组体腺病毒Ad-VEGFl65转染体外培养的EPCs后，在体外能有效高表达目的基因产物，为今后VEGF在肾脏病中的深入研究奠定了基础。     

Experience with the Hemasite Vascular Prosthesis

A 3-year experience with the no-needle vascular prosthesis Hemasite, implanted in 10 patients who underwent hemodialysis and have a long history of multiple vascular access failures, is described. During 182 months of follow-up study, 30 thromboses occurred, while nine of 10 patients did not receive any antiplatelet aggregant treatment. Hemasite was declotted 12 times with a local infusion of urokinase and 12 more times by thrombectomy. A surgical procedure was performed only in the other cases, and the rate of surgical intervention fell from 0.18 interventions per patient per month before Hemasite implantation to 0.027 after implantation.     

血管内皮细胞在动脉粥样硬化中的损伤机理

目的探讨血管内皮细胞在动脉粥样硬化形成过程中的损伤机理。方法复习相关文献。对有关进展进行总结分析。结果各种致病因素造成细胞因子生成增多，激活了控制细胞粘附分子表达的核因子-κB，进而使细胞粘附分子表达增强，促进单核细胞一血管内皮细胞为主的粘附形成增多，释放大量炎性介质(氧自由基及蛋白酶等)，不仅直接损伤血管内皮细胞，并可以通过免疫机理使大量单核细胞及中性粒细胞与血管内皮细胞粘附结合，进一步损伤血管内皮细胞。同时，受损的血管内皮细胞由于膜结构的改变引起抗动脉抗体的产生以致补体系统被激活而加重了血管内皮细胞的损伤，也促进了动脉粥样硬化的发生和发展。结论正确认识血管内皮细胞在动脉粥样硬化发生、发展过程中受损机理，对于动脉粥样硬化的防治具有重要意义。     

体外扩增兔内皮细胞种植异体血管移植的实验研究

目的探讨自体静脉内皮细胞衬里异体血管移植的可行性。方法将体外扩增培养的兔静脉内皮细胞铺植于经处理的异体血管,体外培养9d,然后行异体静脉原位移植术。结果经传代培养收获的内皮细胞总数为(9.47±0.35)×106个。细胞种植3h后异体血管腔面可见有内皮细胞附着,但未形成完整内膜;9d时腔面被一层连续的单层内皮细胞覆盖,细胞形态饱满,平行排列,呈梭形。术后8周所有移植段血管保持畅通。结论预衬自体内皮细胞的异体血管可保持长期的通畅性。     

A New Device for Endothelial Cell Seeding of a Small-Caliber Vascular Prosthesis

Abstract: A device permitting homogeneous endothelial cell seeding of a small-calíber arteríal prosthesís has been developed. The prosthesis is maintained firmly attached to a rotative scaffolding device. This device is activated by an electrical motor at constant and adjustable speed. The whole system is maintained at 37°C in a cell culture incubator. The 4 mm internal diameter polytetrafluoroethylene (PTFE) prosthesis was coated with biological glue and seeded with human saphenous vein endothelial cells obtained by mechanical detachment. Cell seeding density was 2.10⁴ cells/cm² (Group A, n = 6) or 10⁵ cells/cm² (Group B, n = 6). Rotation speed was 8 revolutions per hour (rph) during 90 min. Analysis of the homogeneity of cell seeding was permitted by cell counts on five different segments of the prosthesis. Each longitudinal segment was analyzed at three different subsegments of the circumference. The average adhesion was 43 ± 4% in Group A and 38%± in Group B of seeded cells. No difference could be observed between the different segments and subsegments. In the two groups, cells were spread, and in Group B, a complete endothelial cell layer was obtained on the graft surface. This study permits validation of the device to allow homogeneous cell seeding in an arterial prosthesis.     

Galectin-3对外周血内皮祖细胞源性血管内皮细胞增殖能力的影响

目的观察重组人半乳糖凝集素-3(Galectin-3)对外周血内皮祖细胞源性血管内皮细胞增殖能力的影响。方法提取人外周血内皮祖细胞,将其定向诱导为成熟血管内皮细胞,加入不同终浓度的Galectin-3进行培养,观察不同浓度Galectin-3对外周血内皮祖细胞源性内皮细胞增殖能力的影响。结果 0.1、1.0、2.5、5.0和10.0μg/ml浓度组外周血内皮祖细胞源性内皮细胞的增殖能力均高于0μg/ml组,其中0.1,1.0及2.5μg/ml浓度组与0μg/ml组比较差异无统计学意义(P>0.05),而5.0及10.0μg/ml组细胞增殖能力明显高于0、0.1、1.0及2.5μg/ml组,差异有统计学意义(P<0.05),10.0μg/ml浓度组细胞增殖能力又明显高于5.0μg/ml浓度组,差异有统计学意义(P<0.05)。结论 Galectin-3可促进体外培养的外周血内皮祖细胞源性血管内皮细胞的增殖能力。     

In Vitro and In Vivo Evaluation of a Small Caliber Vascular Prosthesis Fixed with a Polyepoxy Compound

Abstract: A small caliber vascular prosthesis obtained from an ovine internal thoracic artery (3. 8–4. 5 mm ID) fixed with a polyepoxy compound and treated with heparin has been evaluated. Cytocompatibility was evaluated in vitro using human endothelial cells (HEC). HEC were obtained from human saphenous vein and cultivated in culture medium supplemented with 25% human serum. Graft segments were rinsed using a standard protocol proposed by the manufacturer. Tissue reaction was tested on a rabbit model of subcutaneous implantation. The patency rate and healing patterns were evaluated comparatively with polytetrafluorethylene (PTFE) 4 mm ID prosthesis in a canine model of carotid interposition. Cytocompatibility assay showed that there was low adhesion on vascular grafts (20 ± 2% of endothelial cells seeded) and no growth of HEC on the graft surface. The graft patency rate was 55% in both groups, and actuarial freedom from occlusion was not different at 3 months (37. 7 ± 15% in Denacol–fixed grafts versus 38. 1 ± 14% in PTFE). Histological studies on the biological grafts shows a frequent neointimal hyperplasia at the anastomosis (5/12), a lack of endothelial cells lining the graft surface, a good preservation of the media, and a moderate inflammatory response in the adventicia. The Denacolfixed graft has presented excellent surgical properties and preservation of the histological structure. Nevertheless, the patency rate was not improved when compared with the PTFE control graft.     

猪肾小球内皮细胞与系膜细胞的发育过程及相互关系

目的:明确中国实验用小型猪肾小球内皮细胞与系膜细胞的发育过程及相互关系。方法:采集不同时间点(胚胎28～112d及出生后1d、7d、14d、21d)中国实验用小型猪肾组织,应用免疫荧光技术检测胚肾发育不同阶段(帽状间充质、肾小囊体、逗号形体、"S"形体、毛细血管袢期肾小球及成熟肾小球)内皮细胞标志物CD31与系膜细胞标志物α-SMA的表达情况。结果:内皮细胞标志物CD31在胚肾早期呈散在性分布,继而围绕肾小囊体和逗号形体呈"环抱"状分布,然后进入"S"形体下端的血管裂隙内聚集形成无管腔的"前毛细血管束",最后表达于成熟肾小球毛细血管内皮细胞。系膜细胞标志物α-SMA在早期胚肾、肾小囊体和逗号形体阶段均无表达;"S"形体早期分布于"S"形体周围,后期进入"S"形体下端的血管裂隙;毛细血管袢期聚集在肾小球血管极根部;随着肾小球发育逐渐向肾小球内延伸,最终表达于成熟肾小球系膜区。CD31与α-SMA双重染色的结果显示,在毛细血管袢期CD31阳性的内皮细胞聚集形成无管腔的前毛细血管束,而α-SMA阳性的系膜细胞聚集在肾小球血管极的根部;随着肾小球发育,α-SMA阳性的系膜细胞逐渐由血管极根部向肾小球内迁移,同时CD31阳性的内皮细胞逐渐形成带有管腔的毛细血管丛。结论:中国实验用小型猪肾小球内皮细胞的发育开始于后肾间充质阶段,系膜细胞的发育开始于"S"形体阶段,即肾小球系膜细胞发育在内皮细胞之后;在肾小球血管丛形成过程中,内皮细胞与系膜细胞间可能存在重要的相互作用。