尿道细胞外基质修复尿道缺损的研究

目的 寻求一种较理想的尿道修复材料。方法 20只家兔尿道制成尿道细胞外基质(ECM)；另40只随机分3组：尿道ECM移植组(实验组)，对照组Ⅰ及对照组Ⅱ。实验组切除尿道1．O～1．5cm后用尿道ECM修复并于术后10d、3周、6周及24周行组织再生情况观察；另于术后10周、24周各取4只行膀胱尿道造影；24周时实验组及对照组Ⅰ各取4只行尿流动力学检测；24周实验组取4只行尿道镜检查。结果 缺损修复术后10d，基质中见单层上皮细胞且有血管长入ECM；3周时尿道ECM管腔已完全被上皮细胞覆盖；6周时可见平滑肌细胞再生，炎性细胞消失；24周后其组织结构与正常组织相比差异无统计学意义。膀胱尿道造影无尿液外渗，无梗阻及结石形成。尿流动力学检测结果实验组与对照组Ⅰ差异无统计学意义；尿道镜检查证实尿道黏膜完整光滑，尿道内径及其形态正常。结论 尿道细胞外基质是一种理想的尿道修复材料。     

血管细胞外基质和尿道细胞外基质修复兔尿道缺损的比较研究

目的　寻求较理想的尿道修复材料。　方法　切取 10只家兔的腹主动脉和尿道各 3cm ,制备成血管细胞外基质 (VECM)和尿道细胞外基质 (UECM)。另 2 0只分别切除尿道 2 .5cm并随机分为VECM修复组和UECM修复组。两组均于修复术后 10d、3周、6周及 2 4周行组织再生情况研究 ;另于术后 10周、2 4周各取 2只行膀胱尿道造影 ;2 4周两组各取 2只行尿动力学检测和尿道镜检查。　结果　制备的VECM和UECM均为白色透明状 ,但VECM较UECM弹性和机械强度好。缺损修复术后10d ,基质中见单层上皮细胞且有血管长入ECM ,基质和受体尿道连接处有炎性细胞浸润 ;3周时基质管腔已完全被上皮细胞覆盖 ;6周时可见平滑肌细胞再生 ,炎性细胞消失 ;2 4周后其组织结构与正常尿道组织结构一致。VECM修复组和UECM修复组相比其组织再生过程无差异。尿动力学检测VECM修复组和UECM修复组的膀胱容量分别为 (30 .2± 1.6 )ml和 (32 .1± 1.4 )ml、尿道最高压分别为(15 .2 7± 1.36 )mmHg和 (14 .6 8± 1.6 5 )mmHg、尿道最低压分别为 (12 .4 9± 1.2 3)mmHg和 (11.96±0 .98)mmHg ,差异无统计学意义 (P >0 .0 5 )。膀胱尿道造影可见尿道壁完整光滑通畅 ,不能分辨移植区与正常组织 ,无尿液外渗 ,无梗阻及结石形成 ;尿道镜检查证实VECM修复组和U     

异体真皮细胞外基质重建尿道的实验和临床研究

目的：寻求理想的尿道修复材料。方法: 2只犬真皮脱细胞处理后制成真皮细胞外基质框架;18只犬分为2组;异体真皮细胞外基质（ECM）移植组（实验组）15只,异体真皮移植组（对照组）3只,全麻下将尿道中段切除5cm,将同样长度真皮ECM或异体真皮缝成直径0．3cm管状替代缺损尿道,8F硅塑管留置4周,实验组分别于术后1,3,6,12,24周每次3只取材行光镜,电镜及免疫组化检查。结果：组织学显示：1周时移植物边缘腔内面有尿上皮,管壁可见少量肌束,6周时移植物区尿路上皮达4－5层,肌层分布均匀,24周后移植物已无法与宿主尿道辨别,对照组术后8－10d移植物坏死出现尿瘘,在实验基础上用于临床2例均获成功。结论ECM为理想的尿道修复材料。     

尿道细胞外基质在兔尿道重建中的应用

目的 评价尿道细胞外基质作为一种生物材料重建尿道的效果. 方法 切取新西兰兔的尿道制备尿道细胞外基质.手术切除实验组1～1.5 cm的尿道片段后用细胞外基质修复缺损,采用ELISA法检测术前、术后12、24及48 h血清TNFα的水平,评估术后兔的免疫反应状态.术后10d和3、6、24周取修复段尿道,行组织学观察并做尿道造影、尿道镜及尿流动力学检查. 结果 术后实验组血清TNFα水平较对照组略有升高,但无统计学意义(P>0.05).术后10 d,上皮细胞开始从边缘向细胞外基质移行并出现新生小血管;3周上皮细胞覆盖细胞外基质的整个管腔;6周出现排列不规则的平滑肌纤维;24周平滑肌数量明显增多,成束状排列.尿道造影、尿道镜及尿流动力学检测检查显示,尿道基质管壁光滑,排尿通畅. 结论 尿道细胞外基质是一种安全有效的尿道重建材料.     

尿道细胞外基质的研制

目的 探讨尿道细胞外基质(ECM)的制备方法。方法 采用4因素3水平，9次实验的正交设计[L9(3^4)]。切取37只兔的尿道，27条按正交设计随机分为9组行尿道脱细胞处理，试验重复3次。采用HE染色及计算机图像分析对尿道基质进行残余细胞成分计数，统计分析获得最佳方法：0．4％胰蛋白酶、l％甲醛加0．2％戊二醛、40U／ml DNA酶(A3B2C3方案)。另l0条兔尿道制备成尿道ECM并用于同种异体尿道缺损修复实验；分别于修复术后l0d，3、6及24周取材观察缺损修复处组织再生情况。结果 各组脱细胞后残余细胞成分量均不相同，第7、9组未发现细胞残余成分；制备的尿道ECM经扫描电镜分析未发观细胞残片。缺损修复实验术后10d，基质中见单层上皮细胞，且有血管长入ECM中，但管径较小，基质和受体尿道连接处有炎性细胞浸润；3周时尿道ECM管腔已完全被上皮细胞覆盖；6周时可见平滑肌细胞再生，炎性细胞消失。24周后基本结构近似正常。结论 制备尿道细胞外基质中3个关键条件的最佳水平为A3B2C3，即在尿道脱细胞处理过程中采用0．4％胰蛋白酶、1％甲醛加0．2％戊二醛、40U／ml DNA酶。     

应用异体脱细胞尿道基质修复尿道缺损

目的探讨应用同种脱细胞尿道基质修复尿道缺损的可行性。方法将14只雄性新西兰兔分为两组，切除实验组长约1.0～1．5cm的尿道，用相应长度脱细胞尿道基质修复；对照组行假手术。术后行尿道造影并取尿道标本作病理检查。结果12只实验兔的脱细胞基质移植物没有移位。除2例狭窄、2例尿瘘外，其余满意效果。病理检测示，术后3周尿道管腔上皮化，6个月基质中平滑肌及血管再生明显。结论同种脱细胞尿道基质材料可以修复兔尿道部分缺损。     

脱细胞胆管细胞外基质替代胆总管缺损的研究

目的 探讨脱细胞胆管细胞外基质（ECM）的制备方法及其替代胆总管缺损的可行性。方法 采用4因素3水平9次实验的正交设计[L9（3^4）]，切取活体犬的肝外胆管39段，每段长1cm，其中27段按正交设计随机分为9组行脱细胞处理，试验重复3次。采用苏木素．伊红（HE）染色及计算机图像分析对ECM进行残余细胞成分计数，进行极差分析及方差分析，获得制备ECM的最佳方案。另12段胆管按最佳方案制备成ECM并用于同种异体胆总管缺损修复实验。分别于术后2、4、12及24周取材观察缺损修复处组织再生情况。结果脱细胞后第7、9组未发现残余细胞，ECM富含胶原，第9组胶原呈立体交叉交联。移植术后2周，ECM内膜面见少量单层上皮细胞，有血管长入其中，ECM中有少量成纤维和平滑肌细胞，吻合口处有较多炎性细胞浸润；4周ECM管腔被上皮细胞覆盖，排列整齐，ECM中有较多成纤维和少量平滑肌细胞，吻合口有少量炎性细胞浸润；12周吻合痕迹消失，ECM中有较多成纤维、平滑肌细胞和少量纤维细胞；24周ECM中有以平滑肌细胞和纤维细胞为主，少量成纤维细胞，结构近似正常。结论制备ECM中3个关键条件的最佳水平为A383C3，即在胆管脱细胞处理过程中采用0．4％胰蛋白酶、1．0％叠氮二苯基磷（DPPA）、40U／ml DNA酶。所得ECM应用于犬同种异体胆总管端端吻合修补，效果满意。     

脱细胞尿道及其海绵体基质制备的实验研究

目的探索脱细胞尿道及其海绵体基质的制备方法.方法取健康壮年兔完整尿道及其海绵体组织,以Triton-X 100与NH3H2O联合提取法进行脱细胞处理.标本做HE染色,组织学观察分析脱细胞效果.结果脱细胞处理11天后,成功获得脱细胞及其海绵体基质,所得基质外观良好.HE染色观察无细胞存在,弹力纤维排列规整,间隙较大,结构无破坏.结论利用Triton-X 100与NH3H2O联合提取法可成功制备完整无细胞尿道及其海绵体基质,为尿道再造修复提供崭新思路.     

异体颗粒状脱细胞真皮基质与自体刃厚皮复合移植修复大鼠皮肤缺损创面效果观察

目的 观察异体颗粒状脱细胞真皮基质(PADM)与自体刃厚皮复合移植修复大鼠皮肤缺损创面的效果.方法 采用随机数字表法将12只SD大鼠分为实验组和对照组,每组6只.于2组大鼠背部制作全层皮肤缺损创面,实验组创面复合移植SD大鼠异体PADM(扩张比10:5)及厚度0.20 mm的自体刃厚皮,对照组创面仅移植厚度0.20 mm自体刃厚皮.术后2周起打开敷料观察大鼠创面愈合情况.术后2、3、4、6、8、12、20周计算2组创面移植皮片成活率、收缩率(或扩张率).术后20周取2组创周正常皮肤及创面皮肤标本,采用HE染色法观察胶原纤维束结构,测量胶原纤维束直径和间隙率;用天狼星红染色法观察Ⅰ、Ⅲ型胶原分布情况,测量Ⅰ、Ⅲ型胶原含量及其比值.对实验数据行独立样本t检验、Levene检验、t'检验.结果 (1)术后2周,实验组大鼠创面移植皮片成活率[(76.1±13.1)%]低于对照组[(94.5±1.3)%,t'=3.440,P=0.018].术后3周,实验组创面移植皮片收缩率[(34±8)%]明显大于对照组[(16±12)%,t=-3.211,P=0.009];术后8周,2组移植皮片扩张率接近一致.(2)HE染色和天狼星红染色显示,与大鼠创周正常皮肤比较,对照组移植皮片胶原纤维束呈均质化改变,胶原纤维纤细,排列紊乱;实验组移植皮片胶原纤维束结构、排列更接近创周正常皮肤,可见未完全降解的PADM.与对照组创面皮肤胶原纤维束直径[(7.3±1.4)μm]、间隙率[(17±4)%]、Ⅰ型胶原含量[(68.1±8.4)%]、Ⅲ型胶原含量[(32.0±8.4)%]以及Ⅰ、Ⅲ型胶原比例(2.3±1.0)比较,实验组胶原纤维束更粗[(9.6±0.8)μm,t=-3.562,P=0.005],间隙率更大[(24±5)%,t=-2.760,P=0.020],Ⅰ型胶原含量更高[(80.2±5.4)%,t=-2.981,P=0.014],Ⅲ型胶原含量更低[(19.8±5.4)%,t=2.981,P=0.014],Ⅰ、Ⅲ型胶原比例更高(4.3±1.2,t=-3.204,P=0.009).实验组创面皮肤上述胶原相关指标更接近于创周正常皮肤水平.结论 异体PADM在体内作为真皮再生模板,有助于改善自体刃厚皮所修复的大鼠皮肤缺损创面中真皮胶原纤维束的结构,提高再生真皮组织的成熟度.

Abstract：

Objective To evaluate the effects of mixed grafting of allogeneic PADM and autologous STS on wound healing of full-thickness defect in rats. Methods Full-thickness defects with size of 6 cm×4 cm were produced on the back of 12 SD rats, and they were divided into E group(n =6) and C group ( n = 6) according to the random number table. The wounds in E group were grafted with a mix of allogeneic PADM (expansion rate 10: 5) and autologous STS with thickness of 0.2 mm, while those in C group were grafted with autologous STS in the same thickness. The wound healing rate, survival rate, contraction rate,and expansion rate of transplanted skin were observed at post operation week (POW) 2, 3, 4, 6, 8, 12,20. Tissue samples form wounds and surrounding normal skin were harvested at POW 20 for histopathological observation as follows. The structure of collagen fiber bundle was observed by HE staining, the diameter and gap rate of collagen fiber bundle were also measured. The distribution of type Ⅰ and Ⅲ collagen was observed by sirsus red staining, and the contents of type Ⅰ , Ⅲ collagen and their ratio were also examined.Data were processed with independent samples t test, Levene test, and t' test. Results Survial rate of transplanted skin in E group at POW 2 [(76. 1 ± 13. 1)%] was obviously lower than that in C group [(94.5 ± 1.3)%, t' =3.440, P =0.018]. Contraction rate of transplanted skin in E, C groups at POW 3 showed significant difference [(34±8)%vs. (16 ±12)%, t = -3.211, P =0.009]. Compared with those in peri-wound normal skin, collagen fiber bundles in C group showed signs of homogenization, and collagen fibers were thin with irregular arrangement. Collagen fiber structure and arrangement of composite skin in E group were similar to those surrounding normal skin with incomplete degradation of PADM. Diameter of collagen fiber bundle [( 9.6 ± 0.8) μm] , gap rate between collagen bundle [( 24±5) %] , content of type Ⅰ collagen [( 80.2 ± 5.4) %] and the ratio of typeⅠto type Ⅲ collagen(4.3 ± 1.2) in E group were all increased as compared with those inC group [(7.3±1.4) μm (t = -3.562, P =0.005), (17±4)%( t =-2.760, P =0.020), (68.1 ±8.4)%(t = -2.981, P =0.014), 2.3±1.0(t = -3.204, P =0. 009)], while content of type Ⅲ collagen [( 19.8 ± 5.4) %] in E group was lower than that in C group [(32.0 ±8.4)% , t = 2. 981, P = 0. 014]. Above-mentioned indexes of collagen in wound of E group were similar to those of normal skin surrounding the wound. Conclusions Allogeneic PADM used as dermal regeneration template is beneficial in improving collagen fiber bundle structure in dermis layer of rats with fullthickness skin wounds when repaired with autologous STS, and it accelerates maturation of regenerative dermal tissue.     

膀胱细胞外基质移植物修复膀胱缺损的实验研究

目的：探讨膀胱细胞外基质修复膀胱缺损的可行性。方法：应用生物化学方法制备兔膀胱细胞外基质。实验组27只兔膀胱分别切除20％（9只），30％－40％（9只），用异体膀胱细胞外基质以缺损区1.5倍大小植于缺损区；对照组A（6只）仅行膀胱部分（40％－50％）切除；对照组B（6只）行膀胱部分（40％－50％）切除后用未经脱细胞处理的异体膀胱进行移植。结果：实验组术后10d可见移植物内有膀胱成分出现，移植物内表面完全被尿路移行上皮覆盖；术后1个月所有膀胱壁成分均在移植物内存在；术后3个月平滑肌成分再生良好，生化检查及尿动力学检查均正常。对照组A的膀胱容量较术前缩小（P＜0.05），对照组B膀胱移植补片均于术后3－5d坏死。结论：膀胱细胞外基质是修复膀胱缺损较理想的材料。     

组织工程学材料替代输尿管缺损的临床应用

目的总结新型替代材料修复输尿管狭窄的临床经验。方法采用组织工程学材料同种异体脱细胞输尿管细胞外基质修补治疗输尿管长段狭窄4例。男1例，女3例。平均年龄43岁。病程12～37个月，平均21个月。其中肾盂输尿管移行区狭窄1例，输尿管中上段狭窄1例，输尿管下段狭窄2例。狭窄长度3～10cm。结果4例随访11～22个月，平均17．5个月。术后4周拔除输尿管支架管1例，术后8周拔除支架管3例，无狭窄、漏尿，排尿良好。术后6个月IVU检查：输尿管造影剂通过正常，无狭窄，无渗漏。结论同种异体脱细胞输尿管细胞外基质为机体提供了良好的供宿主细胞“爬行替代”的框架，在修补后可有效地与宿主组织融合。修补后的输尿管愈合良好。     

Tissue-engineering applications for phallic reconstruction

Pathologic penile conditions often require reconstructive surgery. Due to the limited amount of autologous tissues available for reconstruction, other tissue substitutes have been used. Phallic reconstruction using engineered autologous genital tissue, i.e., tissue derived from the patient's own cells, may be preferable. In this article we describe tissue-engineering approaches that may be applicable to genital reconstruction.     

组织工程学材料替代输尿管缺损的动物实验及临床应用

目的　寻求理想的输尿管替代材料。　方法　将２０ 只兔双侧输尿管脱细胞处理后制成输尿管细胞外基质（ＥＣＭ） ；另６０ 只兔分为３ 组：输尿管ＥＣＭ 移植组（ 实验组） ，对照组１ 及对照组２ 。　结果　实验组术后组织学检查显示１ 周时ＥＣＭ 边缘部腔内面已见移行上皮细胞，管壁有肌细胞长入。６ 周时取材ＥＣＭ 区已修复近正常输尿管组织结构。１２ 、２４ 周时ＩＶＵ 检查；输尿管显影良好，通畅。对照组１ 术后７ ～１０ 天，移植的异体输尿管均坏死。对照组２ 输尿管缺损区６ 周时均被纤维包裹而明显增粗。在实验取得满意结果的基础上成功应用于临床。　结论　异体脱细胞输尿管细胞外基质为理想的输尿管替代材料。     

Penis covering and simultaneous urethral replacement by scrotal skin for severe penile and urethral necrosis

Objectives  Penile strangulation with severe necrosis and concomitant significant tissue loss is a challenging problem for reconstructive surgery. Final reconstruction and tissue augmentation of the resected penis often require multi-step plastic reconstruction. Methods  Severe damage of the phallus with gangrene and septic complication caused by a bronze ring were first conservatively treated to achieve dry gangrene. The glans of the penis, the whole penile skin, and distal third of the corpus spongiosum with the urethra were lost to necrosis, resulting in a missing urethral segment on the penis. Surgery was performed 3 months after removing the metal ring; a one-step skin and urethral reconstruction was done. Results  Good functional and esthetic results could be achieved by mid-term follow-up. Conclusion  In long-lasting penile strangulation with damaged blood supply, further surgery and treatment are needed in addition to immediate decompression. Most commonly in these situations, amputation seems to be the best and easiest action to be taken. However, our case shows that even in a most severe situation, careful conservative treatment allows preserving the phallus, and subsequently reconstruction enables reaching the best functional and esthetic results.     

Effect of catheter material on the incidence of urethral strictures.

The aetiology of urethral strictures in patients catheterised for short periods at the time of surgery is the subject of some debate. The occurrence of epidemics of strictures associated with certain batches of latex catheters, and the demonstrable in vitro cytotoxicity of latex in general, have supported the suggestion that catheter material (and in particular latex) has an important role. We have looked retrospectively at 299 patients undergoing transurethral resection of the prostate gland by 1 urologist, 135 of whom had a latex catheter and 164 a plastic one; 19 patients developed strictures over the next 12 months with no proven difference attributable to the catheter material. The study provides no evidence that the short-term use of latex catheters is associated with stricture formation.     

Comparison of partial urethral replacement with acellular matrix versus spontaneous urethral regeneration in a canine model

OBJECTIVE: To determine whether acellular matrix could be used for partial urethral replacement and to compare regeneration over acellular matrix versus normal spontaneous urethral regeneration. MATERIALS AND METHODS: The study included 21 male mongrel dogs in which a 3-cm segment including half of the urethral circumference was excised. In 13 dogs (study group), the defect was covered by acellular matrix of the same length and width obtained from female mongrel dogs and prepared to have complete cell lysis with keeping of the fiber framework. In 8 dogs (control group), the urethral defect was not covered by any urethral tissue. In both groups, an 8F feeding tube was kept inside the urethra for a mean duration of 2 weeks. In the study group, dogs were sacrificed at 1 week, 2 weeks, 3 weeks and then one dog every month for 10 months. In the control group, one dog was sacrificed every month for 8 months. RESULTS: All dogs survived the procedure. In the study group, 10 dogs underwent urethrogram; 8 were normal, 1 had diverticulum and 1 had relative narrowing. In the control group, 6 dogs underwent urethrogram; 5 were normal and 1 showed relative narrowing. Histopathological examination of the study group showed gradual regeneration over the acellular matrix with normal appearance at 20 weeks. In the control group, normal healing was observed at 2 months and thereafter. CONCLUSION: Regeneration of all components of the urethra can occur gradually over acellular matrix and is complete at 20 weeks. Regeneration of a urethral defect 3-cm long including half of the urethral lumen is possible with or without acellular matrix.     

髋关节置换术患者侧卧位导尿效果观察

目的探讨髋关节置换术患者侧卧位留置导尿管操作流程及其效果。方法将60例髋关节置换术患者按手术时间的单双日分为对照组和观察组各30例。对照组按常规方法于椎管内麻醉后置平卧位行导尿术,导尿后摆放手术体位;观察组椎管内麻醉后(患肢在上)不改变体位(侧卧位)实施导尿术。结果观察组皮肤损伤、尿道损伤、一次性导尿成功率与对照组比较,差异无统计学意义(均P>0.05);切口标记移位和手术准备时间与对照组比较,差异有统计学意义(P<0.05,P<0.01)。结论髋关节置换术患者麻醉后采取侧卧位导尿,有利于缩短手术准备时间,避免手术切口标记移位。     

Bladder replacement by entero-cystoplasty after radical cystectomy for bladder cancer--the urethral Kock pouch

From September 1989 to March 1990, 6 male patients with invasive bladder cancer, 49 to 70 years old in age, underwent bladder replacement with the ileum (the urethral Kock pouch) after radical cystectomy. Follow up ranged between 3 and 9 months. Urodynamic evaluation showed the ileal bladder to be a low pressure reservoir with a capacity that increased to more than 250 ml. The ileal bladder was emptied by straining without significant residual urine in all patients except one who was performing intermittent self-catheterization. All patients were continent in the daytime. However, all patients required pads at night because of occasional loss of a little urine. Excretory urograms revealed excellent upper tract function. The procedure is suitable whenever the urethra can be preserved after cystectomy for cancer.