髂内动脉结扎对大鼠阴茎海绵体组织nNOS神经纤维及eNOS表达的影响

目的 研究髂内动脉结扎对大鼠阴茎海绵体组织神经型一氧化氮合酶(nNOS)神经纤维及内皮型一氧化氮合酶(eNOS)表达的影响。方法 45只Wistar雄性成年大鼠随机分为假手术组(21只)和手术组(24只)。其中手术组采用双侧髂内动脉结扎的方法制备动脉性勃起功能障碍(ED)大鼠模型,假手术组作为对照组只切开腹腔不结扎髂内动脉。于术后1周、3周、6周末分别取各组1/3大鼠,观察其勃起功能,并采用免疫组化SP法检测大鼠阴茎组织中nNOS神经纤维的数量及eNOS的表达。结果 手术组与假手术组相比,1周末、3周末和6周末在阴茎勃起次数和阴茎组织nNOS神经纤维数量及eNOS表达上均有显著差异(P<0.01)。手术组大鼠阴茎勃起次数3周末高于1周末,6周末高于3周末;eNOS的表达3周末明显低于1周末,而6周末高于3周末,差异均有显著性意义(P<0.001),并呈增加趋势。手术组大鼠阴茎组织nNOS神经纤维数量三者之间差异均有显著性(P<0.001),呈进行性下降。结论 髂内动脉结扎影响阴茎勃起功能和nNOS、eNOS的表达。阴茎组织中nNOS阳性神经纤维数量的减少,可能是髂内动脉结扎术后阴茎勃起功能障碍发生的原因之一。     

阴茎海绵体平滑肌细胞的鉴定分析进展

勃起功能障碍(erectile dysfunction,ED)是一种多因素引起的男科难治性疾病[1],严重影响患者的整体健康和生活质量[2].阴茎海绵体平滑肌细胞(CSMC)是组成阴茎海绵体的主要功能成分,约占整个阴茎组织成分的40%～50%,是阴茎神经调控的主要效应器部位[3,4],因此也成为调节阴茎勃起及维持勃起的重要因素.从这意义上来讲,CSMC的研究显得尤为重要,而CSMC的培养和鉴定是这研究的基础,因此CSMC纯化鉴定在细胞水平上研究ED的机制颇受到关注,本文就体外培养的CMSC鉴定分析做一综述.     

尼古丁对大鼠阴茎海绵体内源性CO浓度及NOS活性的影响

目的:观察尼古丁对成年雄性大鼠阴茎海绵体内源性一氧化碳(CO)浓度及一氧化氮合酶(NOS)活性的影响,探讨吸烟对勃起功能损害的可能机制。方法:40只成年雄性Wistar大鼠分为4组,尼古丁注射1个月组、2个月组、3个月组和对照组,尼古丁注射组尼古丁0.5 mg/(kg.d)皮下分别注射1、2、3个月,对照组注射生理盐水。处理后,取阴茎海绵体,用改良双波长分光光度法检测CO浓度,改良Griess法检测NOS活性。结果:对照组CO浓度为(13.66±0.40)μmol/mg prot,NOS活性为(9.72±0.47)U/mg prot。尼古丁注射1个月,CO浓度和NOS活性分别下降为(12.43±0.56)μmol/mg prot和(8.44±0.69)U/mg prot,显著低于对照组(P均<0.01);尼古丁注射2个月,CO浓度和NOS活性分别下降为(11.41±0.52)μmol/mg prot和(7.53±0.24)U/mg prot,显著低于对照组和尼古丁注射1个月组(P<0.01);尼古丁注射3个月,海绵体CO浓度和NOS活性分别下降为(10.52±0.59)μmol/mg prot和(6.64±0.31)U/mg prot,均显著低于对照组和尼古丁注射1个月、2个月组(P均<0.01)。结论:尼古丁可导致成年雄性大鼠阴茎海绵体内源性CO浓度及NOS活性下降,提示内源性CO及NOS参与吸烟引起勃起功能障碍的病理生理过程。     

轻度血尿酸升高对大鼠肾小球内皮细胞及血管平滑肌细胞功能的影响

目的 通过观察轻度血尿酸升高对大鼠肾小球内皮细胞功能损伤及血管平滑肌细胞增殖的影响,探讨轻度血尿酸升高是否能导致肾脏损害及降尿酸治疗对肾脏的保护作用.方法 用雄性SD大鼠为研究对象,随机分为4组:对照组(n=15)、氧嗪酸组(n=15)、别嘌呤醇组(n=12)和氧嗪酸+别嘌呤醇组(n=12).予以低盐饮食,每隔10天监测各组大鼠的动脉血压.于试验后20 d及40 d用ELISA法分别测定各组大鼠内皮细胞功能受损的指标[一氧化氮(NO)、内皮素(ET)1、纤溶酶原激活物(PAI)1]、血管平滑肌细胞增殖的指标[血小板衍生因子(PDGF)、环氧化酶(COX)2、单核细胞趋化蛋白(MCP)1的含量]以及炎性反应指标[白细胞介素(IL)18、肿瘤坏死因子(TNF)α];同时观察各组大鼠肾功能及肾脏组织病理变化;免疫组化法检测各组大鼠肾组织中PDGF、一氧化氮合酶(NOS)的表达.结果 与对照组相比,氧嗪酸组大鼠血浆NO浓度显著降低(P＜0.05),ET-1、PAI-1、PDGF、MCP-1、COX2、TNF-α、IL-18浓度均显著升高(均P＜ 0.05).光镜下,各组大鼠肾组织均未见尿酸结晶形成,氧嗪酸组肾小血管管壁增厚,内膜增生,管腔狭窄;免疫组化结果显示,与对照组相比,氧嗪酸组NOS的表达显著减少(7.33％±2.11％比25.75％±2.33％,P＜0.05),PDGF的表达显著增多(31.18％±2.83％比8.09％±1.81％,P＜ 0.05).经别嘌呤醇降尿酸干预治疗后大鼠血清中内皮细胞损伤指标NO上调(P＜0.05),而ET-1及PAI-1均下调(均P＜0.05);而血管平滑肌增殖指标及炎性指标均下调(均P＜ 0.05).结论 轻度血尿酸升高可导致肾小球内皮细胞功能受损、血管平滑肌细胞增殖;降尿酸治疗能改善内皮细胞功能,减轻血管平滑肌细胞的增殖.     

大鼠阴茎海绵体NOSⅠ、NOSⅢ mRNA和蛋白表达研究

目的 :探讨正常大鼠阴茎海绵体中一氧化氮合酶 (NOS) Ⅰ和 Ⅲ mRNA和蛋白的表达。　方法 :取雄性SD大鼠阴茎组织 ,应用高度特异性抗体经免疫沉淀、Western印迹分析技术分别检测NOSⅠ 、NOS Ⅲ 蛋白的表达 ;RT PCR方法检测NOSⅠ和NOS Ⅲ mRNA的表达。　结果 :正常大鼠阴茎海绵体组织中有NOSⅠ 和NOS ⅢmRNA和蛋白的表达 ,并且NOSⅢmRNA和蛋白的表达明显多于NOSⅠmRNA和蛋白的表达。　结论 :不同的NOS亚型可能共同参与阴茎勃起的调控。     

衰老对大鼠阴茎海绵体内皮细胞功能的影响

目的 :探讨衰老对大鼠阴茎海绵体内皮细胞功能的影响。　方法 :利用YH 4压力传感器分别检测了青龄(5个月 )和老龄 (2 0个月 )两组大鼠阴茎海绵体内压 (ICP)在乙酰胆碱 (Ach)、硝普钠 (SNP)和A2 3187作用下的变化 ;并测定了两组大鼠阴茎海绵体一氧化氮合酶 (NOS)的活性。　结果 :青龄组基础ICP为 (9.4± 2 .3)mmHg(1mmHg=0 .133kPa) ,老龄组为 (7.2± 1.7)mmHg,二者间差异无显著性 (P >0 .0 5 )。在浓度分别为 10 -6、10 -5、10 -4mol/L的Ach作用下 ,两组大鼠ICP值间差异均有显著性 (P <0 .0 1)。当Ach浓度为 10 -4mol/L时 ,两组大鼠ICP值达到最高 ,青龄组为 (5 4 .8± 4 .2 )mmHg ,老龄组为 (40 .3± 2 .8)mmHg。A2 3187(10 μmol/L)可以进一步提高老龄组ICP值 ,由(40 .3± 2 .8)mmHg上升到 (5 6 .2± 4 .1)mmHg ,两者间差异有显著性 (P <0 .0 1) ;青龄组提高不明显 ,由 (5 4 .8± 4 .2 )mmHg上升到 (5 5 .8± 4 .7)mmHg ,两者间差异无显著性 (P >0 .0 5 )。在SNP(10 -4mol/L)作用下青龄组ICP值为(5 8.9± 4 .7)mmHg ,老龄组为 (5 1.7± 5 .3)mmHg ,两者间差异无统计学意义 (P >0 .0 5 )。两组大鼠阴茎海绵体内NOS的活性差异无统计学意义 (P >0 .0 5 )。　结论 :大鼠阴茎海绵体内皮细胞对内皮细胞激动剂     

高脂血症对大鼠阴茎海绵体中NOS及CO含量影响的研究

目的 探讨高脂血症对大鼠阴茎海绵体中一氧化氮合成酶(NOS)活性及一氧化碳(CO)浓度的影响及两者之间的关系.方法 将32只雄性wistar大鼠随机分为2组:高脂血症组和正常对照组,每组各16只大鼠.通过建立大鼠高脂血症模型,于实验开始后10d和20d,分别随机取各组1/2大鼠观察其勃起功能;用分光光度法分别测定海绵体匀浆中NOS活性和CO浓度.结果 在第10天及第20天,高脂血症组和正常对照组相比,大鼠阴茎海绵体内NOS活性、CO浓度及阴茎勃起次数差异均有统计学意义(P<0.05);而且第20天高脂血症组NOS活性、CO浓度及阴茎勃起次数,均明显低于第10天,两两比较差异均有统计学意义(P<0.05).结论 高脂血症大鼠阴茎海绵体中NOS及CO下降,这可能是高脂血症引起勃起功能障碍的发病机制之一.     

Hhcy大鼠阴茎海绵体内NOS及CO含量的研究

目的:检测高同型半胱氨酸血症(Hhcy)大鼠阴茎组织内一氧化氮合酶(NOS)和一氧化碳(CO)的含量并探讨其对阴茎勃起功能的影响。方法:取成年雄性Wistar大鼠20只,随机均分为2组:正常对照组和Hhcy组。Hhcy组给予3%高蛋氨酸饲料喂养,正常对照组给予普通饲料喂养,4周后取血,测定血清Hhcy含量;取阴茎海绵体组织、匀浆,测定NOS和CO的含量。结果:Hhcy组血清中同型半胱氨含量[(22.32±1.65)μmol/L]显著高于正常对照组[(4.90±1.73)μmol/L],阴茎组织中的NOS[(6.45±1.12)nmol/(g·min)]和CO[(10.60±0.92)μmol/L]含量明显低于正常对照组[(10.77±0.60)nmol/(g·min)和(13.36±0.44)μmol/L]。结论:给予4周高蛋氨酸饮食的Wistar大鼠能引起Hhcy。Hhcy大鼠阴茎海绵体组织中内源性NOS和CO降低。Hhcy是大鼠阴茎勃起功能障碍的独立危险因素。     

大鼠前列腺增生模型阴茎海绵体内nNOS、eNOS表达研究

目的 分析引起阴茎勃起的主要神经递质NO合成的限速酶nNOS和eNOS在BPH模型大鼠阴茎海绵体中的表达.试探讨BPH引起ED的可能因素.方法 20只雄性SD大鼠随机分为正常对照组和BPH模型组.通过去势后丙酸睾酮注射建立BPH模型,10只老年大鼠为老年对照组.4周后处死大鼠,分析不同组前列腺湿质量与前列腺指数,并在光镜下观察前列腺组织病理学改变.应用免疫组化方法研究不同组间大鼠阴茎海绵体内nNOS和eNOS阳性表达情况并比较组间差异.结果 BPH模型组中前列腺湿质量和前列腺指数与正常对照组比较差异有统计学意义;显微镜下BPH模型组前列腺组织呈明显增生表现,大鼠BPH造模成功.免疫组化方法显示BPH模型组、老年对照组大鼠阴茎海绵体内的nNOS及eNOS阳性细胞表达率明显降低;nNOS及eNOS阳性表达与正常对照组的比较差异均有统计学意义.结论 nNOS及eNOS是调控阴茎勃起的神经递质NO合成的关键酶,BPH模型组的阴茎海绵体内nNOS、eNOS表达明显减少或活性降低,导致NO释放减少可能是BPH引起勃起功能障碍的重要原因.     

衰老对大鼠阴茎海绵体NOSⅠ的表达和NOS活性的影响

目的 :探讨衰老对大鼠阴茎海绵体一氧化氮合酶Ⅰ (NOSⅠ)mRNA、蛋白的表达和NOS活性的影响。　方法 :30只雄性SD大鼠按不同月龄分为成年组、老年组和衰老组 ,应用Western印迹、RT PCR方法分别检测不同年龄组阴茎海绵体NOSⅠ蛋白及mRNA的表达 ;用紫外分光光度计测定不同年龄组阴茎海绵体NOS的活性。　结果 :成年组NOSⅠ 蛋白的表达量最高 ,老年组和衰老组显著降低 ,分别为成年组的 75 .6 %和 6 1.2 % ;NOSⅠmRNA的表达与蛋白表达的变化一致 ;老年组NOS活性与成年组差异无显著性 (P >0 .0 5 ) ,衰老组NOS活性明显降低 ,是成年组的70 .4 % ,并且差异非常显著 (P <0 .0 1)。　结论 :衰老引起NOSⅠ 蛋白及mRNA的表达降低和NOS活性的显著降低 ,可能是老年性阴茎勃起功能障碍的主要机制之一。     

人参皂甙对去势大鼠阴茎海绵体组织NO含量与细胞凋亡的影响

目的研究人参皂甙对去势大鼠阴茎海绵体组织细胞凋亡、NO含量的影响,探讨人参皂甙壮阳功效的可能机制。方法40只成年雄性大鼠随机分为去势组、对照组及不同剂量(25mg/kg、100mg/kg)人参皂甙组共4组,1周后取阴茎海绵体,放免法检测血清睾酮含量(ng/ml),全自动生化分析仪比色法测定海绵体NO含量(μg/g),末端脱氧核糖核酸转移酶介导的duTP缺口末端标记法测定细胞凋亡。结果对照组血清睾酮水平浓度为(1.51±0.86),在去势组、人参皂甙治疗组(25mg/kg、100/mg/kg)均未测到。去势组阴茎海绵体NO浓度(14.45±2.38)较对照组(39.8±3.28)显著降低(P<0.01),25mg/kg人参皂甙组阴茎海绵体NO水平(16.02±2.67)与去势组(14.45±2.38)接近(P>0.05),100mg/kg人参皂甙组NO水平(37.88±7.06)较去势组细胞凋亡数(14.45±2.38)明显升高(P<0.05),与对照组(39.8±3.28)接近(P>0.05)。大剂量100mg/kg人参皂甙组(12.51±1.81)较去势组(26.02±5.25)低(P<0.05)。25mg/kg人参皂甙组凋亡细胞积分光密度(27269.60±4920.42)与去势组比较(33931.50±2459.36)差异无统计学意义,大剂量100mg/kg人参皂甙组(18766.36±3040.42)较去势组(33931.54±2459.36)低,两者比较差异有统计学意义(P<0.05)。结论100mg/kg剂量的人参皂甙不能增加去势大鼠血清睾酮含量,但可以提高去势大鼠阴茎海绵体组织NO水平,减少海绵体细胞凋亡。人参皂甙对去势大鼠阴茎海绵体细胞凋亡的抑制作用可能与其增加NO水平有关。     

腺病毒介导的激活大鼠阴茎海绵体平滑肌细胞中NO／cGMP通路的实验研究

目的：探讨靶向大鼠iNOS基因的shRNA重组腺病毒载体对大鼠阴茎海绵体平滑肌细胞iN0s基因的激活作用,为阴茎勃起功能障碍（ED）的基因治疗提供实验依据。方法：将前期构建的重组腺病毒AdS—iN—OSrshRNA-EGFP（AdU6／shiNOS）和对照病毒AdU6／shControl,分别转染大鼠阴茎海绵体平滑肌细胞,分别在不同病毒MOI（25,50,75）值下72小时后采样检测。采用realtimeRT-PCR半定量检测AdU6／shiNOS对细胞iNOS基因mRNA表达影响;Western—blot法检测海绵体平滑肌细胞iNOS蛋白表达变化。然后培养基中加L—Arg（10mmol／L）,用酶联免疫法检测病毒转染72小时后海绵体平滑肌细胞内cGMP的浓度变化,记录AdU6／shiNOS对平滑肌细胞内cGMP的影响。结果：AdU6／shiNOS转染大鼠阴茎海绵体平滑肌细胞72小时后,和空白对照组、阴性对照组相比iN0s基因在mRNA和蛋白表达水平均显著上调（P〈O．05）,呈剂量依赖性,MOI一75时RNAa效果最好。而且转染72小时后,实验组原代平滑肌细胞内cGMP水平显著高于对照组及空白组（Pd0．05）。结论：利用腺病毒介导的RNAa技术,提高海绵体平滑肌细胞iN0s基因表达获得成功,可以增加阴茎海绵体平滑肌细胞cGMP水平,激活了NO／cGMP通路,这为勃起功能障碍的基因治疗研究开辟了新的方向。     

2型糖尿病大鼠血浆同型半胱氨酸与阴茎海绵体内NOS和内源性CO的相关性研究

目的:研究2型糖尿病性大鼠血浆同型半胱氨酸(Hcy)与阴茎海绵体内NOS和内源性CO的相关性。方法:选取3月龄雄性Wistar大鼠50只,随机选取10只为对照组(A组);高糖高脂饲料饲养4周后从其他40只大鼠中筛选出30只构建成功的糖尿病(DM)大鼠模型,随机分成3组:DM大鼠组(B组);胰岛素治疗组(C组)和叶酸+维生素B12治疗组(D组)。8周及12周后注射阿朴吗啡观察各组大鼠阴茎勃起情况。12周后测各组大鼠血浆总Hcy含量及阴茎海绵体内NOS活性和CO含量。结果:与A组比较,B组大鼠血浆Hcy浓度明显升高,阴茎勃起功能明显降低,阴茎海绵体NOS活性和CO含量均下降,差异有显著性(P<0.01)。2型DM大鼠中高Hcy血症发生率为55%。与B组比较,C组和D组中大鼠血浆Hcy浓度显著下降,阴茎勃起功能、阴茎海绵体NOS活性均升高(P<0.01),Hcy与NOS(rA=-0.89,rB=-0.76,rC=-0.91,rD=-0.91)及CO含量(rA=-0.82,rB=-0.77,rC=-0.93,rD=-0.81)均呈负相关。结论:2型DM大鼠血浆中的高Hcy可能是引起阴茎海绵体NOS活性下降、CO含量下降,进而导致DM ED发病的分子机制之一。胰岛素、叶酸和维生素B12可以改善DM大鼠的勃起功能,提高阴茎海绵体NOS活性和CO含量。     

兔阴茎海绵体平滑肌细胞的快速分离及鉴定

目的探讨兔阴茎海绵体平滑肌细胞快速分离方法,为应用膜片钳技术研究阴茎勃起机制提供实验材料.方法采用木瓜蛋白酶和胶原酶两步酶解消化法,快速分离出新西兰大白兔阴茎海绵体平滑肌细胞并应用免疫组化鉴定.结果分离的细胞成活率较高,贴壁呈长梭形,胞膜光滑完整,胞浆均匀,可用于膜片钳记录.免疫组化鉴定为兔阴茎海绵体平滑肌细胞.结论酶解消化快速分离兔阴茎海綿体平滑肌细胞为全细胞膜片钳技术研究阴茎勃起功能障碍的电生理机制提供了较好的实验材料.     

Intracellular calcium concentration of corpus cavernosum smooth muscle cells is decreased by the overexpression of PnNOS gene in adipose tissue‐derived stem cells

The study investigated the effects of adipose tissue‐derived stem cells (ADSCs) modified with penile neuronal nitric oxide synthase (PnNOS) gene on intracellular calcium concentration in rat corpus cavernosum smooth muscle cells (CCSMCs). ADSCs and CCSMCs of Sprague–Dawley (SD) rats were isolated and cultured in vitro respectively. The rat PnNOS gene was transferred into the ADSCs mediated by a recombinant adenovirus vector. The expression of the PnNOS gene was detected. At the same time, the concentration of nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) was assayed. After coculturing with the CCSMCs of SD rats, which were isolated and expanded ex vivo, the cGMP and NO levels of ADSCs and CCSMCs were measured. Intracellular calcium concentration ([Ca²⁺]_i) in rat CCSMCs was measured with Fluo‐3/AM by flow cytometer after cocultured with ADSCs overexpressing PnNOS gene. The mRNA and protein expression of PnNOS gene mediated by recombinant adenovirus vector significantly overexpressed and lasted at least 2 weeks. Meanwhile, the concentration of NO and cGMP in ADSCs was greatly increased. The concentration of cGMP was significantly increased, and [Ca²⁺]_i was obviously decreased in CCSMCs compared with the control groups (P < 0.05) after cocultured with ADSCs for 3 days. These findings demonstrated that ADSCs overexpressing PnNOS gene might decrease [Ca²⁺]i in CCSMCs by up‐regulating NO–cGMP signalling pathway.     

Nitric oxide mediates relaxation in rabbit and human corpus cavernosum smooth muscle

Summary We investigated in vitro the relaxant effect of exogenous acetylcholine (ACh) and electric-field stimulation (EFS) on rabbit and human corpus cavernosum smooth muscle strips (CC) precontracted with phenylephrine. The effects of EFS and ACh were monitored alone, after muscarinic receptor blockade and after inhibition of nitric oxide (NO) formation with l-N-nitroarginine (l-NOARG). In rabbit und human CC, both atropine and l-NOARG abolished the relaxant effects of ACh. The relaxant effects of EFS, however, were only slightly reduced by atropine to 97.5±17.5% in human CC and to 89.0±6.1% in rabbit CC. l-NOARG further reduced the EFS effects to 0.8±1.7% in human CC and to 16.2±8.7% in rabbit CC. In strips obtained from impotent patients with diabetes mellitus, the relaxant effects appeared to be significantly less than in strips from nondiabetic impotent men. Tetrodotoxin blocked the relaxant EFS effects in human and rabbit strips completely. The data indicate the important role of NO in cholinergically induced relaxation of cavernous smooth muscle in rabbits and humans. Our findings support the idea of NO as the nonadrenergic noncholinergic neurotransmitter in penile erection in both species. Rabbit erectile tissue might serve as an in vitro animal model for further investigation.     

Activation of the iNOS/NO/cGMP pathway by Revactin® in human corporal smooth muscle cells

BackgroundThe combination of the nutraceuticals, Paullinia cupana, ginger rhizome, muira puama, and the amino acid L-citrulline (COMP-4) has been shown to stimulate the production of inducible nitric oxide synthase (iNOS), nitric oxide (NO), and cGMP in rat corpora cavernosa smooth muscle cells (CSMC). When administered to middle-aged rats, long-term treatment with COMP-4 resulted in both an increase in the number of CSMC and an improvement in erectile function. We, therefore, aimed to determine whether a commercial formulation of COMP-4, Revactin^®, could have a similar stimulatory effect on human CSMC.MethodsPrimary human CSMC cultures (HCSMC) were grown and incubated with Revactin® for up to 24 hours. cGMP generation and nitrite formation were determined by ELISA and Griess reaction, respectively. IBMX (1 mM), sildenafil (0.4 mM), and L-NIL (4 µM) were utilized as modulators of the NO-cGMP pathway. iNOS, endothelial NOS (eNOS), and neuronal NOS (nNOS) expressions were determined by Western blot.ResultsRevactin^® up-regulated both nitrite formation and cGMP expression, achieving the highest expression at 24 hours in the HCSMC. These effects were completely blocked by L-NIL. Revactin^® up-regulated iNOS expression, but not that of eNOS or nNOS.ConclusionsThe results presented in this study confirmed that Revactin^® activated the iNOS-NO-cGMP pathway intracellularly in HCSMC. It still needs to be determined whether the upregulation of this pathway would be an effective approach for counteracting the fibrosis and apoptosis of the corporal smooth muscle cells associated with aging.     

Construction of cavernosum smooth muscle using umbilical artery smooth muscle cells seeded on acellular corporal collagen matrices

The objective of this study was to investigate the feasibility of tissue engineering of corpus cavernosal smooth muscle. Acellular corporal collagen matrices (ACCMs) were obtained from the penis of adult rabbits by a cell removal procedure. ACCMs were implanted into the back muscles of allogenic rabbits to investigate the resulting immunological reaction. Human umbilical artery smooth muscle cells (HUASMCs) were isolated from human umbilical arteries through explant techniques and expanded in vitro . Subsequently, third and fifth passage HUASMCs were seeded to ACCMs at a concentration of 30 × 10⁶ cells/mL. Then, seeded ACCMs were implanted subcutaneously in athymic mice. The implants were retrieved at 10, 20 and 40 days after implantation. Histochemistry, immunohistochemistry and scanning electron microscopy were performed to analyse the morphological characteristics of the engineered tissues. Additionally, organ bath studies were performed to address the contractility of the engineered tissues. The decellularization process successfully extracted all cellular components while preserving the original collagen fibers. The immunological reaction to ACCMs consisted of only a transient nonspecific inflammatory response. Light and scanning electron microscopy demonstrated that HUASMCs extended onto the three-dimensional ACCMs scaffolds in vitro . Histological analyses of the explants from all time points demonstrated a progressive regeneration of smooth muscle, with structures very similar to native corpus cavernosum smooth muscle. The maximum contraction force induced by phenylephrine and electrical stimulation were 3.64 ± 0.18 g/100 mg and 2.50 ± 0.21 g/100 mg, respectively. Our study demonstrates that HUASMCs can be seeded on three-dimensional ACCM scaffolds and will develop tissues similar to that of the native corpus cavernosum smooth muscle.