人脐动脉平滑肌细胞复合脱细胞基质构建海绵体平滑肌的动物实验研究

目的 探讨脐动脉平滑肌细胞(HUASMC)与阴茎海绵体脱细胞基质(ACCM)复合构建海绵体平滑肌的可行性.方法 以1%的Triton-X100与0.1%NH3H2O混合液对兔阴茎海绵体进行脱细胞处理,制备ACCM.采用贴块法分离、培养、扩增HUASMC.HUASMC以30×10<'6>/ml密度接种ACCM,细胞-ACCM体外复合10 d后将复合物植入9只5周龄BALB/C裸鼠背部皮下,术后10、20和40 d分别对移植物进行HE染色、免疫组织化学染色和器官浴槽实验,评价其裸鼠体内构建情况.结果 ACCM为白色圆柱状,镜下为富含胶原的疏松多孔结构,不含细胞成分.HUA-SMC与ACCM相容性良好,HUASMC在与ACCM接触部位充分伸展,并沿ACCM窦隙活跃生长.9只裸鼠均存活,植入部位无感染,无植入物排斥发生.随培育时间延长,裸鼠体内ACCM逐渐降解,植入的HUASMC分化形成结构良好、交错排列的平滑肌组织.器官浴槽实验显示,构建组织对去氧肾上腺素和电刺激均表现出收缩功能,去氧肾上腺素和电刺激所诱导的最大收缩力分别为(3.64±0.18)和(2.50±0.21)g.结论 HUASMC作为种子细胞与ACCM复合可构建出具有一定形态和功能的组织工程海绵体平滑肌.     

男科学组织工程研究前景

利用组织工程技术进行细胞移植的设想 ,已经为男科领域的组织再建工作提供了许多可能。为改善、修复或替代现有组织的功能 ,组织工程的应用研究 ,已经在睾丸间质细胞、睾丸假体、阴茎海绵体、阴茎假体等方面开展。虽然大多数再建工作仍然停留在实验阶段 ,但是有些技术也被用于临床 ,并取得满意的结果。本文简要地综述了组织工程在男科学的应用     

Peyronie''''s病的外科治疗进展

Peyronie's病常并发阴茎弯曲和勃起疼痛,内科治疗不理想,须外科治疗,主要治疗方式有三类:包括阴茎白膜缩短术、斑块切开或切除补片修复术和阴茎假体植入术,因阴茎组织结构的特殊性,治疗效果均不理想,脱细胞细胞外基质和阴茎假体植入技术兴起与发展可望为Peyronie's病患者阴茎修复重建提供较为有前景的治疗模式.     

一种新的脱细胞阴茎海绵体基质的制备方法

目的探索一种新的脱细胞阴茎海绵体基质的制备方法。方法取健康壮年兔完整阴茎海绵体组织，以Triton-X100与NH3·H2O（氨水）联合提取法进行脱细胞处理。标本作HE染色，组织学观察分析脱细胞效果。结果脱细胞处理25天后，成功获得脱细胞海绵体基质。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-X100与NH3·H2O联合提取法可成功制备完整无细胞阴茎海绵体基质。     

兔阴茎海绵体脱细胞基质的构建

组织工程支架--脱细胞海绵体基质筛选方法有待于进一步研究[1].本研究旨在检测脱细胞过程中形态学和组织学的变化,探讨阴茎海绵体脱细胞基质制备.     

阴茎再造勃起功能重建的研究现状

勃起功能重建作为再造阴茎功能重建的重要组成部分,近年来在材料选择、手术方法及组织工程学等方面不断发展,阴茎再造术也获得了不断地改进和完善。本文对近几年阴茎再造勃起功能重建的进展,以及常用的阴茎支撑材料,包括自体组织支撑物、人工假体支撑物及组织工程材料等研究进展进行综述。     

泌尿外科组织工程技术的研究现状和展望

泌尿外科组织工程技术为泌尿道的组织、器官的微创修复和功能重建带来了新的希望。近年来，有关泌尿系组织工程的种子细胞和支架材料的研究以及组织工程化组织如肾脏、输尿管、膀胱、尿道、阴茎海绵体等的实验和临床研究报道层出不穷。其基本方法是应用细胞生物学和工程学的原理，将体外培养扩增后具有生物学活力及特定功能的细胞（种子细胞）与可降解生物支架材料复合，然后将该细胞-支架复合物（组织工程化组织）植入体内组织缺损部位，     

Kallistatin基因在大鼠阴茎海绵体中的表达研究

目的:探讨人组织激肽释放酶结合蛋白(kallistatin)在阴茎海绵体中的表达情况,为勃起功能调控和ED治疗寻找新的分子靶点。方法:采用qRT-PCR、Western印迹和免疫荧光技术检测kallistatin在大鼠阴茎海绵体组织的表达情况,同时检测主动脉中的表达水平进行对照。结果:qRT-PCR和Western印迹检测结果显示,Kallistatin mRNA和蛋白在大鼠阴茎海绵体组织中均有表达,且其表达水平与主动脉相比均无统计学差异(P>0.05);免疫荧光检测显示,Kallistatin表达于阴茎海绵体内皮细胞和平滑肌细胞,且定位于细胞胞质,与主动脉相比,两者间表达亦无明显差异(P>0.05)。结论:Kallistatin基因高表达于阴茎海绵体且定位于海绵体内皮细胞和平滑肌细胞内,提示kallistatin可能在海绵体内皮细胞和平滑肌细胞的细胞功能中发挥作用,从而参与阴茎勃起功能调控。     

国产可膨胀型单根圆柱体阴茎假体再植入术体会(附1例报道)

<正>勃起功能障碍(erectile dysfunction,ED)严重影响着患者的生活质量与心身健康。对于使用一线、二线治疗无效的或者希望永久解除困扰的ED患者,阴茎假体植入术(penile prosthesis implantation,PPI)已成为其临床治疗的金标准[1,2]。目前报道以双侧海绵体内可膨胀性阴茎假体植入术居多[3,4]。但当阴茎海绵体空间过度狭小、无法双侧同时植入阴茎海绵体时,可否设计成单根圆柱体,实施单根圆柱体的可膨胀阴茎假体植入术?我们近来收治1例外伤性器质性ED患者,行国产可膨胀型单根圆柱体阴茎假体再植     

阴茎海绵体纤维化：海绵体假体植入前直视下选择性海绵体组织切除

阴茎异常勃起、阴茎海绵体炎或假体感染引起的纤维化增加了海绵体假体植入术的困难，因不能充分扩张海绵体或需要行增加感染率的海绵体成形术。作者采用直视下海绵体多处切开后选择性切除纤维化组织，创造假体圆柱体所需管腔内径的手术方法。     

Human neural crest stem cells transplanted in rat penile corpus cavernosum to repair erectile dysfunction

OBJECTIVE

To investigate the feasibility of applying neural crest stem cells (NCSCs), with multipotent capacity, to repair injury in the penile cavernosum, the HNC10.K10 (K10) immortalized NCSC line was transplanted into the penile cavernosum of adult rats, as one of the causes of erectile dysfunction is damaged penile cavernous smooth muscle cells and sinus endothelial cells.

MATERIALS AND METHODS

The K10 human NCSC line was generated via transfection of primary cultured NCSC with a retroviral vector encoding v‐myc. K10 NCSCs were transplanted into the cavernosum of adult rats. The expression of cell type‐specific markers for endothelial cells (CD31 and von Willebrand factor), and specific markers for smooth muscle cells (smooth muscle cell actin, calponin, and desmin) was determined immunohistochemically in the penile cavernosum of rats 2 weeks after transplantation.

RESULTS

In the rat cavernosum, transplanted K10 NCSCs identified by human nuclear antigen labelling expressed cell type‐specific markers for endothelial cells (CD31 and von Willebrand factor), and specific markers for smooth muscle cells (smooth muscle cell actin, calponin, and desmin) 2 weeks after transplantation. Human NCSCs transplanted into the rat penile corpus cavernosum differentiated into endothelial cells or smooth muscle cells, as shown by their expression of cell type‐specific markers for the cell types.

CONCLUSION

It appears that NCSCs are an ideal cell source for reconstructing endothelial and smooth muscle cells in the corpus cavernosum in cell therapy for patients with erectile dysfunction.     

Evoked cavernous activity.

PURPOSE: Corpus cavernosum electromyography has been widely done to evaluate autonomic dysfunction in patients with erectile dysfunction. We assessed the value of corpus cavernosum electromyography, evoked cavernous activity and penile sympathetic skin responses for their accuracy in determining autonomic involvement in cases of erectile dysfunction. MATERIALS AND METHOD: We evaluated 75 men with erectile dysfunction by corpus cavernosum electromyography, evoked cavernous activity and penile sympathetic skin response tests at our neurourology laboratory. The etiology of dysfunction was vascular, neurogenic, psychogenic or mixed based on a detailed medical and sexual history, physical examination, electrophysiological and laboratory studies, penile color Doppler ultrasonography, and cavernosography and/or cavernosometry. Autonomic involvement was clinically assessed by systemic findings, such as orthostatic hypotension, impaired gastrointestinal motility, sinus dysrhythmia and secretomotor changes. A concentric electromyography needle placed in the right cavernous body was used to record corpus cavernosum electromyography and evoked cavernous activity. The right median nerve was stimulated electrically with 13 to 16 mA. to determine evoked cavernous activity and the penile sympathetic skin response. The latter response was recorded with silver disc electrodes placed on the left cavernous body. All tests were performed using an electromyography/evoked potential machine. We determined the relationships among corpus cavernosum electromyography, evoked cavernous activity and penile sympathetic skin response tests in respect to etiological factors. RESULTS: The 56 patients with normal corpus cavernosum electromyography activity had also evoked cavernous activity and a penile sympathetic skin response except for 1 with no penile sympathetic skin response but evoked cavernous activity. None of these patients had autonomic neuropathy. Of the 19 patients without corpus cavernosum electromyography activity 11 had evoked cavernous activity, including 10 with no autonomic neuropathy. The remaining 8 patients had no evoked cavernous activity, of whom 7 had autonomic neuropathy. A penile sympathetic skin response was recorded in 18 men with absent corpus cavernosum electromyography. CONCLUSIONS: Due to false-negative results on corpus cavernosum electromyography and penile sympathetic skin response testing evoked cavernous activity seems more reliable for determining autonomic involvement in the pathophysiology of erectile dysfunction.     

Expression of functional prostaglandin D (DP) receptors in human corpus cavernosum smooth muscle

Prostaglandin D(2) (PGD(2)) binds to specific G-protein coupled receptors (DP) and induces smooth muscle relaxation by stimulating the synthesis of intracellular cAMP. In this study, we examined the role of PGD(2) and DP receptors in regulating human penile smooth muscle contractility. We determined that human corpus cavernosum tissue and smooth muscle cells in culture expressed functional DP receptor and lipocalin-like prostaglandin D synthase by reverse-transcribed polymerase chain reaction (RT-PCR). Functional PGD synthase activity was confirmed by the synthesis of PGD(2) in human corpus cavernosum smooth muscle cells upon addition of exogenous arachidonic acid. Organ bath preparations of human corpus cavernosum tissue strips, contracted with phenylephrine, relaxed in a dose-dependent fashion to either PGD(2) or the DP selective agonist BW245C. Cultures of human corpus cavernosum smooth muscle cells treated with BW245C showed a two-fold increase in cAMP synthesis. These data are consistent with the expression of functional DP receptors in human corpus cavernosum. This suggests the presence of an intact prostanoid autocrine system that may play a role in regulating penile erectile function.     

Standardized evaluation of erectile dysfunction in 95 consecutive patients

We investigated 95 patients referred for erectile dysfunction by penile blood pressure measurement, the intracavernous papaverine test and Doppler investigation of the penile arteries. Furthermore, penile cutaneous perception threshold, bulbocavernosus reflex latency and somatosensory cortical evoked potentials of the pudendal nerve were measured. In selected cases cavernosometry, cavernosography and corpus cavernosum electromyography were performed. Doppler investigation of the cavernous arteries after papaverine injection was more reliable than penile blood pressure measurement in the diagnosis of arteriogenic erectile dysfunction. Decreased sensibility of the penis may be the sole factor responsible for inability to sustain an erection. Erectile dysfunction may be provoked by impaired function of the pudendal nerve. Penile cutaneous perception threshold measurement and corpus cavernosum electromyography are mandatory in the evaluation of neurogenic etiology. Cavernosometry and cavernosography are reliable methods in the determination of abnormal drainage from the corpus cavernosum.     

Activators of soluble guanylate cyclase for the treatment of male erectile dysfunction

Soluble guanylate cyclase (sGC) is an important enzyme in corpus cavernosum smooth muscle cells as it is one of the regulators of the synthesis of cGMP. The efficacy of sildenafil (Viagra) in the treatment of male erectile dysfunction indicates the importance of the cGMP system in the erectile response as the increased levels of cGMP induce relaxation of the corpus cavernosum. sGC is physiologically activated by nitric oxide (NO) during sexual stimulation, and its activity can be pharmacologically enhanced by several NO-donors. Agents like YC-1 can also activate sGC after binding to a novel allosteric site in the enzyme, a site different from the NO binding site. YC-1 can relax rabbit cavernosal tissue and it facilitates penile erection in vivo. This review summarizes the enzymology, biochemistry and pharmacology of this novel allosteric site and its relevance for the regulation of penile function. This type of sGC activators represent a new class of compounds with a different pharmacological profile in comparison to the classical NO-donors and they could be beneficial for the treatment of male erectile dysfunction.     

Electrical activity of corpus cavernosum during flaccidity and erection of the human penis: a new diagnostic method?

We investigated 7 normal men and 1 diabetic patient with erectile dysfunction. Electromyography electrodes were placed in the corpus cavernosum of the penis and electrical activity was recorded during flaccidity. With sexual arousal the activity decreased and tumescence was initiated. During tumescence and full erection the electrical activity of the corpus cavernosum almost ceased but in the diabetic patient (neurogenic impotence) an increase was observed. This discoordination might be the cause of the erectile dysfunction. Recording the electrical activity of the corpus cavernosum in patients with suspected neurogenic erectile dysfunction could become clinically valuable, since this is the first test possible to study the function of the autonomic motor system that normally regulates penile function.     

Extracorporeal shock wave therapy combined with engineered mesenchymal stem cells expressing stromal cell-derived factor-1 can improve erectile dysfunction in streptozotocin-induced diabetic rats

BackgroundFor erectile dysfunction (ED) in diabetes mellitus (DM) patients who have poor response to drugs, extracorporeal shock wave therapy (ESWT) and engineered mesenchymal stem cell (MSC) therapy have been studied as alternative treatment options. The objective of this study is to investigate whether ESWT in combination with stromal cell-derived factor-1 expressing engineered mesenchymal stem cell (SDF-1 eMSC) therapy can have synergistic effects on ED in streptozotocin-induced diabetic rats.MethodsFifty 8-week-old male Sprague-Dawley rats were randomly divided into five groups (N=10 per group): (I) Normal group, (II) DM ED, (III) DM ED + ESWT group, (IV) DM ED + SDF-1 eMSC group, and (V) DM ED + ESWT + SDF-1 eMSC group. Each groups were treated with bilateral injections of SDF-1 eMSC or ESWT following the experiment protocol for eight weeks.ResultsThe ratio of ICP/MAP was distinctly higher in the DM ED + ESWT + SDF-1 eMSC group than that in the DM ED group. Concentration of α-smooth muscle actin (α-SMA) was elevated the highest in the DM ED + ESWT + SDF-1 eMSC group. Additionally, ESWT increased the intensity of SDF-1 expression in the corpus cavernosum. ESWT + SDF-1 eMSC treatment also induced neuronal nitric oxide synthase (nNOS) and NO/cGMP expression in the corpus cavernosum. Furthermore, numbers of penile progenitor cells were increased in DM ED rats.ConclusionsCombined treatment of ESWT with SDF-1 eMSC treatment is more effective than by a single therapy. It could be used as a potential and effective synergistic treatment for DM ED.     

Adipocyte accumulation in penile corpus cavernosum of the orchiectomized rabbit: a potential mechanism for veno-occlusive dysfunction in androgen deficiency

Androgens are deemed to be critical for the development, growth, and maintenance of penile tissue as well as for erectile function. Androgens are also reported to inhibit differentiation of stroma progenitor cells into adipocytes and promote differentiation into smooth muscle. The objective of this study was to investigate whether androgen deprivation results in accumulation of adipocytes in the corpus cavernosum. Mature, New Zealand white male rabbits were subjected to sham surgery (control) or orchiectomy. Two weeks after surgery, erectile function was assessed by monitoring changes in intracavernosal blood pressure (ICP) in response to pelvic nerve stimulation. All ICP measurements were normalized to the mean systemic arterial blood pressure. In parallel studies, penile cross sections from control and orchiectomized rabbits were fixed and stained with either Masson's trichrome or hematoxylin and eosin to assess smooth muscle and connective tissue content. Alternatively, tissue sections were stained with Toluidine blue to assess accumulation of fat-containing cells. Orchiectomy resulted in loss of erectile function and penile atrophy, associated with reduced trabecular smooth muscle and increased connective tissue content. Most strikingly, tissue from orchiectomized animals exhibited accumulation of fat-containing cells (adipocytes) in the subtunical region of the corpus cavernosum. We hypothesize that androgen deprivation promotes differentiation of progenitor stroma cells into an adipogenic lineage producing fat-containing cells, thus altering erectile function.     

干细胞治疗阴茎勃起功能障碍的研究进展

阴茎勃起功能障碍(ED)是指男性反复或者持续性的难以达到和维持充分的阴茎勃起,无法完成性交或满意性活动的病理现象。海绵体神经(CN)损伤引起的勃起神经反射中断,是患者出现ED的直接原因,此外,CN损伤后,阴茎海绵体组织平滑肌细胞和内皮细胞凋亡增加,海绵体平滑肌纤维数量减少加重了ED的发生。因此,尽早干预CN损伤的病理过程,促进CN再生是治疗CN损伤性ED的关键。近年来,干细胞在ED治疗中的应用日益成为临床研究热点。现对胚胎干细胞(ESC)、间充质干细胞(MSCs)、肌源性干细胞(MDSCs)、脂肪干细胞(ADSCs)在ED治疗中的研究综述如下。     

Intracavernous indurations after injuries to the external genital parts caused by a motorcycle tank

We have experienced 2 cases of intracavernous induration after injuries to the external genital parts caused by a motorcycle tank. Case 1. A 28-year-old male was admitted to our department complaining of painless indurations of the penile radix. Cavernosography showed segmental filling defect in left corpus cavernosum. Because erectile disturbance was noted, resection of the induration was carried out. Microscopic section of the excised tissue showed only fibrosis. Case 2. A 20-year-old male visited our clinic with chief complaints of induration of the penile radix and erectile disturbance. Corpus cavernosography demonstrated filling defect in bilateral corpus cavernosum. We recommended the resection of the indurations, but the patient refused. A brief review on etiology and therapy of intracavernous fibrosis was made.