PLGA生物细胞支架种植自体成纤维细胞对阴茎增粗的安全性有效性研究

目的观察可吸收性PLGA细胞支架(Maxpol-T)种植、自体成纤维细胞移植行阴茎增粗的安全性和有效性。方法雄性成年新西兰兔分为假手术对照组(A组)、单纯Maxpol-T移植组(B组)、种植自体成纤维细胞的Maxpol-T移植组(C组)。麻醉下切取阴囊皮肤,分离、鉴定和扩增成纤维细胞种植到Maxpol-T后,移植于兔阴茎深筋膜下,连续观察6个月,评估阴茎增粗的效果及其安全性。结果C组和B组阴茎直径较A组显著增加(P<0.01),C组术后1、3和6个月阴茎直径增加稳定,无显著变化(P>0.05),未发生明显排斥及炎症反应,局部组织病理学观察发现皮下组织增厚,新生结缔组织增生和新生血管,无炎症细胞显著浸润。但是,B组阴茎直径在术后第3个月和6个月有显著降低的趋势(P<0.01)。结论应用自体成纤维细胞种植Maxpol-T细胞支架较单纯Maxpol-T支架行新西兰兔阴茎增粗更为安全有效,无明显排斥反应和炎症反应,推荐前者作为安全有效的组织工程细胞支架材料。     

应用PLGA生物支架种植自体成纤维细胞进行阴茎增粗的临床研究

目的 观察PLGA支架(Maxpol-T)种植自体成纤维细胞移植进行阴茎增粗的临床安全性和有效性.方法 筛选知情同意下自愿参加临床研究的成年阴茎短小患者,局麻下获取阴囊真皮组织,分离、培养、鉴定、扩增成纤维细胞与Maxpol-T支架共培养后植入阴茎深筋膜下增粗阴茎.分别在手术后1,3,6个月随访,利用量尺和彩色超声比较手术前后阴茎周径变化,利用IIEF5评分表和副作用观察表评估其临床安全性,进行统计学分析.结果 参加临床实验患者24名,完成临床实验22名(91.67%),平均年龄(31.1±8.4)岁.手术前阴茎疲软状态平均周径(8.18±0.83)cm, 阴茎勃起状态平均周径(10.26±1.22)cm.手术后1,3,6个月后阴茎疲软状态平均周径分别为(11.79±1.18)cm,(12.20±1.24)cm,(12.19±1.27)cm,与术前相比差异具有统计学意义(P<0.001).阴茎周径增加2cm以上的临床有效率为95.45%.术后6个月患者满意度调查结果非常满意68.18%,很满意27.27%,满意4.55%,与术前相比差异具有统计学意义(P<0.001).2名患者术后切口乙,I期愈合,其他患者甲,I期愈合.所有患者手术前后IIEF5评估勃起功能无变化.结论 利用生物可降解细胞支架(Maxpol-T)种植体自体成纤维细胞后,手术植入阴茎深筋膜下行阴茎增粗临床安全有效,临床有效率95.45%.手术后迟发性阴茎皮下少量积液发生率8.70%,可能与个别患者对PLGA过敏反应有关.     

硅胶片置入及自体颗粒脂肪注射阴茎延长增粗术

目的探讨防止阴茎延长后回缩的阴茎延长增粗手术。方法在阴茎根部背侧V形切开,将全部阴茎浅悬韧带及部分深悬韧带切断,为了防止切断的阴茎韧带与耻骨联合重新粘连而回缩,根据阴茎悬韧带附着于耻骨联合的面积,采用宽1.5～2.5cm,长2.3～3.6cm,厚2～3mm的硅胶片置入,缝合于其表面,采用自体颗粒脂肪30～48ml,行阴茎背侧深筋膜表面注射。结果16例患者,阴茎静态下延长长度为1.8～5.1cm,平均2.91cm;增粗直径0.6～1cm,平均0.85cm,观察期为3个月～2年,回缩率小于8%,阴茎增粗后缩小率低于10%。结论硅胶片置入结合自体颗粒脂肪注射是一种理想的阴茎延长增粗术,可有效防止阴茎延长后的回缩。     

阴茎增粗术的研究进展

近20年来,人们对精神生活以及性生活质量的要求有所提高,由于广告和媒体对阴茎大小重要性的渲染,使阴茎增大手术的市场需求量持续增长.仅在美国,从1991年至1998年就有10000人行阴茎增大术[1].目前,用于阴茎增粗的填充材料主要包括自体组织移植和各种人工合成材料的替代物,但都存在不同程度的缺陷,应用受到了一定限制.     

真皮脂肪复合组织条移植在阴茎增粗延长术中的应用

目的 探讨应用真皮脂肪复合组织条游离移植行阴茎增粗延长术的方法及临床疗效.方法 2004年5月至2010年12月,应用自体真皮脂肪组织条游离移植加阴茎根部切口阴茎延长术,对24例患者行阴茎增粗延长.采用阴茎根部切口,切断阴茎浅悬韧带及部分深悬韧带进行阴茎延长,同时在阴茎深筋膜与海绵体白膜间游离移植长6.0～9.5 cm,宽1.2～1.5 cm,厚度0.6～0.8 cm真皮脂肪复合组织条,自耻骨前区腔隙直至阴茎冠状沟,以增粗阴茎.结果 24例中23例一期愈合,1例发生切口脂肪液化,换药后愈合.阴茎常态及勃起后形态良好,勃起时延长长度为2.5 ～4.8 cm,平均3.2 cm;阴茎周径延长长度为1.8～3.0 cm,平均2.4 cm.本组18例获得3个月至5年的随访,所有患者对术后阴茎形态满意,对性生活质量满意度高,未见手术并发症.结论 真皮脂肪复合组织条移植加阴茎悬韧带切断术是一种较好的阴茎增粗延长术式,手术安全,效果肯定.     

阴茎延长同期行脱细胞异体真皮基质补片双平面植入阴茎增粗术

目的探讨阴茎延长同期行脱细胞异体真皮基质（acellular dermal matrix,ADM）补片双平面植入阴茎增粗术的方法与效果。方法采用阴茎根部倒V形切口。离断阴茎浅悬韧带后,沿阴茎纵轴切开Dartos筋膜,在其深面向远端分离。距冠状沟1．5～2．0cm处环形切开Buck筋膜,将补片前部植于Buck筋膜与白膜间,后部植于Dartos筋膜与Buck筋膜间。缝合Dartos筋膜切口,V—Y成形术闭合阴茎根部切口。结果35例术后无1例发生阴茎皮肤坏死、补片外露并发症。25例获随访6～24个月,对阴茎外形均感满意;无1例出现补片移位或皱褶、阴茎头感觉异常;其中21例已婚者均感性生活满意。结论经阴摹根部切口行脱细胞异体真皮基质补片双平面植入阴茎增粗术,通过调整补片植入层次,在确保补片足够的组织覆盖及刚茎皮肤血供情况下,在I期内行延长并增粗阴：拳术,具有并发症少、疗效满意的优点。     

应用脱细胞异体真皮移植增粗阴茎的临床研究

目的：研究应用脱细胞异体真皮移植增粗阴茎的效果、手术并发症及移植物应植入的最佳解剖层次。方法：应用脱细胞异体真皮植入增粗阴茎25例,A组13例,移植物植入Buck’s筋膜深面,白膜浅面;B组12例,移植物植入Dartos筋膜深面,Buck’s筋膜浅面。结果：术后阴茎中段周径增大1．1～3．2cm,A、B两组均无阴茎畸形、勃起功能障碍、脱细胞真皮外露等并发症发生,A组4例早期出现龟头麻木,3个月后恢复正常,B组无类似病例。结论：应用脱细胞异体真皮移植增粗阴茎效果明显,手术创伤小,无供区损伤,但植入Buck’s筋膜深面可引起术后早期龟头麻木。     

自体脂肪间充质干细胞移植治疗肛瘘的实验研究

为探讨自体脂肪间充质干细胞(ADMSCs)移植在肛瘘(FIA)创面愈合中的作用,是否可分化为多种细胞类型.通过橡皮筋结扎手术建立了日本家兔FIA模型,实验组移植自体ADMSCs,对照组移植磷酸盐缓冲液(PBS);治疗观察14 d,取FIA组织进行病理检查.结果显示,ADMSCs移植组所有瘘管均愈合,无明显瘢痕;对照组3...     

兔自体骨髓间充质干细胞体内复合移植的成骨研究

目的 观察兔自体骨髓间充质干细胞（MSCs)体内复合移植的成骨能力，以寻求理想的MSCs载体。方法将10只新西兰大白兔随机分成A、B两组，从自体骨髓中分离出MSCs，体外培养并扩增后分别与相同大小的小牛脱钙骨（DBCB），自固化磷酸钙人工骨（CPC）复合移植于两组大白兔左侧骶棘肌中，同时右侧骶棘肌分别植入相同大小的DBCB，CPC作空白对照；16周后取出标本，观察成骨情况并行组织学检查。结果 A组5例：MSCs DBCB侧均发现有新骨组织形成，单纯DBCB侧3例被完全吸收，降解，2例大部分吸收，降解；B组5例；MSCs CPC侧和单纯CPC侧均未见骨组织形成，植入物也无明显吸收，结论 自体骨髓MSCs在适合载体负载下可在体内自动分化成骨，DBCB是MSCs的良好载体之一。     

应用腹膜管联合皮瓣行兔阴茎再造的初步研究

目的：建立皮瓣包绕游离腹膜管再造阴茎的动物模型，探索在阴茎再造术中使用腹膜再造尿道的新方法。方法：成年雄性新西兰大白兔24只，随机分成腹膜组和腹膜皮片组各12只。再造阴茎时，腹膜组中将腹膜浆膜面向内缝合成腹膜管替代尿道，腹膜皮片组则在腹膜管前端吻接一段1cm长的阴囊皮片，形成腹膜皮片复合管，并以此管替代尿道，两组均以腹壁浅血管筋膜蒂皮瓣包绕再造尿道形成阴茎体，观察阴茎成活和尿道通畅情况。结果：再造阴茎全部成活，无尿道狭窄发生，作为尿道衬里的腹膜浆膜面光滑、湿润、富有光泽，未见溃疡及纤维化。腹膜组8只兔尿道口闭合，同时伴有尿瘘。腹膜皮片组3只兔各1处尿瘘。结论：使用皮瓣包绕游离腹膜管再造阴茎的方法可以成功构建阴茎再造的动物模型，为游离腹膜联合皮瓣转移阴茎再造术的临床应用提供了实验依据。     

Tissue engineering penoplasty with biodegradable scaffold Maxpol-T cografted autologous fibroblasts for small penis syndrome

In this study, we investigated the safety and efficacy of a poly acid-co-glycolide biodegradable scaffold (Maxpol-T) coated by autologous fibroblasts (AF) for penile girth enlargement in small penis syndrome (SPS). Eighty patients with SPS were enrolled in a clinical study at 2 medical centers; 69 patients completed the study protocol. Scrotal skin was harvested under local anesthesia, and AFs were cultured and seeded on a Maxpol-T scaffold; the cografted scaffold was implanted under the Buck's fascia of penile shaft via a circumcising incision. Patients were followed up at 1, 3, and 6 months to evaluate penile girth changes. Patient satisfaction was assessed via Visual Analogue Scale and scored on the International Index of Erectile Function-5 (IIEF-5). Mean preoperative penile girth in the flaccid and erect state was 8.18 ± 0.83 cm and 10.26 ± 1.22 cm, respectively. At the 6-month postoperative follow-up, mean penile girth in the flaccid and erect state was increased to 12.19 ± 1.27 cm and 13.18 ± 1.31 cm, respectively (P < .001 for change in both flaccid and erect state). Sixty-five patients (94.2%) reported satisfaction with the procedure. Among them, 4 cases (5.8%) were dissatisfied, 7 cases (10.1%) were satisfied, 26 cases (37.7%) were very satisfied, and 32 cases (46.4%) were extremely satisfied. All men maintained IIEF-5 scores of more than 22. Complications included prolonged subcutaneous edema in 3 patients (4.3%) and pinpoint erosion at the suture area in 3 patients (4.3%). Implantation of autologous fibroblasts seeded on a Maxpol-T collagen scaffold holds promise as a safe and novel technique for penile girth enhancement in patients with SPS.     

自体脂肪移植阴茎整形术

目的：自体脂肪移植技术已经广泛用于整形美容手术，探讨自体脂肪移植在阴茎延长和增粗术中的临床应用方法和效果。方法：用注射器在患者的腹部、 大腿内侧、臀部吸取脂肪，处理吸取的脂肪后，环绕阴茎，用注射器均匀注射于阴茎皮下。结果：45例患者，单纯脂肪移植29例；脂肪移植＋阴茎延长15例。手术后观察1～6个月。效果良好44例；脂肪液化、感染1例。结论：自体脂肪移植用于阴茎增大整形的手术的效果良好，手术后良好的包扎和塑形是手术成功的关键因素。     

Tissue engineered heart valves: autologous cell seeding on biodegradable polymer scaffold

We previously reported on the successful creation of tissue-engineered valve leaflets and the implantation of these autologous tissue leaflets in the pulmonary valve position. Mixed cell populations of endothelial cells and fibroblasts were isolated from explanted ovine arteries. Endothelial cells were selectively labeled with an acetylated low-density lipoprotein marker and separated from fibroblasts using a fluorescent activated cell sorter. A synthetic biodegradable scaffold consisting of polyglycolic acid fibers was seeded first with fibroblasts then subsequently coated with endothelial cells. Using these methods, autologous cell/polymer constructs were implanted in 6 animals. In 2 additional control animals, a leaflet of polymer was implanted without prior cell seeding. In each animal, using cardiopulmonary bypass, the right-posterior leaflet of the pulmonary valve was resected completely and replaced with an engineered valve leaflet with (n = 6) or without (n = 2) prior cultured cell seeding. After 6 h and 1, 6, 7, 9, and 11 weeks, the animals were sacrificed and the implanted valve leaflets were examined histologically, biochemically, and biomechanically. Animals receiving leaflets made from polymer without cell seeding were sacrificed and examined in a similar fashion after 8 weeks. In the control animals, the acellular polymer leaflets were degraded completely leaving no residual leaflet tissue at 8 weeks. The tissue-engineered valve leaflet persisted in each animal in the experimental group; 4-hydroxyproline analysis of the constructs showed a progressive increase in collagen content. Immunohistochemical staining demonstrated elastin fibers in the matrix and factor VIII on the surface of the leaflet. The cell labeling experiments demonstrated that the cells on the leaflets had persisted from the in vitro seeding of the leaflets. In the tissue-engineered heart valve leaflet, transplanted autologous cells generated proper matrix on the polymer scaffold in a physiologic environment at a period of 8 weeks after implantation.     

A biodegradable composite scaffold for cell transplantation.

Cell transplantation is rapidly becoming a therapeutic option to treat disease and injury. However, standard techniques for cell seeding on non-woven polymer meshes or within gels may not be suitable for immediate implantation or surgical manipulations of freshly isolated cells. Therefore, a biodegradable composite system was developed as a way to rapidly entrap cells within a support of predefined shape to potentially facilitate cell delivery into a target site (e.g. meniscal tears in the avascular zone). The composite construct consisted of freshly isolated cells, in this case pig chondrocytes, entrapped in a fibrin gel phase and dispersed throughout the void volume of a polyglycolic acid (PGA) non-woven mesh. Composites were cultured for up to 4 weeks. In vitro degradation of fibrin gel was evaluated via gel-entrapped urokinase. At 28 days in culture, glycosaminoglycan (GAG) content per cell in the composite scaffolds was 2.6 times that of the PGA-only cell construct group and 88% that of native pig cartilage. Total collagen content per cell in the composite scaffolds was not significantly different from the PGA-only cell construct group (P > 0.02) and represented 40% of the value determined for native cartilage. Varying the concentration of entrapped urokinase could effect controlled degradation of fibrin gel.     

骨髓基质干细胞片层复合聚乳酸羟基乙酸支架构建组织工程骨

目的 制备骨髓基质干细胞片层,探讨其对组织工程骨成骨的作用.方法 培养骨髓基质干细胞(BMSCs)并向成骨方向诱导,利用温度反应性培养皿制备骨髓基质干细胞片层,光学及电子显微镜下观察其形态.将经过预处理的三维多孔聚乳酸羟基乙酸(PLGA)支架注射BMSCs悬液并复合干细胞片层后植入犬左侧下颌骨缺损,为实验侧;将未复合细胞片层的支架植入右侧,为对照侧.16周后,对新生骨行影像学及组织学观察分析.结果 片层中细胞排列紧密,生长活跃.实验侧吸光度值高于对照侧(P＜0.05),并可见较多哈弗系统及板层骨样结构.结论 经温度反应性培养皿制备的干细胞片层结合PLGA支架可形成具有板层状结构的组织工程骨.     

新型多孔PLGA/HA复合支架体外细胞相容性的实验研究

目的研究新型多孔聚乳酸乙醇酸/羟基磷灰石(PLGA/HA)支架材料的体外细胞相容性。方法采用贴壁法对兔骨髓基质细胞(BMSCs)进行体外矿化诱导培养,扩增后与实验A组(含5%HA的PLGA/HA),实验B组(含10%HA的PLGA/HA)及对照C组(仅含PLGA)分别进行体外复合培养;并通过定性及定量法检测BMSCs在材料表面的粘附能力、增殖活力,验证细胞材料复合体的成骨活性。比较分析各组之间的差异。结果兔BMSCs在每组材料的表面均能生长,经体外诱导后在支架材料的表面形成钙结节,A、B组细胞的粘附及增殖能力均强于C组(P<0.05),A、B组之间无差异。结论兔BMSCs与新型多孔PLGA/HA支架材料有良好的相容性。     

新型多孔PLGA/HA复合支架体外细胞相容性的实验研究

目的研究新型多孔聚乳酸乙醇酸／羟基磷灰石（PLGA/HA）支架材料的体外细胞相容性。方法采用贴壁法对兔骨髓基质细胞（BMSCs）进行体外矿化诱导培养，扩增后与实验A组（含5％HA的PLGA／HA），实验B组（含10％HA的PLGA／HA）及对照C组（仅含PLGA）分别进行体外复合培养；并通过定性及定量法检测BMSCs在材料表面的粘附能力、增殖活力，验证细胞材料复合体的成骨活性。比较分析各组之间的差异。结果兔BMSCs在每组材料的表面均能生长，经体外诱导后在支架材料的表面形成钙结节，A、B组细胞的粘附及增殖能力均强于C组（P〈0．05），A、B组之间无差异。结论兔BMSCs与新型多孔PLGA／HA支架材料有良好的相容性。     

Abdominal wall repair using a biodegradable scaffold seeded with cells

Background/Purpose

The repair of large abdominal wall defects is still a challenge for pediatric surgeons. Synthetic materials, however, may lead to high complication rates. This study was aimed at applying tissue-engineering methods to abdominal wall repair.

Methods

3T3 mouse fibroblasts were expanded in vitro. In the next step, a biodegradable material—polyglycolic acid (PGA)—was actively seeded with 10⁷ cells/scm of PGA scaffold. Culture medium (Dulbecco's Modified Eagle's Medium with 10% fetal bovine serum) was changed every 6 hours after seeding cells on PGA fibers. Under general anaesthesia, C57BL/6J black mice underwent creation of a 2 × 3-cm abdominal wall defect (60%-70% of abdominal surface). The defect was repaired in the experimental group with the fibroblast-seeded PGA scaffold. In the first control group, the defect was covered with acellular PGA, and in the second control group, by skin closure. Animals were killed after 30 days to assess the histologic and gross findings.

Results

No abdominal hernia was found in animals repaired with cell-seeded and acellular scaffolds. All animals with skin closure died within 7 days. In every case, tissue-engineered construct was thicker then in controls. Histologic and gross examination revealed a good neovascularisation in tissue-engineered abdominal walls comparing to the acellular matrix. There was no intensive scar formation between abdominal wall and skin.

Conclusions

Engineered soft tissue constructs can provide structural replacement of severe and large abdominal wall defects. Tissue engineering in the near future will possibly enter clinical practice.