Decellularized porcine conjunctiva as an alternative substrate for tissue-engineered epithelialized conjunctiva

PurposeThe long-term success of visual rehabilitation in patients with severe conjunctival scarring is reliant on the reconstruction of the conjunctiva with a suitable substitute. The purpose of this study is the development and investigation of a re-epithelialized conjunctival substitute based on porcine decellularized conjunctiva (PDC).MethodsPDC was re-epithelialized either with pre-expanded human conjunctival epithelial cells (PDC + HCEC) or with a human conjunctival explant placed directly on PDC (PDC + HCEx). Histology and immunohistochemistry were performed to evaluate epithelial thickness, proliferation (Ki67), apoptosis (Caspase 3), goblet cells (MUC5AC), and progenitor cells (CK15, ΔNp63, ABCG2). The superior construct (PDC + HCEx) was transplanted into a conjunctival defect of a rabbit (n = 6). Lissamine green staining verified the epithelialization in vivo. Orbital tissue was exenterated on day 10 and processed for histological and immunohistochemical analysis to examine the engrafted PDC + HCEx. A human-specific antibody was used to detect the transplanted cells.ResultsFrom day-14 in vitro onward, a significantly thicker epithelium and greater number of cells expressing Ki67, CK15, ΔNp63, and ABCG2 were noted for PDC + HCEx versus PDC + HCEC. MUC5AC-positive cells were found only in PDC + HCEx. The PDC + HCEx-grafted rabbit conjunctivas were lissamine-negative during the evaluation period, indicating epithelial integrity. Engrafted PDC + HCEx showed preserved progenitor cell properties and an increased number of goblet cells comparable to those of native conjunctiva.ConclusionPlacing and culturing a human conjunctival explant directly on PDC (PDC + HCEx) enables the generation of a stable, stratified, goblet cell-rich construct that could provide a promising alternative conjunctival substitute for patients with extensive conjunctival stem and goblet cell loss.     

人胚雪旺细胞组织工程神经修复坐骨神经缺损的实验研究

目的：探索人胚雪旺细胞作为组织工程的种子细胞修复周围神经缺损的可行性。方法：通过组织工程方法用PLGA导管和polyglactin 910纤维负载人胚雪旺细胞预先构置好人工神经，然后用于修复大鼠20mm的坐骨神经缺损，并与神经切断后原位缝合以及用单纯的PLGA导管进行修复的实验组进行对照。通过活体肢体功能观察、靶器官肌肉测量、电生理检测、辣根过氧化物酶示踪、连续组织切片图像分析以及透射电镜等检查神经再生情况。结果：人工神经修复组神经再生良好，效果接近于神经原位缝合组，明显优于单纯的PLGA导管修复组。结论：人胚雪旺细胞构建的人工神经可以修复20mm的周围神经缺损。     

趾蹼血管分型的临床认识

目的分析120例足趾移植再造手指病例，对趾蹼间的血管形态和交通情况进行观察，并阐述其临床意义。方法临床观察120例156侧趾蹼，对第一跖骨背动脉和第一趾底动脉在趾蹼间的走向、口径以及两者的交通支情况进行记录并进行分型。结果按孙博的分型方法：Ⅰ型72侧，占46．2％；Ⅱ型57侧，占36．5％；Ⅱ型7侧，占4．5％；Ⅳ型3侧，占1．9％；Ⅴ型15侧，占9．6％；另有2侧（占1．3％）第一跖骨背血管口径〉1mm，虽发出趾背动脉供养蹲趾胫侧和第二趾腓侧，但在趾蹼内与趾底血管无吻合，故称之为孙博Ⅵ型。结论（1）从解剖形态看应增加孙博Ⅵ型较全面；（2）从临床实用角度出发，趾蹼间血管可以简单分为三型：Ⅰ型交通支口径在0．5mm以上。Ⅱ型交通支在0．5mm以下。Ⅲ型无交通；（3）手术中采用第一套或第二套供血系统，首先要看趾蹼间血管的交通支，即按本组的分型属Ⅰ型采用第一套供血系统，Ⅱ、Ⅲ型采用第二套供血系统。     

树枝状静脉移植在手部创伤中的应用

目的探讨手掌严重碾挫伤应用血管移植手术方法的选择。方法对12例手掌碾挫伤伴血管损伤的患者，采用树枝状浅表静脉倒转后多级移植修复缺损的手掌动脉，同时对缺损皮肤一期行植皮或皮瓣修复。结果术后12例创面全部Ⅰ期愈合，手指血循环好。术后随访3～22个月，手功能评定： 优5例，良4例，可2例，差1例，优良率为75％。手掌恢复了良好的外形和功能，能完成手指的捏、握、对掌和对指功能。结论对严重的手掌毁损伤，利用树枝状静脉移植修复缺损的血管，能获得较好的手功能和外形。     

大鼠异体肢体移植术后急性排斥反应阶段血管内皮细胞ICAM-1表达的动态变化

Objective To investigate the dynamic changes of intercellular adhesion molecule-1 (ICAM-1) expression in endothelial cells (EC) during hindlimb allograft acute rejection in rats and the inhibitory effect of cyclospofine A (CsA) on the acute rejection. Methods The rat model of hindlimb allograft was developed. The rats were randomly divided into following groups: control group (Wistar→Wistar), rejection group ( Sprague-Dawley→Wistar) and CsA-treated group (Sprague-Dawley→Wistar). At postoperative day 1, day 4 and day 7samples of the femoral artery from the allograft limb were harvested to observe the pathologic changes. ICAM-1 expression in EC was quantified using immunohistochemical assay. Results Slight swelling and weak ICAM-1expression of EC were observed in the control group. In the rejection groups, obvious EC swelling and massive lymphocyte infiltration were seen. ICAM-1 expression in EC was significantly stronger and elevated. In CsA-group only mild infiltration of lymphocytes was seen and ICAM-I expression in EC was also much weaker.Conclusion During rat hindlimb allograft acute rejection the expression of ICAM-1 in EC was closely related to the occurrence and development of acute rejection. CsA could reduce the expression of ICAM-1 in EC and hence inhibit acute rejection of composite allograft.     

微囊化异种甲状旁腺组织移植的研究

目的探讨微囊化异种甲状旁腺(PTG)组织移植对Wistar大鼠甲状旁腺功能低下的治疗作用及微囊的通透性。方法 40只去甲状旁腺Wistar大鼠随机分为微囊组、非微囊组、空囊组、空白对照组。用海藻酸-钡交联微囊包裹兔甲状旁腺组织,移植至Wistar大鼠肾包囊。移植后每隔2周取血测血钙,移植后第16周取移植物进行透射电镜检查及T淋巴细胞、大鼠IgG抗体的免疫组织化学染色。结果微囊组移植后第4周血清钙由(1．62±0．04)mmol／L恢复至正常水平 (2．2～2．6)mmol／L,9例维持至观察期结束(P<0．01),非微囊组、空囊组及空白对照组血清钙差异无统计学意义(P>0．05)。第16周取出移植物检测显示移植物活性良好,微囊周围可见较多T 淋巴细胞浸润,囊壁及囊内IgG抗体染色阳性。结论海藻酸-钡交联微囊可对甲状旁腺组织起到有效的保护作用,使甲状旁腺组织较长时间存活并发挥正常功能。但海藻酸-钡交联微囊并未减轻受体免疫排斥反应的激活且未有效的免疫隔离IgG抗体。     

尿激酶溶栓对急性心肌梗死患者单核细胞组织因子表达的影响

目的 :了解冠心病患者外周循环单核细胞组织因子 (TF)表达与冠心病病情的关系 ;观察溶栓治疗对急性心肌梗死患者单核细胞TF表达的影响。方法 :测定了 2 2例稳定型心绞痛 (SA)患者 ,2 1例不稳定型心绞痛(UA)患者 ,1 5例急性心肌梗死 (AMI)患者用尿激酶溶栓前及溶栓后 ,1 8例健康人外周循环单核细胞组织因子阳性率。单核细胞组织因子阳性率测定用免疫荧光染色及流式细胞术。结果 :SAP组 ,UAP组和AMI组单核细胞TF阳性率高于对照组 ,UAP组和AMI组单核细胞TF阳性率高于SAP组 ,单核细胞TF阳性率与冠心病病情显著相关。急性心肌梗死患者溶栓后 2h单核细胞TF阳性率升高。结论 :外周血单核细胞TF的表达与冠心病病情严重程度密切相关 ;急性心肌梗死患者溶栓治疗后单核细胞TF的表达进一步增强 ,这可能是溶栓后凝血酶活化的原因之一。     

应用皮瓣移植修复足跟部皮肤软组织缺损

目的通过8种不同皮瓣的临床应用,总结足跟部皮肤软组织缺损的修复效果。方法采用了游离足背皮瓣、游离背阔肌皮瓣、游离肩胛皮瓣、桥式胫后动脉岛状皮瓣、桥式携带背阔肌皮瓣或桥式携带肩胛皮瓣、胫后动脉岛状皮瓣、胫后动脉皮支岛状皮瓣、跖底内侧动脉岛状皮瓣,对足跟部皮肤软组织缺损进行修复的临床应用。结果95块皮瓣修复足跟部皮肤软组织缺损,成活93块,成功率为97.9%。随访1～15年,治疗效果满意。结论应用皮瓣移植的方法,才能使足跟部皮肤软组织缺损得到有效的修复。     

齿槽裂修复方法的研究进展

目的 探讨齿槽裂修复治疗的目的、方法以及治疗时机的选择。方法 查阅1950年至2006年有关齿槽裂修复的文献，归纳文献中报道的不同方法，并评价其各自的优缺点。结果 齿槽裂修复的主要目的：关闭口鼻瘘；建立稳定、连续的上颌骨牙弓；为牙齿萌出提供基础；为上唇和鼻底提供稳定支架。主要治疗方法：植骨术；牵引成骨技术；组织工程骨和生长因子应用；引导骨再生技术。患者最佳的手术治疗时机是9～11岁时混合牙列期。结论 在9～11岁混合牙列期手术，以髂骨松质骨为移植材料被认为是修复齿槽裂的主要手段。牵引成骨技术、组织工程技术和引导骨再生技术，将是齿槽裂修复的新方向。