组织工程化人工神经构建的方法研究

目的：探索用组织工程方法构建人工神经的可行性。方法：用鼠尾胶和层粘蛋白共同修饰的polyglytin 910纤维以及生物膜作为支架与成年兔雪旺细胞培养2周。观察细胞生长迁移情况。结果：雪旺细胞能在共聚物纤维上迁移包裹，均匀分布于支架之间及吸附在支架表面。雪旺细胞在生物膜上能吸附生长。结论：成年雪旺细胞可以在修饰后的polylytin910纤维和生物膜上得到扩增，二者共同形态的三维立体结构具有人工神经的基本特性。     

许旺细胞在聚羟基乙酸纤维上三维定向培养

目的 为制备组织工程化的神经桥接物。方法 将体外扩增的许旺细胞（ＳＣｓ）接种在纵向排列的、可吸收材料ＰＧＡ纤维支架上，并对这种立体培养的细胞进行光镜、电镜和免疫组化观察，结果 发现许旺细胞可与聚羟基乙酸（ＰＧＡ）纤维良好吸附。并在其上边分裂边迁移从而形成了类似Ｂｕｅｎｇｎｅｒ带的纵向排列的细胞链，且其间含有一定的间隙，同时细胞大量分泌的层粘蛋白（ＬＮ）沉积在纤维表面形成了纵向的、类似变性神经的结构     

用组织工程方法桥接周围神经缺损的实验研究

目的 寻找修复周围神经缺损的新的有效替代材料。方法 将体外培养扩增的许旺细胞接种在生物可吸收材料聚羟基乙酸纤维支架上培养２周,形成一种新的神经桥接物,并对其进行了观察和动物移植试验。结果 发现接种的许旺细胞可在聚羟基乙酸纤维上良好吸附、迁移形成一种新型的、非管状的、具有三维纵向细胞排列和导层粘蛋白通道的组织工程化神经桥接物,用其修复近交的Ｗｉｓｔａｒ大鼠１５ｍｍ坐骨神经缺损时,取得了接近自体神经移     

乙交酯-丙交酯共聚物支架-细胞复合体移植对大鼠损伤脊髓的修复作用

目的 观察神经干细胞、雪旺细胞和组织工程材料乙交酯-丙交酯共聚物共移植后对大鼠损伤脊髓形态和功能的修复作用.方法 36只Wistar大鼠,随机数字法分为乙交酯-丙交酯共聚物移植组、神经干细胞/乙交酯-丙交酯共聚物绀和神经干细胞+雪旺细胞/乙交酯-丙交酯共聚物组.体外培养、鉴定胚胎脊髓源神经干细胞和雪旺细胞,制备和构建乙交酯-丙交酯共聚物支架细胞复合体并移植到大鼠脊髓T9半横断损伤部位,应用BBB行为评分和电生理技术在术后4、12周评价大鼠脊髓功能的恢复情况;应用透射电镜、HE染色和免疫组织化学染色方法在形态结构上观察轴突和髓鞘再生情况,以及神经干细胞在脊髓内的存活、迁移和分化情况.结果术后4、12周,细胞移植组的BBB评分较对照组明显提高(P＜0.05);细胞移植组的体感诱发电位和运动诱发电位波幅较对照组都有所好转.术后12周移植材料正中横断面透射电镜可见新生的无髓及有髓神经纤维;脊髓标本免疫组织化学染色显示移植的神经十细胞呵以在宿主脊髓内存活、迁移并分化成神经元和少枝胶质细胞,未分化成星形胶质细胞.结论 神经干细胞、雪旺细胞和组织工程材料乙交酯-丙交酯共聚物共移植可以促进半横断损伤的大鼠脊髓轴突再生,改善肢体的运动功能.     

雪旺细胞种于医用组织引导再生胶原膜构建自体人工神经的实验研究

目的;设计并构建一种新型的神经引导导管。方法：将兔雪旺细胞种到用成粘蛋白多孔医用组织引导的再生胶原膜支架培养2周后,用倒置显微镜、扫描电镜观察雪旺细胞吸附、生长迁移情况;以雪旺细胞胶原膜管的形式植入体内,观察它诱导神经再生的能力。结果：成年兔雪旺细胞在医用组织引导再生胶原膜上生长良好,并均匀分布于支架表面,且基质分泌旺盛。体内模型发现神经已通过移植物长入远端,胶原膜已吸收。结论：雪旺细胞可以在多聚医用组织引导再生胶原膜上得到扩增,种有雪旺细胞的医用组织引导再生胶原膜导管可以形成一个诱导神经轴突再生的微环境,形成的三维立体结构具有人工神经的基本特性,为组织工程方法修复长段神经缺损提供了研究基础。     

包埋后的聚羟基乙酸与软骨细胞体外培养实验研究

目的:通过卵磷脂和多聚赖氨酸分别和共同包埋以聚乳酸固定成形的聚羟基乙酸支架与软骨细胞体外培养,来观察其亲水性和对细胞吸附力的改变以及对细胞功能的影响。方法:将软骨细胞种于上述支架体外培养,通过倒置显微镜及扫描电镜观察支架的亲水性、对细胞的吸附力及基质产生情况。结果:支架以卵磷脂包埋后细胞悬液易浸入到支架内;以多聚赖氨酸包埋后细胞易吸附在支架纤维表面;以卵磷脂和多聚赖氨酸共同包埋后细胞均匀分布于支架纤维之间及吸附在支架纤维表面,且基质产生旺盛。结论:卵磷脂具有增强支架亲水性作用;而多聚赖氨酸除增加支架对细胞的吸附力外,还具有促进细胞功能的作用。以卵磷脂和多聚赖氨酸共同包埋支架可能是组织工程技术中较理想的支架之一。     

组织工程化人工神经内部支架及其生物相容性研究

目的 研制组织工程化人工神经内生长雪旺细胞的内部支架结构。方法 用６－０的生物可吸收Ｖｉｃｒｙｌ或ＰＤＳ纤维，在硅胶圈上纺织成三维立体网架，经Ｍａｔｒｉｇｅｌ蛋白胶涂层，与１０＾６／ｍｌ新生或成年鼠雪旺细胞共同培养３ｄ。Ｌｉｖｅ／Ｄｅａｄ荧光染色和扫描电镜观察细胞贴附情况。将１０根经Ｍａｔｒｉｇｅｌ蛋白胶涂层的Ｖｉｃｒｙｌ或ＰＤＳ纤维，植入１．５ｃｍ长的胶原神经管内，然后将其桥接于Ｌｅｗｉｓ鼠从骨神经缺损处。术后５周取材作组织学透射电镜观察。结果 在三维立体网架上，雪旺细胞沿纤维纵轴贴附伸展，并围绕纤维形成多条细胞链。荧光染色显示在纤维上有纵行的活细胞带，并见到多层重叠现象。组织工程倾人工神经在体内５周，含有涂层纤维的胶原神经管表面为为有丰富的新生血管和软组织所取供。神经束已沿生物纤维越过神经管中点。结论 培养     

神经轴突延长模型的建立

目的：建立体外延长人类神经轴突的方法，寻找合适的支持周围神经移植的基质材料。方法：将人胚背根神经节（dorsal root gangim,DRG)神经元细胞种在股原膜支架上，用自制的神经轴突延长装置对其延长至10cm以上。观察神经元细胞及轴突的生长情况，测定轴突延长后轴膜的渗透性。结果：人胚神经元细胞经过延长后可保持其正常形态及维持轴膜的功能。结论：该模型的建立为研究延长神经轴突构建人工神经提供了依据。     

经胶原包埋、修饰的聚羟基乙酸与软骨细胞体外培养的实验研究

目的：探讨经胶原包埋、修饰后的聚羟基乙酸（poloyglycolic acid简称PGA）作为组织技术中细胞培养支架的可行性；方法：将用胶原包埋的PGA和没有包埋的PGA分别与软骨细胞共同置于二氧化碳培养箱中培养，观察二者的亲水性，对细胞的吸附能力和细胞分泌基质的能力进行比较；结果：以胶原包埋的PGA和没有包埋的PGA亲水性没有明显的差异，二者亲水性均不强，而前者对细胞的吸附能力和细胞在其上面生长、分泌基质的能力明显增强；结论：以胶原包埋、修饰的PGA作为组织工程技术中细胞生长的支架，具有很大的应用前景。     

对应用嗅神经鞘细胞移植治疗脊髓损伤的看法

近年来国内外在实验性细胞移植治疗脊髓损伤上，对雪旺细胞（SCs）、胚胎干细胞、神经干细胞及嗅神经鞘细胞（OECs）的研究从细胞培养、分离、纯化到实验性脊髓损伤的移植修复都取得了一定的进展。动物实验显示，OECs能分泌多种神经营养因子和粘附分子，具有细胞粘附及促轴突再生的功能。OECs移植用于治疗成年动物脊髓损伤能维持神经元存活和轴突再生．促进下行传导通路纤维的再生和运动功能的恢复：OECs还能穿过星形胶质细胞形成的瘢痕环境．为受损轴突提供有利于其迁移、生长的支架．成为神经再生的桥梁。     

To fabricate artificial nerves with tissue engineering methods

OBJECTIVE: To fabricate artificial nerves with tissue engineering methods in vitro. METHODS: Schwann cells (SCs) were cultured and seeded on polyglactin 910 fibers wrapped by biomembrane coated with rat tail glue and laminin for 2 weeks. The absorbability on the scaffolds, growth and migration of SCs were assessed with a light microscope, a scanning electron microscope and a transmission electron microscope. RESULTS: SCs could migrate and proliferate on polyglactin 910 fibers. They were well distributed between scaffolds and absorbed on surface of scaffolds and formed a bungner band, on which SCs produced more matrices. SCs seeded on the biomembrane could also grow well. Axon regeneration in the distal nerve stump was observed at 8 weeks. CONCLUSIONS: Adult SCs can be expanded on coated fibers and biomembrane. Three-dimensional scaffold of SCs has the basic characteristics of artificial nerves. These findings offer a novel method to fabricate artificial nerves with tissue engineering methods for repairing defected long nerves.     

Fabricating autologous tissue to engineer artificial nerve

This study reports on the successful fabrication of artificial nerves with tissue engineering methods. Schwann cells were cultured for 2 weeks, seeded on polyglactin 910 scaffolds, and biomembrane-coated with rat-tail glue and laminin. Observation of the scaffolds' adsorptivity to Schwann cells, and of the growth and migration of Schwann cells, was made using a light microscope, and by scanning and transmission electron microscopy. The Schwann cells were able to migrate and proliferate on the polyglactin 910 fiber. Schwann cells were well-distributed, and formed a Büngner band on which the Schwann cells produced more matrices. Schwann cells on the biomembrane also grew well. We investigated the role of the tissue engineering conduit guide in vivo, using an established rabbit peripheral nerve regeneration model. At 8 weeks, axonal regeneration was observed in the distal nerve stump. Adult Schwann cells can be produced on the coated fiber and the biomembrane. Three-dimensional scaffolds with Schwann cells had the basic characteristics of the artificial nerve. These findings will provide a practical method for fabricating tissue-engineered artificial nerve to repair long nerve defects.     

运用组织工程学原理构建许旺细胞三维培养体系

目的 探讨普通重力和模拟微重力条件下构建许旺细胞三维培养体系的方法。方法 对聚羟基乙酸细丝支架进行化学改性处理后再以层粘连蛋白进行生物学修饰;分别在普通重力和模拟微重力条件下把原代许旺细胞悬液接种在支架材料上,观察许旺细胞的粘附生长情况。结果 两种重力条件下许旺细胞在聚羟基乙酸支架表面都能良好贴壁生长,培养2d时细胞生长稳定、密度适中;模拟微重力条件下许旺细胞在支架表面的分布更均匀。结论 普通重力和模拟微重力环境都适宜构建许旺细胞的三维培养体系,模拟微重力环境可能为有利。     

Experimental study on the adhesion, migration and three-dimensional growth of Schwann cells on absorbable biological materials

Objective: To study the adhesion, migration and three-dimentional growth of Schwann cells on PLA(polylactic acid) nonspinning fibre cloth and polyglycolic/polylactic acid (PLGA) fibres. Methods: Schwann cells/ECM gel solution and PLA nonspinning fibre cloth and PLGA fibres pretreated by collagen, polylysine and ECM were co-cultured. Then the migration and three-dimensional growth of Schwann cells on the fibres were observed under phase contrast microscope and laser scanning confocal microscope. Results : Schwann cell/ECM solution was compounded with PLA nonspinning fibre cloth. With formation of gel, most Schwann cells resided in the fibre net holes, and adhered to the fibres to form a multiplayerarranged Schwann cell column like Biingner band.Schwann cells could adhere to PLGA fibres and grew and migrated along the fibres. ECM gel could significantly increase the adhering and migrating cell number. Conclusions: ECM gel can facilitate the adhesion,growth and migration of Schwann cells on the seteroframe.It is a good integrating material for tissue engineering bioartificial nerve.     

Prevention of postoperative wound dehiscence in high risk patients. A randomized comparison of internally applied resorbable polyglactin 910 mesh and externally applied polyamide fiber mesh

This randomized prospective study was undertaken to compare the effectiveness of two methods used to prevent post-operative wound dehiscence in 100 high risk patients. Fifty patients were treated with a polyamide fiber mesh applied externally to the skin using a topical glue, while the other 50 patients were treated with a resorbable intraperitoneal mesh of polyglactin 910. Each patient had at least one of the following risk factors for wound dehiscence: undernutrition, malignancy, ascites, peritoneal infection, immunodepression, abdominal irradiation, repetitive surgical procedures through the same incision, or pulmonary disease. Judging criteria consisted of the development or absence of dehiscence and the frequency of abdominal wall complications. The percentage of wound dehiscence was significantly increased in patients with externally applied polyamide fiber mesh as compared to those with intraperitoneal polyglactin 910 mesh (p less than 0.01). Abdominal wall complications and the duration of healing did not vary significantly in frequency between the two groups. The intraperitoneal placement of a polyglactin mesh was more effective and better tolerated than external application of a polyamide fiber mesh.     

Intraoperative handling and wound healing: controlled clinical trial comparing coated VICRYL plus antibacterial suture (coated polyglactin 910 suture with triclosan) with coated VICRYL suture (coated polyglactin 910 suture)

BACKGROUND: Coated polyglactin 910 suture with triclosan was developed recently in order to imbue the parent suture, coated polyglactin 910, with antibacterial activity against the most common organisms that cause surgical site infections (SSI). Because such alterations could alter the physical properties of the suture, this study sought to compare the intraoperative handling and wound healing characteristics of coated polyglactin 910 suture with triclosan and traditional coated polyglactin 910 suture in pediatric patients undergoing various general surgical procedures. METHODS: This was a prospective, randomized, controlled, open-label, comparative, single-center study. Pediatric patients (age 1-18 years) undergoing various surgical procedures were randomized in a 2:1 ratio to treatment with either coated polyglactin 910 suture with triclosan or coated polyglactin 910 suture. The primary endpoint was the surgeon's assessment of the overall intraoperative handling of coated polyglactin 910 suture with triclosan and traditional coated polyglactin 910 suture without triclosan. The secondary endpoints included specific intraoperative suture handling measures and wound healing assessments. The suture handling measures were (1) ease of passage through tissue; (2) first-throw knot holding; (3) knot tie-down smoothness; (4) knot security; (5) surgical handling; (6) surgical hand; (7) memory; and (8) suture fraying. Assessment of wound healing included the following: Healing progress, infection, edema, erythema, skin temperature, seroma, suture sinus, and pain. Adverse events were recorded. RESULTS: Scores for intraoperative handling were favorable and not significantly different for both sutures, although coated polyglactin 910 suture with triclosan received more "excellent" scores (71% vs. 59%). Wound healing characteristics were comparable for both sutures except for pain on postoperative day 1. Significantly fewer patients treated with polyglactin 910 suture with triclosan reported pain on day 1 than patients who received the other suture (68% vs. 89%, p = 0.01). The overall incidence of adverse events was 18%; none was devicerelated. CONCLUSIONS: Coated polyglactin 910 suture with triclosan performed as well or better than traditional coated polyglactin 910 suture in pediatric patients undergoing general surgical procedures. The incidence of postoperative pain was significantly less in patients treated with coated polyglactin 910 suture with triclosan than the traditional suture. We speculate that polyglactin 910 suture with triclosan, by inhibiting bacterial colonization of the suture, reduced pain that can be an indicator of "subclinical" infection. Coated polyglactin 910 suture with triclosan may be a useful alternative in patients at increased risk of developing SSI.     

PLGA生物膜和延长的神经轴突束构建人工神经的体外研究

目的 探索新的构建人工神经的方法，寻找合适的支持周围神经移植的基质材料。方法 将鼠胚皮质神经元细胞种在PLGA生物膜支架上，用自制的神经轴突延长装置对其延长。观察神经元细胞及轴突生长情况，测定轴突延长后轴膜的渗透性。同时用免疫组化方法对神经微丝蛋白进行染色。结果 鼠胚的神经元和神经轴突束可以在PLGA生物膜上延长和继续培养。这些神经元和延长后的轴突柬可以保持正常形态和功能。结论 这种复合神经元轴突束的三维支架结构具有人工神经的基本特征。本研究为用延长的神经轴突束修复神经缺损打下基础。     

Parietal reinforcement prostheses: an original intraperitoneal experimental study

Summary The use of prosthetic materials is widely accepted for incisional and complex hernias, but the type of prosthesis in the abdominal wall still arouses acute controversy. We report an original experimental protocol testing three material placed intraperitoneally in the rat: a polyester mesh, a compound prosthesis (juxtaposition of a polyester mesh and a mesh of polyglactin 910) and a composite prosthesis (where fibers of polyester and polyglactin 910 were woven in the same mesh). There were two main criteria for assessment: the biologic tolerance to the material on the one hand, characterized histologically by the ratio of the surface of fibrosis to the surface of the inflammatory granuloma in contact with the material, and the nature of the adhesions between prosthesis and abdominal wall and the intraperitoneal viscera on the other. Statistical analysis of the results led to a preference for the homogeneous polyester prosthesis, compared with compound and composite prostheses (polyester and polyglactin 910) and to abandonment of the intraperitoneal site for insertion of such materials.