人半月板纤维骨细胞在纤维蛋白胶中体外培养

目的：研究纤维软骨细胞的分离培养及在纤维蛋白胶中三维培养时的表现。方法：（1）分离培养人半月板纤维软骨细胞;（2）制备纤维蛋白胶;（3）在纤维蛋白中培养细胞,观察其表现并测定生长曲线、倍增时间;（4）通过透射电镜、AB－PAS染色、Ⅱ型胶原免疫组化检查等,了解细胞生长及软骨基质的分泌情况。结果：（1）原代培养细胞呈线形,能在纤维蛋白中保持;（2）细胞在纤维蛋白中倍增时间为第1－4d:7.0d,第5－7d:2.3d;（3）AB－PAS染色（＋）、Ⅱ型胶原免疫组化检查（＋）。结论：人半月板纤维软骨细胞能在纤维蛋白胶中增殖,保持其表型和分泌胞外基质。     

兔关月板纤维软骨细胞的培养及生物学特性研究

目的：通过半月板纤维软骨细胞的分离、培养、鉴定,观察其生长特点,研究半月板生物学特性及其损伤修复的细胞学基础。方法：机械分离兔半月板软骨,胰蛋白酶、胶原酶联合消化,１０％ＦＢＳＥ的ＤＭＥＭ中原代和传代培养,例置显微镜动态观察细胞形态及生长情况,ＧＡＧ、Ⅱ型胶原免疫组化染色,电镜观察细胞超微结构。结果：培养细胞呈多角形,有突起,富含分泌颗粒,ＧＡＧ、Ⅱ型胶原染色阳性,细胞线粒体和内质网发达。结论：培养的细胞保持了体内纤维软骨细胞的基本特性。     

兔半月板纤维软骨细胞的培养及生物学特性研究

目的 :通过半月板纤维软骨细胞的分离、培养、鉴定 ,观察其生长特点 ,研究半月板生物学特性及其损伤修复的细胞学基础。方法 :机械分离兔半月板软骨 ,胰蛋白酶、胶原酶联合消化 ,10 ?S的DMEM中原代和传代培养 ,倒置显微镜动态观察细胞形态及生长情况 ,GAG、Ⅱ型胶原免疫组化染色 ,电镜观察细胞超微结构。结果 :培养细胞呈多角形 ,有突起 ,富含分泌颗粒 ,GAG、Ⅱ型胶原染色阳性 ,细胞线粒体和内质网发达。结论 :培养的细胞保持了体内纤维软骨细胞的基本特性。     

Ⅰ型胶原半月板支架负载纤维软骨细胞体外培养

[目的]观察Ⅰ型胶原半月板支架对纤维软骨细胞的吸附作用及对细胞生物学性状的影响，评价其作为半月板组织工程支架的可行性及价值。[方法]构建Ⅰ型胶原半月板支架，将体外培养的兔半月板纤维软骨细胞吸附于该支架上三维立体培养，通过相差倒置显微镜、组织学、扫描电镜及免疫组织化学检测支架对半月板纤维软骨细胞的表型、增殖及功能的影响。[结果]Ⅰ型胶原能制成理想大小、形状的三维立体多孔半月板支架，纤维软骨细胞在支架孔壁贴附良好，维持表型稳定，分泌胞外基质。[结论]Ⅰ型胶原半月板支架细胞相容性良好，但力学性能相对较差，通过与其它生物材料复合以提高其机械力学强度，可望制得理想的半月板组织工程支架。     

人胎关节软骨细胞体外培养的生物学特性

目的 研究软骨组织工程中传代软骨细胞与原代的差异。方法 用５个月人胎关节软骨分离培养细胞,观察细胞存活率、贴壁率、生长曲线和组织形态学的改变。结果 ⑴软骨块在４℃下,３ｄ内细胞存活率可达９３．４％～９７．６％。⑵原代细胞为圆形或三角形;第４代有一半转变成梭形,到第６代全部变为长梭形。⑶传代细胞贴壁时间（２～３ｈ）短于原代（４～７ｈ）。玻璃瓶内贴壁率传代细胞为７８．７％～８５．５％,原代８．８％。⑷     

人负重关节软骨细胞的体外分离与培养

目的　研究人负重关节软骨细胞的分离培养技术。方法　对 10例手术切除的负重关节软骨进行0 2 %Ⅱ型胶原酶分离、10 %DMEM中培养 ,观察软骨细胞获得率、贴壁率、增殖状况及形态学改变。结果　① 0 2 %Ⅱ型胶原酶消化尽 2 0 0～ 70 0mg软骨标本需 4～ 14h ,细胞获得率 (1 82± 0 5 7)× 10 6/g ,存活率(79 4± 9 9) %。②细胞可传 6～ 12代 ,前 4代培养时间平均 (33± 10 )d ,生长倍数平均 198倍 ,倍增时间(4 13± 1 34)d。③第 3、4代细胞呈多角形、部分梭形 ,8、9代后细胞梭状铺平。结论　 30 0～ 5 0 0mg软骨标本 ,经该方法培养 3～ 4周后可达第 4代 ,细胞数可达 (0 6～ 1 0 )× 10 8个 ,可满足临床上修复 15～ 2 5cm2软骨缺损的需要。     

纤维蛋白凝胶和脱钙骨基质支架材料复合软骨细胞修复兔膝关节软骨缺损的实验研究

目的：探讨纤维蛋白凝胶和脱钙骨基质支架材料复合软骨细胞作为软骨组织工程支架的可行性及有效性,并为后续研究可注射性材料做基础。方法：体外分离培养软骨细胞后接种到纤维蛋白凝胶和脱钙骨基质支架材料体外培养4周,然后植入兔膝关节软骨缺损区继续培养4、8、12周后取材,分别行大体、组织学、Ⅱ型胶原免疫组织化学观察。并进行Wakitani评分,观察其体内修复关节缺损效果。结果：大体观察4周后,实验组软骨缺损区可有乳白色组织修复,12周可修复完全,并无明显凹凸感。光镜下8周可见大量软骨细胞修复,并在TB染色下见Ⅱ型胶原比4周时明显增多。12周时软骨陷窝结构形成,细胞形态排列及Ⅱ型胶原与正常软骨组织相近。结论：纤维蛋白凝胶和脱钙骨基质支架材料复合软骨细胞可以作为软骨组织工程支架材料,能够用于再生修复软骨的缺损。并为构建可注射性修复材料提供途径。     

可注射性纤维蛋白胶对兔骨髓基质细胞增殖和分化的影响

目的观察纤维蛋白胶（Fs）对体外培养的兔骨髓基质细胞（MSCs）增殖和分化的影响。方法实验分两组：含有Fs的实验组和不加任何材料的对照组。采用细胞培养、组织化学及电镜等方法对两组MSC的增殖活性、贴壁率、碱性磷酸酶（ALP）表达、Ⅰ型胶原表达、超微结构及细胞在材料中的生长情况进行研究。结果①实验组MSCs的增殖活性和贴壁率明显高于对照组（P〈0．05）。②实验组MSCs的ALP活性显著高于对照组（P〈0．05）。实验组第7天时的ALP活性显著高于第5天时本组的ALP活性（P〈0．05）。实验组MSCs的Ⅰ型胶原表达水平显著高于对照组（P〈0．05）。③扫描电镜观察：实验组MSCs与材料融合生长。透射电镜观察：实验组的MSCs细胞增殖活性好,向成骨细胞方向分化水平高,而对照组MSCs的细胞增殖活性相对较低,分化相对较差。结论FS生物相容性好,对MSCs增殖和向成骨细胞方向的分化均有明显促进作用,可作为骨组织工程材料的注射型载体进行进一步研究。     

兔腱细胞体外培养及其生物学特性的研究

了解腱细胞的生长，形态，合成胶原的能力及冻存对腱细胞的生长的影响。方法：应用显微外科技术剥除兔屈肌腱外膜组织后，经胰蛋白酶及胶原酶分步消化分离出腱细胞。细胞经含２０％新生小牛血清的Ｆ－１２培养液传代培养６代后全部冻存。腱细胞合成的胶原型以免疫组织化学法进行鉴定。     

The Use of Fibrin Glue in the Treatment of Fistula-In-Ano: A Prospective Study

Purpose This prospective study was done to analyze the efficacy of commercial fibrin glue application in the healing of patients with fistulas-in-ano. Methods This clinical trial of 36 patients was performed during the period from November 2003 to May 2004. Thirty men and six women were treated for a fistula-in-ano with commercial fibrin glue application. None of the patients had undergone prior attempts to correct fistulas-in-ano surgically. All patients received preoperative mechanical bowel preparation and prophylactic intravenous antibiotics. In the operating room, the patients underwent an anorectal examination under spinal anesthesia. The external and internal fistula tract openings were then identified. The fistula tract was curetted. Fibrin glue was injected into the external fistula opening until fibrin glue could be seen coming from the internal opening. No dressing was applied over the external opening. Thereafter, the patient was discharged. A 1-week course of oral antibiotics was prescribed. The patients were followed up for 70 weeks with a mean of 54 weeks (range 40–70 weeks). Results The cause of the fistula-in-ano was cryptoglandular in all patients. The overall initial success rate was 77.8% (28/36). No complications were encountered related to the application. Two patients underwent a reapplication with fibrin glue and the fistulas of these patients were all closed. The overall success rate was 83.3% (30/36). Conclusion Fibrin glue application was thus found to be an easy, safe, effective, and useful alternative treatment in the management of fistulas-in-ano. However, our findings need substantiation by increasing the number of patients and prolonging the follow-up duration, as well as carrying out comparative studies.     

Combined Use of Hair and Fibrin Glue for Soft Tissue Augmentation: Experimental Study

A vast variety of alloplastic materials together with autogenous grafts have been used for the correction of contour deformities related to aesthetic and reconstructive purposes. Despite a number of well-tolerated materials, the ideal has not yet been reached, although well-tolerated autogenous grafts have higher rates of resorption and distortion. The limited volume of autogenous grafts and concerns about donor-site morbidity may obligate the surgeon to use implantation materials. Covering almost every property of an ideal implantation material, hair, an organic autogenous nonvital tissue, was used for soft tissue augmentation in this experimental study. Hair pellets of 10 New Zealand rabbits were prepared and shaped as 1 × 1 × 1-cm cubes with the help of fibrin sealant, then inserted subcutaneously. The materials were evaluated 4 months later. No complications such as infection or extrusion were seen. The materials were intact, with no signs of resorption, but the shapes were distorted due to the pressure of the surrounding tissues. Histopathologic findings also demonstrated that the hair was well tolerated by the adjacent tissues. Different processes may enable hair to be used as a filler material in clinical practice. Well-tolerated, nonresorbable injectable materials or shaped implants may be obtained at low cost using hair.     

The Wound-Healing Effect of Fibrin Glue for Tracheal Anastomosis in Experimental Pulmonary Surgery

The leakage of tracheal anastomoses is one of the major complications that occurs after tracheal reconstruction. Improved reinforcing methods for anastomoses would thus be clinically useful. To find a better technique, we examined the postoperative would-healing effect of fibrin glue on tracheal anastomosis in the rat. Experimental rats were divided into two groups. In the control group (n = 21), the trachea was anastomosed by interrupted absorbable sutures. In the fibrin glue group (n = 21), the trachea was anastomosed in the same manner as the control group, with the addition of fibrin glue around the area of anastomosis. In the two groups, we studied the amount of hydroxyproline and histological findings on the seventh, 14th, and 21st postoperative day. The amount of hydroxyproline and collagen fibers in the fibrin glue group was more than in the control group on the seventh postoperative day. These results suggest that fibrin glue has a promotive effect in the healing of tracheal anastomosis. Received: August 24, 2000 / Accepted: May 15, 2001     

Laparoscopic Incisional Hernia Repair With Fibrin Glue in Select Patients

Background and Objective:

Laparoscopic treatment of incisional hernias can be performed using different types of fixation devices and prosthesis. We present a case series of 19 patients with incisional hernias with a diameter of <6cm, who underwent laparoscopic repair using Hi-tex dual-side mesh, positioned intraperitoneally, fixed to the abdominal wall by fibrin glue (Tissucol).

Methods:

Nineteen patients with incisional hernias <6cm in diameter were enrolled in this study and treated laparoscopically with Hi-tex and Tissucol. Surgical complications and patient outcomes were assessed with a clinical follow-up.

Results:

Laparoscopic repair of incisional hernias by using Hi-tex mesh affixed to the parietal wall with fibrin glue was feasible and easy in patients with parietal defects <6cm in diameter. Mean operating time was 30 minutes. Mean hospital stay was 1.5 days. Almost no postoperative pain, major surgical complications, seroma formation, relapses, or prosthesis infection occurred during a mean follow-up of 20 months.

Conclusions:

In select patients, Hi-tex mesh affixed using fibrin glue allows laparoscopic repair of incisional hernias with very good patient outcomes, especially in terms of postoperative pain and seroma formation.     

生物蛋白胶预防鸡趾鞘管区屈肌腱粘连的实验研究

目的 研究生物蛋白胶预防鸡趾鞘管区屈肌腱粘连的作用。方法 种禽褐鸡60只，随机分为生物蛋白胶组（FG）和生理盐水组（NS），每组各30只动物。将第三、四趾趾深屈肌腱切断，作改良Kessler法缝合，不修复腱鞘，然后在腱鞘内肌腱周围及皮下按分组注入相应药物。术后第2、4、6周取材，分别进行大体观察、组织学检查、生物力学测定。结果 术后2、4、6周，组织学检查示两组动物肌腱吻合处的炎症细胞浸润情况及肌腱的愈合进程无明显差别。缝合处粘连半定量评分、将肌腱拉出鞘管所需最大力量、耗功，FG组与NS组相比，差异均无显性意义（P＞0．05）。结论 局部应用FG不能有效预防术后肌腱粘连，但不影响肌腱的愈合进程，也不引起额外的炎症反应。     

兔阴茎海绵体平滑肌细胞的体外培养及其生物学特性

目的 :研究体外培养的新西兰白兔阴茎海绵体平滑肌细胞的生物学特性。　方法 :采用组织块培养法 ,对兔阴茎海绵体平滑肌细胞进行活细胞观察 ,并测定其细胞生长曲线、贴壁率、细胞分裂指数。　结果 :①兔阴茎海绵体平滑肌细胞为梭形 ,呈长轴平行排列 ,具有明显的方向性 ;②体外贴壁快 ,生长迅速 ,体外培养的阴茎海绵体平滑肌细胞在合适的传代条件和比例下能够生存并保持其稳定的生物学特性。　结论 :体外培养的兔阴茎海绵体平滑肌细胞模型可用于检测某些药物对阴茎勃起的影响。     

新型复合纤维蛋白胶的磷酸钙骨水泥的生物安全性研究

目的 研究新型复合纤维蛋白胶的磷酸钙骨水泥的生物相容性和生物安全性,探讨其用于临床修复骨缺损的可行性。方法 制备新型复合纤维蛋白胶的磷酸钙骨水泥,获取材料浸提液。选择急性毒性试验、溶血试验、微核试验、细胞毒性试验,对新型复合人工骨材料进行生物相容性和安全性评价。结果 该磷酸钙骨水泥材料浸提液未引起小鼠急性毒性反应;各实验组肉眼下未见明显溶血反应,溶血率0.05);浸提液对小鼠MC3T3成骨细胞的生长分化无明显影响,细胞毒性分级为Ⅰ级。结论新型复合纤维蛋白胶的磷酸钙人工骨材料具有良好的生物相容性和生物安全性。     

Cauterization Versus Fibrin Glue for Aerostasis in Precision Resections for Secondary Lung Tumors

Background: Aerostasis control in multiple resections for metastatic pulmonary diseases is a real problem. Long-term air leaks prolong postoperative hospitalization and result in an additional financial burden on the healthcare system. We focused our study on the evaluation of fibrin glue (Tissucol) as an effective means to minimize or prevent air leaks.Methods: We initiated a case-control study whereby 100 patients underwent precision resections for lung metastases. The subjects were divided into 2 groups, both with 50 patients: group 1 was treated with fibrin glue and group 2 with cauterization. Evaluation parameters consisted of the following: air leak duration, expected complications, drain time, and in-hospital stay.Results: In group 1, air-leak time was 2.68 ± 1.72 days, versus 7.80 ± 8.52 for group 2 (P < .001). In group 1, there were 2% complications, whereas in group 2 there were 28% (P < .001). Drain time was 4.54 ± 1.83 days for group 1 and 9.54 ± 8.35 for group 2 (P < .001). In-hospital stay was 6.54 ± 1.83 days for group 1 and 11.54 ± 8.35 for group 2 (P < .001).Conclusions:In the group treated with fibrin glue, we observed significant advantages. Our experience shows that the use of fibrin glue can improve aerostasis control in nonanatomical resections with high risk of air leak.Presented at the 55th Annual Cancer Symposium of the Society of Surgical Oncology, Denver, Colorado, March 14–17, 2002.     

纤维蛋白凝胶复合骨形态发生蛋白和庆大霉素缓释药物治疗感染性骨缺损的实验研究

目的 探讨将纤维蛋白凝胶(FG)作为骨形态发生蛋白(BMP)及庆大霉素的共同载体,一期治疗感染性骨缺损。方法 48只青紫兰兔,制作慢性骨髓炎模型,清创后造成胫骨干骺端内侧1.5 cm长半环形骨缺损,采用三种方法进行处理:A组,植入FG、BMP和庆大霉素复合物;B组植入FG/BMP复合物;C组作为空白对照。术后观察动物一般情况,作骨细菌培养及其计数,X线摄片及组织学检查。结果 A组感染控制及骨修复均良好,感染控制率、再生骨量明显优于B组;B、C两组在感染控制率上无显著差异;C组动物骨修复差。结论 FG、BMP及庆大霉素复合物具有抗感染及促进成骨的双重作用,可用于感染性骨缺损的治疗,也可用于污染严重的开放性损伤造成的骨缺损的治疗,方法简便、易行。