应用胶原凝胶构建组织工程化皮肤的研究

目的探讨以胶原凝胶为支架材料构建组织工程化皮肤的可行性。方法体外分离、培养人皮肤表皮细胞和成纤维细胞;利用自制的胶原蛋白制成胶原凝胶作为组织工程支架材料;在成功构建人工真皮的基础上种植表皮细胞,构建复合人工皮肤;采用HE染色与免疫组织化学的方法对复合人工皮肤进行组织学检测。结果HE染色可见,构建的复合人工皮肤具有表皮和真皮双层结构;免疫组织化学染色显示,Ⅳ型胶原、纤维连接蛋白和层粘连蛋白阳性,在形态结构上与正常皮肤相似。结论培养的人表皮细胞和成纤维细胞种植于胶原凝胶支架上,气-液界面培养可构建出具有类似正常皮肤结构的组织工程化皮肤。     

强脉冲光对大鼠皮肤胶原蛋白影响的定量分析

目的　定量分析强脉冲光对大鼠皮肤胶原蛋白含量的影响,进一步明确强脉冲光在治疗皮肤光老化中的作用机制。方法　选择15只SD大鼠,每只选3个部位用强脉冲光治疗,采用同一治疗参数,能量密度34J/cm2 ,分为3个脉冲,脉宽各为4ms、5ms、6ms,脉冲延时为2 0ms及2 5ms。分别切取治疗后第1、3、5、7、15、30天皮肤和未治疗部位皮肤,进行组织学观察。结果　治疗后,第1、3、5、7天皮肤胶原蛋白染色与未治疗部位比较无明显差别,第15天强于未治疗部位,第30天明显强于未治疗部位,免疫组化染色Ⅰ、Ⅲ型胶原明显增多(P <0 .0 0 1)。结论　强脉冲光治疗大鼠皮肤后可引起胶原含量的增加,在实验基础上为强脉冲光的临床应用提供了理论依据。     

胶原凝胶复合骨胶原基质(BCM)负载关节软骨细胞的实验研究

目的 探讨胶原凝胶包埋软骨细胞接种BCM支架的三维培养对软骨细胞生长及功能的影响.方法 将胶原凝胶包埋的关节软骨细胞接种BCM支架并在体外培养,应用倒置相差显微镜和扫描电镜观察软骨细胞的粘附、生长和增殖情况,培养14d,行苏木精-伊红、甲苯胺蓝染色观察软骨组织形成情况.结果 软骨细胞在支架上粘附、生长和增殖良好,体外培养14d能形成较成熟的软骨组织.结论 胶原凝胶复合BCM支架具有良好的细胞相容性,可作为负载生长因子的载体.     

实验性肝纤维化胶原变化的研究

目的：研究肝纤维化过程中胶原成分的变化规律。方法：制作四氯化碳肝纤维化模型，测定肝组织羟脯氨酸含量，用ＶＧ、ＲＦ染色显示Ⅰ、Ⅱ型胶原并进行计算机图像分析，放免法测定肝匀浆及血清Ⅲ型前胶原水平，应用上述指标反映肝纤维化过程中胶原的变化规律。结果：随实验时间延长，肝内胶原含量逐渐增加。肝纤维化早、中期以Ⅲ型胶原增生为主，肝纤维化后期及肝硬化期以Ⅰ期胶原为增生为主。血清ＰＣⅢ与肝内胶原变化基本同步。结论     

High Cell-Density Culture System of Hepatocytes Entrapped in a Three-Dimensional Hollow Fiber Module with Collagen Gel

Abstract: A compact three-dimensional (3D) module is needed for hepatocyte culture in order to develop an effective hybrid artificial liver system that can retain hepa-tocellular structure and differentiated functions. We treated the 3D module with collagen gel to entrap rat hepatocytes. This method yielded a high hepatocellular density (2 times 10⁷ cells/ml) over a period of 14 days and maintained the secretion of albumin and ureogenesis at the same level as the control monolayer method. The ammonia removal remained at 43% of the Day 0 value over 8 days of perfusion. Our data show that this approach may be useful for liver support therapy in an ex-tracorporeal circuit.     

NO2和LPG燃烧产物对大鼠肺脂质过氧化,胶原含量和尿HOP的影响

本研究分别观察了ＮＯ２染毒和ＬＰＧ燃烧产物自然暴露对Ｗｉｓｔａｒ雄性成年大鼠肺脂质过氧化、肺胶原含量和尿ＨＯＰ排泄的影响及其相互关系。结果发现７．５２ｍｇ／ｍ＾３的ＮＯ２对三者均有一定影响，染毒剂量不同大鼠尿ＨＯＰ排泄也有相应的变化。尿ＨＯＰ的变化早于肺胶原含量，且两者有一定的相关关系。ＬＰＧ燃烧产物暴露期间，尿ＨＯＰ含量一直高于对照组，但未见肺胶原含量的变化，８周时尿ＨＯＰ与肺脂质过氧化物之间有     

An ultrastructural study on anterior cruciate ligament reconstruction using autogenous patellar tendon grafts in dogs

An ultrastructural study was undertaken concerning morphological changes within the autografted patellar tendon (PT) after being transplanted to the anterior cruciate ligament (ACL) in mongrel dogs. After 4 weeks, the arrangements of both large and small collagen fibrils in the PT graft became disordered, and the number of inflammatory cells increased. However, the same PT grafts revealed postsurgical signs of newly-produced collagen fibrils around activated fibroblasts at 12 to 24 weeks. At 52 weeks after the transplantation, small collagen fibrils increased in both number and density, showing a remarkable morphological similarity to the collagen fibrils of normal ACL. These data indicate that the characteristics of the PT graft eventually resemble those of a normal ACL. This paper was presented in part at the Combined Meeting of the Orthopedic Research Societies of the USA, Japan and Canada in Banff, Alberta, Canada, October, 1991.     

Transurethral collagen injection for treatment of stress urinary incontinence

Summary Minimally invasive treatment of urinary incontinence has become a subject of major interest in recent years. We examined the use of transurethral collagen injection for incontinence treatment. A total of 48 patients were selectet for this procedure from April 1993 to February 1997 – 26 male patients (19 post-RPX incontinence and 7 post-TUR incontinence) and 22 female patients (all after previous incontinence surgery) were treated by injection of collagen into the continence region. The whole group underwent an average of 1.8 sessions, and a mean collagen injection volume of 14.5 ml was delivered per session. Mean follow-up was 9.2 months. Of the female population, 68.2 % were cured or greatly improved. In the male population only 47.3 % of the post-RPX patients and 6/7 of the post-TUR patients benefited from the procedure. In males, treatment outcome depends on the degree of pretreatment incontinence, because all grade III incontinence patients did worse. Therefore we conclude: transurethral collagen injection is an interesting method in the treatment of urinary incontinence if proper patient selection is assured.      

Isolated human hepatocytes in culture display markedly different gene expression patterns depending on attachment status

In vitro human hepatocyte cultures are a key tool in the investigation of xenobiotic toxicity and metabolism. In most in vitro hepatocyte studies, the cells are allowed to adhere to an extracellular matrix, such as collagen. Unfortunately, the ability of freshly isolated hepatocytes to adhere to collagen varies from donor to donor. We used microarray analysis to determine what gene expression differences exist between hepatocytes in suspension and hepatocytes attached to collagen. Results from different donors showed a considerable difference in gene expression patterns between the two hepatocyte populations. In addition, we also compared the gene expression profiles of hepatocytes in culture with liver tissue. The results showed that both hepatocytes in suspension and hepatocytes attached to collagen display significant gene expression differences compared with liver tissue. Finally, we show that both populations of hepatocytes are responsive to dexamethasone and regulate some of the same genes. Overall, our results suggest that either significant gene expression changes occur in isolated hepatocytes or that suspended and attached cells represent different populations of hepatocytes found in intact livers.