低流速流体剪切力、炎性因子和P38MAPK抑制剂基于微流控芯片对于软骨细胞表型的影响

发布时间：2018-06-15 19:17

本文选题：关节镜 + 乆窝囊肿　；参考：《大连医科大学》2015年硕士论文

【摘要】：乆窝囊肿是指乆窝深部滑囊肿大或者向后突出的总称,是关节骨科常见的一种临床病理症候群。近些年来,随着科学技术的不断创新和发展,治疗乆窝囊肿的手术方式发生了日新月异的变化。在当下对于此病的临床治疗中,关节镜渐渐取代了传统的手术方式。本论文中将选取一定数量的病例来评价关节镜技术对于乆窝囊肿的临床治疗效果。目的:评价关节镜对于乆窝囊肿的临床效用。方法:采集2013-2014年大连医科大学附属第一医院关节外科30名经过MRI检查并确诊为乆窝囊肿的患者的临床病例资料。其中,男性患者20人,女性患者10人。年龄为25到65岁。平均52岁。所选取的所有病例只有一侧患有疾病,对侧未有累及。其中,左膝出现病理变化者17例,右膝出现病理变化者13例。乆窝处的局部隆起是手术前的主要症状。疾病所累及的关节处有轻重不一的疼痛感。通过B超技术的使用,手术前确诊囊肿均与关节腔相连通。术前根据Rauschning和Lindgren分级法:Ⅰ级8例,Ⅱ10例,Ⅲ12例。对比术前术后Rauschning和Lindgren的评分,测评手术后患者的疾病改善情况。结果:所选取的所有患者的关节内疾病都已经被确诊。其中,半月板撕裂的患者占有17人,骨性关节炎者9人,前交叉韧带撕裂的患者则有4人。经过术后一段时间的观察,出血、切口裂开、乆窝内部结构损伤的现象没有出现。手术后3天之内出院。随访时间8至24个月。平均12个月。绝大部分患者术后恢复情况良好。Rauschning和Lindgren评分的结果:治疗后Ⅱ级患者2例,再次复发者一例。结论:关节镜下对于乆窝囊肿的治疗操作简单,对机体的损伤小,易于患者的恢复。关节软骨为透明软骨,具有弹性,能够减低外力的冲击,维持局部组织的稳定性。创伤、炎症等因素可以使关节软骨损伤,造成软骨细胞细胞外基质的合成和分解失去平衡,进而为骨性关节炎的发生提供基础。软骨组织中不存在血管、神经和淋巴,因此其再生功能非常局限。关节软骨主要由软骨细胞和细胞外基质组成.其中,Ⅱ型胶原、蛋白多糖、无机盐和水构成了胞外基质的有形成分。作为软骨细胞胞外基质的特异性胶原,Ⅱ型胶原占胞外基质胶原总量的90%以上,随着软骨细胞代数、密度和培养时间的增加,Ⅱ型胶原的表达量逐渐减少,Ⅰ型胶原的表达量逐渐增加。同时,软骨细胞的外形在此过程中发生了变化,由圆形或多角形逐渐变为成纤维细胞样的长梭形。生理条件下,关节软骨浸泡在关节滑液中,当关节运动时,其产生的流体剪切力对于维持软骨细胞正常功能的维持具有重要作用。由于骨性关节炎是力学因素和生物化学因素等多种因素共同作用的结果。因此,研究模拟流体剪切力和其他复合因素对于软骨细胞表型的维持作用的影响显得格外重要。目的:流体剪切力对于软骨表型的变化至关重要。本研究以微流控芯片技术为基础,建立了一个能够同时产生四个大小剪切力芯片的平台来研究低流速流体剪切力对于软骨细胞表型的影响。同时,还加入了分解性细胞因子和P38MAPK抑制剂SB203580,来考察多种不同因素的共同作用下软骨细胞表型的维持和变化的情况。方法:1.提取和培养大鼠的原代软骨细胞。2.用光刻胶技术制作芯片的模板,以PDMS为原材料制作成芯片。将PDMS芯片和玻璃片用等离子照射后进行不可逆封接。3.通过CFD模拟和电路类比的方法对芯片结构进行数值模拟。4.应用免疫荧光染色的方法对经过剪切力作用后的软骨细胞进行Ⅰ型胶原,Ⅱ型胶原和Glut-1染色。结果:1.Ⅱ型胶原随着流体剪切力的增强表达量逐渐增加,软骨细胞的去分化作用随着剪切力的增加而作用明显。2.P38MAPK抑制剂能够有效的阻断机械力信号通路,使软骨细胞的表型持续表达。3.分解性细胞因子TNF-α与流体剪切力的共同作用下,软骨细胞的特征性表型随着剪切力的增加而破坏加重。同时,去分化效果也显著增加。4.低流速流体剪切力对于葡萄糖转运蛋白-1(Glut-1)的表达起促进作用。结论:低流速流体剪切力对于软骨细胞表型的影响是一把双刃剑,一方面,随着剪切力的增大流体剪切力能够对软骨细胞的特异性表型起到维持作用。另一方面,流体剪切力的增大也加速了软骨细胞的去分化作用。P38MAPK抑制剂SB203580能够有效抑制机械力信号通路,阻断剪切力对于软骨细胞表型维持和去分化的作用。炎性因子TNF-α对于软骨细胞的表型维持起到相反作用。
[Abstract]:The cyst of the fossa is the general name of the deep or backward protrusion of the fossa fossa. It is a common clinical pathological syndrome in the joint Department of orthopedics. In recent years, with the continuous innovation and development of science and technology, the surgical methods for the treatment of the cyst of the fossa have changed rapidly. A certain number of cases were selected to evaluate the clinical efficacy of arthroscopy in the treatment of cyst of the fossa cyst. Objective: To evaluate the clinical efficacy of arthroscopy for the cyst of fossa fossa. Methods: 2013-2014 years of 2013-2014 years of joint surgery in the first hospital of Dalian Medical University were examined and confirmed as the diagnosis. There were 20 male patients and 10 female patients. The average age was 25 to 65 years old. The average age was 52 years. All cases were only one side of the disease and the opposite side was not involved. Among them, there were 17 pathological changes in the left knee and 13 cases with pathological changes in the right knee. The local bulge at the fossa was before the operation. Main symptoms. There was a different sense of pain in the joints involved in the disease. Through the use of B-ultrasound technique, all the diagnosed cysts were connected with the articular cavity before operation. Before operation, 8 cases, 10 cases, and 12 cases were classified according to the Rauschning and Lindgren classification method. The scores of Rauschning and Lindgren were compared before and after operation, and the improvement of the disease after operation was evaluated. Results: all of the patients' intra-articular diseases were confirmed. Among them, 17 patients with meniscus tear, 9 patients with osteoarthritis and 4 patients with anterior cruciate ligament tears were observed after a period of time, bleeding, incision split, and internal structural injury in the fossa. 3 days after the operation. The follow-up time was 8 to 24 months. An average of 12 months. The recovery of most patients was good.Rauschning and Lindgren score: 2 cases of second class patients after treatment and one recurrence. Conclusion: the treatment of the cyst under arthroscopy is simple, the injury to the body is small, the recovery of the patient is easy. The cartilage, with elasticity, can reduce the impact of external force, maintain the stability of the local tissue. Trauma, inflammation and other factors can cause articular cartilage damage, cause the synthesis and decomposition of the extracellular matrix of cartilage cells to lose balance, and thus provide the basis for the occurrence of osteoarthritis. The regeneration function is very limited. Articular cartilage consists mainly of chondrocytes and extracellular matrix. Among them, type II collagen, proteoglycan, inorganic salt and water constitute the tangible components of the extracellular matrix. As the specific collagen of the extracellular matrix of cartilage cells, type II collagen accounts for more than 90% of the total extracellular matrix of the extracellular matrix, with the cartilage cell algebra and density. The expression of type II collagen decreased gradually and the expression of type I collagen increased gradually. At the same time, the shape of cartilage cells changed from round or polygonal to fibroblast like long spindle. Under physiological conditions, the articular cartilage was soaked in synovial fluid, when joint movement, The shear force produced by the fluid plays an important role in maintaining the normal function of cartilage cells. Osteoarthritis is the result of a variety of factors such as mechanical and biochemical factors. Therefore, the study of the influence of simulated fluid shear force and other complex factors on the maintenance of chondrocyte phenotype appears to be a lattice. Objective: Objective: fluid shear force is very important for the change of cartilage phenotype. Based on microfluidic chip technology, a platform which can produce four shear force chips at the same time was established to study the effect of low velocity fluid shear force on chondrocyte phenotype. Meanwhile, the decomposable cytokine and P38MA were added. PK inhibitor SB203580, to investigate the maintenance and change of chondrocyte phenotype under the common action of a variety of different factors. Methods: 1. extract and cultivate the primary chondrocytes of the rat.2. using photoresist technology to make the chip template, and make the chip with PDMS as the raw material. The PDMS chip and the glass slice are irradiated by plasma. Reverse sealing and connecting.3. through CFD simulation and circuit analogy method to simulate the structure of chip.4. application immunofluorescence staining method for collagen type I, type II collagen and Glut-1 staining after shear force action. Results: 1. type II collagen increased gradually with the increase of fluid shear force, cartilage cell Dedifferentiation effect with the increase of shear force obviously the.2.P38MAPK inhibitor can effectively block the mechanical signal pathway, which makes the phenotype of chondrocytes continuously express the.3. decomposed cytokine TNF- alpha and the fluid shear force, and the characteristic phenotype of chondrocytes is aggravated with the increase of shear force. The effect of.4. low velocity fluid shear force on the expression of glucose transporter -1 (Glut-1) was also promoted. Conclusion: the effect of low velocity fluid shear force on chondrocyte phenotype is a double-edged sword. On the one hand, with the increase of shear force, the fluid shear force can maintain the specific phenotype of cartilage cells. On the other hand, the increase of fluid shear force also accelerates the dedifferentiation of cartilage cells..P38MAPK inhibitor SB203580 can effectively inhibit the mechanical signaling pathway and block the effect of shear force on the phenotype maintenance and dedifferentiation of chondrocytes. Inflammatory factor TNF- alpha plays the opposite role in the maintenance of chondrocyte phenotype.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R687.4

【相似文献】