EFFECTS OF STROMAL DERIVED FACTOR-1 (SDF-1) BLOCKADE

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 在先前的研究中，SDF-1已被证明在肩部炎症、肩峰下滑囊炎和肩袖疾病中发挥重要作用。 为了确定SDF-1抑制剂在治疗肩部炎症中的潜在生物学功效，这些研究将利用大鼠肩袖损伤模型。 本项目的具体目的是检查SDF-1通路抑制剂AMD 3100、T140类似物（4F-苯甲酰基-TN 14003）和CTCE-9908在减轻炎症方面的生物学作用及其对肩袖肌腱愈合的影响。 我们假设SDF-1抑制剂将减少肩峰下囊的炎症，但不会抑制肩袖肌腱愈合。 具体目标 1. SDF-1/CXCR 4抑制剂对大鼠肩部局部炎症的影响：将在患有肩袖损伤诱导的肩峰下炎症的大鼠中检查SDF-1/CXCR 4受体抑制剂（AMD 3100、T140类似物和CTCE-9908）和CXCR 4抗体（MAB 171和12 G5）的形态学和生物化学影响。 雄性刘易斯大鼠将进行手术以模拟冈下肌腱的旋转袖损伤并在相邻肩峰下囊中诱导炎症（滑囊炎）。 抑制剂将通过植入式渗透微型泵全身给药。 处死动物后，收获肩峰下囊，并通过定量PCR和酶联免疫吸附试验（ELISA）测量SDF-1、白细胞介素-1（IL-1）和肿瘤坏死因子（TNF-α）的表达。 还将进行肩峰下炎症的组织学评价和免疫组织化学，以评估SDF-1阻断对局部炎症反应的影响。 2. SDF-1/CXCR 4抑制剂对大鼠肩关节肩袖肌腱愈合的影响：将在肩袖损伤大鼠中检查SDF-1/CXCR 4受体抑制剂（AMD 3100、T140类似物和CTCE-9908）和CXCR 4抗体（MAB 171和12 G5）对肩袖肌腱愈合的形态学、生物化学和生物力学影响。(see在从损伤部位获取肩袖肌腱后，将通过组织学和I型和III型胶原的局部表达来评估肌腱愈合。 将使用MTS 810伺服液压载荷测试架（MTS Corp，伊甸园草原，MN）评估肌腱强度。冈上肌腱的插入和连接部位将刚性嵌入聚氨酯（Smooth On，伊斯顿，PA）中，并在恒定位移下承受轴向拉伸的单调载荷，直至失效。 根据MTS载荷-位移跟踪，将计算屈服载荷、最大失效载荷、线性刚度、屈服位移和失效位移。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In prior studies, SDF-1 has been shown to play a significant role in shoulder inflammation, subacromial bursitis and rotator cuff disease. In order to determine the potential biologic efficacy of SDF-1 inhibitors in treating shoulder inflammation, these studies will utilize a rotator cuff injury model in the rat. The specific aims of this project are to examine the biologic effect of SDF-1 pathway inhibitors AMD 3100, T140 analog (4F-Benzoyl-TN14003) and CTCE-9908, on reducing inflammation and their effects on rotator cuff tendon healing. We hypothesize that SDF-1 inhibitors will decrease inflammation in the subacromial bursa, but will not inhibit rotator cuff tendon healing. Specific Aims 1. Effect of SDF-1/CXCR4 Inhibitors on Local Inflammation in the Rat Shoulder: The morphological and biochemical effects of SDF-1/CXCR4 receptor inhibitors (AMD3100, T140 analog and CTCE-9908), and antibodies to CXCR4 (MAB171 and 12G5) will be examined in rats with subacromial inflammation induced by rotator cuff injuries. Male Lewis rats will undergo surgery to mimic a rotator cuff injury of the infraspinatus tendon and to induce inflammation (bursitis) in the adjacent subacromial bursa. Inhibitors will be systemically administered via implanted osmotic minipumps. After sacrificing the animals, subacromial bursa will be harvested and SDF-1, interleukin-1 (IL-1), and tumor necrosis factor (TNF-alpha) expression will be measured by quantitative PCR and enzyme linked immunosorbent assay (ELISA). Histologic evaluation for subacromial inflammation and immunohistochemistry will also be performed to assess the effect of SDF-1 blockade on the local inflammatory response. 2. Effect of SDF-1/CXCR4 Inhibitors on Rotator Cuff Tendon Healing in the Rat Shoulder: The morphological, biochemical, and biomechanical effects of SDF-1/CXCR4 receptor inhibitors, (AMD3100, T140 analog and CTCE-9908), and antibodies to CXCR4 (MAB171 and 12G5) on rotator cuff tendon healing will be examined in rats with rotator cuff injuries. (see above) After harvesting rotator cuff tendons from the site of injury, tendon healing will be assessed histologically and by local expression of type I and III collagen. Tendon strength will be assessed using an MTS 810 servohydraulic load frame (MTS Corp, Eden Prairie, MN). The insertion and attachment sites of the supraspinatus tendon will be embedded rigidly in urethane (Smooth On, Easton, PA) and subjected to monotonic loading in axial tension at a constant displacement until failure. From the MTS load-displacement tracing, the yield load, maximum failure load, linear stiffness, displacement to yield and displacement to failure will be calculated.