MOLECULAR BASIS OF GROWTH FACTOR ENHANCED TENDON REPAIR

本研究的重点是明确肌腱损伤早期生长因子和基质基因表达的变化。它还将评估增强愈合反应的药物手段，以实现更接近正常肌腱的机械性能的肌腱修复组织。最终目标是愈合肌腱，不太可能持续再损伤。 我们的具体目标包括文件的时间和空间表达的胶原蛋白I型和III型，生长因子胰岛素样生长因子I（IGF-I）和转化生长因子β（TGF-β），和IGF结合蛋白在屈肌腱愈合过程中胶原酶损伤的马。 生长因子的表达和随后的胶原蛋白表达变化之间的关系是中心的兴趣。 这些信息将为本次和未来研究中生物制剂给药的选择、时间和持续时间提供科学依据。 先前的体外研究记录了IGF-I对肌腱愈合的有益作用，本提案中概述的体外研究将提供β-氨基丙腈（BAPN）的初步数据。 这些体外研究的信息将与颞叶愈合研究的信息相结合，以进行概述的两项体内研究，一项单独使用IGF-I，另一项使用IGF-I和BAPN的组合。 将通过组织学染色、原位杂交、免疫染色、放射自显影和生物化学试验分析组织的DNA含量和基质组成。 将根据具体研究设计进行统计分析，包括配对t检验、单因素方差分析、Tukey事后比较、个体回归分析和Pearsons相关性。 显著性将设定为p小于或等于0.05。

The focus of this research proposal is to define the growth factor and matrix gene expression changes in early tendon injury. It will also evaluate pharmaceutical means of enhancing the healing response to achieve tendon repair tissue that more closely resembles the mechanical properties of normal tendon. The ultimate goal is healed tendons that are less likely to sustain reinjury. Our specific aims include documentation of the temporal and spatial expression of collagen types I and III, the growth factors insulin-like growth factor I (IGF-I) and transforming growth factor beta (TGF-beta), and the IGF binding proteins during the healing response of flexor tendon following collagenase injury in the horse. The relationship between growth factor expression and subsequent changes in collagen expression are of central interest. This information will provide the scientific basis for decisions regarding selection, timing, and duration of administration of biologicals in this and future investigations. Previous in vitro studies document the beneficial effects of IGF-I on tendon healing and the in vitro study outlined in this proposal will provide preliminary data on beta-aminopriopionitrile (BAPN). The information from these in vitro studies will be combined with that from the temporal healing study to perform the two in vivo studies outlined, one using IGF-I alone and the other using a combination of IGF-I and BAPN. Tissues will be analyzed by histologic stains, in situ hybridization, immunostaining, autoradiography, and biochemical assays for DNA content and matrix composition. Statistical analyses will be performed as dictated by the specific study design, including paired t-tests, one-way analysis of variance, Tukey's post hoc comparisons, individual regression analysis, and Pearsons's Correlation. Significance will be set at p less than or equal to 0.05.