Does FAK mediate implant microtopography

描述（由申请人提供）：随着人口老龄化的增加，越来越多的人被定义为部分无牙。护理标准已经扩大到包括使用牙种植体。为了成功种植体必须与周围的硬组织结合，这一过程被称为骨整合。由于微地形在分子水平上对细胞的影响，骨整合的定义是模糊的。这些表面如何影响骨整合过程中成骨细胞分化的分子途径尚不清楚。将可预测的植入物表面形貌与临床结果相结合，取决于对细胞和分子水平三维生物学的良好理解。我们的假设是牙种植体表面微形貌对成骨细胞分化和运动的影响是通过局灶黏附激酶（FAK）信号通路调节的。FAK是一种与整合素受体相关的细胞内酪氨酸激酶。我们的总体策略是在一系列表征良好的钛种植体表面微地形（600g沟槽与50um氧化铝喷砂）上对人类腭间充质前成骨细胞进行细胞粘附、分化和迁移研究。我们将确定微地形效应是否通过FAK调制。Real - time PCR和Western blotting策略将用于从野生型对照中分离的RNA和蛋白质，或使用siRNA-FAK、FRNK显性阴性构建体或Y925F ha标记的FAK突变体降低FAK基因和蛋白质表达水平的细胞。这将使我们能够确定FAK、FAK-磷酸酪氨酸或FAK- mapk通路是否受到种植体表面微形貌的调节。扫描电子显微镜将使我们能够评估静态表型变化。标记细胞的实时成像将允许实时评估动态变化。这项工作的意义在于，它将深入了解如何通过改变植入物表面微形貌，在生物材料表面的导电和感应效应方面，以可控和可预测的特定部位方式设计骨组织。这将允许一种控制的、局部的、特定部位的组织工程方法与牙科种植治疗相结合。

DESCRIPTION (provided by applicant): As the aging population increases more individuals are being defined as partially edentulous. Standards of care have expanded to include the use of Dental implants. To be successful the implant must integrate with the surrounding hard tissues, a process known as osseointegration. The definition of osseointegration is vague with respect to cellular influences of the microtopography at the molecular level. How these surfaces influence molecular pathways of osteoblast differentiation during osseointegration is not known. Integration of predictable implant surface topography with clinical outcomes depends on a well defined understanding of the three dimensional biology at the cellular and molecular level. Our hypothesis is that the effect of Dental implant surface microtopographies on osteoblast differentiation and motility is regulated through focal adhesion kinase (FAK) signaling pathways. FAK is an intracellular tyrosine kinase that is associated with integrin receptors. Our overall strategy is to perform cell adhesion and differentiation, and migration studies with human palatal mesenchymal pre-osteoblast cells on a series of well characterized titanium implant surface microtopographies 600g grooved versus 50um Al2O3 sandblasted). We will determine if microtopography effects are modulated through FAK. Real time PCR and Western blotting strategies will be used on RNA and protein isolated from wild type controls, or cells whose level of FAK gene and protein expression has been reduced using siRNA-FAK, FRNK dominant negative constructs, or Y925F HA-tagged FAK mutants. This will allow us to determine if FAK, FAK-phosphotyrosine, or FAK-MAPK pathways are modulated by implant surface microtopographies. Scanning electron microscopy will allow us to evaluate static phenotypic changes. Live imaging of labeled cells will allow for evaluation of dynamic changes in real time. The significance of this work is that it would yield insight into how osseous tissues may be engineered in a controlled and predictable site specific manner with respect to conductive and inductive effects of the biomaterial surface by alteration of the implant surface microtopography. This would allow for a controlled, localized, site specific tissue engineering approach in association with Dental implant therapy.