Does FAK mediate implant microtopography

FAK 是否介导种植体微形貌

• 批准号: 6910952
• 负责人: GALEN B SCHNEIDER
• 金额: $ 22.13万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6910952
• 关键词: RNA interference; biological signal transduction; biomaterial interface interaction; cell differentiation; cell motility; endosseous dental implant; enzyme activity; focal adhesion kinase; medical implant science; osteoblasts; polymerase chain reaction; scanning electron microscopy; surface property; tissue /cell culture; titanium; western blottings

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.

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缺牙。护理标准已扩大到包括使用牙科植入物。为了获得成功，种植体必须与周围的硬组织整合，这一过程称为骨整合。骨整合的定义在分子水平上的微观形貌的细胞影响方面是模糊的。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。可预测的种植体表面形貌与临床结果的整合取决于在细胞和分子水平上对三维生物学的明确理解。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黏着斑激酶（FAK）信号通路调节的。FAK是与整联蛋白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充分表征的钛种植体表面显微形貌（600 g开槽与50 um Al 2 O3喷砂）上，使用人腭间充质前成骨细胞进行细胞粘附和分化以及迁移研究。我们将确定微形貌效应是否通过FAK调制。真实的时间PCR和蛋白质印迹策略将用于从野生型对照或FAK基因和蛋白质表达水平已经使用siRNA-FAK、FRNK显性阴性构建体或Y 925 F HA标记的FAK突变体降低的细胞分离的RNA和蛋白质。这将使我们能够确定FAK、FAK-磷酸酪氨酸或FAK-MAPK通路是否受到植入物表面微观形貌的调节。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真实的评价动态变化。这项工作的意义在于，它将产生洞察骨组织如何可以在一个可控的和可预测的网站特定的方式工程化的生物材料表面的导电和诱导效应，通过改变植入物表面的微观形貌。这将允许与牙科种植体治疗相关的受控的、局部的、位点特异性的组织工程方法。