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无牙颌。护理标准已扩大到包括牙种植体的使用。为了成功，种植体必须与周围的硬组织整合，这一过程称为骨整合。就分子水平上微形貌的细胞影响而言，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。可预测的种植体表面形貌与临床结果的整合取决于对细胞和分子水平上的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粘着斑激酶（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特征明确的钛种植体表面微形貌（600g 凹槽与 50um Al2O3 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。我们将确定微地形效应是否通过 FAK 进行调节。实时 PCR 和蛋白质印迹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 显性失活构建体或 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，以受控和可预测的位点特定方式对生物材料表面的传导和感应效应进行工程化。这将允许与牙种植体治疗相关的受控、局部、特定部位的组织工程方法。