Osteoblast Response to Amorphous Calcium Phosphate

成骨细胞对无定形磷酸钙的反应

• 批准号: 6741234
• 负责人: BRIAN J LOVE
• 金额: $ 7.18万
• 依托单位: VIRGINIA POLYTECHNIC INST AND ST UNIV
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6741234
• 关键词: bioengineering /biomedical engineering; biomaterial development /preparation; biomaterial interface interaction; bone regeneration; calcium phosphate; laboratory rat; medical implant science; oral facial restoration; oral facial restoration material; osteoblasts; polymers; tissue /cell culture; tissue engineering; tissue support frame

DESCRIPTION (provided by applicant): The broad, long-term objective is to evaluate how osteoblast response is mediated by nutrient transport provided by dissolving amorphous calcium phosphate (ACP) particulates. We hypothesize that the attraction of ACP lies in its higher dissolution rate, and its ready interaction with osteoblasts leading to more rapid mineralization. We propose that ACP mineral immobilized membranes could be the first step in creating viable scaffolds for structural repair and regeneration. The proposed effort is distributed among three linked Aims. Aim 1 will perform targeted cell culture studies of stabilized forms of ACP both separated by neighboring osteoblasts through a membrane in the Transwell plate and by direct contact experiments under cell culture conditions between cells and ACP. We will evaluate the release rate of target ions in solution and track cell viability and function with incubation time in growth media deprived of calcium phosphate. Protein expression will be evaluated by Northern blot and other gel and assay techniques. Aim 2 will construct model porous scaffolds incorporating and immobilizing stabilized ACP within them by a photopolymerizable pre-polymer dispersion. Aim 3 will evaluate osteoblast cell response to the immobilized ACP composites from cell culture using Transwell plates to compare cell viability and function in this different mode. Similar assays are planned in Aim 3 as in Aim 1 with the distinction that other controls will be required to separate the matrix effect from the mineral effect. Thus Aim 3 leverages off of the experience gained in cell culture during Aim 1 with the ability to fabricate mineral immobilized membrane matrices in Aim 2. At the end of the project, we will have compiled strong evidence linking ACP solubility, neighboring osteoblast viability, and tissue growth and proliferation. We believe that these foundation elements will provide the basis for a successful R-01 proposal concept to be submitted as this project is completed.

描述（由申请方提供）：广泛的长期目标是评价成骨细胞反应如何通过溶解无定形磷酸钙（ACP）颗粒提供的营养转运介导。我们推测ACP的吸引力在于其较高的溶解速率，以及其与成骨细胞的快速相互作用，从而导致更快的矿化。我们建议，ACP矿物固定膜可能是第一步，在创造可行的支架结构修复和再生。拟议的努力分布在三个相互关联的目标中。目标1将对ACP的稳定形式进行靶向细胞培养研究，这些稳定形式通过Transwell板中的膜与相邻成骨细胞分离，并通过细胞和ACP之间在细胞培养条件下的直接接触实验分离。我们将评估靶离子在溶液中的释放速率，并跟踪细胞活力和功能，以及在缺乏磷酸钙的生长培养基中的孵育时间。将通过北方印迹和其他凝胶和测定技术评价蛋白质表达。目的2将构建模型多孔支架，通过光聚合预聚物分散体将稳定的ACP掺入并固定在其中。目的3将使用Transwell板评估成骨细胞对来自细胞培养的固定化ACP复合物的反应，以比较在这种不同模式下的细胞活力和功能。在目标3中计划了与目标1类似的试验，区别在于需要其他对照来将基质效应与矿物质效应分开。因此，目标3利用了目标1期间在细胞培养中获得的经验，具有在目标2中制造矿物质固定化膜基质的能力。在项目结束时，我们将收集到强有力的证据，将ACP溶解度、邻近成骨细胞活力、组织生长和增殖联系起来。我们相信，这些基础要素将为在该项目完成时提交的成功的R-01提案概念提供基础。