Novel Nano Calcium Phosphate for Pulp Gene Delivery

描述（由申请人提供）：基因治疗有望治疗多种疾病。然而，使用生物载体的转移技术并非没有严重的问题。因此，使用非病毒方法的基因递送是高度优选的，这是由于递送方便、易于制造、成本效益和生物相容性。然而，这些技术受到低转染效率的限制。为了解决这个问题，我们已经开发了纳米尺寸的磷酸钙颗粒，NanoCaPs，作为一种新的质粒DNA转染的传递系统。我们假设NanoCaPs将作为一种上级转染系统，因为减小的晶体尺寸将导致增加的表面活性DNA结合和更大的细胞摄取以及增强的转染效率。这种优雅的非病毒基因传递系统可以单独使用或掺入合成或天然聚合物中，以持续的方式传递基因。 为了证明我们的假设，我们建议合成纳米CaP掺入生物可降解的纤维蛋白基质，以提高质粒DNA（pDNA）的体外和体内转染效率。该技术将增加标记基因（荧光素酶和/或β-半乳糖苷酶）以及治疗性转染物的摄取和表达。牙本质再生治疗模型将用于局部施用复合纳米级磷酸钙载体基质中的编码大鼠骨形态发生蛋白-7（BMP-7）的质粒DNA载体，以增强牙本质再生。我们初步的体外和体内数据与我们设计和开发这种有效的质粒基因治疗系统的目标非常一致。这些结果表明，我们的方法用于牙本质再生的潜在成功应用。 本研究将为进一步开展牙本质质粒基因治疗相关的应用和基础科学研究提供具体的基础，也将为研究牙本质修复和再生的生物学提供模型。已经制定了具体目标，以提供与体外和体内pDNA转染效率相关的基本问题的解决方案，以及包埋在纤维蛋白基质中的pDNA结合的NanoCaP用于牙本质治疗的具体应用，从而提供目前缺失的关键信息。

DESCRIPTION (provided by applicant): Gene therapy holds promise for treatment of a variety of diseases. However, transfer techniques using biological vectors is not without serious problems. Therefore, gene delivery using non-viral approach is highly preferable due to convenience of delivery, ease of manufacturing, cost effectiveness, and biocompatibility. However, these techniques are limited by low transfection efficiency. To address this problem, we have developed nano-sized calcium phosphate particles, NanoCaPs, as a novel delivery system for plasmid DNA transfection. We hypothesize that NanoCaPs will serve as a superior transfection system because the reduced crystalline dimensions will result in increased surface-active DNA binding and greater cellular uptake and enhanced transfection efficiency. This elegant non-viral gene delivery system could be used alone or incorporated into synthetic or natural polymers to deliver genes in a sustained manner. To prove our hypothesis we propose to synthesize NanoCaPs incorporated in a biodegradable fibrin matrix to enhance both the in vitro and in vivo transfection efficiency of plasmid DNA (pDNA). This technology will increase the uptake and expression of marker genes (luciferase and/or beta-galactosidase) as well as therapeutic transfectents. A dentin regenerative therapy model will be used to locally administer plasmid DNA vectors encoding rat bone morphogenetic protein-7 (BMP-7) in a composite nano-sized calcium phosphate carrier matrix to enhance dentin regeneration. Our preliminary in vitro and in vivo data is in excellent agreement with our objective to design and develop this efficient plasmid gene therapy system. These results suggest the potential successful application of our approach for dentin regeneration. The present study will provide a concrete foundation for conducting further applied and basic science research related to dentin plasmid gene therapy and will also provide a model to study the biology for dentin repair and regeneration. The specific aims have been formulated to provide solutions to fundamental questions related to in vitro and in vivo pDNA transfection efficiency and the specific application of the pDNA bound NanoCaPs embedded in a fibrin matrix for dentin therapy thus providing key information currently missing.