A Novel RNA Based Biomaterial for Bone Regeneration

一种基于 RNA 的新型骨再生生物材料

基本信息

批准号：
8819172
负责人：
Satheesh Elangovan
金额：
$ 18.39万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-12-01 至 2016-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8819172
关键词：
Aftercare Alkaline Phosphatase Animals Area Biocompatible Biocompatible Materials Biological Response Modifiers Biomimetic Materials Bone Development Bone Marrow Bone Regeneration Bone Tissue Calvaria Cells Charge Chemicals Clinical Collagen Collagen Type I Control Groups Core-Binding Factor Defect Dentistry Development Effectiveness Extrinsic asthma Fracture Fracture Healing Future Gene Proteins Genes Goals Histological Techniques Histology Human Imaging Techniques Immune Immunoprecipitation Implant In Vitro Inbred BALB C Mice Intramuscular Intraperitoneal Injections Knowledge Left Lung diseases Measures Medicine Messenger RNA Methods Minerals Modeling Modification Mus Nodule Non-Viral Vector Osteocalcin Osteogenesis Physical condensation Preparation Proteins RNA RNA Stability Rattus Recombinant Proteins Recombinants Regenerative Medicine Safety Serum Site Staining method Stains Stromal Cells Surface Surface Properties System Testing Therapeutic Time Tissue Engineering Transfection Translations Viral Vector Work X-Ray Computed Tomography base bone bone morphogenetic protein 2 chemical release chemical synthesis clinically significant cost effective cytotoxicity dosage gene therapy immunogenicity implantation in vitro Model in vivo innovation nanoparticle new technology novel novel strategies osteoblast differentiation prevent primary outcome public health relevance recombinant human bone morphogenetic protein-2 regenerative scaffold targeted cancer therapy targeted delivery uptake

项目摘要

DESCRIPTION (provided by applicant): Chemical modification of bases in mRNA (cmRNA) has been shown recently to significantly enhance its cellular uptake and intra-cellular stability. This strategy was shown to be safe and efficient when used for reprogramming cells, in targeted cancer therapy or for therapeutic in vivo delivery of target molecules to treat lethal lung disease or to prevent allergic asthma. This study is directed at harnessing this novel technology for regenerative applications. Specifically, this study aims to develop a biomaterial based delivery system that releases cmRNA of bone morphogenetic protein-2 (BMP-2) in a controlled fashion to induce bone regeneration. The specific aims are: (1) to synthesize and characterize a biomaterial-based cmRNA (BMP-2) delivery system and (2) to determine the in vivo efficacy of a biomaterial-based cmRNA (BMP-2) delivery system to induce bone formation. For aim 1, polyethylenimine (PEI)-cmRNA (encoding BMP-2) nanoplexes will be synthesized using established methods and characterized for size, surface charge and cmRNA condensation. The cytotoxicity and transfection efficiency of synthesized nanoplexes will be evaluated in human bone marrow stromal cells (BMSC) and murine pre-osteoblastic cells (MC3T3-E1). Expression levels of bone specific genes (osteocalcin and type I collagen), core binding factor (Cbfa)-1 and Osterix in the transfected cells will be determined. The functionality of transfection will be examined by assessing mineral nodule formation and measuring alkaline phosphatase activity in transfected cells. For aim 2, the optimized preparations will be implanted in calvarial bone defects (CBD) in Fisher 344 rats. After four and eight weeks, animals will be euthanized and new bone volume and bone area (%) at the implanted sites will be assessed using imaging and histologic techniques.

描述（由申请人提供）：最近已显示MRNA（CMRNA）中碱基的化学修饰，可显着增强其细胞摄取和细胞内稳定性。该策略在用于重编程细胞，靶向癌症治疗或在体内递送靶分子的治疗以治疗致死性肺疾病的治疗方面被证明是安全有效的或防止过敏性哮喘。这项研究旨在利用这项新技术用于再生应用。具体而言，这项研究旨在开发一种基于生物材料的递送系统，该系统以受控方式释放骨形态发生蛋白-2（BMP-2）的CMRNA以诱导骨骼再生。具体目的是：（1）综合和表征基于生物材料的CMRNA（BMP-2）输送系统和（2）确定基于生物材料的CMRNA（BMP-2）递送系统的体内功效以诱导骨形成。对于AIM 1，将使用已建立的方法合成聚乙基亚胺（PEI）-CMRNA（编码BMP-2）纳米词，并针对尺寸，表面电荷和CMRNA缩合进行表征。合成纳米插曲的细胞毒性和转染效率将在人骨髓基质细胞（BMSC）和鼠前构成细胞前细胞（MC3T3-E1）中进行评估。将确定转染细胞中骨特异性基因（骨钙素和I型胶原蛋白），核心结合因子（CBFA）-1和Osterix的表达水平。转染的功能将通过评估矿物结节的形成和测量转染细胞中的碱性磷酸酶活性。对于AIM 2，优化的制剂将植入Fisher 344大鼠的颅骨缺损（CBD）中。四个和八周后，将使用成像和组织学技术评估动物在植入部位的新骨体积和骨面积（％）。