Scaffold-Mediated Gene Delivery for Engineering of Osteochondral Tissues

用于骨软骨组织工程的支架介导的基因传递

• 批准号: 9069429
• 负责人: Charles A. Gersbach
• 金额: $ 19.96万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9069429
• 关键词: Address; Adult; Affect; Architecture; Binding; Biochemical; Biocompatible Materials; Biological; Bone Marrow; Cartilage; Cell Culture Techniques; Cell Differentiation process; Cell Separation; Cells; Chondrogenesis; Clinical; Complex; Conditioned Culture Media; Cues; Defect; Development; Diffusion; Disease; Doxycycline; Engineering; Fiber; Filament; Gene Delivery; Gene Expression; Gene Transfer; Genes; Genetic Engineering; Geometry; Goals; Growth; Guided Tissue Regeneration; Healed; Health; Heterogeneity; Histocompatibility Testing; Histologic; Histology; Human; Immobilization; Immunohistochemistry; Implant; In Situ; In Vitro; Injury; Joints; Lead; Lentivirus Vector; Life; Lysine; Mechanics; Mediating; Mesenchymal Stem Cells; Methods; Modeling; Musculoskeletal; Natural regeneration; Operative Surgical Procedures; Orthopedics; Oryctolagus cuniculus; Osteogenesis; Pain; Pattern; Phenotype; Physiological; Polymers; Procedures; Property; Regenerative Medicine; Research Proposals; Role; Signaling Molecule; Skin; Specific qualifier value; Stem cells; Structure; Subfamily lentivirinae; Surface; Techniques; Technology; Testing; Thick; Tissue Engineering; Tissues; Transgenes; Translations; United States; Viral; Work; autocrine; base; bone; bone morphogenetic protein 2; cartilage regeneration; cell type; cellular engineering; conditioning; disability; healing; implant material; in vivo; innovation; microCT; novel; osteochondral repair; osteochondral tissue; osteogenic; paracrine; particle; preclinical study; regenerative; scaffold; skeletal tissue; spatiotemporal; stem cell differentiation; three dimensional structure; tissue regeneration; transcription factor; transgene expression; vector; viral gene delivery

DESCRIPTION: The regeneration of damaged or diseased skeletal tissues remains a significant clinical challenge and cause for human disability and discomfort. Tissue engineering and regenerative medicine are emerging as promising strategies for treating these conditions by creating living tissue substitutes composed of cells, bioactive factors, and a biodegradable scaffold. Tissue engineering has been particularly successful in creating uniform tissues, such as skin and cartilage. However, many injuries and diseases affect the interfaces between tissues, such as the transition between bone and cartilage. Many regenerative medicine studies that address these more complex tissues have been successful in engineering tissues in vitro with combinations of progenitor cells and biomaterials. However, these approaches typically require elaborate, laborious, and expensive procedures for cell isolation, expansion, and in vitro cell conditioning for proper cell differentiation and tissue formation. A primary challenge has been the translation of these promising preclinical studies into methods that are not only straightforward to practice but also effectively direct cell differentiation and tissue formation tat recapitulates the complexity of natural tissues. Technologies that capitalize on advances in cell engineering but minimize in vitro cell manipulation can greatly enhance the promise of regenerative medicine. This project develops a method to deliver bioactive factors to cells in a precise spatial pattern within a three dimensional scaffold. As a result, stem cells seeded onto these scaffolds or endogenous progenitor cells that infiltrate the scaffold will be stimulated to produce different tissue types in predefined patterns. In this study, we will first generate osteochondral tissues with human mesenchymal stem cells through spatially controlled gene transfer of differentiation factors. Precise spatial control will be enabled by innovative methods of biomaterial-mediated gene delivery and a microscale weaving technique for creating 3D polymer scaffolds. We will then expand this in vitro approach to a rabbit model of microfracture surgery, where the scaffold is populated by endogenous progenitor cells from the bone marrow that are instructed to form tissues in situ based on viral transgene delivery from the scaffold. This work is significant to developing tissue engineering strategies for creating complex structures that recapitulate the heterogeneity and function of native tissue interfaces and minimize in vitro cell manipulations.

产品说明：受损或患病骨骼组织的再生仍然是一个重大的临床挑战，也是人类残疾和不适的原因。组织工程和再生医学正在成为通过创建由细胞、生物活性因子和可生物降解支架组成的活组织替代物来治疗这些病症的有前景的策略。组织工程在创造均匀的组织，如皮肤和软骨方面特别成功。然而，许多损伤和疾病会影响组织之间的界面，例如骨和软骨之间的过渡。许多针对这些更复杂组织的再生医学研究已经成功地在体外用祖细胞和生物材料的组合来工程化组织。然而，这些方法通常需要精细、费力和昂贵的程序来进行细胞分离、扩增和体外细胞调节以进行适当的细胞分化和组织形成。一个主要的挑战是将这些有前途的临床前研究转化为不仅易于实践而且有效地指导细胞分化和组织形成的方法，达特重现天然组织的复杂性。利用细胞工程的进步，但最大限度地减少体外细胞操作的技术可以大大提高再生医学的前景。该项目开发了一种在三维支架内以精确的空间模式将生物活性因子递送到细胞的方法。因此，接种到这些支架上的干细胞或渗入支架的内源性祖细胞将被刺激以产生预定模式的不同组织类型。在本研究中，我们将首先通过空间控制的分化因子基因转移，用人骨髓间充质干细胞生成骨软骨组织。精确的空间控制将通过生物材料介导的基因递送的创新方法和用于创建3D聚合物支架的微尺度编织技术来实现。然后，我们将这种体外方法扩展到微骨折手术的兔模型，其中支架由来自骨髓的内源性祖细胞填充，这些细胞被指示基于从支架递送的病毒转基因原位形成组织。这项工作是重要的，以开发组织工程策略，创造复杂的结构，概括的异质性和功能的天然组织接口，并尽量减少在体外细胞操作。