Bone Scaffolds for Heat Shock Protein Induced Regeneration and Healing

主要研究者：Rylander，Nicole提案编号：1067654项目总结：与癌症、损伤、发育异常和退行性疾病相关的骨相关疾病极大地降低了数百万人的健康和生活质量。这些疾病可通过骨或其功能丧失导致严重残疾，从而需要骨替代（每年进行超过300万例骨科手术）或有效的再生策略。使用热应力和张应力的调节可以上调ECM的产生、细胞增殖和称为热休克蛋白（HSP）的分子伴侣。已显示上调的HSP与增强的细胞增殖和ECM形成所需的胶原蛋白生物合成之间的联系。最终目标是通过应力调节和HSP递送开发具有增强体内伤口愈合和骨再生能力的变革性上级骨支架。将确定理想的应力调节策略和外源性HSP递送方案以创建更多功能性骨支架，并将测试这些支架在啮齿动物颅面缺损模型中促进愈合的功效。本研究的目的是：1）构建一种新型的微生物反应器系统，2）使用微生物反应器系统向骨支架单独施加组合的热和拉伸应力以及与HSP递送组合施加组合的热和拉伸应力，并确定用于增强骨形成的理想条件，和3）评估用热+张应力和HSP递送预处理的骨支架在大鼠颅面缺损模型中愈合骨缺损的有效性。智力优势：这将是第一个研究重点是利用HSP的骨再生的潜力，通过组合应力调节和外源性HSP交付功能性骨支架的发展。另一个新颖的方面将是热应力和张应力的组合使用，以增强骨支架内和体内颅面骨缺损模型中的细胞增殖和骨ECM形成。将创建并利用第一种微型生物反应器系统来组合施加热应力和张应力，以确定促进骨形成的最佳应力调节方案。我们试图创造一种上级骨支架，由于使用了新的制造方法，该方法具有变革性，该方法包括与整合的HSP释放微球偶联的共静电纺丝聚合物，用热应力+拉伸应力调节，以及微球的表面封装，用于将HSP从支架释放到周围组织。能够在空间上和时间上控制HSP在支架内递送并递送至周围伤口部位的支架将提供独特的途径来刺激骨愈合并促进支架在具有损伤或患病组织的患者中的现有骨内的成功整合。更广泛的影响：这项研究将建立一种新的方法来促进骨生长和再生，以开发更有活力的骨替代品和刺激患者骨再生的策略。从本研究中获得的知识将直接转化为恢复骨组织的功能，并消除与骨相关损伤相关的现有残疾。最终，应力调节策略和HSP递送方法可用于开发多种工程组织替代物，例如韧带、肌腱、肌肉和神经，以允许刺激患者中任何类型的损伤或患病组织的愈合。 这项研究将使学生在研究生，本科和高中阶段获得组织工程，生物运输，成像和细胞生物学方面的经验。将资助两名代表性不足的研究生。少数民族本科生将融入研究的各个方面，以促进学生感知其教育与研究的相关性的机制，从而激励他们在学习中脱颖而出，促进研究生院的追求。残疾高中生将体验与该项目相关的第一手研究技术，如支架和微球的制造，使用Instron测试材料性能，调节支架和测量支架反应。这个机会将创造生物医学工程的魅力，并鼓励学生，尽管他们的挑战，在研究的未来是可以实现的。

PI: Rylander, NicoleProposal Number: 1067654Project Summary: Bone related disorders associated with cancer, injury, abnormal development, and degenerative conditions dramatically diminish the health and quality of life of millions of people. These disorders can cause significant disability through loss of bone or its functionality, creating a need for bone replacements (over 3 million orthopaedic procedures performed annually) or effective regenerative strategies. Conditioning using thermal and tensile stress can up-regulate ECM production, cell proliferation, and molecular chaperones called heat shock proteins (HSPs). A link has been shown between up-regulated HSPs and enhanced cell proliferation and collagen biosynthesis needed for ECM formation. The ultimate goal is to develop a transformative, superior bone scaffold through stress conditioning and HSP delivery with the capability to enhance wound healing and bone regeneration in vivo. Ideal stress conditioning strategies and exogenous HSP delivery protocols will be identified to create more functional bone scaffolds and the efficacy of these scaffolds to promote healing in a rodent craniofacial defect model will be tested. Study objectives are to 1) Construct a novel microbioreactor system to apply combinatorial (thermal+tensile) stress and create a scaffold capable of exogenous HSP delivery and wound healing, 2) Apply combinatorial thermal and tensile stress alone and in combination with HSP delivery to bone scaffolds using the microbioreactor system and determine ideal conditions for enhancing bone formation, and 3) Evaluate effectiveness of bone scaffolds preconditioned with thermal+tensile stress and HSP delivery to heal bone defects in a rat craniofacial defect model. Intellectual Merit: This will be the first study focused on harnessing the potential of HSP based bone regeneration through combinatorial stress conditioning and exogenous HSP delivery in development of functional bone scaffolds. Another novel aspect will be the combined use of thermal and tensile stress to enhance cell proliferation and bone ECM formation within bone scaffolds and in an in vivo craniofacial bone defect model. A first-of-its-kind microbioreactor system will be created and utilized to apply thermal and tensile stress in combination to allow determination of optimal stress conditioning protocols to promote bone formation. We seek to create a superior bone scaffold which is transformative due to the use of novel fabrication methods comprised of co-electrospinning polymers coupled with integrated HSP releasing microspheres, conditioning with thermal+tensile stress, and surface encapsulation of microspheres for HSP release from the scaffold to the surrounding tissue. Scaffolds capable of controlled HSP delivery spatially and temporally within the scaffold and to the surrounding wound site will provide a unique avenue to stimulate bone healing and promote successful integration of the scaffold within existing bone in patients with injured or diseased tissue. Broader Impacts: This research will establish a new methodology for enhancing bone growth and regeneration for development of more viable bone replacements and strategies for stimulating bone regeneration in patients. Knowledge gained from this study will directly translate to restoring functionality of bone tissue and eliminating the existing disabilities associated with bone-related impairments. Ultimately, stress conditioning strategies and HSP delivery methods can be utilized for development of a wide array of engineered tissue replacements such as ligaments, tendons, muscles, and nerves to permit stimulation of healing of any type of injured or diseased tissue in patients. This research will enable students to gain experience in tissue engineering, biotransport, imaging, and cell biology at the graduate, undergraduate, and high school level. Two underrepresented graduate students will be supported. Minority undergraduate students will be integrated into every aspect of the research to facilitate a mechanism for students to perceive the relevance of their education to research thereby inspiring them to excel in their studies and promote pursuance of graduate school. High school students with disabilities will experience first-hand research techniques related to this project such as fabrication of scaffolds and microspheres, testing of material properties using the Instron, conditioning scaffolds, and measuring scaffold response. This opportunity will create fascination with biomedical engineering and encourage students that despite their challenges a future in research is attainable.