Calvarial Regeneration using Biomatrix-Encapsulated Skeletal Progenitors

描述（由申请人提供）：由于先天性疾病，创伤，癌症和手术干预，颅面骨骼缺损经常发生，是重建中最具挑战性的问题之一。虽然婴儿有能力治愈大而复杂的颅骨缺损，但一岁以上的婴儿即使是小的颅骨缺损也没有足够的愈合反应。虽然在过去的一个世纪中已经开发了大量的策略，但目前可用的方法拼凑反映了每种治疗技术的不足之处。在这方面，骨骼干细胞生物学在未来的组织工程应用中具有巨大的潜力。该项目的目标是使基于自体骨骼祖细胞的颅面骨骼再生成为临床现实。采用跨学科的方法，我们将汇集我们在干细胞生物学，生物工程和颅面外科方面的专业知识，以解决以细胞为基础的骨骼组织工程翻译的障碍。在Specific Aim 1中，我们将从不同组织中分离纯化人类骨骼祖细胞群体，并确定其扩增和分化所需的关键祖细胞生态位因子。在Specific Aim 2中，我们将开发一种新型的高通量，基于微流体的FACS （MF-FACS）设备，用于在水凝胶为基础的微环境中同时分离和封装单个骨骼祖细胞，从而促进其再生能力。在具体目标3中，我们将评估移植的包封骨祖细胞在临界尺寸颅骨缺损模型中的再生潜力。补充生态位因子的作用将使用可调节的宏观水凝胶支架材料进一步评估。我们相信这个高度创新的项目最终会产生一个有效的基于细胞的颅面骨骼再生团，适合临床试验。

DESCRIPTION (provided by applicant): Debilitating craniofacial skeletal defects, which occur frequently as a result of congenital disorders, trauma, cancer, and surgical interventions, are some of the most challenging problems for reconstruction. While infants demonstrate the ability to heal large and complex calvarial defects, those older than one year of age have an insufficient healing response to even small skull defects. Although a plethora of strategies have been developed over the past century, the patchwork of methods currently available reflects the inadequacies of each therapeutic technique. In this regard, skeletal stem cell biology holds enormous untapped promise for future tissue engineering applications. The goal of this project is to make autogenous skeletal progenitor cell-based craniofacial skeletal regeneration a clinical reality. Using an interdisciplinary approach, we will bring together our expertise in stem cell biology, bioengineering, and craniofacial surgery to tackle the roadblocks to translation of cell-based skeletal tissue engineering. In Specific Aim 1, we will prospectively isolate pure populations of human skeletal progenitor cells from different tissues and identify key progenitor cell niche factors necessary for their expansion and differentiation. In Specific Aim 2, we will develop a novel high throughput, microfluidics-based FACS (MF-FACS) device for simultaneous isolation and encapsulation of individual skeletal progenitor cells in a hydrogel-based, microenvironment conductive to their regenerative capabilities. In Specific Aim 3, we will assess the regenerative potential of transplanted encapsulated skeletal progenitors in critical-sized calvarial defect models. The role of supplementary niche factors will be further assessed using a tunable macroscale hydrogel scaffolding material. We believe this highly innovative project will ultimately produce an effective cell-based craniofacial skeletal regeneration regiment suitable for clinical testing. PUBLIC HEALTH RELEVANCE: The current tools available to doctors to repair bone structures of the head and face damaged from accidents, birth defects or cancer are inadequate. Using cells that have been removed from the patient, this project seeks to identify, protect, and return only those cells capable of growing into new bone structures. We seek to improve public health by developing a point-of-care method for surgical reconstruction of living bone.