Tissue Engineering a Suture Replacement for Children with Craniosynostosis

描述（由申请人提供）： 颅缝早闭定义为一条或多条颅缝过早融合。目前的治疗涉及广泛的手术干预，包括额眶前移和根治性颅骨复位。虽然目前的技术通常成功地恢复正常的脑生长载体，增加颅内容量，降低颅内压，手术干预是广泛的，结果是可变的。此外，颅骨再生迅速，手术后不久就可以拒绝。当发生再融合时，需要进行二次手术，这增加了患者的发病率和死亡率。我们的小组已经着手从根本上改善颅缝早闭的治疗，尽量减少手术干预的范围，改善结果，并抑制再融合的发生。我们处于独特的位置，可以设计一种新的治疗方法，可以量身定制，以改善治疗儿童谁目前与颅缝早闭，无论分子或遗传病因。为了确定适当的生长因子线索、剂量和细胞分数，我们将使用新型喷墨打印技术进行干细胞表征，以评估从人非综合征冠状缝骨性结合的兔模型中分离的骨骼肌来源和脂肪来源细胞的成骨潜力（目标#1A）。基于我们在目标#1A中的发现，我们将使用我们的喷墨打印来创建缝线置换支架，以创建固定化生长因子的空间限定图案，其将在体外可重复地形成由交错的非骨区域分开的两个骨区域（目标#1B）。然后，我们将测试我们的缝线置换支架在体内改善患有颅缝早闭的兔的手术治疗的有效性（目标#2）。为了证明这种方法在人类中的临床相关性，我们将采用与目标#1中类似的祖细胞表征，对来自患有颅缝早闭儿童的骨骼肌和脂肪的细胞进行表征，并评估这些细胞对我们的缝线替代支架的体外反应性（目标#3）。通过实现这些目标，我们将在改善颅缝早闭的外科治疗方面取得重大进展。此外，可用于表征干细胞分化潜能和空间控制多能细胞分化的技术的发展将在组织工程领域产生广泛影响。 公共卫生相关性声明（由申请人提供）：项目叙述颅缝早闭是一个或多个颅骨缝线过早融合的术语。缝线融合是由于骨和非骨组织之间未能形成正确的图案。我们已经着手通过组织工程来改善颅缝早闭儿童的手术治疗。我们将开发新兴的喷墨打印技术，以表征祖细胞和可重复的模式组织形成体内。通过细胞分化的模式化，有可能创造出与正常的、未融合的缝合线功能相似的组织。在执行拟议的工作中开发的技术将对干细胞生物学领域、组织工程的应用以及颅缝早闭儿童的治疗产生广泛的影响。

DESCRIPTION (provided by applicant): Craniosynostosis is defined as the premature fusion of one or more of the cranial sutures. Current therapy involves extensive surgical intervention including fronto-orbital advancement and radical calvarial bone repositioning. Although the current techniques often successfully restore normal brain growth vectors, increase intracranial volume, and decrease intracranial pressure, the surgical intervention is extensive and the outcomes are variable. Furthermore, the calvarial bone regenerates quickly and can refuse shortly after surgery. When refusion occurs, secondary surgeries are required which increases patient morbidity and mortality. Our group has set out to radically improve the treatment of craniosynostosis by minimizing the extent of surgical intervention, improving outcomes, and inhibiting the occurrence of refusion. We are in a unique position to design a novel therapy that could be tailored to improve the treatment of children who present with craniosynostosis, regardless of the molecular or genetic etiology. To determine the proper growth factor cues, doses, and cell fraction, we will employ stem cell characterization using a novel inkjet printing technology to assess the osteogenic potential of skeletal muscle-derived and adipose-derived cells isolated from a rabbit model of human nonsyndromic coronal suture synostosis (Aim #1A). Based on our findings from Aim #1A, we will create a suture replacement scaffold using our inkjet printing to create spatially-defined patterns of immobilized growth factors that will reproducibly form two regions of bone separated by an interdigitating non- bone region in vitro (Aim #1B). We will then test the effectiveness of our suture replacement scaffolds to improve the surgical treatment of rabbits with craniosynostosis in vivo (Aim #2). To demonstrate the clinical relevance of this approach in humans, we will employ similar progenitor cell characterization as in Aim #1 on cells derived from skeletal muscle and adipose of children with craniosynostosis and assess these cells' responsiveness to our suture replacement scaffolds in vitro (Aim #3). By completing these Aims, we will make significant progress in improving the surgical treatment of craniosynostosis. Furthermore, the development of technologies that can be used to both characterize stem cell differentiation potential and to spatially control the differentiation of multipotent cells will have broad impact in the field of tissue engineering. Public Health Relevance Statement (Provided by Applicant): Project Narrative Craniosynostosis is the term given to the premature fusion of one or more of the calvarial sutures. Fusion of a suture results from a failure of correct pattern formation between bone and non-bone tissues. We have set out to improve the surgical management of children with craniosynostosis through tissue engineering. We will develop emergent inkjet printing technology to characterize progenitor cells and to reproducibly pattern tissue formation in vivo. By patterning cell differentiation, it may be possible to create tissues that function similarly to normal, unfused sutures. The technology that will be developed in performing the proposed work will have broad impact on the field of stem cell biology, the application of tissue engineering, and on the treatment of children with craniosynostosis.