Tissue Engineering in Congenital Craniofacial Defects

这项研究的长期目标是开发组织工程学策略，以了解和 颅面畸形的治疗。这一提议既是假设的，也是设计驱动的。我们会 研究颅面畸形Apert综合征小鼠模型细胞的组织发育。 我们推测，由co-Pi建立的FGFR2敲入Apert小鼠模型的突变细胞将 与正常细胞相比，发育组织和对生长因子的反应方式明显不同 当生长在单层或三维水凝胶支架中时。此外，我们假设 当正常细胞和突变细胞在多层水凝胶支架中共培养时，增殖，组织 发展，增长因素的反应也会发生变化。将制定提案的设计驱动部分 修复医源性和疾病小鼠颅面缺损的一种临床实用策略 模型，使用实验室开发的细胞封装技术。为了检验这些假设并设计 维修系统将调查以下具体目标： 具体目的1.研究FGFR2基因敲除细胞的细胞行为和组织发育 艾伯特综合征小鼠模型。具体而言，这一目标将涉及1。)兔关节软骨的形成和成骨 来自Apert综合征FGFR2+/S252W小鼠模型的细胞与来自正常同胞和 永生化克隆化的小鼠间充质干细胞系。来自A.)长骨和b。)头盖骨 将在体外对2D单层和3D水凝胶支架进行评估。 特定目标2.研究突变细胞和正常细胞对 生长因子和共培养。具体而言，这一目标将涉及1。)突变细胞和正常细胞对 2D和3D水凝胶培养中的肌肉骨骼相关生长因子，以及2.应用一种新型的3D双层结构 水凝胶体系共培养正常和突变细胞以确定共培养对细胞行为的影响 以及正常和突变细胞的组织发育。 具体目的3.在颅面模型中检测突变细胞和正常细胞的体内功能。具体来说， 1.组织发育。)医源性小鼠颅骨缺损和2。)Apert综合征的先天缺陷将 使用嵌入水凝胶中的细胞进行评估。正常和突变的小鼠细胞系将用于 添加到克隆来源的小鼠间充质干细胞系中。组织发育将通过以下方式进行评估 放射学和组织学分析

The long term objective of this research is to develop tissue engineering strategies for the understanding and treatment of craniofacial abnormalities. This proposal is both hypothesis and design driven. We will investigate tissue development of cells from mouse models of the craniofacial abnormality Apert syndrome. We hypothesize that mutant cells from the FGFR2 knock-in Apert mouse model developed by the co-Pi will develop tissue and respond to growth factors in a significantly different manner compared to normal cells when grown in monolayer or in three dimensional hydrogel scaffolds. Furthermore, we hypothesize that when the normal and mutant cells are cocultured in multilayered hydrogel scaffolds, proliferation, tissue development, and growth factor response will change. The design driven portion of the proposal will develop a clinically practical strategy for repair of craniofacial defects, in iatrogenic defects and in the disease mouse model, using cell encapsulation technologies developed by the lab. To test these hypotheses and design the repair system the following specific aims will be investigated: Specific aim 1. Investigate cell behavior and tissue development of cells isolated from FGFR2 knock-in mouse model of Apert Syndrome. Specifically this aim will address 1.) chondrogenesis and osteogenesis of cells from a Fgfr2+/S252W mouse model of Apert Syndrome, compared to cells from normal siblings and an immortalized, clonal mouse mesenchymal stem cell line. Cells from a.) the long bone and b.) the cranium will be evaluated in 2D monolayer and 3D hydrogel scaffolds in vitro. Specific aim 2. Investigate cell behavior and tissue development of mutant and normal cells in response to growth factors and co-culture. Specifically this aim will address 1.) response of mutant and normal cells to musculoskeletal-related growth factors in 2D and 3D hydrogel culture, and 2.) apply a novel 3D bilayered hydrogel system to coculture normal and mutant cells to determine the effects of coculture on cell behavior and tissue development of normal and mutant cells. Specific aim 3. Determine in vivo function of mutant and normal cells in craniofacial models. Specifically, tissue development in 1.) iatrogenic mouse cranial defects and 2.) congenital defects in Apert Syndrome will be evaluated using cells embedded in hydrogels. Normal and mutant mouse cell lines will be used in addition to a clonally-derived mouse mesenchymal stem cell line. Tissue development will be assessed by radiographic and histological analyses