Application of Tubular Perfusion System (TPS) Generated Prevascularized Bone Tiss

管状灌注系统（TPS）产生预血管化骨组织的应用

基本信息

批准号：
8512532
负责人：
John P Fisher
金额：
$ 31.61万
依托单位：
UNIV OF MARYLAND, COLLEGE PARK
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-16 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8512532
关键词：
Alginates Bioreactors Bone Injury Bone Regeneration Bone Tissue Calcium Cell Culture Techniques Cell Proliferation Cells Clinical Treatment Coculture Techniques Culture Media Defect Deposition Development Devices Diffusion Disease Encapsulated Endothelial Cells Engineering Environment Excision Fostering Goals Growth Healed Human Implant In Vitro Investigation Laboratories Mechanical Stress Mechanics Mesenchymal Stem Cells Methods Nutrient Osteocalcin Osteogenesis Oxygen Perfusion Polymers Regenerative Medicine Stem cells Structure System Technology Therapeutic Tissue Engineering Tissue Grafts Tissues Translating Trauma Tubular formation Umbilical vein Up-Regulation Vascularization base bone cell growth clinical application clinically relevant design fluid flow healing implantation improved in vivo innovation novel osteoblast differentiation osteopontin oxygen transport repaired response scaffold shear stress success tissue culture treatment strategy tumor

项目摘要

DESCRIPTION (provided by applicant): In vitro and in vivo nutrient transfer limits must be overcome in order to increase the feasibility of cell based therapeutic strategies. To enhance in vitro nutrient transport, the tubular perfusion system (TPS), a novel bioreactor recently developed by our laboratory, will dynamically culture human mesenchymal stem cells (hMSCs) in three dimensional scaffolds. This system utilizes an elegant design to create an effective cell culture environment without the drawbacks often associated with more complicated perfusion systems. The TPS design consists of hMSCs encapsulated in alginate beads which are tightly packed in a tubular growth chamber. Perfusing media through this growth chamber enhances nutrient transfer while exposing the cells to shear stress. To enhance in vivo vascularization, a prevascular network will be templated within the engineered tissue prior to implantation. To accomplish this, the TPS bioreactor will be optimized to support a coculture of endothelial cells and hMSCs. To examine this strategy of enhanced in vitro nutrient transport and in vivo vascularization, we propose first to investigate the TPS culture environment, particularly alginate bead size, bead composition, and media perfusion rate, that promotes hMSC proliferation and subsequent osteoblastic differentiation. Second, we propose to investigate the impact of endothelial cell coculture parameters, specifically coculture ratio, on the development of a prevascular network as well as the proliferation and differentiation of hMSCs. Third, we propose to implement a synthetic polymer sleeve system to support successful implantation of the in vitro cultured tissue. This strategy allows for the in vitro culture of functional engineered tissue, provides an elegant method for the in vivo implantation of the tissue, and fosters rapid integration of the implanted tissue into the host vasculature. Successful completion of these studies will demonstrate the feasibility of this fundamental technology for enhanced in vitro and in vivo nutrient transfer within cell based devices.

描述（由申请人提供）：必须克服体外和体内营养转移限制，以提高基于细胞的治疗策略的可行性。为了增强体外养分的运输，我们实验室最近开发的一种新型生物反应器的管状灌注系统（TPS）将在三维支架中动态培养人间质干细胞（HMSC）。该系统利用一种优雅的设计来创建一个有效的细胞培养环境，而没有通常与更复杂的灌注系统相关的缺点。 TPS设计由封装在藻酸盐珠中的HMSC组成，后者紧紧地包装在管状生长室中。通过这种生长室培养培养基可以增强营养转移，同时暴露细胞剪切应激。为了增强体内血管形成，植入之前将在工程组织中模板进行前血管网络。为此，将优化TPS生物反应器以支持内皮细胞和HMSC的共培养。为了研究这种增强体外营养转运和体内血管化的策略，我们首先建议研究TPS培养环境，尤其是藻酸盐珠的大小，珠子组成和培养基灌注率，从而促进HMSC增殖以及随后的成骨分化分化。其次，我们建议研究内皮细胞共培养参数，特别是共培养比的影响，对前血管网络的发展以及HMSC的增殖和分化。第三，我们建议实施一个合成的聚合物套筒系统，以支持成功植入体外培养的组织。该策略允许在功能工程组织的体外培养，为组织的体内植入提供了一种优雅的方法，并促进了将植入组织的快速整合到宿主脉管系统中。这些研究的成功完成将证明这种基本技术在基于细胞的设备中增强体外和体内营养转移的可行性。