Engineered Cartilage Storage and Transport Solution

工程软骨储存和运输解决方案

• 批准号: 6991750
• 负责人: Kelvin G.M. Brockbank
• 金额: $ 16.73万
• 依托单位: CELL AND TISSUE SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-07 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6991750
• 关键词: biomaterial development /preparation; biomaterial evaluation; biotechnology; cartilage; cell proliferation; chondrocytes; cryopreservation; growth factor; hypothermia; physiology; tissue /cell preparation; tissue /organ preservation; tissue engineering; tissue resource /registry

DESCRIPTION (provided by applicant): The overall objective of this proposal is to define the hypothermic storage conditions and shelf life for tissue engineered cartilage constructs (TECCs) required for commercial manufacturing and distribution. Preliminary studies employing TECCs have demonstrated that chondrocyte viability and construct biochemistry were significantly impacted by hypothermic storage in chondrocyte culture medium. The use of hypothermia as the principal means to suppress metabolism in a reversible way is the foundation of most of the effective methods for tissue and organ storage. In this proposal two solution designs (Unisol base formulations) that mimic the intracellular environment and the extracellular environment of cells will be tested at two temperatures (25 degrees and 4 degrees C) and compared with commercially available organ preservation solutions. These temperatures were selected on the basis of the two contrasting degrees of hypothermia and practical convenience in the clinical arena (ambient vs. refrigeration). Furthermore, the potential benefits of addition of chondrocyte growth factors (trophic factors) to the Unisol solutions will be assessed. Growth factors are often required for survival, maintenance and proliferation of specific cell types in vitro and some growth factors have been shown to have dramatic effects upon survival of organs under hypothermic conditions. The hypotheses for this study are that 1) the Unisol base formulations preserve the quality of stored TECCs better than commercially available organ preservation solutions, and 2) the quality of low temperature stored TECCs is improved in the presence of growth factors known to play a role in the normal physiology of chondrocytes. TECCS will be manufactured using polyglycolic acid felt and human chondrocytes. They will be employed to compare two organ preservation solutions with Unisol base formulations for up to eight weeks of hypothermic storage. Subsequently the effects of three chondrocyte growth factors upon short (days) and longer term (weeks) TECC hypothermic storage will be investigated. The quality of the TECCs will be determined by returning them to physiologic conditions and studying metabolic activity, viability, apoptosis and histology. Chondrocyte Phenotypic stability will also be assessed by analysis of relative matrix component mRNA expression following hypothermic exposure to growth factors. These studies will lead to the development and commercialization of Unisol formulations specifically designed for cold storage of tissue engineered cartilage constructs.

描述（由申请人提供）：本提案的总体目标是定义商业生产和分销所需的组织工程软骨结构（TECC）的低温储存条件和有效期。采用TECs的初步研究已经证明，软骨细胞活力和构建体生物化学受到软骨细胞培养基中低温储存的显著影响。使用低温作为以可逆方式抑制代谢的主要手段是大多数组织和器官储存的有效方法的基础。在本提案中，将在两种温度（25 ℃和4 ℃）下测试模拟细胞的细胞内环境和细胞外环境的两种溶液设计（Unisol基础制剂），并与市售器官保存溶液进行比较。这些温度的选择是基于两种对比度的低温和临床竞技场中的实际便利性（环境温度与冷藏温度）。此外，将评估向Unisol溶液中添加软骨细胞生长因子（营养因子）的潜在获益。生长因子通常是体外特定细胞类型的存活、维持和增殖所必需的，并且已经显示某些生长因子在低温条件下对器官的存活具有显著影响。本研究的假设是：1）Unisol基质制剂比市售器官保存溶液更好地保存了储存的TECC的质量，2）在已知在软骨细胞正常生理学中起作用的生长因子存在下，低温储存的TECC的质量得到改善。TECCS将使用聚乙醇酸毡和人软骨细胞制造。他们将被用来比较两种器官保存溶液与Unisol基础配方长达8周的低温储存。随后将研究三种软骨细胞生长因子对短期（数天）和长期（数周）TECC低温储存的影响。将通过将TECC恢复到生理条件并研究代谢活性、存活力、凋亡和组织学来确定TECC的质量。还将通过分析低温暴露于生长因子后的相对基质组分mRNA表达来评估软骨细胞表型稳定性。这些研究将导致专门设计用于组织工程软骨构建物冷藏的Unisol制剂的开发和商业化。

项目成果

Protocol Development for Vitrification of Tissue-Engineered Cartilage.

组织工程软骨玻璃化的协议开发。

• DOI: 10.12665/j84.brockbank
• 发表时间: 2009
• 期刊: Bioprocessing
• 作者: Farooque,TanyaM;Chen,Zhenzhen;Schwartz,Zvi;Wick,TimothyM;Boyan,BarbaraD;Brockbank,KelvinGM
• 通讯作者: Brockbank,KelvinGM