INSTRUMENT TO CHARACTERIZE CELL-FREE 0XYGEN CARRIERS

发明了一种表征无细胞血红蛋白和/或无细胞血红蛋白与红细胞的混合物的O2转运的新仪器，并已申请专利。该仪器使用小样本测量扩散O2运输，并且适用于无细胞血红蛋白溶液的开发和监测已接受它们的患者。本提案的目的是优化设计的仪器，以测量O2运输的无细胞血红蛋白溶液在体外使用人工毛细管系统。在具体目标1下，我们将建立一个人工毛细血管系统，以评估体外无细胞血红蛋白溶液的O2转运。我们将优化我们现有的人工毛细管仪器的实验板设计。将使用无细胞血红蛋白溶液（包括PEG-血红蛋白和交联四聚体血红蛋白的模型溶液）和RBC悬液进行O2转运的测量。在具体目标2下，我们将使毛细管仪器自动化，并进一步开发数据解释软件。 数据分析模型使用基于Krogh圆柱体模型的数学概念，用于将O2输送出限定几何形状的圆柱形管（即，人造毛细血管）。该程序需要1）优化运行时间效率和2）针对来自人造毛细管装置中的血红蛋白溶液和/或红细胞的实验数据进行测试。这一建议的假设是，一个新的方法来评估O2运输的无细胞氧载体之前，候选产品可以准确地进行评估。这是由于在无细胞载体存在下O2的易化扩散现象，其可导致自动调节性血管收缩。拟议的商业应用：在无细胞氧载体存在下，O2转运（Hb，Het）的临床评价方法不足。我们已经发明了一种新的仪器来表征O2运输的无细胞血红蛋白，包括评估促进扩散。该仪器具有很大的潜力，可用于接受血红蛋白产品的患者的临床测量，以及新候选溶液的检测。

A new instrument to characterize O2 transport by cell-free hemoglobin and/or mixtures of cell-free hemoglobin and red blood cells has been invented, and a patent has been applied for. This instrument measures diffusional O2 transport using a small sample, and is applicable both to the development of cell-free hemoglobin solutions and to monitoring patients who have received them. The goal of this proposal is to optimize the design of this instrument to measure O2 transport by cell-free hemoglobin solutions in vitro using an artificial capillary system. Under Specific Aim 1, we will build an artificial capillary system to assess O2 transport by cell- free hemoglobin solutions in vitro. We will optimize the breadboard design of our present artificial capillary instrument. Measurements of O2 transport will be made using cell-free hemoglobin solutions, including model solutions of PEG-hemoglobin and cross-linked tetrameric hemoglobin, and RBC suspensions. Under Specific Aim 2, we will automate the capillary apparatus and further develop software for data interpretation. The data- analysis model uses mathematical concepts based on the Krogh cylinder model for O2 transport out a cylindrical tube of defined geometry (i.e., the artificial capillary). This programs requires 1) optimization for run time efficiency and 2) testing against experimental data from hemoglobin solutions and/or red blood cells in the artificial capillary device. The hypothesis underlying this proposal is that a new method to assess O2 transport by cell-free oxygen carriers is required before candidate products can be evaluated accurately. This is due to the phenomenon of facilitated diffusion of O2 in the presence of cell-free carriers, which can lead to autoregulatory vasoconstriction. PROPOSED COMMERCIAL APPLICATIONS: Methods of clinical evaluation of O2 transport (Hb, Het) in the presence of cell-free oxygen carriers are inadequate. We have invented a new instrument to characterize O2 transport by cell-free hemoglobin that includes evaluation of facilitated diffusion. This instrument has great potential for clinical measures in patients who have received hemoglobin- based products, as well as potential in the testing of new candidate solution.