PDF(Pubmed)
Abstract：
BACKGROUND: Quality assessment of modified or processed red blood cell (RBC) components, such as pathogen-reduced RBCs, using only in vitro testing may not always be predictive of in vivo performance. Mouse or rat in vivo models are limited by a lack of applicability to certain aspects of human RBC biology. Here, we used a guinea pig model to study the effects of riboflavin combined with UV light on the integrity of RBCs in vitro and following transfusion in vivo.
METHODS: Guinea pig RBCs were collected from whole blood (WB) treated with varying UV doses (10, 20, 40 or 80 J/mL) in the presence of riboflavin (UVR-RBCs). In vitro tests for UVR-RBCs included hemolysis, osmotic fragility, and cellular morphology by scanning electron microscopy. Guinea pigs transfused with one-day post-treatment UVR-RBCs were evaluated for plasma hemoglobin (Hb), non-transferrin bound iron (NTBI), total iron and Perls-detectable hemosiderin deposition in the spleen and kidney, and renal uptake of Hb.
RESULTS: Acute RBC injury was dose dependently accelerated after treatment with UV light in the presence of riboflavin. Aberrant RBC morphology was evident at 20, 40, and 80 J/mL, and membrane lysis with Hb release was prominent at 80 J/mL. Guinea pigs transfused with 40 and 80 J/mL UVR-RBCs showed increased plasma Hb levels, and plasma NTBI was elevated in all UVR-RBC groups (10-80 J/mL). Total iron levels and Perls-hemosiderin staining in spleen and kidney as well as Hb uptake in renal proximal tubules were increased 8 hours post-transfusion with 40 and 80 J/mL UVR-RBCs.
CONCLUSIONS: UVR-RBCs administered to guinea pigs increased markers of intravascular and extravascular hemolysis in a UV dose-dependent manner. This model may allow for the discrimination of RBC injury during testing of extensively processed RBCs intended for transfusion.
METHODS: Guinea pig RBCs were collected from whole blood (WB) treated with varying UV doses (10, 20, 40 or 80 J/mL) in the presence of riboflavin (UVR-RBCs). In vitro tests for UVR-RBCs included hemolysis, osmotic fragility, and cellular morphology by scanning electron microscopy. Guinea pigs transfused with one-day post-treatment UVR-RBCs were evaluated for plasma hemoglobin (Hb), non-transferrin bound iron (NTBI), total iron and Perls-detectable hemosiderin deposition in the spleen and kidney, and renal uptake of Hb.
RESULTS: Acute RBC injury was dose dependently accelerated after treatment with UV light in the presence of riboflavin. Aberrant RBC morphology was evident at 20, 40, and 80 J/mL, and membrane lysis with Hb release was prominent at 80 J/mL. Guinea pigs transfused with 40 and 80 J/mL UVR-RBCs showed increased plasma Hb levels, and plasma NTBI was elevated in all UVR-RBC groups (10-80 J/mL). Total iron levels and Perls-hemosiderin staining in spleen and kidney as well as Hb uptake in renal proximal tubules were increased 8 hours post-transfusion with 40 and 80 J/mL UVR-RBCs.
CONCLUSIONS: UVR-RBCs administered to guinea pigs increased markers of intravascular and extravascular hemolysis in a UV dose-dependent manner. This model may allow for the discrimination of RBC injury during testing of extensively processed RBCs intended for transfusion.
摘要：
背景：改良或加工的红细胞（RBC）成分的质量评估，例如病原体减少的红细胞，仅使用体外测试可能并不总是可以预测体内性能。小鼠或大鼠体内模型受限于缺乏对人RBC生物学的某些方面的适用性。这里,我们使用豚鼠模型研究了核黄素联合紫外线对体外和体内输血后红细胞完整性的影响。
方法:在核黄素(UVR-RBC)的存在下,从用不同UV剂量(10、20、40或80J/mL)处理的全血(WB)收集豚鼠RBC。UVR-RBC的体外测试包括溶血，渗透脆弱性，扫描电镜和细胞形态。用治疗后一天的UVR-RBC输注的豚鼠评估血浆血红蛋白（Hb），非转铁蛋白结合铁（NTBI），总铁和Perls-可检测的含铁血黄素在脾脏和肾脏中的沉积，和肾吸收Hb。
结果：在核黄素存在下用紫外光治疗后，急性红细胞损伤呈剂量依赖性加速。在20、40和80J/mL时，红细胞形态异常明显，Hb释放的膜溶解在80J/mL时显著。输入40和80J/mLUVR-RBC的豚鼠显示血浆Hb水平升高，所有UVR-RBC组（10-80J/mL）的血浆NTBI均升高。在输注40和80J/mLUVR-RBC后8小时，脾脏和肾脏中的总铁水平和Perls-含铁血黄素染色以及肾脏近端小管中的Hb摄取增加。
结论：给予豚鼠UVR-RBC以紫外线剂量依赖性方式增加血管内和血管外溶血的标志物。该模型可以允许在用于输血的广泛处理的RBC的测试期间区分RBC损伤。
方法:在核黄素(UVR-RBC)的存在下,从用不同UV剂量(10、20、40或80J/mL)处理的全血(WB)收集豚鼠RBC。UVR-RBC的体外测试包括溶血，渗透脆弱性，扫描电镜和细胞形态。用治疗后一天的UVR-RBC输注的豚鼠评估血浆血红蛋白（Hb），非转铁蛋白结合铁（NTBI），总铁和Perls-可检测的含铁血黄素在脾脏和肾脏中的沉积，和肾吸收Hb。
结果：在核黄素存在下用紫外光治疗后，急性红细胞损伤呈剂量依赖性加速。在20、40和80J/mL时，红细胞形态异常明显，Hb释放的膜溶解在80J/mL时显著。输入40和80J/mLUVR-RBC的豚鼠显示血浆Hb水平升高，所有UVR-RBC组（10-80J/mL）的血浆NTBI均升高。在输注40和80J/mLUVR-RBC后8小时，脾脏和肾脏中的总铁水平和Perls-含铁血黄素染色以及肾脏近端小管中的Hb摄取增加。
结论：给予豚鼠UVR-RBC以紫外线剂量依赖性方式增加血管内和血管外溶血的标志物。该模型可以允许在用于输血的广泛处理的RBC的测试期间区分RBC损伤。
