Microhemodynamics after Infusion of an Artificial O_2 Carriers Using Laser Flash Irradiation.

使用激光闪光照射人工 O_2 载体输注后的微血流动力学。

• 批准号: 12480268
• 负责人: SAKAI Hiromi
• 金额: $ 9.47万
• 依托单位: Waseda University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12480268/
• 关键词: Microcirculation; Artificial Oxygen Carrier; Hemoglobin-Vesicles; Con focal Laser Scanning Microscopy; Metabolism; Non-invasive Method; Diagnostics; Methemoglobin; 生体機能材料; 生体情報・計測; 医用・生体画像; 出血ショック; 細網内皮系; 過渡吸収スペクトル

(1)Hemoglobin-vesicles (HbV) has been developed to substitute the function of RBCs. We developed new methods to analyze the microhemodynamics after infusion of HbV into rodents, and evaluated the safety and efficacy of HbV. Fluorescence-labeled HbV was infused into hamsters and the liver was observed in situ with a laser con focal microscopy. This allowed visualization of HbV flowing in the sinusoid, and the particles were finally captured and accumulated in the Kupffer cells, which could be observed as intensified fluorescence. Histopathological analysis using anti-human Hb antibody and serum clinical chemistry demonstrated the prompt metabolism and excretion of HbV without any irreversible changes.(2)Capillary RBC flow in the skin of rat tail during an extreme hemodilution with albumin was microscopically observed with a noninvasive transcutaneous technique using flash epi-illumination. The RBC flow rates monotonously increased up to 294 ± 110% of the basal value at the 60% level of … More blood exchange, which was in contrast to the abdominal aortic blood flow increase to 148 ± 20%. Capillary hematocrit did not appear to fall in the same proportion as the systemic hematocrit, which coincided with the previous observations. These results indicate that microhemodynamic properties during hemodilution can be transcutaneously analyzed with this new technique without surgical preparation of the skin.(3)The O_2 release from flowing HbVs was examined using an O_2-permeable polymer tube (inner diameter, 28 μm) exposed to a deoxygenated environment. Measurement of O_2 release was performed using an apparatus that consisted of an inverted microscope and a scanning-grating spectrophotometer with a photon-count detector, and the rate of O_2 release was determined based on the visible absorption spectrum in the Q band of Hb. The rate of O_2 release from the HbV/RBC mixtures was similar to that of RBCs alone. On the other hand, the addition of acellular Hb solution to RBCs significantly enhanced the rate of deoxygenation. This should mainly be due to the difference in the particle size (250 vs. 7 nm) that affects their diffusion for the facilitated O_2 transport. Less

（1）已经开发出血红蛋白宇宙（HBV）来替代RBC的功能。我们开发了新的方法来分析HBV输注啮​​齿动物后的微动力学，并评估了HBV的安全性和效率。将荧光标记的HBV注入仓鼠中，并用激光局部显微镜原位观察到肝脏。这允许可视化正弦曲线中流动的HBV，并最终捕获并积聚在库普弗细胞中，可以将其视为强烈的荧光。使用抗人类HB抗体和血清临床化学的组织病理学分析表明，HBV的迅速代谢和极端变化而没有任何不可逆的变化。（2）使用粉蛋白的极端血液动作在极端血液动作的情况下，大鼠皮肤中的毛细血管RBC流动使用了非影响力的经闪光性的闪光性闪光技术。 RBC流量单调增加到基本值的294±110％，在60％的水平…更多的血液交换水平，这与腹主动脉血流增加到148±20％相反。毛细血管血细胞比容似乎没有与系统性血细胞比容的比例相同，这与先前的观察结果相吻合。这些结果表明，可以通过这种新技术经过手术而无需外科手术制备皮肤准备血液稀释过程中的微型动力学特性。（3）使用O_2渗透聚合物管（内部直径，28μm，28μm）暴露于脱氧环境中的O_2流动HBV的O_2释放。使用由倒置显微镜组成的设备和带有光子计数检测器的扫描详细信息分光光度计进行了O_2释放的测量，并根据HB Q带中的可见吸收光谱确定O_2释放速率。 O_2从HBV/RBC混合物中释放的速率与单独的RBC相似。另一方面，在RBC中添加细胞HB溶液可显着提高脱氧速率。这应该主要是由于粒径（250 vs. 7 nm）的差异会影响其准备好的O_2传输的差异。较少的

项目成果

H.Sakai, H.Horinouchi, Y.Masada, S.Takeoka, M.Takaori, K.Kobayashi, E.Tsuchida.: "Metabolism of hemoglobin-vesicles (artificial oxygen carriers) and their influence on organ functions in a rat model."Biomaterials. 25. 4317-4325 (2004)

H.Sakai、H.Horinouchi、Y.Masada、S.Takeoka、M.Takaori、K.Kobayashi、E.Tsuchida.：“大鼠模型中血红蛋白囊泡（人工氧载体）的代谢及其对器官功能的影响

Sakai H, Takeoka S, Intaglietta M, Tsuchida E, et al.: "Systemic and Microvascular responses to the hemorrhagic shock and resuscitation with Hb-vesicles"American Journal of Physiology Heart and Circulatory Physiology. 283. H1191-H1199 (2002)

Sakai H、Takeoka S、Intaglietta M、Tsuchida E 等人：“Hb 囊泡对失血性休克和复苏的全身和微血管反应”美国生理学心脏和循环生理学杂志。

Y, Teramura, H.Kanazawa, H.Sakai, S.Takeoka, E.Tsuchida.: "The prolonged oxygen -carrying ability of Hb vesicles by coencapsulation of catalase in vivo."Bioconjugate Chem.. 14. 1171-1176 (2003)

Y、Teramura、H.Kanazawa、H.Sakai、S.Takeoka、E.Tsuchida.：“通过体内过氧化氢酶的共封装延长 Hb 囊泡的携氧能力。”Bioconjugate Chem.. 14. 1171-1176 (2003

土田英俊, 宗慶太郎, 酒井宏水, 小松晃之, 武岡真司, 堀之内宏久, 末松誠, 小林紘一: "酸素輸液(人工赤血球)の安全度と体組織への酸素輸送"麻酔. 53(増刊号). S55-S66 (2003)

Hidetoshi Tsuchida、Keitaro So、Hiromizu Sakai、Teruyuki Komatsu、Shinji Takeoka、Hirohisa Horinouchi、Makoto Suematsu、Koichi Kobayashi：“输氧（人工红细胞）和向身体组织输送氧气的安全性”麻醉 53。 S55-S66 (2003)

C.Contaldo, S.Schramm, R.Wettstein, H.Sakai, S.Takeoka, E.Tsuchida, M.Leunig, A.Banic, D.Erni.: "Improved oxygenation in ischemic hamster flap tissue is correlated with increasing hemodilution with Hb vesicles and their O_2 affinity."Am. J. Physiol. Heart

C.Contaldo、S.Schramm、R.Wettstein、H.Sakai、S.Takeoka、E.Tsuchida、M.Leunig、A.Banic、D.Erni.：“缺血性仓鼠皮瓣组织氧合的改善与血液稀释的增加相关