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）血红蛋白囊泡（HbV）已被开发以替代RBC的功能。我们开发了新的方法来分析输注HbV后啮齿动物的微血流动力学，并评估HbV的安全性和有效性。将标记的HbV注入仓鼠体内，用激光共聚焦显微镜原位观察肝脏。这允许可视化在窦状隙中流动的HbV，并且颗粒最终被捕获并积聚在枯否细胞中，这可以被观察为增强的荧光。使用抗人Hb抗体和血清临床化学的组织学分析表明，HbV的迅速代谢和排泄没有任何不可逆的变化。（2）采用无创性经皮闪光照明技术，在显微镜下观察了白蛋白极度血液稀释时大鼠尾部皮肤毛细血管内红细胞的流动。在60%水平下，RBC流速单调增加至基础值的294 ± 110%。 ...更多信息 血液交换，与腹主动脉血流量相比增加到148 ± 20%。毛细血管红细胞压积的下降比例似乎与全身红细胞压积不同，这与先前的观察结果一致。这些结果表明，在血液稀释过程中的微血流动力学特性可以transmitted分析与这种新技术，而无需手术准备的皮肤。（3）用内径为28 μm的透氧聚合物管在缺氧环境中检测HbVs的氧释放。用倒置显微镜和带光子计数检测器的扫描光栅分光光度计组成的装置测量O_2释放，根据Hb在Q带的可见吸收光谱确定O_2释放速率。HbV/RBC混合物的O_2释放速率与单独RBC的O_2释放速率相似。另一方面，添加脱细胞血红蛋白溶液的红细胞显着提高脱氧速率。这主要是由于颗粒尺寸的差异（250与7 nm）影响了它们的扩散以促进O_2的传输。少

项目成果

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

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)