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)已开发出可替代红细胞功能的血红蛋白-囊泡(HBVs)。我们发展了一种新的方法来分析啮齿动物注射乙肝病毒后的微血流动力学，并评价了乙肝病毒的安全性和有效性。将荧光标记的乙肝病毒注入仓鼠体内，用激光共聚焦显微镜对肝脏进行原位观察。这使得可以看到乙肝病毒在肝窦中流动，颗粒最终被捕获并积累在库普弗细胞中，这可以观察到增强的荧光。用抗人Hb抗体和血清临床化学方法进行组织病理学分析，证实了病毒的迅速代谢和排泄，没有任何不可逆转的变化。(2)采用闪光照射无创经皮技术，观察了白蛋白极端血液稀释过程中大鼠尾部皮肤内的毛细血管红细胞流动。60%…水平时，红细胞流率单调增加至基础值的294±110%更多的血液交换，腹主动脉血流量增加到148±20%。毛细血管红细胞压积与全身性红细胞压积下降的比例不同，这与先前的观察结果一致。这些结果表明，利用这一新技术可以在不进行皮肤手术准备的情况下对血液稀释过程中的微血流动力学特性进行经皮分析。(3)使用内径为28μm的透氧聚合物管在低氧环境中检测流动的HbV的O_2释放。用倒置显微镜和带有光子计数探测器的扫描光栅仪组成的装置测量O2释放，并根据Hb在Q波段的可见吸收光谱确定释放O2的速率。HBVRBC混合物释放O2的速率与单纯RBC相似。另一方面，向红细胞中加入无细胞HB溶液可显著提高红细胞的脱氧速率。这主要是由于颗粒大小的不同(250 nm和7 nm)影响了它们的扩散，从而促进了O2的传输。较少

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)