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系統識別號 U0026-0812200915254782
論文名稱(中文) 利用高頻超音波對比劑影像強化評估在心肌梗塞後心室結構與功能之變化
論文名稱(英文) Comprehensive Structural and Functional Evaluation of Post-Infarcted Hearts Using Contrast-Enhanced Echocardiography
校院名稱 成功大學
系所名稱(中) 醫學工程研究所碩博士班
系所名稱(英) Institute of Biomedical Engineering
學年度 97
學期 2
出版年 98
研究生(中文) 張昌華
研究生(英文) Chang-Hua Chang
學號 p8696419
學位類別 碩士
語文別 中文
論文頁數 87頁
口試委員 指導教授-張憲彰
指導教授-謝清河
口試委員-孫永年
口試委員-葉秩光
口試委員-鄭國順
中文關鍵字 心肌梗塞  沉積式對比劑  強心劑Dobutamine  超音波擊破對比劑/回沖模型 
英文關鍵字 High resolution ultrasound  Myocardial infarction  Microbubbles  Dobutamine  Myocardial perfusion 
學科別分類
中文摘要 在生物醫學領域中,藉由小動物模擬有關人類疾病如癌症、心血管疾病等,為了觀察所建立起的動物模型,必須有一套高解析度、高靈敏度和非侵入式的影像系統以進行活體影像觀察。本研究即藉由高頻超音波系統搭配沉積式對比劑的使用以獲取較高的空間與流速解析度,使我們能觀察到小動物細微的組織結構以及血流情況,因此可以用來評估小鼠之心肌梗塞。本研究內容分作三大部分:實驗前的參數設定、心臟結構性評估和心臟功能性評估; 在實驗參數設定中,製作仿體測試找出對比劑的最佳注射濃度,並對強心劑Dobutamine作藥物活性測試,還有影像處理的基本流程;而第二部分的結構性評估中,為了比較梗塞面積的大小,利用對比劑加強心劑之超音波影像訊號,並且與組織切片,利用兩種不同的方式評估,其初步結果呈現高度的相關性;而最後一個部份於心臟的功能性評估時,以文獻中超音波擊破對比劑/回沖模型,藉由影像強度的變化,比較正常心肌灌流與梗塞區域的灌流曲線變化,也都有初步的成果。這項可以對心臟作一個整體的結構性和功能性評估,對於研究者在開發新藥上或基因治療上,均會有極大的幫助。
英文摘要 Background:
Real-time quantification of the myocardial structure-function relationship is essential to evaluate cardiac pathophysiology. We hypothesize that high resolution ultrasound accompanied with contrast agents can be employed to evaluate cardiac structures and functions after myocardial infarction (MI) in vivo.

Methods and Results:
Using high resolution ultrasound (VisualSonics Vevo 770) and microbubbles (MBs, DEPO or SonoVue), real-time echocardiography was continuously monitored after bolus injection of 50 ul MBs into mouse tail veins at 7 days, or 1 or 3 months after MI. While both DEPO and SonoVue MBs enhanced structure-function evaluation in the LV cavity, only DEPO MBs enabled the myocardial imaging and thus the differentiation of the infarcted and the remote areas. An excellent correlation on myocardial area and volume was observed between the current echnocardiographic method and conventional histological examination (ratio bias = 0.94 on Bland-Altman analysis). DEPO MB administration also improved the quantification of dobutamine (1.5 ug/g, i.p.)-mediated myocardial perfusion. Furthermore, myocardial flow velocity was successfully deduced from the relationship of the rates of MB destruction and replenishment at the border zone.

Conclusions:
We demonstrate that DEPO MB-enhanced echocardiography can be utilized to quantify (1) infarct area/volume, (2) myocardial perfusion and (3) myocardial flow velocity. Our established myocardial structure-function analytical tool can therefore provide an excellent platform to assess the effects of therapeutic strategies in treating MI and heart failure.
論文目次 中文摘要 ………………………………………………………………………………… I
英文摘要………………………………………………………………………………… II
誌謝………………………………………………………………………………………III
目錄 ………………………………………………………………………………………IV
圖索引 ………………………………………………………………………………… VI
表索引 …………………………………………………………………………………VIII
第一章 緒論
1.1前言…………………………………………………………………………………… 1
1.2心肌梗塞到心肌纖維化的過程……………………………………………………… 2
1.3臨床上診斷心肌梗塞的方法………………………………………………………… 3
1.4利用超音波對比劑診斷心肌梗塞…………………………………………………… 4
1.5小動物高頻超音波之介紹- VisualSonicsTM………………………………………… 6
1.6沉積式超音波對比劑………………………………………………………………… 8
1.7 研究動機與目的 ……………………………………………………………………11
第二章 實驗前之參數設定及準備
2.1 研究目的 ……………………………………………………………………………12
2.2 實驗方法與結果 ……………………………………………………………………12
2.2.1高頻超音波之影像擷取與量測 …………………………………………………12
B-mode (Brightness mode)成像原理及影像格式 ………………………………13
B-mode影像處理流程 …………………………………………………………13
M-mode ( Motion mode)介紹及量測方法 ………………………………………17
隨機單盲試驗(Random-Single blinding) ………………………………………18
老鼠生理參數之量測與維持 ……………………………………………………21
2.2.2 沉積式對比劑DEPOTM之體內與體外測試 ……………………………………24
仿體試驗(Phantom test) …………………………………………………………26
仿體試驗結果與討論 ……………………………………………………………29
DEPOTM之體內試驗(in vivo test) ………………………………………………31
DEPOTM之體內試驗方法 ………………………………………………………32
DEPOTM之體內試驗結果與討論 ………………………………………………35
2.2.3 Dobutamine之體內活性試驗 ……………………………………………………40
強心劑Dobutamine之體內試驗方法 ……………………………………………41
強心劑Dobutamine之體內活性試驗結果與討論 ………………………………42
內容 頁數
第三章 心肌梗塞後心臟的結構性評估
3.1 研究目的……………………………………………………………………………45
3.2 實驗材料與方法……………………………………………………………………46
3.2.1動物model建立 …………………………………………………………………46
麻藥配置…………………………………………………………………………46
手術步驟…………………………………………………………………………47
3.2.2超音波影像取樣流程 ……………………………………………………………48
3.2.3病理組織切片 ……………………………………………………………………52
切片方法 …………………………………………………………………………53
Masson’s Trichrome染色方法 …………………………………………………54
3.2.4 超音波與組織切片的關聯性 ……………………………………………………56
3.3實驗結果與討論 ……………………………………………………………………58
3.3.1動物model建立 …………………………………………………………………58
3.3.2病理組織切片分析 ………………………………………………………………60
3.3.3超音波與組織切片的關聯性之結果與討論 ……………………………………64
超音波影像與組織切片影像……………………………………………………64
Bland-Altman分析法和簡單線性迴歸…………………………………………67
3.4結論 …………………………………………………………………………………69

第四章 心肌梗塞後心臟的功能性評估
4.1 研究目的 ……………………………………………………………………………71
4.2 實驗材料方法 ………………………………………………………………………73
4.2.1對比劑之擊破/回沖模型 ………………………………………………………73
4.2.2 找出心肌灌流異常區域和計算心肌血流速度 …………………………………74
4.2.3 強心劑Dobutamine對心臟的功能性改善 ………………………………………78
4.3實驗結果與討論 ……………………………………………………………………80
4.4結論 …………………………………………………………………………………82
參考文獻 …………………………………………………………………………………83
自述 ………………………………………………………………………………………87



圖索引
內容 頁數
Fig.1.1 心肌梗塞後心室重塑化 …………………………………………………………3
Fig. 1.2 (A)、(B)冠狀動脈阻塞之病人之心臟超音波圖 ………………………………4
Fig 1.3 (A) 小動物高頻超音波系統-VisualSonicsTM Vevo 770
(B) 動物載台和整合型軌道系統(C) 氣體麻醉系統 ……………………………7
Fig. 1.4 (A)使用Vevo 770內建影像系統重組成三維立體結構
(B)將MeWo癌細胞植入小鼠後腳(Hind leg)所生成的瘤 ………………………8
Fig 1.5(A)(C)為小鼠注射顯影劑前之心臟超音波圖
(B)影像呈現混濁的狀態
(D)為過高濃度的對比劑導致超音波訊號衰減 …………………………………9
Fig 1.6活體顯微鏡所可以知道沉積式超音波對比劑穩定停留在
心肌微血管內的機制 ……………………………………………………………10
Fig 2.1 影像刻度與像素點轉換 …………………………………………………………14
Fig 2.2 ROI影像圈選(A) 原始影像(B) ROI圈選結果…………………………………14
Fig 2.3影像強化 …………………………………………………………………………15
Fig 2.4動態輪廓圈選 ……………………………………………………………………15
Fig 2.5心肌面積計算定義 ………………………………………………………………16
Fig 2.6圈選結果與心肌面積計算結果 …………………………………………………16
Fig 2.7正常老鼠心臟之M-mode 影像 …………………………………………………19
Fig 2.8心肌梗塞老鼠心臟之M-mode 影像 ……………………………………………20
Fig 2.9可量測小鼠生理參數和具有加熱功能之載台 …………………………………22
Fig 2.10小動物生理參數偵測器 …………………………………………………………23
Fig 2.11沉積式對比劑DEPOTM的組成…………………………………………………25
Fig 2.12仿體試驗系統架構模擬圖 ………………………………………………………27
Fig 2.13超音波探頭與仿體的相對位置圖(A)上方鳥瞰圖(B)側面圖 …………………28
Fig 2.14仿體試驗結果 ……………………………………………………………………30
Fig 2.15 FVB白色小鼠 …………………………………………………………………31
Fig 2.16小鼠尾靜脈注射方式傳遞DEPOTM對比劑……………………………………32
Fig 2.17 SonoVueTM與DEPOTM之比較…………………………………………………34
Fig 2.18 DEPOTM沉積式對比劑體內實驗 ………………………………………………36
Fig 2.19 DEPOTM沉積式對比劑於體內訊號強度的變化 ………………………………37
Fig 2.20 SonoVueTM對比劑體內實驗 ……………………………………………………38
Fig 2.21 SonoVueTM對比劑於體內訊號強度的變化 ……………………………………39
Fig 2.22強心劑Dobutamine之包裝外觀 ………………………………………………40
內容 頁數
Fig 2.23注射強心劑之老鼠其左心室射出比率的變化曲線 …………………………43
Fig 2.24注射強心劑之老鼠其心跳速率的變化曲線 ………………………………44
Fig 3.1利用DEPOTM對比劑評估心肌結構之流程圖 ………………………………45
Fig 3.2舒泰50和2% Rompun之包裝外觀 ……………………………………………46
Fig 3.3利用LAD冠狀動脈結紮建立小鼠之心肌梗塞模型 …………………………47
Fig 3.4 LAD冠狀動脈結紮後,從心尖開始會因為缺氧變成淡紅色…………………48
Fig 3.5 超音波探頭(白色)和固定器(黑色) ……………………………………………49
Fig 3.6 超音波影像為線性方式作取樣 …………………………………………………49
Fig 3.7 Long axis view影像取樣…………………………………………………………50
Fig 3.8 Short axis view影像取樣…………………………………………………………51
Fig 3.9經4%福馬林組織固定後的小鼠心臟……………………………………………52
Fig 3.10切片機可以調整每次切片的厚度………………………………………………53
Fig 3.11每個玻片約可以收下3張組織切片……………………………………………53
Fig 3.12控制組與對照組(心肌梗塞)之%EF的比較 ……………………………………59
Fig 3.13其心臟未結紮的小鼠之切片……………………………………………………61
Fig 3.14心肌梗塞後一個月之小鼠心臟切片……………………………………………62
Fig 3.15心肌梗塞後三個月之小鼠心臟切片……………………………………………63
Fig 3.16乳突肌與心肌之接合處的位置…………………………………………………64
Fig 3.17心肌梗塞後之一個月的小鼠,其心臟超音波影像和切片影像………………65
Fig 3.18心肌梗塞後之三個月的小鼠,其心臟超音波影像和切片影像………………66
Fig 3.19 Bland-Altman分析法之結果……………………………………………………68
Fig 3.20簡單線性迴歸分析結果…………………………………………………………68
Fig 4.1心肌灌流實驗架構圖 ……………………………………………………………72
Fig 4.2超音波擊破對比劑/回沖模型曲線 ………………………………………………73
Fig 4.3 心肌灌流實驗以syringe pump連續提供流體 …………………………………75
Fig 4.4心肌舒張期之輪廓模型 …………………………………………………………76
Fig 4.5心肌舒張期之灌流影像 …………………………………………………………76
Fig 4.6心肌灌流影像加以套色之結果 …………………………………………………77
Fig 4.7從套色後的影像定義,找出灌流異常之影像強度值……………………………77
Fig 4.8強心劑作用下的心臟超音波影像 ………………………………………………78
Fig 4.9使用強心劑前後左心室收縮功能的差異 ………………………………………79
Fig 4.10超音波擊破對比劑/回沖模型 …………………………………………………81




表索引
內容 頁數
Table 1.1心肌梗塞後心肌纖維化的過程 ………………………………………………2
Table 1.2在美國臨床上較常使用的超音波對比劑………………………………………5
Table 2.1注射強心劑之老鼠在各個時間點中,其左心室射出比率的
平均值與標準差 ………………………………………………………………43
Table 2.1注射強心劑之老鼠在各個時間點中,其心跳速率的
平均值與標準差 ………………………………………………………………44
Table 3.1控制組(Sham)與對照組(心肌梗塞)之%EF的比較……………………………59
Table 3.2切片與超音波影像之正常心肌的面積 ………………………………………65
Table 3.3切片與超音波影像之正常心肌的面積 ………………………………………66
Table 4.1使用強心劑前後左心室收縮功能之數據 ……………………………………79
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