適用於生物實驗之超音波刺激裝置設計與應用

Abstract

近年興起低強度超音波治療研究之風潮，不像傳統的熱治療(高強度)超音波以破壞病灶為手段，低強度超音波主要以機械力刺激細胞的機械敏感通道，影響離子之進出，進一步影響生物訊號之傳遞，此應用可能包含神經調節、藥物傳遞、基因治療、組織發育等。 為探索低強度超音波治療於不同領域應用之可行性，建立一套穩定的生物實驗系統是必要的。本實驗室已在相關領域深耕多年，開發出多種適用於不同情境之超音波刺激裝置，並投入生物實驗探討超音波刺激之生物機制。本研究將針對其中三種常用的裝置：超音波即時影像載台、超音波玻璃微管以及聚焦超音波，進一步發展與改良他們的設計，並開發新的生物實驗應用以探索其實用性。 本研究設計出適用於刺激培養皿內細胞與組織的超音波即時影像載台Dish-LIC，並利用其觀察超音波刺激對細胞或組織鈣離子反應之影響，也可用於刺激組織影響其ERK磷酸化反應。將原本用於刺激細胞引起鈣離子反應之超音波玻璃微管，利用其聲流對細胞或組織產生剪應力的特性，應用於特定組織剝離與細胞的adhesion GPCR分離。另外根據聚焦曲面的數學模型，發現了聚焦探頭存在極限聚焦深度的性質，並利用此性質開發出了迷你聚焦探頭；本研究將聚焦超音波應用於開啟小鼠血腦屏障、刺激組織影響其ERK磷酸化反應以及刺激小鼠腦部影響其聽力反應。

In recent years, there has been a wave of research on low-intensity ultrasound therapy. Unlike the traditional thermotherapy (high-intensity) ultrasound that destroys the lesion, low-intensity ultrasound mainly stimulates the mechanosensitive channels of the cells with mechanical stimulation, which affects the ionic influx and efflux, and then further affects the transmission of biological signals. The applications may include neuromodulation, drug delivery, gene therapy, and tissue development. To explore the feasibility of low-intensity ultrasound therapy in different fields, it is necessary to establish a stable biological experimental system. Our laboratory has been working in this field for years, and has developed various ultrasonic stimulation devices and conducted experiments to investigate the biological mechanism of ultrasonic stimulation. In this study, we further developed and improved the design of three commonly used devices, namely, ultrasound live image chamber, ultrasound micropipette, and focused ultrasound, and developed new biological applications to explore their utilities. In this study, we designed the Dish-LIC, an ultrasound live image chamber for stimulating cells and tissues in culture dishes, to observe the effects of ultrasonic stimulation on the calcium ion response of cells and tissues, as well as to stimulate tissues to affect their ERK phosphorylation response. The ultrasound micropipette, originally used to stimulate cells to induce calcium ion response, applied to tissue separation and cellular adhesion GPCR separation by utilizing its acoustic flow to generate shear stress on cells or tissues. In addition, based on the mathematical model of the focusing surface, we discovered the minimal focal distance of a focused ultrasound probe, and developed a mini focusing probe accordingly. In this study, focused ultrasound was applied to open the blood-brain barrier, stimulate tissues to affect ERK phosphorylation, and stimulate the brain to affect the auditory response in mice.