A Study on Acoustic Cavitation Assisted Gene Delivery
Date Issued
2005
Date
2005
Author(s)
Lai, Chun-Yen
DOI
zh-TW
Abstract
Gene therapy is a promising therapeutic technique, and it is defined as correcting defective genes responsible for disease development. The key of gene therapy is gene delivery (also known as transfection). In this research, ultrasound and ultrasound contrast agents are used to induce cavitation, and gene delivery is assisted with cavitation effect. Cavitation is the process of microbubble formation, resonance, and destruction. When microbubbles experience time-varying acoustic field, cavitation may occur. Cavitation effects induced by ultrasound have been found beneficial in the non-thermal treatment, and the advantages of ultrasound are non-invasiveness, focusing and real-time imaging capacities. Although quantitative success of acoustic cavitation assisted gene delivery has been reported in the literature, the relations between gene delivery rate and cavitation effect are still unknown. In this research, methods for effective induction of acoustic cavitation are first established. Then, the quantitatively relations of gene delivery and cavitation dose are analyzed. A 1MHz ultrasound transducer and the commercial ultrasound contrast agent Levovist® are used for cavitation induction. HeLa cells and FITC labeled short sequence DNA are used in the gene delivery experiments. The parameters under investigation are acoustic pressure, microbubble concentration and ultrasound pulse length. The effect of cavitation (microbubble destruction) is quantitatively analyzed using the inertial cavitation dose (ICD) method. After gene delivery experiments, the gene delivery rate and cell viability are calculated under the fluorescent microscope. The curves of gene delivery rate (and cell viability) versus ICD are plotted, and the relations between gene delivery rate/viability and cavitation are examined. The results show that ICD increases with the acoustic pressure, microbubble concentration and pulse length. In the gene delivery experiments, there are high correlations between gene delivery rate (or cell viability) and ICD. Combining the results of gene delivery rate (or cell viability rate) versus ICD of the three parameters, high correlations also exist between gene delivery results and ICD, indicating that gene delivery results may be potentially predicted using cavitation measurements before gene delivery experiments.
Subjects
超音波
微氣泡
穴蝕效應
基因傳遞
慣性穴蝕劑量
ultrasound
microbubbles
cavitation
transfection
gene delivery
inertial cavitation dose
Type
thesis
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