近年來，超音波在生物醫學應用上有廣泛研究，如應用於腫瘤治療 (tumor therapy)、血腦屏障破壞 (disruption of blood-brain barrier)、經皮給藥 (Transdermal Drug Delivery, TDD)等。超音波結合微氣泡之穴蝕效應 (US-MB)，已被證實可以改變皮膚通透度，進而增強經皮給藥效果。
首先，在本研究先探討了超音波(US)結合微氣泡(MBs)對於matriptase之影響。matriptase又屬於第二型穿膜絲胺酸蛋白酶 (Type Ⅱ Transmembrane Serine Protease, TTSP)，和Prostasin在表皮的發育、分化、屏障和傷口癒合扮演重要的角色，而且matriptase受其抑制蛋白：肝細胞生長因子活化酶抑制蛋白type Ⅰ (hepatocyte growth factor activator inhibitor type Ⅰ, HAI-1) 調節。根據西方墨點法(western blotting)實驗結果，於角質細胞HaCaT中看到在120 kDa形成matriptase-HAI-1的複合體，證明US-MB成功打開細胞間隙活化matriptase；在免疫細胞染色 (Immunocytochemistry, ICC) 結果中也可以直接證明超音波介導微氣泡穴蝕效應確實可以有效地促進人類皮膚細胞通透性。
進一步於 ex vivo 人類皮膚實驗結果中，因超音波具有安全性、非侵入性 (non-invasive) 特點，其皮膚組織在進行超音波結合微氣泡實驗後，無任何受損或是水泡產生，並且其細胞間距離約在12小時後可以復原跟剛施打超音波時相同。在蘇木紫與伊紅染色 (Hematoxylin & Eosin stain, H&E stain) 結果中看到US-MB可以將人體皮膚中之角質層掀開，有效地打開皮膚通透度。另外，在本研究發現，在條件：3 W/cm2, 1min, 3 times +MBs下，能量可以從角質層穿到至基底層，於免疫組織染色 (Immunohistochemistry, IHC) 結果中，在0小時之基底層中觀察到活化態matriptase產生，並且隨著時間增加到3小時，其活化態matriptase，由基底層向上延伸至顆粒層，於6小時和12小時下都還有觀察到活化態matriptase存在，而在6小時左右活化態matriptase表現低於3小時，主要是因為HAI-1會抑制matriptase的活化；施打完超音波後於0小時可以發現 prostasin的表現範圍開始往下延伸至棘狀層細胞質中或細胞膜上，並隨著時間增加至3小時於表皮層中的顆粒層表現更加明顯，至6小時亦在棘狀層細胞質中或細胞膜上可以觀察其存在。
由實驗結果看來US-MB扮演重要的角色在matriptase活化機制中。另一方面，其增加反應時間也能提升活化態的matriptase蛋白表現。US-MB可以成功開啟人類皮膚細胞通透性，提升皮膚通透度，對於發展非侵入性之經皮給藥有很大的潛力。

Recently, ultrasound has been widely studied in biomedical applications, such as tumor therapy, blood-brain barrier destruction, transdermal drug delivery (TDD). Ultrasound (US) mediated microbubbles (MBs) cavitation has been confirmed to change the skin permeability, thereby significantly enhancing the effect of TDD.
In this thesis, the role of matriptase activity for US mediated MBs cavitation (US-MB) enhanced TDD was demonstrated. The membrane-associated serine protease matriptase regulated by its inhibitor hepatocyte growth factor activator inhibitor (HAI-1) and prostasin, playing a major role in the development, differentiation, skin barrier and wound healing of the epidermis.
According to our results, western blotting showed a 120 kDa complex of matriptase-HAI-1 in immortalized human keratinocytes cell (HaCaT), proving that US+MBs successfully changed the structure of cell-cell junctions to activate matriptase; US-MB improved skin permeability and reduced thermal effects in immunocytochemistry results.
Furthermore, in the results of ex vivo human skin experiments, since the characteristics of ultrasound are safe and non-invasive, the skin tissue was not damaged or blistered after US-MB experiment. The distances between the cells after US-MB are increased and recovered after 12 hours.
Hematoxylin & Eosin stain result indicated that the structure of stratum corneum has been lifted and effectively opened the skin permeability. In addition, the optimal US-MB parameter is 3 W/cm2, 1 min, 3 times + MBs (total 3 times of US-MB, each time is 1 min). Under this condition, the response of matriptase-HAI-1 from the stratum corneum to the basal layer can be observed significantly.
In immunohistochemistry, the activated matriptase was observed in the basal layer, and prolonged to 3 hours. At 3th hour, the activated matriptase extended from the basal layer to the granular layer. Besides the 6 and 12 hours, the presence of activated matriptase was still observed. The amount of activated matriptase decreased at 6th hour because mainly activated matriptase was inhibited by HAI-1; prostasin expression was observed in the epidermal granular layer to spinous layer and became more obvious in granular layer at the 3th hour. Prostasin was also detected in the cytoplasm or on the cell membrane at the 6th hour.
In this thesis, the result suggests an important role for US-MB regulates the activation mechanism of matriptase. On the other hand, increasing the times of US-MB can also enhance the expression of activated matriptase protein. The permeability of human skin cells skin can be successfully increased by US-MB which is a great potential method for non-invasive TDD in human.

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