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真空注型技術製程優化之研究

為了解決soft99蠟推薦的問題，作者劉學安 這樣論述：

快速製造作為在全球貿易高速擴張下的關鍵技術，在研發階段獲得了廣泛應用。從此基礎延伸出快速模具搭配真空注型技術，可快速實現小批量生產，但真空注型中仍有許多由於人為或製程因素導致的尺寸誤差、幾何變形、氣泡或短射等產品缺陷。本研究於真空注型機中架設可提供穩定密合壓力之合模機構，藉由模穴特徵為長方體與圓柱體的模具進行人工與機構合模之真空注型。透過注型件之幾何比較，證明比起傳統合模方式，注型件之幾何尺寸穩定度分別提升了24.47% 與27.37%。並在維持注型件幾何穩定度的條件下，可使模具材料成本減少23%。另外還探討真空注型製程參數與注型件機械強度之關係，以注型件之抗拉強度作為品質特性，運用田口實驗

設計與變異數分析找出真空注型最佳參數，研究得知模穴溫度為影響最大之參數。當真空注型製程參數為模穴溫度35°C、材料混合時間 40 秒、壓差時間 8 秒以及混合室進氣閥角度 60°時可獲得機械強度優於於其他參數組合之注型件。本研究研製之合模機構與參數組合可使注型件品質保持穩定的幾何尺寸與機械性質，擁有良好的工業實用性與產業參考價值。

視黃酸之聚乳酸/聚乳酸乙醇酸奈米載體應用於雷射輔助經皮吸收以抑制皮膚光老化

為了解決soft99蠟推薦的問題，作者羅珮琦 這樣論述：

總目錄指導教授推薦書..............................................................................................................口試委員會審定書..........................................................................................................誌謝...............................................................

............................................................... iii中文摘要.......................................................................................................................ivAbstract....................................................................................................

.....................vi總目錄........................................................................................................................ viii圖目錄..........................................................................................................................xv表目錄...................

................................................................................................... xviii第一章、緒論................................................................................................................1第一節、皮膚簡介....................................................................

............................11-1 皮膚之生理功能 ....................................................................................11-2 表皮層 ....................................................................................................21-3 真皮層 .........................................................

...........................................31-4 皮下組織 ................................................................................................3第二節、藥物經皮吸收途徑及促進方式............................................................42-1 經皮吸收 ...............................................................

.................................42-2 藥物經皮吸收路徑 ................................................................................42-3 影響經皮吸收因素 ................................................................................52-3-1 藥物特性....................................................................

.................52-3-2 藥物載體與配方.........................................................................62-3-3 皮膚生理狀態.............................................................................62-4 藥物穿透皮膚的促進方式 ....................................................................72-4-1 化學性促進方式.......

..................................................................72-4-2 物理性促進方式.........................................................................7第三節、雷射........................................................................................................83-1 雷射簡介 ..........................

......................................................................83-2 雷射作用原理 ........................................................................................83-3 雷射在醫療的應用及雷射光譜 ............................................................93-4 雷射於皮膚應用上的分類 ..................................

................................103-4-1 汽化式雷射...............................................................................103-4-2 非汽化式雷射...........................................................................113-4-3 飛梭雷射...........................................................................

........113-5 鉺雅鉻雷射 ..........................................................................................123-6 飛梭二氧化碳雷射 ..............................................................................12第四節、皮膚老化.......................................................................................

.......134-1 內在因素 ..............................................................................................134-2 外在因素 ..............................................................................................134-3 光老化 .........................................................................

.........................154-3-1 光老化分子機制.......................................................................154-3-2 膠原蛋白...................................................................................164-3-3 彈性蛋白..................................................................................

.174-3-4 透明質酸...................................................................................184-3-5 基質金屬蛋白酶.......................................................................18第五節、對抗皮膚光老化的策略......................................................................205-1 光保護預防策略 ..................

................................................................205-2 外用藥物的投予來改善光老化 ..........................................................215-2-1 視黃酸 (retinoic acid, RA).......................................................215-2-2 RA 改善皮膚老化之作用與機制.............................................225-

2-3 RA 在使用上的限制.................................................................24第六節、奈米藥物載體......................................................................................256-1 常見奈米粒子種類 ..............................................................................256-1-1 微脂體...................

....................................................................266-1-2 奈米乳劑...................................................................................266-1-3 聚合物奈米粒子.......................................................................276-2 聚乳酸 (polylactic acid, PLA) 及聚乳酸乙醇酸 (poly(lact

ic-coglycolic acid), PLGA) 奈米粒子................................................................286-2-1 PLA 及其刺激膠原蛋白增生之效果.......................................286-2-2 PLGA .........................................................................................296-3 奈米粒子投予途徑 ........................

......................................................296-3-1 口服...........................................................................................306-3-2 腸胃外給藥...............................................................................306-3-3 皮膚投予.........................................

..........................................306-3-4 眼部投予...................................................................................31第二章、研究動機與實驗設計..................................................................................32第一節、研究背景與實驗動機...................................................

.......................32第二節、實驗設計與流程..................................................................................33第三章、實驗材料與方法..........................................................................................34第一節、試劑與儀器....................................................................

......................341-1 藥物、試劑與材料 ..............................................................................341-2 儀器設備 ..............................................................................................37第二節、實驗方法......................................................................

........................392-1 奈米劑型製備 ......................................................................................392-1-1 包覆 RA 之奈米劑型製備......................................................392-1-2 標定尼羅紅 (Nile red) 之奈米劑型製備 ...............................402-1-3 標定玫瑰紅 B (rhodamine B) 之奈米劑型製備.

...................402-2 奈米劑型之物化性質測定 ..................................................................402-2-1 粒徑大小及粒徑分散係數.......................................................402-2-2 表面電位...................................................................................412-2-3 疏水性測定...............

................................................................412-2-3 RA 包覆率.................................................................................412-2-4 RA 之高效液相層析儀分析.....................................................422-3 Free RA 製備...................................................

.....................................432-4 體外釋放試驗裝置及方法 ..................................................................432-5 穿透障壁種類及處理 ..........................................................................442-5-1 豬皮...............................................................................

............442-5-2 去角質豬皮...............................................................................442-5-3 經雷射豬皮...............................................................................452-6 雷射後之微孔洞觀察 ..........................................................................472-7 體外釋放試驗藥物

累積穿透量及皮內含量測定 ..............................472-8 毛囊內藥物含量測定 ..........................................................................472-9 奈米粒子經皮吸收之皮內分布 ..........................................................482-9-1 正立雷射共軛焦顯微鏡之觀察...............................................482-9-2 冷凍切片製備與螢光顯微鏡之

觀察.......................................482-10 體外細胞試驗 ....................................................................................492-10-1 冷凍細胞活化與繼代.............................................................492-10-2 細胞對於奈米粒子之攝取.....................................................492-10-3

細胞對於 free RA 及奈米劑型 RA 之存活率試驗...........502-10-4 細胞對於 UVA 之存活率試驗.............................................512-10-5 Free RA 及奈米劑型 RA 對於 Hs68 細胞暴露於 UVA 的存活率試驗..........................................................................................512-10-6 Western blotting 探討奈米劑型 RA 之光老化預防效果....522-

11 動物安全性試驗 ................................................................................552-11-1 相機及手持顯微鏡之影像觀察.............................................552-11-2 皮膚表皮色差計檢測.............................................................552-11-3 皮膚表皮酸鹼度檢測.............................................

................562-11-4 皮膚表皮水分散失檢測.........................................................562-11-5 皮膚組織病理學觀察.............................................................562-12 組織病理學 ........................................................................................562-12-1 石蠟包埋...............

..................................................................562-12-2 蘇木素-伊紅染色 (hematoxylin and eosin stain, H&E stain)572-13 統計方法 ............................................................................................57第四章、實驗結果..............................................................

........................................58第一節、不同處方 PLA 及 PLGA 奈米劑型之物化性質............................581-1 奈米劑型之粒徑、分散係數及表面電位 ..........................................581-2 奈米劑型之安定性 ..............................................................................581-3 奈米劑型之疏水性 ...........................

...................................................601-4 奈米劑型包覆 RA 後之物化性質與包覆率.....................................60第二節、雷射後之微孔洞觀察..........................................................................632-1 雷射施打於感熱紙之結果 ..................................................................632-2 雷射施打於豬皮

之結果 ......................................................................64第三節、體外經皮吸收試驗..............................................................................703-1 包覆 RA 奈米劑型對於正常豬皮之 RA 累積穿透量....................703-2 包覆 RA 奈米劑型對於正常豬皮之 RA 皮內含量........................703-3 不同處理方式之豬皮對於 RA 皮內含量促進

效果.........................723-3-1 Free RA 組 ................................................................................723-3-2 100% PLA 奈米劑型組 ............................................................733-3-3 PLA:PLGA = 3:7 奈米劑型組..................................................733-4 經雷射處理後毛囊

內 RA 含量.........................................................773-5 奈米粒子經皮吸收之皮內分布 ..........................................................783-5-1 共軛焦顯微鏡拍攝之皮膚影像...............................................783-5-2 螢光顯微鏡拍攝之冷凍切片影像...........................................86第四節、體外細胞試驗.............

.........................................................................894-1 細胞對於標定 rhodamine B 奈米劑型之攝取 .................................894-1-1 流式細胞分析儀檢測之細胞攝取狀況...................................894-1-2 共軛焦電子顯微鏡拍攝之細胞攝取影像...............................914-2 細胞對於 free RA 及奈米劑型 RA 之存活率測試 ...........

.............924-3 細胞對於 UVA 之存活率測試 ..........................................................944-4 奈米劑型 RA 對於照射 UVA 之 Hs68 細胞之存活率測試 ........954-5 奈米劑型 RA 對於 Hs68 細胞之預防光老化效果 ........................974-5-1 膠原蛋白 I 之蛋白表現量......................................................974-5-2 彈性蛋白之蛋白表現量..........

.................................................984-5-3 MMP-1 之蛋白表現量 .............................................................98第五節、動物安全性試驗................................................................................1005-1 外觀影像觀察 ..............................................................

......................1005-2 生理數值檢測 ....................................................................................1015-3 組織病理學觀察 ................................................................................102第五章、綜合討論..........................................................................

..........................104第六章、結論............................................................................................................110第七章、參考文獻....................................................................................................112附錄.........................................

...................................................................................122圖目錄圖一、人類皮膚結構...............................................................................................1圖二、表皮層結構..................................................................................................

.3圖三、藥物經皮吸收之皮膚滲透路徑...................................................................5圖四、藥物經皮吸收之物理性促進方法...............................................................7圖五、雷射基本作用原理及裝置...........................................................................9圖六、雷射光譜圖...................................

..............................................................10圖七、汽化式、非汽化式及飛梭雷射示意圖.....................................................11圖八、雷射系統中生物組織的吸收係數.............................................................12圖九、不同紫外線之穿透能力.....................................................................

........14圖十、皮膚老化過程之分子與生理機制示意圖.................................................16圖十一、膠原蛋白結構.........................................................................................17圖十二、彈性蛋白結構.........................................................................................17圖十三、透明質酸之基本結構....

.........................................................................18圖十四、RA 化學結構式......................................................................................21圖十五、類視黃醇代謝及訊號傳遞路徑.............................................................22圖十六、光老化及 RA 作用機制.............................

...........................................23圖十七、奈米粒子種類.........................................................................................25圖十八、微脂體結構.............................................................................................26圖十九、水包油及油包水奈米乳劑示意圖................................

.........................27圖二十、PLA 及其組成單體................................................................................28圖二十一、PLGA 及其組成單體.........................................................................29圖二十二、奈米劑型藥物投予途徑.....................................................................31

圖二十三、RA 高效液相層析儀分析之檢量線..................................................42圖二十四、RA 標準品 (40 μg/ml) 之高效液相層析圖譜.................................43圖二十五、體外直立式擴散玻璃裝置.................................................................44圖二十六、奈米劑型之疏水性測試..............................................................

.......60圖二十七、雷射施打於感熱紙以相機拍攝之影像.............................................63圖二十八、雷射施打於感熱紙以光學顯微鏡拍攝之影像.................................64圖二十九、雷射施打於豬皮皮膚以手持顯微鏡拍攝之影像.............................66圖三十、雷射施打於豬皮皮膚以光學顯微鏡拍攝之影像.................................67圖三十一、雷射施打於豬皮皮膚以掃描式電子顯微鏡拍攝之影像.................68圖三

十二、雷射施打於豬皮皮膚經蘇木素-伊紅染色切片之縱切面................69圖三十三、不同的 RA 奈米劑型經皮吸收之皮內含量....................................71圖三十四、豬皮經不同處理後在經皮吸收試驗中 RA 之皮內含量................75圖三十五、雷射處理後毛囊內 RA 含量............................................................77圖三十六、共軛焦顯微鏡—皮膚空白對照組............................................

.........79圖三十七、共軛焦顯微鏡—100% PLA 於未施予雷射皮膚之分布影像 .........80圖三十八、共軛焦顯微鏡—100% PLA 於鉺雅鉻雷射皮膚之分布影像 .........81圖三十九、共軛焦顯微鏡—100% PLA 於飛梭二氧化碳雷射皮膚之分布影像.............................................................................................................82圖四十、共軛焦顯微鏡—PLA:PLGA = 3:7 於未施予雷射皮膚之分布影像...83圖四十一、

共軛焦顯微鏡—PLA:PLGA = 3:7 於鉺雅鉻雷射皮膚之分布影像...............................................................................................................84圖四十二、共軛焦顯微鏡—PLA:PLGA = 3:7 於飛梭二氧化碳雷射皮膚之分布影像...................................................................................................85圖四十三、冷凍切

片—皮膚空白對照組.............................................................86圖四十四、冷凍切片—100% PLA 投予於皮膚之分布影像 .............................87圖四十五、冷凍切片—PLA:PLGA = 3:7 投予於皮膚之分布影像...................88圖四十六、細胞攝取奈米劑型之流式細胞儀檢測結果.....................................90圖四十七、細胞攝取奈米劑型之共軛焦顯微鏡影像...........................

..............91圖四十八、不同濃度之 free RA 及奈米劑型 RA 對於 Hs68 之存活率.......93圖四十九、不同 UVA 能量對於 Hs68 之存活率.............................................94圖五十、10 J/cm2 UVA 下以奈米劑型 RA 前處理之細胞存活率...................95圖五十一、5 J/cm2 UVA 下以奈米劑型 RA 前、後處理之細胞存活率.........96圖五十二、膠原蛋白 I 之蛋白表現量......................................

..........................97圖五十三、彈性蛋白之蛋白表現量.....................................................................98圖五十四、MMP-1 之蛋白表現量 ......................................................................99圖五十五、動物安全性試驗之外觀影像...........................................................100圖五十六、動物安全性試驗之生理數值

...........................................................102圖五十七、動物安全性試驗之組織病理學切片...............................................103表目錄表一、PLA 及 PLGA 奈米劑型處方比例..........................................................40表二、鉺雅鉻雷射儀器簡介.......................................................................

..........45表三、鉺雅鉻雷射實驗條件.................................................................................45表四、飛梭二氧化碳雷射儀器簡介.....................................................................46表五、飛梭二氧化碳雷射實驗條件.....................................................................46表六、Western blotting 電

泳膠體製備配方.........................................................53表七、PLA 及 PLGA 奈米劑型物化性質..........................................................59表八、PLA 及 PLGA 奈米劑型包覆 RA 之物化性質 ....................................61表九、奈米劑型對於 RA 之包覆率...................................................................

.62表十、豬皮經不同處理後在經皮吸收試驗中 RA 之皮內含量與促進倍數....76