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研究生:劉彥伶
研究生(外文):Low Yan Ling
論文名稱:搭載白藜蘆醇與阿魏酸的奈米脂質載體的製備及功效
論文名稱(外文):Preparation and functions of resveratrol-ferulic acid loaded nanostructured lipid carrier system
指導教授:蔣丙煌蔣丙煌引用關係
指導教授(外文):Been-Huang Chiang
口試委員:陳錦樹陳健生林育蔚
口試委員(外文):Chin-Shuh ChenChien-Sheng ChenYu-Wei Lin
口試日期:2019-01-08
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:食品科技研究所
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2019
畢業學年度:107
語文別:英文
論文頁數:89
中文關鍵詞:綠色保養品阿魏酸白藜蘆醇奈米脂質載體膠帶剝離法
DOI:10.6342/NTU201900152
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近年來消費者對以天然成分為訴求的皮膚保養品的需求愈來愈高,尤其是具有美白效果的天然保養品在亞洲市場的需求量甚大。在亞洲人眼中,皮膚白皙代表的不僅是美,還隱含了社會階層的標誌。天然的植化素中,阿魏酸與白藜蘆醇已被多個研究證實能有效地抑制紫外線照射引起的物質變性、清除自由基、抑制氧化反應和自由基反應、抑制酪氨酸酶的活動,從而抑制黑色素的生成。保養品多直接使用於皮膚,但皮膚不易穿透的特性卻讓保養品中的有效成分的吸收受到限制。因此,如何使有效的活性物質順利穿透表層皮膚組織到達作用位置極其重要。本研究結合多重乳化技術與奈米結構脂質載體的概念,利用超音波探針均質,製造出同時搭載親脂性與親水性植化素的奈米脂質載體 (nanostructured lipid carrier)。 本研究選用的天然來源原料包括以利普脂、癸酸三酸甘油酯、親脂性乳化劑聚甘油-4異硬脂酸酯(polyglyceryl-4 isostearate) 及親水性乳化劑聚甘油-3甲基葡萄糖二硬脂酸酯(polyglyceryl-3 methylglucose distearate)。在全因子實驗設計的優化下,當聚甘油-3甲基葡萄糖二硬脂酸酯的濃度為2.56%與第二階段超音波均質時間為6分鐘時,奈米脂質載體之粒徑為375.37 ± 90.26 nm,穩定度為94.29% ± 1.39, 阿魏酸包埋率為85.49% ± 8.45,白藜蘆醇包埋率為85.57% ± 1.39。將上述優化的載體配方進行經皮吸收試驗(豬背皮),載體組與控制組相比,白藜蘆醇的穿透量增加了29.31%。而利用膠帶撕貼法的人體經皮吸收試驗,證實了載體組中阿魏酸及白藜蘆醇的穿透率比控制組分別高出17.23%及27.19%。後續的細胞實驗也證實了阿魏酸及白藜蘆醇確實對UVA照射誘導小鼠B16F10黑色素瘤細胞生成黑色素有抑制的作用。本研究的結果提供了以天然來源原料製備搭載植化素的的奈米脂質載體的方法,以及提升了其用於美白保養品的可行性。
Surging consumer demand for skin care product that are free from harmful chemicals has encouraged the development of product derived from natural sources. Among all the product benefits skin lightening has gained a lot of attention in the last years, especially in Asian communities. Topically applied product, however, often suffer from low penetration of active compounds into skin due to excellent barrier of stratum corneum. In an attempt to overcome these problems, this study has developed a nanostructured lipid carriers (NLC) system encapsulating both hydrophilic ferulic acid and lipophilic resveratrol. Both of these two compounds are known to be able to inhibit melanogensis in skin. Natural derived ingredients including illipe nut butter, capric triglyceride, polyglyceryl 4 isostearate, and polyglyceryl 3 methylglucose distearate were chosen to fabricate NLC. A full factorial design was used to optimize the concentration of polyglyceryl 3 methylglucose distearate and duration of ultrasonication during second stage of emulsification. Results showed that 2.56% of polyglyceryl 3 methylglucose distearate and 6 minutes sonication time were suitable to produce stable NLC system with mean diameter of 375.37 ± 90.26 nm, 94.29% ± 1.39 stability, 85.49% ± 8.45 ferulic acid encapsulation efficiency, and 85.57% ± 1.39 resveratrol encapsulation efficiency. The ex vivo penetration study revealed that NLC could increase the penetration of resveratrol for about 29.31% in porcine back skin. The in vivo tape stripping penetration study indicated that the NLC increased the cumulative penetration of ferulic acid and resveratrol by 17.23% and 27.19% respectively as compared to the free compounds. The in vitro antimelanogenic effects of ferulic acid and resveratrol was demonstrated by treating B16F10 mouse melanoma cells with different amount of these phytochemicals loaded NLC before and after exposure to UVA radiations. These findings suggested that NLC made by natural derived ingredients is a promising topical delivery system that had a potent enhancement effect for ferulic acid-resveratrol penetration into skin.
Acknowledgement ii
中文摘要 iii
Abstract iv
Table of contents vi
List of tables x
List of figures xii
Chapter 1 Introduction 1
Chapter 2 Literature Review 4
2.1 Pigmentation in human skin 4
2.1.1 Cell and organelles involved in pigmentation 4
2.1.2 Events in melanogenesis 4
2.1.2.1 Synthesis of proteins required for melanogenesis 4
2.1.2.2 Synthesis of melanin 5
2.1.2.3 Melanosome biogenesis 6
2.1.2.4 Transport of melanosomes from melanocytes to keratinocytes 7
2.1.3 Signaling pathways regulating melanogenesis 7
2.1.4 Factors affecting melanogenesis 9
2.1.4.1 Intrinsic factors 9
2.1.4.2 Extrinsic factors 10
2.1.5 Approaches to achieve depigmentation 12
2.1.5.1 Depigmenting agents 12
2.1.5.2 Mechanisms of depigmentation 14
2.2 Topical delivery 16
2.2.1 General background 16
2.2.2 Human skin structure 16
2.2.3 Main functions of topical delivery 17
2.2.4 Possible route of penetration 18
2.2.5 Challenges associated with topical delivery 19
2.2.6 Strategies to facilitate topical delivery 20
2.2.6.1 Active methods 20
2.2.6.2 Passive methods 20
2.3 Nanodelivery system 21
2.3.1 Types of nanodelivery system 21
2.3.1.1 Liposome 21
2.3.1.2 Microemulsion and nanoemulsion 22
2.3.1.3 Solid lipid nanoparticle 23
2.3.1.4 Nanostructured lipid carrier 24
2.3.2 Important component in nanodelivery system 25
2.3.2.1 Solid lipid 25
2.3.2.2 Emulsifier 26
2.3.3 Methods in producing nanodelivery system 29
2.3.3.1 Low energy emulsification 29
2.3.3.2 High energy emulsification 30
Chapter 3 Aim and Experimental Framework 32
3.1 Objectives 32
3.2 Experimental framework 32
Chapter 4 Materials and Methods 34
4.1 List of chemical and reagent 34
4.2 List of apparatus and equipment 34
4.3 Methods 35
4.3.1 Screening of materials 35
4.3.1.1 Determination of solid lipid-to-liquid oil ratio 35
4.3.1.2 Preparation of stock solution 35
4.3.1.3 Cytotoxicity of emulsifiers 36
4.3.1.4 Concentration of polyglyceryl 4 isostearate 36
4.3.2 Optimization of NLC 37
4.3.2.1 Preparation of NLC 37
4.3.2.2 Experimental design 37
4.3.2.3 Particle size analysis 39
4.3.2.4 Stability 39
4.3.2.5 Encapsulation efficiency 40
4.3.2.6 Cumulative release 40
4.3.3 Penetration study 42
4.3.3.1 Ex vivo skin penetration 42
4.3.3.2 In vivo skin penetration 43
4.3.4 Antimelanogenic potential of ferulic acid and resveratrol 44
4.3.4.1 Cell culture 44
4.3.4.2 Preparation of stock solution 44
4.3.4.3 Cell viability assay 44
4.3.4.4 Treatment of cells with active compounds and UVA irradiation 45
4.3.4.5 Determination of melanin content 45
4.3.5 Statistical analysis 46
Chapter 5 Results and Discussions 47
5.1 Screening of materials 47
5.1.1 Determination of solid lipid: liquid oil ratio 47
5.1.2 Preparation of stock solution 48
5.1.3 Cytotoxicity of emulsifier 49
5.1.4 Determination of concentration of lipophilic emulsifier 51
5.2 Optimization of production of NLC 52
5.2.1 Justification of factors 52
5.2.2 Effects of factors on responses 54
5.2.2.1 Particle size 58
5.2.2.2 Stability 59
5.2.2.3 Encapsulation efficiency 60
5.2.3 Response values under optimized settings of NLC 62
5.3 Penetration study of NLC 63
5.3.1 Ex vivo study of penetration 64
5.3.2 In vivo study of penetration 67
5.4 Antimelanogenesis potential of ferulic acid and resveratrol 70
5.4.1 Cell viability study 70
5.4.2 Effects of active compounds against UVA induced melanogenesis 72
Chapter 6 Conclusion 75
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