臺灣博碩士論文加值系統

Thiazolidinedione（TZD）類藥物為現行少數之胰島素增敏劑（insulin sensitizer），用於治療第二型糖尿病。主要的作用機轉是經由刺激細胞核表面接受器 PPARγ（peroxisome proliferator activated receptor gamma），增加脂肪細胞攝取游離脂肪酸及增加骨骼肌攝取葡萄糖。臨床上使用的TZD 類藥物如羅格列酮(rosiglitazone) ，具有減緩發炎，增加胰島素抗性等優點，但同時具有數種的副作用，像是體重上升，水腫，易骨折，也有造成心臟衰竭相關症狀的疑慮，使其在臨床的應用上受到很大的限制。本篇研究利用結合聚合物乳化技術與注射式凝膠傳輸羅格列酮，除了解決羅格列酮水溶性的問題，也期望能避免羅格列酮可能產生的副作用。在動物實驗的評估中，含有乳化羅格列酮顆粒的凝膠對於組織造成預期之外程度的異物發炎反應。調整系統以降低發炎反應為未來首要解決之目標。

Thiazolidinedione drugs (TZDs) are the only current antidiabetic agents that function primarily by increasing insulin sensitivity. TZDs were first reported as insulin-sensitizing drugs in the early 1980s, but their mechanism remained a mystery until the mid-1990s, when they were found to be ligands for the nuclear receptor transcription factor PPARγ. Molecular compounds such as the clinically available rosiglitazone and pioglitazone have been designed to target PPARγ and for the oral treatment of type 2 Diabetes. However, in addition to attenuating inflammation and promoting insulin sensitivity, these compounds are often associated with severe side effects including weight gain, edema, bone fracture, and congestive heart failure, which have severely limited their clinical application. A rosiglitazone delivery strategy composing of polymeric emulsion particles and injectable gel system was established. In this study, Sprague Dawley® rats were used to evaluate the physiological reactions, but all the combinations of hydrogels with rosiglitazone emulsion particles show the unwanted foreign body reaction in a certain degree.

中文摘要 i
Abstract ii
Contents iii
List of Figures v
List of Tables vii
Chapter 1 Introduction 1
1.1. Obesity and Thermogenic Fat 1
1.1.1. Obesity and Metabolic Disorder 1
1.1.2. Adipose Tissues and Metabolic Potential 2
1.1.3. Plasticity of Adipose Tissue 4
1.1.4. Inducing Browning of White Adipose Tissue 6
1.1.5. Fighting Fat with Fat 9
1.2. Drug Delivery Strategies 10
1.2.1. Emulsions for Low Solubility Drugs 10
1.2.2. Hydrogels for Sustained Drug Delivery 13
Chapter 2 Experimental Design 16
Chapter 3 Materials and Methods 17
3.1. Materials 17
3.2. Rosiglitazone Encapsulated Emulsions 18
3.2.1. Fabricating of Rosiglitazone Encapsulated Emulsions 18
3.2.2. Size Measurement and Rosiglitazone Content Determination 19
3.2.3. Cell Viability Assay 20
3.3. Injectable Hydrogels Preparation 20
3.3.1. Preparation of Sterile Chitosan, HA and MC 20
3.3.2. Preparation of Injectable Hydrogels 21
3.3.3. Rheological Tests 22
3.4. In Vivo Test 23
Chapter 4 Results and Discussions 24
4.1. Rosiglitazone Encapsulated Emulsions 24
4.1.1. Particle Characteristics 24
4.1.2. Cell Viability Assay 27
4.2. Hydrogel Characteristics 29
4.2.1. Chitosan-β-GP Hydrogel Characteristics 29
4.2.2. HAMC Hydrogel Characteristics 31
4.2.3. Rheological Characteristics of HAMC Gels 32
4.3. In Vivo Tests 37
4.3.1. Tissue Section Stain 37
Chapter 5 Conclusions 54
Chapter 6 Reference 55

1.Giordano, A., A. Frontini, and S. Cinti, Convertible visceral fat as a therapeutic target to curb obesity. Nat Rev Drug Discov, 2016.
2.Lee, Y.H., E.P. Mottillo, and J.G. Granneman, Adipose tissue plasticity from WAT to BAT and in between. Biochim Biophys Acta, 2014. 1842(3): p. 358-69.
3.Saito, M., Brown adipose tissue as a regulator of energy expenditure and body fat in humans. Diabetes Metab J, 2013. 37(1): p. 22-9.
4.Sidossis, L. and S. Kajimura, Brown and beige fat in humans: thermogenic adipocytes that control energy and glucose homeostasis. J Clin Invest, 2015. 125(2): p. 478-86.
5.Harms, M. and P. Seale, Brown and beige fat: development, function and therapeutic potential. Nat Med, 2013. 19(10): p. 1252-63.
6.Bartelt, A. and J. Heeren, Adipose tissue browning and metabolic health. Nat Rev Endocrinol, 2014. 10(1): p. 24-36.
7.Kajimura, S. and M. Saito, A new era in brown adipose tissue biology: molecular control of brown fat development and energy homeostasis. Annu Rev Physiol, 2014. 76: p. 225-49.
8.Sanchez-Gurmaches, J., C.M. Hung, and D.A. Guertin, Emerging Complexities in Adipocyte Origins and Identity. Trends Cell Biol, 2016. 26(5): p. 313-26.
9.Soccio, R.E., E.R. Chen, and M.A. Lazar, Thiazolidinediones and the promise of insulin sensitization in type 2 diabetes. Cell Metab, 2014. 20(4): p. 573-91.
10.Nesto, R.W., et al., Thiazolidinedione Use, Fluid Retention, and Congestive Heart Failure. Diabetes Care, 2003. 27(1): p. 256-263.
11.Lago, R.M., P.P. Singh, and R.W. Nesto, Congestive heart failure and cardiovascular death in patients with prediabetes and type 2 diabetes given thiazolidinediones: a meta-analysis of randomised clinical trials. The Lancet. 370(9593): p. 1129-1136.
12.He, H., et al., Rosiglitazone Causes Cardiotoxicity via Peroxisome Proliferator-Activated Receptor γ-Independent Mitochondrial Oxidative Stress in Mouse Hearts. Toxicological Sciences, 2014. 138(2): p. 468-481.
13.Ohno, H., et al., PPARgamma agonists induce a white-to-brown fat conversion through stabilization of PRDM16 protein. Cell Metab, 2012. 15(3): p. 395-404.
14.Seale, P., Transcriptional Regulatory Circuits Controlling Brown Fat Development and Activation. Diabetes, 2015. 64(7): p. 2369-75.
15.Wu, J., H. Jun, and J.R. McDermott, Formation and activation of thermogenic fat. Trends Genet, 2015. 31(5): p. 232-8.
16.Pfeifer, A. and L.S. Hoffmann, Brown, beige, and white: the new color code of fat and its pharmacological implications. Annu Rev Pharmacol Toxicol, 2015. 55: p. 207-27.
17.Tadros, T.F., Emulsion Formation, Stability, and Rheology, in Emulsion Formation and Stability. 2013, Wiley-VCH Verlag GmbH & Co. KGaA. p. 1-75.
18.Batrakova, E.V. and A.V. Kabanov, Pluronic block copolymers: evolution of drug delivery concept from inert nanocarriers to biological response modifiers. J Control Release, 2008. 130(2): p. 98-106.
19.Al Khateb, K., et al., In situ gelling systems based on Pluronic F127/Pluronic F68 formulations for ocular drug delivery. Int J Pharm, 2016. 502(1-2): p. 70-9.
20.Ayorinde, F.O., et al., Analysis of some commercial polysorbate formulations using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Rapid Communications in Mass Spectrometry, 2000. 14(22): p. 2116-2124.
21.Brandner, J.D., The Composition of NF-Defined Emulsifiers: Sorbitan Monolaurate, Monopalmitate, Monostearate, Monooleate, Polysorbate 20, Polysorbate 40, Polysorbate 60, and Polysorbate 80. Drug Development and Industrial Pharmacy, 1998. 24(11): p. 1049-1054.
22.Frison-Norrie, S. and P. Sporns, Investigating the Molecular Heterogeneity of Polysorbate Emulsifiers by MALDI-TOF MS. Journal of Agricultural and Food Chemistry, 2001. 49(7): p. 3335-3340.
23.Kerwin, B.A., Polysorbates 20 and 80 used in the formulation of protein biotherapeutics: structure and degradation pathways. J Pharm Sci, 2008. 97(8): p. 2924-35.
24.Hoffman, A.S., Hydrogels for biomedical applications. Advanced Drug Delivery Reviews, 2012. 64: p. 18-23.
25.Ko, D.Y., et al., Recent progress of in situ formed gels for biomedical applications. Progress in Polymer Science, 2013. 38(3-4): p. 672-701.
26.Prestwich, G.D., et al., Controlled chemical modification of hyaluronic acid: synthesis, applications, and biodegradation of hydrazide derivatives. Journal of Controlled Release, 1998. 53(1–3): p. 93-103.
27.Campoccia, D., et al., Semisynthetic resorbable materials from hyaluronan esterification. Biomaterials, 1998. 19(23): p. 2101-2127.
28.Shu, X.Z., et al., Attachment and spreading of fibroblasts on an RGD peptide–modified injectable hyaluronan hydrogel. Journal of Biomedical Materials Research Part A, 2004. 68A(2): p. 365-375.
29.Zheng, P., et al., Sol-gel transition of methylcellulose in phosphate buffer saline solutions. Journal of Polymer Science Part B: Polymer Physics, 2004. 42(10): p. 1849-1860.
30.Arvidson, S.A., et al., Interplay of Phase Separation and Thermoreversible Gelation in Aqueous Methylcellulose Solutions. Macromolecules, 2013. 46(1): p. 300-309.
31.Liang, H.-F., et al., Novel Method Using a Temperature-Sensitive Polymer (Methylcellulose) to Thermally Gel Aqueous Alginate as a pH-Sensitive Hydrogel. Biomacromolecules, 2004. 5(5): p. 1917-1925.
32.Hoenich, N.A., et al., Biocompatibility of membranes used in the treatment of renal failure. Biomaterials, 1995. 16(8): p. 587-592.
33.Gupta, D., C.H. Tator, and M.S. Shoichet, Fast-gelling injectable blend of hyaluronan and methylcellulose for intrathecal, localized delivery to the injured spinal cord. Biomaterials, 2006. 27(11): p. 2370-9.
34.Khaing, Z.Z. and S.K. Seidlits, Hyaluronic acid and neural stem cells: implications for biomaterial design. J. Mater. Chem. B, 2015. 3(40): p. 7850-7866.
35.Vercruysse, K.P. and G.D. Prestwich, Hyaluronate Derivatives in Drug Delivery. 1998. 15(5): p. 43.
36.Baier Leach, J., et al., Photocrosslinked hyaluronic acid hydrogels: Natural, biodegradable tissue engineering scaffolds. Biotechnology and Bioengineering, 2003. 82(5): p. 578-589.
37.Balazs, E.A., Medical Applications of Hyaluronan and its Derivatives, in Cosmetic and Pharmaceutical Applications of Polymers, C.G. Gebelein, T.C. Cheng, and V.C. Yang, Editors. 1991, Springer US: Boston, MA. p. 293-310.
38.Rodell, C.B., A.L. Kaminski, and J.A. Burdick, Rational design of network properties in guest-host assembled and shear-thinning hyaluronic acid hydrogels. Biomacromolecules, 2013. 14(11): p. 4125-34.
39.Barbucci, R., et al., Hyaluronic acid hydrogel in the treatment of osteoarthritis. Biomaterials, 2002. 23(23): p. 4503-4513.
40.Hruska, M.W. and R.F. Frye, Simplified method for determination of rosiglitazone in human plasma. Journal of Chromatography B, 2004. 803(2): p. 317-320.
41.Lavertu, M., et al., A validated 1H NMR method for the determination of the degree of deacetylation of chitosan. Journal of Pharmaceutical and Biomedical Analysis, 2003. 32(6): p. 1149-1158.
42.Yuan, Y., et al., Deacetylation of Chitosan: Material Characterization and in vitro Evaluation via Albumin Adsorption and Pre-Osteoblastic Cell Cultures. Materials, 2011. 4(12): p. 1399-1416.
43.Pandit, N.K. and J. Kisaka, Loss of gelation ability of Pluronic® F127 in the presence of some salts. International Journal of Pharmaceutics, 1996. 145(1): p. 129-136.
44.Pandit, N.K. and D. Wang, Salt effects on the diffusion and release rate of propranolol from poloxamer 407 gels. International Journal of Pharmaceutics, 1998. 167(1–2): p. 183-189.
45.Desai, P.R., et al., Effect of additives on the micellization of PEO/PPO/PEO block copolymer F127 in aqueous solution. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 2001. 178(1–3): p. 57-69.
46.Mao, G., et al., PEO−PPO−PEO Block Copolymer Micelles in Aqueous Electrolyte Solutions:  Effect of Carbonate Anions and Temperature on the Micellar Structure and Interaction. Macromolecules, 2001. 34(3): p. 552-558.
47.Joshi, T., B. Bharatiya, and K. Kuperkar, Micellization and Interaction Properties of Aqueous Solutions of Mixed Cationic and Nonionic Surfactants. Journal of Dispersion Science and Technology, 2008. 29(3): p. 351-357.
48.Seo, H.J. and J.-C. Kim, Effects of additives on phase transitions of Poloxamer 407/Poloxamer 188 mixture and release property of monoolein cubic phase containing the poloxamers. Journal of Industrial and Engineering Chemistry, 2012. 18(1): p. 88-91.
49.Honda, S., T. Yamamoto, and Y. Tezuka, Tuneable enhancement of the salt and thermal stability of polymeric micelles by cyclized amphiphiles. Nat Commun, 2013. 4: p. 1574.
50.Krupka, T.M. and A.A. Exner, Structural parameters governing activity of Pluronic triblock copolymers in hyperthermia cancer therapy. Int J Hyperthermia, 2011. 27(7): p. 663-71.
51.Budkina, O.A., et al., Cytotoxicity of nonionic amphiphilic copolymers. Polymer Science Series A, 2012. 54(9): p. 707-717.
52.Oshiro, A., et al., Pluronics f-127/l-81 binary hydrogels as drug-delivery systems: influence of physicochemical aspects on release kinetics and cytotoxicity. Langmuir, 2014. 30(45): p. 13689-98.
53.Au, W.-S., Pluronic L-81 ameliorates diabetic symptoms indb/dbmice through transcriptional regulation of microsomal triglyceride transfer protein. World Journal of Gastroenterology, 2009. 15(24): p. 2987.
54.Zuidema, J.M., et al., A protocol for rheological characterization of hydrogels for tissue engineering strategies. J Biomed Mater Res B Appl Biomater, 2014. 102(5): p. 1063-73.
55.Gullapalli, R.P. and B.B. Sheth, Effect of methylcellulose on the stability of oil-in-water emulsions: influence of the disperse phase. International Journal of Pharmaceutics, 1997. 151(2): p. 249-253.
56.Anderson, J.M., A. Rodriguez, and D.T. Chang, Foreign body reaction to biomaterials. Semin Immunol, 2008. 20(2): p. 86-100.
57.Wynn, T.A. and T.R. Ramalingam, Mechanisms of fibrosis: therapeutic translation for fibrotic disease. Nat Med, 2012. 18(7): p. 1028-40.
58.Zhang, L., et al., Zwitterionic hydrogels implanted in mice resist the foreign-body reaction. Nat Biotechnol, 2013. 31(6): p. 553-6.
59.DiEgidio, P., et al., Biomedical implant capsule formation: lessons learned and the road ahead. Ann Plast Surg, 2014. 73(4): p. 451-60.
60.Farrugia, B.L., et al., The localisation of inflammatory cells and expression of associated proteoglycans in response to implanted chitosan. Biomaterials, 2014. 35(5): p. 1462-77.
61.Svensson, S., et al., A novel soft tissue model for biomaterial-associated infection and inflammation - bacteriological, morphological and molecular observations. Biomaterials, 2015. 41: p. 106-21.
62.Veiseh, O., et al., Size- and shape-dependent foreign body immune response to materials implanted in rodents and non-human primates. Nat Mater, 2015. 14(6): p. 643-51.
63.Lee, J.M. and Y.J. Kim, Foreign body granulomas after the use of dermal fillers: pathophysiology, clinical appearance, histologic features, and treatment. Arch Plast Surg, 2015. 42(2): p. 232-9.
64.He, X., et al., The production of fully deacetylated chitosan by compression method. The Egyptian Journal of Aquatic Research, 2016. 42(1): p. 75-81.
65.Di Mascolo, D., et al., Rosiglitazone-loaded nanospheres for modulating macrophage-specific inflammation in obesity. J Control Release, 2013. 170(3): p. 460-8.