奈 Stabilization and Controlled Release of Active Agents via a Degrading Nano-Carrier 料 : 年六 1
奈 奈 兩 奈 料 來 奈 理 行 料 數 聯 度 粒 例 數 量 度 度 量 數 數 度 料 良 量 降 數 不 不易 力奈 來 參 ABSTRACT The object of this study was to manipulate the drug permeability and stability of nano Ca-deficient hydroxyapatite/chitosan (CDHA/CS) nanocomposite membrane incorporated with vitamins for drug storage and controlled release. The physical properties and drug permeation behaviors of Ca-deficient hydroxyapatite/chitosan (CDHA/CS) nanocomposite membrane were systematically investigated in terms of different synthetic sequence and inorganic filler amount. It was found that the lowest permeation coefficient was obtained via in-situ process (P-CS-Ca), which was probably due to the higher crosslinking extent, better interface between filler and matrix, and smaller particle size, as well as dispersion. With an increase of CDHA content, both crosslinking and diffusion path would be increased so that the permeation coefficient of CDHA/CS composite membrane was reduced compared to that of pure chitosan membrane. A minimum permeation coefficient occurs at the CDHA content of 90%. In this condition, the drug can be safely stored for a long time. 2
料 了 例 : 療 料 不 連 料 不 奈 料 不 力 烈 C 見 C 療 了便 C 來 療 了 療 奈 量 度 料 類 料 料 料 料 不 良 料 不 率 來 ε 奈 度 了 不 異 奈 領 都 奈 領 奈 料 率 in-situ process 念 來 奈 (carrier)) 3
(Denature) 奈 料 率 歷 都 降 量 β β 來 料 來 來 量 離 不 領 聯 利 離 不 降 料 不 數 理 度 異 度 兩 料 例 量 來 理 度 不 料 度 了 度 不 料來 度 量 狀 類 類 量 類 理 4
兩 力 利 不 聯 蠟 利 利 粒 狀 度 利 來 不 载 率 利 利 不 度 利 來 不 度 料 度 粒 兩 量 料 料 量 量 利 度 來 度 度 度 理 離 料 5
CS 數 不 不 量 A PCSCa IP B CaCSP IC C CDHACS NI 6
立葉 奈 料 度 濾 度 奈 料 兩 都 奈 不 量 不 例 奈 例 理 聯 7
離 量 量测 不 度 度 度 量 聯 兩 連 量 度 錄數 利 量 度 利 數 C 1 V 1 C 2 V 2 [ DH ] C10 2 At ln = C C 1 2 δ V 數 數 兩 兩 度 數 料 來 Å 流 率 立葉 8
料 度 數 利 降 度 率 利 度 度 率 率 狀 流 粒 9
論 料 量 不 數 理 度 異 不 料 2θ 20 o (102) (022)(200)(040)(220)(140)CDHA 26 o 28 o 32 o 40 o 47 o 50 o 度 (002) (210) (211)(112)(300)(310)(222)(213) 不 都 CDHA 粒度 都 異不 量 例 度 料 度都 不 料 FTIR FTIR 數 1652 cm -1 (amide I, C-O)1580 cm -1 (-NH 2 bending)2990 cm -1 (methylene, -CH 2 ) 1100 cm -1 1040 cm -1 600 cm -1 3450 cm -1 CDHA PO 4 3- OH - 5-2 CDHA CDHA 料 1580 cm -1 1100 cm -1 都 度 降 1580 cm -1 -NH 2 bending 1100 cm -1 CDHA (-PO 4 3- )(-NH 2 ) (-NH 3 + ) CDHA (-PO 4 3- ) 理 度 A 兩 度 異 A (-NH 2 ) 離 (-PO 4 3- ) 理 兩 來 度 異 5-3 TGA 量 CDHA 料 料 TGA 量 5% 量 兩 100 300 連 降 300-600 量 ( 見 5-3 5-4) TPP 離 量 10% 留量 TPP 離 10
降 降 都 連 降 度 了 離 連 降 度 TGA 類 TPP 兩 A B C CDHA TPP 度 A 離 A CDHA B C 來 A TGA 不 異 DMTA 異 數 (DH)( 見 5-5 5-6) 度 B C A A DH 1CDHA 良 DH 降 2 度 不易 DH 降 3CDHA 率 TEM 兩 DH TGA 度 不 CDHA 不 DH CDHA 5-7 5-8 料 TEMSEM TEM 料 CDHA 狀 80nm 20nm TEM A 料 B C 5-8 料 狀 CS A BC 不 量 度 數 兩 料 例 量 理 度 5-9 XRD 量 CDHA 度 來 不 量 90% 11
peak CDHA 5-10 FTIR 量 2990 cm -1 (methylene,-ch 2 )1580 cm -1 (-NH 2 bending) peak 來 1580 cm -1 (-NH 2 bending) CDHA 量 更 度 來 5-11 5-12 TGA 兩 量 CDHA 量 了 連 降 2.5 cm CDHA 量 數 CS 量 30% 10% 兩 數 5-13 5-14 數 CS50%CS100% CS70%CS90% CDHA DH 降 CDHA 量 度 CDHA 量 數 CDHA 量 度 數 5-15 不 量 量 率 數 12
6 論 聯 度 粒 來 2. 例 不 數 50%100%70%90% CDHA 量 度 DH 降 CDHA 量 度 數 兩 量 90% 數 數 降 度 奈 粒 量 數 都不行 力 13
參 [1] The World's No.1 Science & Technology News Service, Nanotechnology may create new organs, NewScientist.com news service, 8 July 2003. [2] Liu DM, Troczynski T, Tseng WJ. Water-based sol-gel synthesis of hydroxyapatite : process development. Biomaterials 2001;22:1721-30. [3] Q. Liu, J. R. de Wijn, and C. A. van Blitterswijk, Nono-apatite/polymer composites: characteristics, Biomaterals, 18 (1997) 1263-1270. [4]. L. Liu, E. B. Hunziker, N. X. Randell, K. de Groot, and P. Layrolle, Proteins incorporated into biomimetically prepared calcium phosphate coatings modulate their mechanical strength and dissolution rate, Biomaterials; 2003:24: 65-70. [5] E. Gentleman, A. N. Lay, D. A. Dickerson, E. A. Nauman, G. A. Livesay and K. C. Dee, Mechanical characterization of collagen fibers and scaffolds for tissue engineering, Biomaterials;2003:24:3805-13. [6] Y. Zhang, and M. Zhang, Calcium phosphate/chitosan composite scaffolds for controlled in vitro antibiotic drug release, J. Biomed. Mater. Res. 2002, 378-385. [7] Liu DM, Troczynski T, Tseng WJ. Water-based sol-gel synthesis of hydroxyapatite: process development. Biomaterials 2001;22[11]:1721-30. [8] Yubao L, Klein CPAT, De Wijn J, Van De Meer S, De Groot K. Shape Change and Phase Transition of Needle-like Non-Stoichiometric apatite crystals. J. Mater. Sci: Mater. Med. 1994;5:263-68. [9] Yubao L, De Groot K, De Wijn J, Klein CPAT, Van De Meer S. Morphology and Composition of Nanograde Calcium Phosphate Needle-like Crystals Formed by 14
simple Hydrothermal Treatment. J Mater Sci: Mater in Medicine 1994;5:326-31. [10] Lu HB, Ma CL, Cui H, Zhou LF, Wang RZ, Cui FZ. Controlled crystallization of calcium phosphate under stearic acid monolayers. J Crystal Growth 1995;155: 120-25. [11] N. Kossoveky, A. Gelman, E. E. Sponsler, H. J. Hnatyszyn, S. Rajguru, M. Torres, M. Pham, J. Crowder, J. Zemanovich, A. Chung and R. Shah. Surface-modified nanocrystalline ceramics for drug delivery applications, Biomaterials, 1994:15:1201. [12] Knowles JC, Calluct S, Georgiou G. Characterisation of the rheological properties and zeta potential of a range of hydroxyapatite powders. Biomaterials, 2000;21:1387-92. [13] Kwon IK, Park KD, Choi SW, Lee SH, Lee EB, Na JS, Kim SH, Kim YH. Fibroblast culture on surface-modified poly(glycolide-co-epsilon-caprolactone) scaffold for soft tissue regeneration, J. Biomaterials Science-polymer, 12 (10): 1147-1160 2001. [14] Lenza RFS, Vasconcelos WL, Jones JR, Hench LL,. Surface-modified 3D scaffolds for tissue engineering, J. Mater. Sci. Mater. Med.13 (9): 837-842 SEP 2002 [15] D. G. Shchukin, G. B. Sukhorukov, and H. Mohwald, Biomimetic fabrication of nanoengineered hydroxyapatite/polyelectrolyte composite shell, Chem. Mater. 2003,15, 3947-50. [16] Sz-Chian Liou, San-Yuan Chen,, Hsin-Yi Lee and Jong-Shing Bow, Structural characterization of nano-sized calcium deficient apatite powders, Biomaterials, 25 (2004) 189 15
[17] Sz-Chian Liou and San-Yuan Chen, Transformation mechanism of different chemically precipitated apatitic precursors into β tricalcium phosphate upon calcinations, Biomaterials, 23 (2002) 4541. [18] Sz-Chian Liou, San-Yuan Chen and Dean-Mo Liu, Synthesis and Characterization of Needlelike Apatitic nanocomposite with controlled aspect ratios, Biomaterials, 24 (2003) 3981. [19] Sz-Chian Liou, San-Yuan Chen,, Hsin-Yi Lee and Jong-Shing Bow, Structural characterization of nano-sized calcium deficient apatite powders, Biomaterials, 25 (2004) 189. [20] S. R. Jameela, A. Jayakrishnan, Glutaraldehyde cross-linked chitosan microspheres as a long acting biodegradable drug delivery vehiclestudies on the in vitro degradation of microsphers in rat muscle, Biomaterials 16 (1995)769. [21] X. Qiu, S. Leporatti, E. Donath, H. Möhwald, Studies on the drug release properties of polysaccharide multilayers encapsulated ibuprofen microparticles, Langmuir 17 (2001) 5375-5380. [22] A. Zanina, A. Vilesov, T. Budtova, Shear-induced solvent release form gel particles: application to drug-delivery systems, International Journal of Pharmaceutics 242 (2002) 137-146. [23] C. Wang, C. J. Lin, Preparation and characterization of nano-sized hydroxyapatite particles and hydroxyapatite /chitosan nano-composite for use in biomedical materials, Materials Letters 57 (2002) 858-861. [24] I. Yamaguchi, K. Tokuchi, H. Fukuzaki, Y. Koyama, K. Takakuda, H. Monma, J. Tanaka, Preparation and microstructure analysis of chitosan/hydroxyapatite nanocomposites, Journal of Biomedical Materials Research 55 (2001) 20-27. [25] M. Sivakumar, I. Manjubala, K. P. Rao, Preparation, characterization and in-vitro 16
release of gentamicin from coralline hydroxyapatite-chitosan composite microspheres, Carbohydrate Polymers 49 (2002) 281-288. [26] Y. Zhang, M. Q. Zhang, Calcium phosphate/chitosan composite scaffolds for controlled in vitro antibiotic drug release, Journal of Biomedical Materials Research 62 (2002) 378-386. [27] J. S. Ahn, H. K. Choi, M. K. Chun, J. M. Ryu, J. H. Jung, Y. U. Kim, C. S. Cho, Release of triamcinolone acetonide from mucoadhesive polymer composed of chitosan and poly (acrylic acid) in vitro, Biomaterials 23 (2002) 1411-1416. [28] Y. Hu, X. Jiang, Y. Ding, H. Ge, Y. Yuan, C. Yang, Synthesis and characterization of chitosan-poly(acrylic acid) nanoparticles, Biomaterials 23 (2002) 3193-3201. 17
Intensity (a.u.) CS P-CS-Ca Ca-Cs-P CDHA-CS CDHA 20 30 40 50 60 2θ (degree) 量 18
CS 90% Absorbance (a.u.) CS 100% P-CS-Ca Ca-CS-P CDHA-CS CDHA 4000 3500 3000 2500 2000 1500 1000 500 Wave number (cm -1 ) 量 不 聯 19
Weight % 100 90 80 70 60 50 40 30 CS 90% P-CS-Ca Ca-CS-P CDHA-CS CS 20 100 200 300 400 500 600 700 800 900 Temperature ( o C) 量 不 聯 20
Weight % 100 90 80 70 60 TPP ion-crosslinking CS 90% P-CS-Ca Ca-CS-P CDHA-CS CS 50 40 100 200 300 400 500 600 700 800 900 Temperature ( o C) 量 不 聯 21
Conc. (mg/ml) 0.025 0.020 0.015 0.010 0.005 CS P-CS-Ca CDHA-CS Ca-CS-P 0.000 0 1 2 3 4 5 6 Time (hr) 量 不 聯 22
0.0009 0.0008 DH (cm 2 /hr) 0.0007 0.0006 0.0005 0.0004 0.0003 0.0002 CS P-CS-Ca CDHA-CS Ca-CS-P PROCESS 量 不 聯 數 23
100 nm P-CS-Ca 100 nm Ca-CS-P 100 nm CDHA-CS 量 不 24
20m 4m CS 20m 4m P-CS-Ca 20m 4m Ca-CS-P 20m 4m CDHA-CS 量 不 25
P-CS-Ca, CS content intensity (a. u.) 100% 90% 70% 50% 30% 10% CDHA 20 30 40 50 60 2θ (degree) 不 量 26
P-CS-Ca, CS content Absorbance (a.u.) 100% 90% 70% 50% 30% 10% 4000 3500 3000 2500 2000 1500 1000 500 Wave number (cm -1 ) 不 量 聯 27
100 P-CS-Ca, CS content Weight % 90 80 70 60 50 40 30 10% 30% 50% 70% 90% 100% 100 200 300 400 500 600 700 800 900 Temperature ( o C) 不 量 聯 28
Weight % 100 90 80 70 60 50 40 30 P-CS-Ca,TPP ion-crosslinking CS content 10% 30% 50% 70% 90% 100% 100 200 300 400 500 600 700 800 900 Temperature ( o C) 不 量 聯 29
Conc. (mg/ml) 0.020 0.015 0.010 0.005 50% 70% 90% 100% P-CS-Ca, CS content 0.000 0 1 2 3 4 5 6 TIME (hr) 不 量 聯 30
0.0009 P-CS-Ca 0.0008 DH (cm 2 /hr) 0.0007 0.0006 0.0005 0.0004 0.0003 0.0002 50 60 70 80 90 100 CS content % 不 量 聯 數 31
20m 4m CS50% 20m 4m CS70% 20m 4m CS90% 20m 4m CS 不 量 32