Veyis KARAKOÇ, Elif Hilal ŞEN, Semra İDE, Deniz TÜRKMEN, Rabel SOOMRO, Adil DENİZLİ

Isıya duyarlı poli N-izopropilakrilamit temelli kriyojellerin SAXS çalışması

B u çalışmanın amacı, NIPA’in düşük kritik çözelti sıcaklığı LCST civarındaki sıcaklığın değiştirilmesiyle ısıya duyarlı poli N-izopropilakrilamid-metakriloil-L-histidin , poli NIPA-MAH monolitik kriyojellerinin nano yapısındaki değişimleri küçük açı x-ışını saçılması SAXS yöntemi ile incelemektir. Poli NIPA-MAH kriyojeller, NIPA, MAH fonksiyonel monomeri ve N,N-metilen-bisakrilamit MBAAm çapraz bağlayıcısı kullanılarak plastik şırınga içerisinde serbest radikal kriyopolimerizasyonu ile hazırlanmıştır. Polimerizasyon buz banyosunda sıfır derecenin altında N, N, N, N-tetrametilen diamin TEMED ve amonyum persulfat APS çifti kullanılarak başlatılmıştır. Poli NIPA-MAH kriyojelinin LCST değeri 34 oC olarak bulunmuştur. Poli NIPA-MAH kriyojelleri 60-100 µm gözenek çapına ve 42.6 m2/g polimer spesifik yüzey alanına sahiptir. Sentezlenen gözenekli kriyojeller geleneksel jellerle karşılaştırıldığında sıcaklık değişimine hızlı tepki vermektedir. Poli NIPA-MAH kriyojelinin yığın yapısı ve yüzey morfolojisi taramalı elektron mikroskobu SEM kullanılarak belirlenmiştir. SAXS, poli NIPA-MAH kriyojelinin sıcaklık değişimiyle gözenek hacmi, duvar kalınlığı ve gözenek şekli gibi nano boyuttaki değişimlerin belirlenmesine imkan vermektedir. 30-40oC sıcaklık aralığında doğal form ve su içerisinde saçılma verileri incelenmiştir. Iki farklı sıcaklıkta 34oC ve 35oC gözenek boyutu ve şekli üzerinde kaydedilebilir bir değişikliği gösteren ayrıntılı bir inceleme de gerçekleştirilmiştir

Anahtar Kelimeler:

N-izopropilakrilamid NIPA , ısıl duyarlı, kriyojel, Küçük Açı X-ışını Saçılması SAXS

Thermosensitive poly N-isopropylacrylamide based cryogel: A SAXS study

The purpose of this study is to observe the structural changes of the thermosensitive poly N-isopropylacrylamide- N-methacryloyl-L-histidine poly NIPA-MAH monolithic cryogel with changing temperature around lower critical solution temperature LCST of NIPA by Small Angle X-Ray Scattering SAXS . Poly NIPA-MAH cryogel was prepared by free radical cryocopolymerization of NIPA with MAH as functional comonomer and N,N- methylene-bisacrylamide MBAAm as crosslinker directly in a plastic syringe. Polymerization initiated by N,N,N,N- tetramethylene diamine TEMED and ammonium persulfate APS pair at subzero temperature in an ice bath. LCST of poly NIPA-MAH cryogel was found to be 34oC. Poly NIPA-MAH cryogel with 60–100 μm in pore diameter have a specific surface area of 42.6 m2/g polymer. This porous cryogel form provides fast temperature response comparing to conventional gels. The surface morphology and bulk structure of poly NIPA-MAH cryogel revealed with scanning electron microscopy SEM . SAXS allowed the determination of nanoalteration of poly NIPA-MAH cryogel such as pore volume, wall thickness and pore shape with changing temperature. Scattering data were analysed both native form and in water at temperature range of 300C-400C. At two different temperatures 34oC and 35oC a detailed investigation on shape of pores and pore size; which indicating a recordable change, were also carried out.

Keywords:

N-isopropylacrylamide NIPA, thermosensitive, cryogels, Small Angle X-Ray Scattering SAXS,

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A. Kumar, A. Srivastava, I.Y. Galaev, B. Mattiasson, Smart polymers: Physical forms and bioengineering applications, Prog. Polym. Sci., 32 (2007) 1205.
E.S. Gil, S.M. Hudson, Stimuli-responsive polymers and their bioconjugates, Prog. Polym. Sci., 29 (2004) 1173.
J.R. Zhang, D.K. Misra, Magnetic drug-targeting carrier encapsulated with thermosensitive smart polymer: Core–shell nanoparticle carrier and drug release response, Acta Biomaterialia, 3 (2007) 838.
H. Ichikawa, Y. Fukumori, A novel positively thermosensitive controlled-release microcapsule with membrane of nano-sized poly(N- isopropylacrylamide) gel dispersed in ethylcellulose matrix, J. Controlled Release, 63 (2000) 107.
A. Kondo, H. Fukuda, Preparation of thermo-sensitive magnetic hydrogel microspheres and application to enzyme immobilization, Journal of Ferment and Bioeng., 84 (1997) 337.
A. Kumar, M. Kamihira, I.Y. Galaev, S. Lijima, B. Mattiasson, Binding of Cu(II)-Poly(N- isopropylacrylamide/vinylimidazole) Copolymer to Histidine-Tagged Protein: A Surface Plasmon Resonance Study, Langmuir, 19 (2003) 865.
E. Kalaycıoglu, S. Patır, E. Piskin, Poly(NIPA-co-MAH) Copolymers and Their Interactions with Human Immunoglobulin-G, Langmuir, 19 (2003) 9538.
H. Tokuyama, T. Iwama, Temperature-Swing Solid- Phase Extraction of Heavy Metals on a Poly(N- isopropylacrylamide) Hydrogel, Langmuir, 23 (2007) 13104.
R. Pelton, Temperature-sensitive aqueous microgels, Advances in Coll. and Interf. Sci., 85 (2000) 1.
M. Turk, S. Dincer, E. Piskin, Smart and cationic poly(NIPA)/PEI block copolymers as non-viral vectors: in vitro and in vivo transfection studies, J. Tissue Eng. Regen. Med., 1 (2007) 377.
N. Shamim, H. Liang, K. Hidajat, M.S. Uddin, Adsorption, desorption, and conformational changes of lysozyme from thermosensitive nanomagnetic particles, J. Coll. and Interf. Sci., 320 (2008) 15.
M. Türk, Z.M.O. Rzayev, S.A. Khalilova, Bioengineering functional copolymers. XIV. Synthesis and interaction of poly(N-isopropylacrylamide-co-3,4-dihydro-2H- pyran-alt-maleic anhydride)s with SCLC cancer cells, Bioorg. & Med. Chem., 18 (2010) 7975.
K. Nagase, J. Kobayashi, T. Okano, Temperature- responsive intelligent interfaces for biomolecular separation and cell sheet engineering, J.R. Soc. Interface., 6 (2009) 293.
N. Bereli, M. Andac, G. Baydemir, R. Say, I. Y. Galaev, A. Denizli, Protein recognition via ion-coordinated molecularly imprinted supermacroporous cryogels, J. Chromatog. A, 1190 (2008) 18.
V.I. Lozinsky, I.Y. Galaev, F.M. Plieva, I.N. Savina, H. Jungvid, B. Mattiasson, Polymeric cryogels as promising materials of biotechnological interest, Trends in Biotechnol., 21 (2003) 445.
W. Xue, S. Champ, M.B. Huglin, T.G.J. Jones, Rapid swelling and deswelling in cryogels of crosslinked poly(N-isopropylacrylamide-co-acrylic acid), Europ. Polym. Journal., 40 (2004) 703.
J. Kobayashi, A. Kikuchi, K. Sakai, T. Okano, Aqueous chromatography utilizing pH-/temperature- responsive polymer stationary phases to separate ionic bioactive compounds, Anal. Chem., 73 (2001) 2027.
H. Kanazawa, M. Nishikawa, A. Mizutani, C. Sakamoto, Y. Morita-Murase, Y. Nagata, A. Kikuchi, T. Okanoc, Aqueous chromatographic system for separation of biomolecules using thermoresponsive polymer modified stationary phase, J. Chromatog. A, 1191 (2008) 157.
P. Maharjan, W.B. Woonton, E.L. Bennett, W.G. Smithers, K.D. Silva, T.W.M. Hearn, Novel chromatographic separation — The potential of smart polymers, Innov. Food. Sci. and Emerg. Tech., 9 (2008) 232.
L. Qin, X.W. He, W. Zhang, Y.W. Li, K.Y. Zhang, Macroporous Thermosensitive Imprinted Hydrogel for Recognition of Protein by Metal Coordinate Interaction, Anal. Chem., 81 (2009) 7206.
H. Liu, M. Liu, L. Ma, J. Chen, Thermo- and pH-sensitive comb-type grafted poly(N,N-diethylacrylamide-co- acrylic acid) hydrogels with rapid response behaviors, Euro. Polym. J., 45 (2009) 2060.
Y. Liu, X. Cao, M. Luo, Z. Le, W. Xu, Self-assembled micellar nanoparticles of a novel star copolymer for thermo and pH dual-responsive drug release, J. Colloid and Interf. Sci., 329 (2009) 244.
E.J. Kim, S. Cho, S.H. Yuk, Polymeric microspheres composed of pH/temperature-sensitive polymer complex, Biomaterials, 22 (2001) 2495.
G. Fundueanu, M. Constantin, P. Ascenzi, Preparation and characterization of pH- and temperature- sensitive pullulan microspheres for controlled release of drugs, Biomaterials, 29 (2008) 2767.
J.M. Benns, J.S. Choi, R.I. Mahato, J.S. Park, S.W. Kim, pH-Sensitive Cationic Polymer Gene Delivery Vehicle: N-Ac-poly(l-histidine)-graft-poly(l-lysine)Comb Shaped Polymer, Bioconjugate Chem., 11 (2000) 637.
H. Yavuz, V. Karakoc, D. Turkmen, R. Say, A. Denizli, Synthesis of cholesterol imprinted polymeric particles, Int. J. Biol. Macromol., 41 (2007) 8.
Y. Canak, S. Ozkara, S. Akgol, A. Denizli, Pseudo-specific bioaffinity chromatography of immunoglobulin-G, React. Funct. Polym., 61 (2004) 369.
I. W. Hamley, V. Castelletto, M. P. S. N. Ricardo, E.N. M. Pinho, C. Booth, D. Attwood, Z. Yang, A SAXS study of the structure of gels formed by mixtures of polyoxyalkylene triblock copolymers, Polym. Int., 56 (2007) 88.
M. H. Zareie, S. Dincer, E. Piskin, Observation of Phase Transition of Thermo-Responsive Poly(NIPA)–PEI Block Copolymers by STM, J. Colloid and Interf. Sci., 251 (2002) 424.
M. Chalal, F.E. Dolle, I. Morfin, J.C. Vial, M.R.A. Armas, J.S. Roman, N. Bolgen, E. Piskin, O. Ziane, R. Casalegno, Imaging the Structure of Macroporous Hydrogels by Two-Photon Fluorescence Microscopy, Macromolecules, 42 (2009) 2749.
P. Perez, F. Plieva, A. Gallardo, J.S. Roman, M.R. Aguilar, I. Morfin, F.E. Dolle, F. Bley, S. Mikhalovsky, I.Y. Galaev, B. Mattiasson, Bioresorbable and nonresorbable macroporus thermosensitive hydrogels prepared by cryopolymerization. Role of the cross-linking agent, Biomacromol., 9 (2008) 66.
H.M. Zareie, E.V. Bulmuş, A.P. Gunning, A.S. Hoffman, E. Piskin, V.J. Morris, Investigation of A Stimuli- Responsive Copolymer by Atomic Force Microscopy, Polymer, 41 (2000) 6723.
V. Karakoç, D. Türkmen, H. Shaikh, N. Bereli, C. Andaç, A. Denizli, Synthesis and characterization of Poly(N- isopropyl acrylamide) thermosensitive based cryogel, Hacettepe J. Biol. Chem., 41 (2013) 159.
B. Garipcan, A. Denizli, A Novel Affinity Support Material for the Separation of Immunoglobulin G from Human Plasma, Macromol. Biosci., 2 (2002) 135.
O. Glatter, O. Kratky, Small Angle X-ray Scattering, Academic Press, London, 1982, p,17
R.J. Roe, Methods of X-ray and Neutron Scattering in Polymer Science, Oxford Univ. Press., 2000, 184- 185
A. Owen, A. Bergmann, On the fractal character of polymer spherulites: an ultra-small-angle X-ray scattering study of poly[(R)-3-hydroxybutyrate] , Polym. Int., 53 (2004) 12.
O. Kratky, G. Porod, Diffuse small-angle scattering of X-rays in colloid systems, J. Coll. Interf. Sci., 4 (1949) 35
O. Kratky, G. Porod, Röntgenuntersuchung gelöster Fadenmoleküle, Rec. Trav. Chim., 68 (1949) 1106
R.J. Roe, Methods of X-ray and Neutron Scattering in Polymer Science, 2000, p. 166
D.I. Svergun, Determination of the regularization parameter in indirect-transform methods using perceptual criteria, J. Appl. Crystallogr., 25 (1992) 495.
S. Pikus, A.L. Davidowicz, E. Kobylas, D. Wianowska D, SAXS examination of the water evaporation process from silica materials coated with a polysaccharide– polyimine copolymer layer, Appl. Surf. Sci., 156 (2000) 189.
S.A. Ravion, Z. Ding, A. Pelle, A.S. Hoffman, D. Letourneur, New antibody purification procedure using a thermally responsive poly(N- isopropylacrylamide)-dextran derivative conjugate, J. Chromatogr. B, 761 (2001) 247.
N. Malmstadt, P. Yager, A.S. Hoffman, P.S. Stayton, A Smart Microfluidic Affinity Chromatography Matrix Composed of Poly(N-isopropylacrylamide)-Coated Beads, Anal. Chem., 75 (2003) 2943.
R. B. Fong, Z. Ding, A. S. Hoffman, P. S. Stayton, Affinity Separation Using an Fv Antibody Fragment-”Smart” Polymer Conjugate, Biotech. and Bioengineering., 79 (2002) 271.
Z. Xu, W. Bae, A. Mulchandani, R. K. Mehra, W. Chen, Heavy Metal Removal by Novel CBD-EC20 Sorbents Immobilized on Cellulose, Biomacromolecules, 3 (2002) 462.