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1 Biomaterials Research (006) 10(3) : Biomaterials Research 7 The Korean Society for Biomaterials w w š t Preparation of Gradient Polymer Surface for Biomedical Applications ½ 1 Á ¼ Á y 3 Á w 1 * Moon Suk Kim 1, Gilson Khang, Jin Ho Lee 3, and Hai Bang Lee 1 * 1 Š ŒŠ e tg, h Š BK-1 f BIN fš, 3 Š Š g Š 1 Medicinal Science Division, Korea Research Institute of Chemical Technology, Daejon Korea BK-1 Polymer BIN Fusion Research Team, Chonbuk National University, Jeonju , Korea 3 Department of Advanced Materials, Hannam University, Daejeon , Korea (Received June 0, 006/Accepted July 5, 006) Over last three decades, various biomaterials have been developed for the utilization in biomedical industry. The practical utilization of biomaterials in industry depends on the study about an appropriate physical and biological response of biomaterials referred as its biocompatibility. The surface modification of biomaterials has recently become an interesting topic in the field of surface engineering. We recently developed a gradient surface. The gradient surface is the surface on which a gradually varying chemical composition exists along its length. Such gradient surface is of particular interest for basic and applied studies of the interactions between biological species and surfaces since the effect of a selected property like wettability or chemical composition can be examined in a single experiment on one surface. The present review focuses on the preparation and characterization of gradient surfaces, and their interactions with biological species. Key words: Biomaterials, Corona, Plasma, Wettability, Biocompatibility, Gradient, Surface, Biological interaction d h tf g tg f jd f f Š tg f g Œ h f f. tg f, f f Š g Šhf g l ŠŠ f. Š i f h fd Š tg ~ Š } lf ll f f hš f hš hš thš hf lf il. v Š thš f l eš } ƒ f h v Š f ŠdŠ. hf ( /x ), ŒŠh l, hš, x f tg ƒ f thš f j d f Š. f h tg f hf Šh i f hš f Š eš h f f Š t l(j f) Œ~f Šh i f Œfd lš f. Šh l f Œfdf j lf r, f hr Œ 1), ŠhŒ Š f l f ) 3) f j f. *sf hf: hblee@krict.re.kr Š i f lf dš l f jdš hhf lf ŒŠhÁ h( ŒŠ, hš, x,, h )f jdš fdš f f. Šhf h f Š f eš hf f e ŠŠ eš Š Š ii df f. f Š f f Š Šhf f f ŠŠ f. f g f f hrhf Š ƒ f l f hiš thf f. f Š (gradient surface) f Šh i g ff Œfd Š hf ƒ Š j. f e hf f ~hf l Š Š f Š l f ŠŠ f. 4) f f Š f dš l, Œf f Šhf if Œfd f t ff Œfd, -f 5), Œf f l 6) 7) f fd f eš dš f Š. u f jd df f hi t f ff Š hf f Š f d Š, t f f Œfd fš Š f th fd f h Š. 15
2 16 Á ÁflŒÁfŠ t š q w v (RFPD) l s l f Š f fd f. RFPD Š Š f h f Š ~ f fš (Figure 1). f f xš 8-11) RFPD v f Š dl h f hfš. f f l v f f f hrhf l Š, f f. f Š f (PE), Š (PP), ~ fƒ (PET), ~ } fƒ (PMMA), ~f (PS) f f hd f. Š l, ŠŠ f f ~f Š Š. f s Šf hf f l f v f l Šf f hrhf. l s Š l f f s ~ fiš, f ht f o l l hf hf o hiš f. 9-11) Figure l s fš PE Œf PS f f Š fd Š y lf j f. f l hf ~Œ j ~ xf Š, f f Šf Š hš Œ f e Š. hš Œ f f Œf jf fšf (,, z,, x, x ) Š f Š fd eš Š. f f l 1) f Š fd l s f l l Šf j, hf f Œ~ Š. l s f l ~ Š Š hf f s Š Š hšl f xhf ŒŠf l. 13) g ù f Š z h s fdš f hf Š f Š (Figure Figure 1. Schematic diagram showing preparation of gradient surface by plasma discharge treatment. 9) Figure. Plausible mechanism of the formation of oxygen-based functionalities on a polymer surface by oxygen plasma discharge treatment. 1) Figure 3. Schematic diagram showing the corona discharge apparatus for the preparation of a wettability gradient on polymer surfaces by the corona discharge treatment. 14,15) Biomaterials Research 006
3 tf Šh fdf eš f hi 17 3). j z h hf f hi 14,15) Š eš d. z h f h f ~ eš l Š f f. l, e hr l Š h f eš x f Œ~f h (100 khz 10~35 watts) dlf f ( 1.0 cm/s) Š f. f s Šf, f hh l Š h f z hf v. f f fš, f f hr Œ, Š fd l hf Š. z hf Š tg ƒ f Š ŠŠ eš d. f Š f 16-18) j Š hdš d dfš. t -» tg Š t fd f ff Šh t thš f f ff e Š eš jdš. f Š ff Š d Š l e tœ f ŒŠhf h ~ f. t y PE, PP, PS, PET, PMMA Š Š ff f l l h s fš lš, f f hf f l l h fš l Š, e f f fd Œ f. l l s fš f f ŒŠh i Œ f fdš ht ESCA wh f fdš Š. l l s f Œ f Š eš g Š hf f. 8,19) t s p» x f f dlf ft h l ƒef d ƒ f f Œ~ Š f u fe l f f l (Œf Œ hš f ) l h f. 0,1) Comb-like PEO f PE f ~} fƒ } (PEO-MA)f ƒ jš fš Š. 18-) PEO e 1, 5, 10 } d, comb-like PEO h f hr l Š z s s fš f hr Œ. PEO-MA PEO f l Š PEO-MAf ƒ f PEO-MA } PEO l f ft Š Š. j comb-like PEO f f fdš ht FTIR-ATR, ESCA whf Š ƒlf Š. f Š w h f hr l Š PEOf PEO f PE ƒ ff. t» f fd f z h s ff fd } (AA), sodium p-stryrene sulfonate (NaSS), f fd N,N- Š } f (DMAPAA) f f tf ƒ jš fš Š (Figure 4). z s fš 3) PE Œ f f tf ƒ jšf eš h z s PE ƒ jš. fƒ (Phosphate-buffered saline) Œf ph f l d PE-COOH PE-NaSS ff hš f, PE-DMAPAA f h Š f. f f f f fdš ht, ESCA, FTIR-ATR ƒlf rš. f f f f f PE hrhf l Š f ƒœ ff j., f f f f ƒ f hf f ƒ fd f i fš } ll. t s» Šh Œ hf f fd f h Š f xœ f (PEI) h eš d h. PEI fœ f f. PEIf ŒŠh f ff z s PE x f N- -N'-(3- f Š) x f Œ Œ ~f Š h. ESCA 4,5) PEI C-N if fš ~ 1S } f PE f l Š. l 1S }f Š PE f hr l. PEI PE hrhf l ff Œf. t v» Ishihara f Š Œ hš f l - ~} z ff Š f Š, f t fš hdš. 6,7) f Š, f, (MAPC)f f ƒ f Š eš z h s ~} f ~ z (MAPC) ƒ PE f f Š r f ŠŠ. Figure 4. Gradient surface with chargeable groups. 3) Vol. 10, No. 3
4 김문석 강길선 이진호 이해방 18 구배표면의 특성분석 물을 이용한 접촉각 분석 화학적 구배표면은 거의 항상 접촉각 측정 방법을 사용하여 그들의 젖음성 특성을 알 수 있다. 특히, 접촉각 측정은 표면을 연구하는 가장 훌륭한 기술 중의 하나라고 할 수 있다. 접 촉각은 다양한 방법으로 측정될 수 있다. 구배표면의 길이 에 대해 마이크로 리터의 물방울을 떨어뜨리고 간단한 각도측 정법으로 표면에 떨어뜨린 물방울의 접촉각을 Figure 5에 나타 낸 바와 같이 측정할 수 있다. 8,9) 30,31) 3) Electron Spectroscopy for Chemical Analysis (ESCA, 전자 분광법) 분석 X-ray photoelectron spectroscopy (XPS)이라 불리는 ESCA 는 비록 XPS 기기를 이용 표면 조사로 수 mm 에서 1- µm 까지 비교적 작은 공간의 해상도를 가지고, 표면 특성을 분석하는데 매우 유용하다. ESCA 정보의 중요성은 나노미터 범위 내에 있으며, 적외선 분광기보다는 더 훌륭하다. 대부분 의 구배표면 연구가 고분자 재료를 사용하여 수행하기 때문에 ESCA에서 거의 피할 수 없는 구배 표면의 탄소 오염은 구배 표면의 ESCA 분석에서 심각한 복잡요소로 작용한다. 비록 ESCA는 완전한 정량을 제공하진 않지만, 표면 조성물의 작은 변화에 매우 민감하다. RFPD 처리에 의한 고분자 표면의 화학적 구조에서 변화는 ESCA로 분석되었는데 Figure 6은 코로나 처리된 PE 표면의 탄소 1S core의 scan spectra 정도를 보여준다. 구배 (0.5 cm 부근)의 소수성 부분은 알킬 탄소 (-C-C-) 피크가 ~85 ev의 결합에너지를 가짐을 보여준다. 코로나 전압의 세기가 증 가하는 위치에 따라 탄소-산소 상관관계를 보여주는 높은 결합 에너지를 나타내는 새로운 피크가 확인되었다. 높은 결합에너 지 부분의 피크는 -C-O-가 ~86.6 ev (e.g., 수산화 혹은 에테르그룹)에서, -C=O-가 ~87.9 ev (e.g., 케톤 혹은 알 데히드 그룹)에서, O=C-O-가 ~89.1 ev (e.g., 카프복시산 혹은 에스터 그룹)에서 일치됨을 알았다. 산소를 바탕으로 한 작용기는 코로나 전압의 세기가 증가함에 따라 증가하였다. 게 다가, 조사한 스펙트럼의 결과는 공기 중의 모든 전자 (수소와 헬륨을 제외한)에 대해 표면의 원소성분에 대한 정보를 제공한 다. 원소는 각 원소의 고유의 결합에너지를 통해 확인하였다. 코로나 처리의 경우, 수산기 혹은 에테르그룹 (-C-O-)은 다른 33) 9,34) Figure 5. Contact angle of gradient PE surface. Biomaterials Research 006 ESCA carbon 1S core level spectra of a corona-treated PE surface along the sample length. Numbers labeled on the spectra (0.5 to 4.5) represent the positions from the untreated end of the gradient surface. Figure 6. 34) 작용기보다 표면에 더 많이 생성되었다. Infrared Spectroscopy 분석 비록 적외선분광기의 표면에 대한 자극감응성은 종종 구배 표면에 흡착된 단일분자 층이 너무 깊어서 감지할 수 없는 마 이크로미터 범위 내에 있을지라도 Infrared spectroscopy in attenuated total reflection (ATR) mode는 종종 구배 표면을 특성분석하기 위해 사용 가능하다. 본 연구팀은 코로나 방전 처리한 PE 표면에 과잉의 산소를 도입시켰고, 연속적으로 FTIR-ATR을 이용하여 적외선 스펙트럼을 결정하였다. 코로 나 방전을 통해 장기간 노출된 구배 부분은 강한 C=O 흡착 부분을 지녔으며, 강도가 점차적으로 증가하는 피크를 보였다 (Figure 7). 이와 같이, 일반적으로 적외선 분광학은 구배 표면 을 특성화하기 위한 적절한 방법 중의 하나이다. Atomic Force Microscopy (AFM) 분석 AFM은 고해상도의 표면 이미지를 위한 방법으로 확립되었 다. 또한 표면 위의 나노미터 해상도의 분자특성을 분석하기 위한 훌륭한 기기이다. 코로나 처리 후, AFM 데이터는 0.5 cm 위치의 10 nm에서 4.5 cm 위치의 100 nm까지 코로나 처리한 PE 표면의 거침성이 증가하는 것을 보여준다. 또한, 코로나 전압의 세기를 PE 길이에 따라 증가시켰을 때 코로나 처리된 PE 표면의 거침성은 변한다는 것을 보여준다. 이와 같 이, AFM은 구배 표면 이미지화에 대한 훌륭한 방법으로 이용 될 수 있다. 14-7) 35) 4) Time-Of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS) 분석 TOF-SIMS는 1 µm 미만의 공간적 해상도를 가지며, 원소와
5 생체의료학적 응용을 위한 고분자 구배표면 제조 Figure FTIR-ATR spectra along the PE length after corona treatment. 14) 분자의 종류를 감지 할 수 있는 표면분석기술이다. 이러한 TOF-SIMS는 특성과 이미지화 가능성 때문에, 분석방법이 표면 의 구성 성분을 분석하는데 이상적이다. 본 연구팀에서는 구배 표면의 표면성분을 Figure 8과 같이 분석하였다. 기타 분석 구배표면을 1-anilinoaphthalene-8-sulfonate (ANS)의 존재 하 에 total internal reflection fluorescence (TIRF) spectroscopy 로 분석 가능하다. ANS 형광의 높은 강도는 구배의 소수성 부분에서 발견되고, 친수성 부분에서 형광 강도는 나타나지 않 았다. 이와 같이 TIRF는 구배 실리카 혹은 유리 표면의 젖음 성을 분석하기 위해 사용될 수 있다. 36) 37) 구배표면의 생체 분자와의 상호 작용 생체적합성 재료의 최근 개발 동향은 세포외 기질 (ECM)이 나 특유의 세포반응으로부터 유래된 생체분자를 인식할 수 있 는 생체모방성 재료에 초점이 맞춰지고 있다. 생체적합성 재료 의 구배 개질은 생체분자를 인식할 수 있는 특성을 지닌 생 체모방성 재료를 만들기 위한 가장 단순한 방법 중의 하나이 다. 이와 같이 구배 표면은 단백질과 세포와 같은 생물학적 종의 생물학적 상호작용과 거동을 시험하기 위해 매우 큰 잠 재력을 제공할 수 있다. 구배표면으로 생물학적 종의 흡착은 작용기의 표면밀도가 증가할수록 점진적으로 감소함이 관찰되 었다. 젖음성 뿐만 아니라, 그들의 전하특성, 그리고 표면의 작용기는 생물학적 종의 흡착에 중요한 역할을 하는 것처럼 보인다. 이것은 표면의 친수성과 소수성의 관계에 연관되어 있 을 수도 있다. 이와 같이 생체적합성 재료로 특별한 위치에서 혹은 를 통한 생물학적 종의 빠르고 정확한 인식과 결합반응에 대한 연구는 도전할만한 목표이다. 본 장에서는 구배 표면에 단백질 흡착과 세포점착 연구를 소개하 고자 한다. in vivo in vitro 39-41) TOF-SIMS images of negative ion (A) (C H O ) and (B) (C H O ) obtained at different positions of a corona-treated PE surface and (C) plot of TOF-SIMS intensities of characteristic carboxyl secondary negative ions versus length treated by corona. The image analysis area was µm. Figure ) 단백질과의 상호작용 본 연구팀에서는 코로나 처리한 PE 구배 표면에서 단백질 Vol. 10, No. 3
6 130 Á ÁflŒÁfŠ rf f f f Šƒ f i Š. 15,4) f r f ESCA Š f l ~ Œ l rf s d. f l rf Šf j e. ESCA f l } ( Š l, ~399 ev) hf hr ff j, rf f f l Š hrhf l Šf ~. f f f l ff Š Œfdf l Š f. f r f Š FTIR-ATR Š. f r l f II ( cm 1 ) bands Š hf hrhf Šf rš. Œ h d f r f vf l f hf xœf Š. l f fd Šl z s PE f of lf r ~rf l f fxš. comb-like PEO ƒ PE r lf hf. i f Š PE r lf PEO-MA ƒ Ž f, PE f f lf r ff ~. f f Š f ~ PE l ff Š Œfd f. Š lf Œ ~ f hiš. f Š, f hf Š g } Š f avidin-biotin Œfd (K D ~ M)f fdš f PE h rhf l Š Œ r Š. 5) s y f hf PE Chinese Š ovary (CHO), e, f ƒ f Š. f x f ex hf 34) f h Š x f l ex f r f r. f l Š Figure 9 fš r s hf Œ. Š Š d, d f f x f Š h Š x f f l f h. h Š x 43) f hrš, Š, gš f h f f l Š f l e, HeLa S3, e Š f fš 7,8,0,) r. x f l f hr f hr z hs fš j hf PE f fdš Š. 44) u Š t l Œ~f l Figure 9. SEM pictures of different types of cells adhered on wettability gradient PE surfaces after a 1 day culture (original magnification, 400). C. A. means water contact angle (degree). 34) f Œfdf Š eš hf (ƒ h f ) thf v. hf f l h s z h s fdš Š f l hi. t l l r/~r f hr/l f Š eš f hf f dš j ghf Š f ƒlf l hf f Š f f ehf Š f f. f f f h Š f d Š f. hf t lf hf Š Œ ~f Šh if Œfdf t hf i Š dš f. ƒ Š fd hš f l z h s ƒ ff Š Š ff, ŒŠ, hš, fhš t lf hf Šh Œf if Œfd Š edš h d j f. hf PE f hf l lf l f f dš Š hf Šh if edš f. f 1987 g l Š ŒŠ e jhff vlš t f Š l Š. f f 1985 / f Œ xœ d (Tissue culture dish) fš 86 Š h Š Šff l e~ Š Biomaterials Research 006
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