国际生物医学工程杂志
國際生物醫學工程雜誌
국제생물의학공정잡지
INTERNATIONAL JOURNAL OF BIOMEDICAL ENGINEERING
2012年
2期
65-69,后插1-后插2
,共6页
贺庆%敖强%韩大庆%王臻%刘伟强%公衍道%张秀芳
賀慶%敖彊%韓大慶%王臻%劉偉彊%公衍道%張秀芳
하경%오강%한대경%왕진%류위강%공연도%장수방
琼脂糖%壳聚糖%正电荷%蛋白吸附%生物相容性
瓊脂糖%殼聚糖%正電荷%蛋白吸附%生物相容性
경지당%각취당%정전하%단백흡부%생물상용성
Agarose%Chitosan%Positive charge%Protein adsorption%Biocompatibility
目的 以琼脂糖/壳聚糖共混凝胶为模型,研究壳聚糖材料生物相容性的可能机理.方法 通过共混法,制备出一系列不同壳聚糖含量的琼脂糖/壳聚糖共混凝胶.利用傅里叶变换红外光谱(FTIR)分析共混凝胶的化学基团,利用荧光素-4-异硫氰酸酯(FITC)标记法观察琼脂糖和壳聚糖之间的可共混性.通过Zeta电势测量共混凝胶的电荷,利用二喹啉甲酸(BCA)法分别测定胎牛血清(FBS)总蛋白和牛血清白蛋白(RSA)在共混凝胶上的吸附,利用酶联免疫吸附(ELISA)法测定纤黏连蛋白(FN)在共混凝胶上的吸附.细胞实验以人微血管内皮细胞系(HMEC-1)为模型,通过观测细胞的黏附、增殖和形态来评价共混凝胶的细胞相容性.结果 琼脂糖/壳聚糖共混凝胶含有壳聚糖特征性的化学基团.琼脂糖和壳聚糖之间存在着良好的可共混性,壳聚糖的氨基基团在共混凝胶中呈均匀分布.在pH酸性条件下(pH 3.0)共混凝胶带有较强的正电荷,然而在pH中性条件下(pH 7.4)所有共混凝胶的Zeta电势均降低至0 mV附近.各组共混凝胶之间对FBS总蛋白以及BSA的吸附差异无统计学意义,但是共混凝胶对FN的吸附却随着壳聚糖含量的升高而显著升高.细胞实验结果显示:随着壳聚糖含量的提高,共混凝胶的细胞相容性有明显改善,HMECs在壳聚糖含量较高的凝胶上表现出良好的黏附、铺展和增殖.结论 相对于血清中的其他蛋白,壳聚糖组分对FN存在优先吸附,从而能够促进细胞在共混凝胶表面的黏附铺展.与传统观点不同,本研究发现壳聚糖的生物相容性与其所携带的正电荷无关.
目的 以瓊脂糖/殼聚糖共混凝膠為模型,研究殼聚糖材料生物相容性的可能機理.方法 通過共混法,製備齣一繫列不同殼聚糖含量的瓊脂糖/殼聚糖共混凝膠.利用傅裏葉變換紅外光譜(FTIR)分析共混凝膠的化學基糰,利用熒光素-4-異硫氰痠酯(FITC)標記法觀察瓊脂糖和殼聚糖之間的可共混性.通過Zeta電勢測量共混凝膠的電荷,利用二喹啉甲痠(BCA)法分彆測定胎牛血清(FBS)總蛋白和牛血清白蛋白(RSA)在共混凝膠上的吸附,利用酶聯免疫吸附(ELISA)法測定纖黏連蛋白(FN)在共混凝膠上的吸附.細胞實驗以人微血管內皮細胞繫(HMEC-1)為模型,通過觀測細胞的黏附、增殖和形態來評價共混凝膠的細胞相容性.結果 瓊脂糖/殼聚糖共混凝膠含有殼聚糖特徵性的化學基糰.瓊脂糖和殼聚糖之間存在著良好的可共混性,殼聚糖的氨基基糰在共混凝膠中呈均勻分佈.在pH痠性條件下(pH 3.0)共混凝膠帶有較彊的正電荷,然而在pH中性條件下(pH 7.4)所有共混凝膠的Zeta電勢均降低至0 mV附近.各組共混凝膠之間對FBS總蛋白以及BSA的吸附差異無統計學意義,但是共混凝膠對FN的吸附卻隨著殼聚糖含量的升高而顯著升高.細胞實驗結果顯示:隨著殼聚糖含量的提高,共混凝膠的細胞相容性有明顯改善,HMECs在殼聚糖含量較高的凝膠上錶現齣良好的黏附、鋪展和增殖.結論 相對于血清中的其他蛋白,殼聚糖組分對FN存在優先吸附,從而能夠促進細胞在共混凝膠錶麵的黏附鋪展.與傳統觀點不同,本研究髮現殼聚糖的生物相容性與其所攜帶的正電荷無關.
목적 이경지당/각취당공혼응효위모형,연구각취당재료생물상용성적가능궤리.방법 통과공혼법,제비출일계렬불동각취당함량적경지당/각취당공혼응효.이용부리협변환홍외광보(FTIR)분석공혼응효적화학기단,이용형광소-4-이류청산지(FITC)표기법관찰경지당화각취당지간적가공혼성.통과Zeta전세측량공혼응효적전하,이용이규람갑산(BCA)법분별측정태우혈청(FBS)총단백화우혈청백단백(RSA)재공혼응효상적흡부,이용매련면역흡부(ELISA)법측정섬점련단백(FN)재공혼응효상적흡부.세포실험이인미혈관내피세포계(HMEC-1)위모형,통과관측세포적점부、증식화형태래평개공혼응효적세포상용성.결과 경지당/각취당공혼응효함유각취당특정성적화학기단.경지당화각취당지간존재착량호적가공혼성,각취당적안기기단재공혼응효중정균균분포.재pH산성조건하(pH 3.0)공혼응효대유교강적정전하,연이재pH중성조건하(pH 7.4)소유공혼응효적Zeta전세균강저지0 mV부근.각조공혼응효지간대FBS총단백이급BSA적흡부차이무통계학의의,단시공혼응효대FN적흡부각수착각취당함량적승고이현저승고.세포실험결과현시:수착각취당함량적제고,공혼응효적세포상용성유명현개선,HMECs재각취당함량교고적응효상표현출량호적점부、포전화증식.결론 상대우혈청중적기타단백,각취당조분대FN존재우선흡부,종이능구촉진세포재공혼응효표면적점부포전.여전통관점불동,본연구발현각취당적생물상용성여기소휴대적정전하무관.
Objective To investigate the possible mechanisms for biocompatibility of chitosan material using agarose/chitosan blended hydrogels as a model.Methods A series of agarose/chitosan blended hydrogels with different chitosan content were prepared by the blending method.The chemical groups of the blended hydrogels were analyzed by the Fourier transform infrared (FTIR) spectroscopy.The blending compatibility between the agarose and chitosan was evaluated with the fluorescein-4-isothiocyanate (FITC) staining method.The charge of the blended hydrogels was determined by the zeta potential measurement.The adsorption of total fetal bovine serum (FBS) proteins and bovine serum albumin (BSA) on the blended hydrogels was measured by the bicinchoninic acid (BCA) method.The adsorption of fibronectin (FN) on the blended hydrogels was measured with ELISA.Cell culture experiment adopted human microvascular endothelial cell line (HMEC-1) as the model.The cytocompatibility was studied by evaluating adhesion,proliferation,and morphology of the cells on the blended hydrogels.Results Characteristic chemical groups of chitosan could be detected in the agarose/chitosan blended hydrogels.The chitosan had a good blending compatibility with the agarose.The amino groups of chitosan were uniformly distributed in the blended hydrogels.The blended hydrogels were strongly positively charged at acidic pH (pH 3.0),however,the zeta potentials of all the hydrogels were reduced to nearly 0 mV at neutral pH (pH 7.4).There were no significant differences in the adsorption of total FBS proteins and BSA between the blended hydrogel groups.However,the adsorption of FN on the hydrogels significantly increased with the increase of chitosan content.Cell culture experiment indicated that the cytocompatibihty of the blended hydrogels was significantly improved with the increase of chitosan content.The HMECs exhibited higher levels of adhesion,spreading,and proliferation on the hydrogels with higher chitosan content.Conclusion Results in this study indicated that the chitosan component preferentially adsorbed FN compared to the other serum proteins,leading to adhesion and spreading of the cells on the blended hydrogels.In contrast to prevailing views,it was found in the present study that the biocompatibility of chitosan did not relate to its positive charge.