角膜修复材料表面等离子体改性与表面性能研究
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摘要
壳聚糖和氟硅丙烯酸脂广泛应用于角膜修复材料的研究，角膜修复材料的发展对材料表面特性提出了更高的要求，但壳聚糖膜表面亲水性较差，表面自由能尤其是其极性分量较低，缺乏细胞识别位点，不利于细胞在材料表面的黏附与增殖。氟硅丙烯酸树脂（FSA）是新一代高透氧性硬性角膜接触镜材料，但材料表面憎水，不利于泪液在镜片表面的铺展与润湿；且表面非极性基团与蛋白质分子间的非极性相互作用容易导致蛋白与脂质等在镜片表面的黏附，影响镜片佩戴舒适性和佩戴时间。为改善两种角膜修复材料的表面特性，本论文在对材料表面性能进行综合表征的基础上，采用低温等离子体进行表面处理和表面接枝改性，改善其表面物化特性和生物学性能。
采用角分辨XPS对壳聚糖膜表面化学基团的空间分布进行分析表征。结果表明，壳聚糖膜随着在空气中放置时间延长，其表面极性基团-OH等逐渐转移到膜的内部；壳聚糖膜表面N元素主要以-NH_2形式存在，其分子链上的-CONH_2主要伸向壳聚糖膜内部。表面极性基团的这种空间取向是造成壳聚糖膜表面亲水性较差，表面自由能较低的主要原因。
实验分别采用非反应性O2和N_2低温等离子体对壳聚糖膜进行表面处理。壳聚糖膜在O_2等离子体处理下，表面引入-OH、-COO、H_2N-C=O等极性基团，表面亲水性得到改善；采用O_2等离子体进行改性的最佳工艺条件为100W下处理60s，如功率过大或处理时间过长，则容易引起壳聚糖膜表面刻蚀；O2等离子体对壳聚糖膜进行表面处理未造成明显的表面形貌变化，但表面粗糙度有一定增加。采用N_2等离子体处理，也能改善壳聚糖膜表面亲水性，但效果不如O_2明显，采用100W等离子体功率和60s处理时间，其表面接触角下降为65°；N_2等离子体处理可在表面引入一定量的N元素，引入的N元素主要是以O=C-NH_2的形式存在；等离子体处理后暴露于大气中，膜表面活性自由基与空气中的氧气、水汽等进行反应，在表面引入含氧极性基团是等离子体处理样品表面极化的主要过程。
等离子体处理在壳聚糖膜表面引入的极性基团会随着时间的推移而逐渐转移到膜的内部，这种极性基团的翻转内迁，导致了等离子体处理效果的时效性，其亲水性逐渐下降，接触角增大，表面自由能尤其是其极性分量逐渐下降，壳聚糖膜表面等离子体处理的时效性大约为10天。
实验采用Ar等离子体诱导接枝在壳聚糖膜表面引入聚乙二醇（PEG）分子链；表面接枝层厚度约10nm；PEGMA单体浓度10%、Ar等离子体功率100W、时间60s时表面接枝量最大；采用PEGMA单体比采用具有相同分子量的PEGDA单体接枝率高；将PEGMA单体接枝到壳聚糖膜表面后，其表面水接触角下降，亲水性改善，表面自由能和粗糙度增加。表面接枝壳聚糖膜具有较好的牛血清蛋白吸附特性；细胞试验结果表明等离子体处理和表面接枝壳聚糖膜表面的细胞黏附率高于未经处理的壳聚糖膜，表面改性壳聚糖膜具有较好的表面细胞相容性。
采用O_2、N_2和Ar等离子体对硬性透气性（RGP）FSA角膜接触镜片进行表面处理均能极大地改善镜片表面的亲水性。等离子体处理在表面引入了含O基团，其中表面Si-CH3发生分子键的断裂，形成Si-O，硅烷转变为亲水性的无机硅氧基团（Si-O），是表面亲水性改善的主要原因。在等离子体处理下，表面分子中C-O发生部分断键，生成O-C=O。N2等离子体处理能够在RGP表面引入含N基团，N元素主要是以O=C-NH_2的形式存在。等离子体处理后表面活性自由基与空气中的氧气反应形成含氧基团是表面活化的主要过程，其中表面引入的O主要是与Si结合形成Si-O。等离子体处理对RGP表面形貌的影响主要取决于等离子体功率的大小，Ar等离子体相比O2和N2等离子体对表面的刻蚀更为明显，等离子体处理工艺以100W和120s为宜。三种气氛等离子体相比较，N2相比Ar和O_2等离子体较为温和，而Ar等离子体在功率较大时，容易导致表面刻蚀，表面粗糙度增加，表面亲水性反而下降，这与等离子体中的粒子能量大小有关。
采用Ar等离子体辅助接枝能有效地将PEGMA单体分子接枝聚合到RGP表面，所得PEGMA接枝层的厚度大约10纳米；结果表明：表面接枝量大小与单体浓度有关，采用去离子水作为溶剂，PEGMA单体浓度10%时具有最大的表面接枝量，RGP表面接枝PEG后，表面亲水性改善，RGP镜片表面溶菌酶蛋白质的吸附量减少，表面改性RGP镜片具有较好的表面细胞相容性。
采用低温等离子体技术对壳聚糖膜和角膜接触镜材料进行表面改性，能有效改善材料表面的亲水性和生物学性能，在角膜修复材料领域具有广泛的应用前景。
Chitosan and fluorosilicone acrylates (FSA) have been widely used for cornea repair studyand application. But chitosan’s poor surface hydrophilicity and low surface free energy,together with the lack of cell-recognition sites restrict the adhesion and propagation of cell onits surface. The peculiar molecular structure of FSA makes it a promident material for makingrigid gas permeable contact lens (RGP). But the surface hydrophobicity makes the lensunwettable for the tears, and the non-polar interaction between the surface unpolar groupswith the tear protein will lead to the surface protein and lipid adsorption. The study focusedon the surface modification and characterization of chitosan membrane and RGP contact lensin order to improve surface hydrophilicity and biological properties by plasma surfacetreatment and graft polymerization.
The surface properties of chitosan membrane were characterized by angle-resolved X-rayphotoelectron spectroscopy (XPS). The results indicated that the polar groups (-OH) onchitosan membrane migrated into the membrane bulk, which made the surface hydrophobicwith low surface free energy. The surface nitrogen mainly exists in the form of–NH_2. The-CONH_2groups on chitosan molecule also distributed into the membrane bulk.
In this study, chitosan membranes were surface modified by oxygen and nitrogen plasmarespectively. The surface hydrophilicity of chitosan membrane was greatly improved byoxygen plasma treatment. The mechanism for surface hydrophilicity improvement was thesurface oxidation after plasma treatment and exposure to air. Through this kind of surfaceoxidation, polar groups, such as–OH,-COO, H_2N-C=O were incorporated onto chitosanmembrane surface. The optimal plasma condition for chitosan surface treatment was100Wand60s. Higher plasma power or longer exposure time will bring about surface etching. Noevident morphology change was observed, but surface roughness increased slightly afterplasma treatment. Nitrogen plasma can also improve the surface hydrophilicity of chitosanmembrane. But the effect was not so evident compared with oxygen plasma. The surfacecontact angle decreased to about65°when the plasma power was100W and exposure timewas60s. Some kind of nitrogen can be incorporated onto chitosan membrane surface with theform of O=C-NH_2. The surface oxidation of chitosan membrane after exposing to air was still the main mechanism of surface activitation, which induced the improvement of surfacehydrophilicity and surface free energy.
For the ageing effect of plasma treatment, the incorporated polar groups on chitosanmembrane surface will migrated into the membrane bulk gradually. This kind of redistributionof polar groups induced the recovery of surface hydrophobicity. The aging effect of chitosanmembrane after plasma treatment lasted for about10days before the surface contact anglereaching the value of untreated sample.
Ar plasma induced surface graft of PEG onto chitosan membrane surface was carried out inthe study. The thickness of the grafted PEG layer was in the order of10nm indicated by XPSresults. The surface graft ratio depends on the monomer concentration and argon plasmacondition. The optimal monomer concentration was10%under the plasma power of100Wand plasma exposure time of60s. Methoxy polyethylene Glycol Monomethacrylate (PEGMA)was more favorable for surface graft of chitosan membrane than Polyethylene GlycolDimethacrylate (PEGDA) for its higher graft ratio under the same condition. The surfacehydrophilicity and surface free energy of chitosan membrane were improved by the graft ofPEGMA on membrane surface. The surface roughness was increased by the grafted PEGchain. The surface modified chitosan membrane has a good protein adsorption property. Andthe human corneal epithelial cell (HCEC) cell adhesion on chitosan membrane surface wasstrengthened.
Oxygen, nitrogen and argon plasma were applied in the study for RGP lens surfacetreatment respectively. They all could improve the surface hydrophilicity of RGP lens. Themain reason for the improvement of surface hydrophilicity was the incorporation ofoxygen-containing groups on RGP lens surface. The chemical state of Si was changed fromsilixane (Si-CH3) into hydrophilic Si-O through the broken of Si-C bond. The formation of theinorganic Si-O was the main reason for the surface hydrophilicity improvement. Some C-Obond on RGP surface was also broken by plasma treatment, and transformed into O-C=O,which was another reason for the surface hydrophilicity improvement. Nitrogen plasma canintroduce O=C-NH_2group onto RGP lens surface. The reaction between the radicalsgenerated by plasma treatment and air oxygen was the main mechanism for surface oxidation.And the introduced oxygen was mainly in the state of Si-O. The effect of plasma treatment on surface morphology depends on the plasma power. Compared with oxygen and nitrogenplasma, argon plasma has a more strong influence on surface morphology by surface etching.The optimal condition for plasma treatment is100W and1_20s. Ar plasma with higher powerwill induced the increase of surface roughness by surface etching.
PEGMA can be grafted successfully onto RGP surface through Ar plasma induced graftpolymerization. The thickness of the grafted PEG layer is in the order of several nanometers.The surface graft ratio depends on the monomer concentration. And the graft ratio reaches themaximum when the monomer concentration is10%with water as the solvent. After the graftof PEGMA, the surface hydrophilicity of RGP was improved, and the surface adsorption ofprotein on RGP surface was decreased greatly compared with the untreated sample. Thesurface modified RGP lens showed good cell compatibility.
采用角分辨XPS对壳聚糖膜表面化学基团的空间分布进行分析表征。结果表明，壳聚糖膜随着在空气中放置时间延长，其表面极性基团-OH等逐渐转移到膜的内部；壳聚糖膜表面N元素主要以-NH_2形式存在，其分子链上的-CONH_2主要伸向壳聚糖膜内部。表面极性基团的这种空间取向是造成壳聚糖膜表面亲水性较差，表面自由能较低的主要原因。
实验分别采用非反应性O2和N_2低温等离子体对壳聚糖膜进行表面处理。壳聚糖膜在O_2等离子体处理下，表面引入-OH、-COO、H_2N-C=O等极性基团，表面亲水性得到改善；采用O_2等离子体进行改性的最佳工艺条件为100W下处理60s，如功率过大或处理时间过长，则容易引起壳聚糖膜表面刻蚀；O2等离子体对壳聚糖膜进行表面处理未造成明显的表面形貌变化，但表面粗糙度有一定增加。采用N_2等离子体处理，也能改善壳聚糖膜表面亲水性，但效果不如O_2明显，采用100W等离子体功率和60s处理时间，其表面接触角下降为65°；N_2等离子体处理可在表面引入一定量的N元素，引入的N元素主要是以O=C-NH_2的形式存在；等离子体处理后暴露于大气中，膜表面活性自由基与空气中的氧气、水汽等进行反应，在表面引入含氧极性基团是等离子体处理样品表面极化的主要过程。
等离子体处理在壳聚糖膜表面引入的极性基团会随着时间的推移而逐渐转移到膜的内部，这种极性基团的翻转内迁，导致了等离子体处理效果的时效性，其亲水性逐渐下降，接触角增大，表面自由能尤其是其极性分量逐渐下降，壳聚糖膜表面等离子体处理的时效性大约为10天。
实验采用Ar等离子体诱导接枝在壳聚糖膜表面引入聚乙二醇（PEG）分子链；表面接枝层厚度约10nm；PEGMA单体浓度10%、Ar等离子体功率100W、时间60s时表面接枝量最大；采用PEGMA单体比采用具有相同分子量的PEGDA单体接枝率高；将PEGMA单体接枝到壳聚糖膜表面后，其表面水接触角下降，亲水性改善，表面自由能和粗糙度增加。表面接枝壳聚糖膜具有较好的牛血清蛋白吸附特性；细胞试验结果表明等离子体处理和表面接枝壳聚糖膜表面的细胞黏附率高于未经处理的壳聚糖膜，表面改性壳聚糖膜具有较好的表面细胞相容性。
采用O_2、N_2和Ar等离子体对硬性透气性（RGP）FSA角膜接触镜片进行表面处理均能极大地改善镜片表面的亲水性。等离子体处理在表面引入了含O基团，其中表面Si-CH3发生分子键的断裂，形成Si-O，硅烷转变为亲水性的无机硅氧基团（Si-O），是表面亲水性改善的主要原因。在等离子体处理下，表面分子中C-O发生部分断键，生成O-C=O。N2等离子体处理能够在RGP表面引入含N基团，N元素主要是以O=C-NH_2的形式存在。等离子体处理后表面活性自由基与空气中的氧气反应形成含氧基团是表面活化的主要过程，其中表面引入的O主要是与Si结合形成Si-O。等离子体处理对RGP表面形貌的影响主要取决于等离子体功率的大小，Ar等离子体相比O2和N2等离子体对表面的刻蚀更为明显，等离子体处理工艺以100W和120s为宜。三种气氛等离子体相比较，N2相比Ar和O_2等离子体较为温和，而Ar等离子体在功率较大时，容易导致表面刻蚀，表面粗糙度增加，表面亲水性反而下降，这与等离子体中的粒子能量大小有关。
采用Ar等离子体辅助接枝能有效地将PEGMA单体分子接枝聚合到RGP表面，所得PEGMA接枝层的厚度大约10纳米；结果表明：表面接枝量大小与单体浓度有关，采用去离子水作为溶剂，PEGMA单体浓度10%时具有最大的表面接枝量，RGP表面接枝PEG后，表面亲水性改善，RGP镜片表面溶菌酶蛋白质的吸附量减少，表面改性RGP镜片具有较好的表面细胞相容性。
采用低温等离子体技术对壳聚糖膜和角膜接触镜材料进行表面改性，能有效改善材料表面的亲水性和生物学性能，在角膜修复材料领域具有广泛的应用前景。
Chitosan and fluorosilicone acrylates (FSA) have been widely used for cornea repair studyand application. But chitosan’s poor surface hydrophilicity and low surface free energy,together with the lack of cell-recognition sites restrict the adhesion and propagation of cell onits surface. The peculiar molecular structure of FSA makes it a promident material for makingrigid gas permeable contact lens (RGP). But the surface hydrophobicity makes the lensunwettable for the tears, and the non-polar interaction between the surface unpolar groupswith the tear protein will lead to the surface protein and lipid adsorption. The study focusedon the surface modification and characterization of chitosan membrane and RGP contact lensin order to improve surface hydrophilicity and biological properties by plasma surfacetreatment and graft polymerization.
The surface properties of chitosan membrane were characterized by angle-resolved X-rayphotoelectron spectroscopy (XPS). The results indicated that the polar groups (-OH) onchitosan membrane migrated into the membrane bulk, which made the surface hydrophobicwith low surface free energy. The surface nitrogen mainly exists in the form of–NH_2. The-CONH_2groups on chitosan molecule also distributed into the membrane bulk.
In this study, chitosan membranes were surface modified by oxygen and nitrogen plasmarespectively. The surface hydrophilicity of chitosan membrane was greatly improved byoxygen plasma treatment. The mechanism for surface hydrophilicity improvement was thesurface oxidation after plasma treatment and exposure to air. Through this kind of surfaceoxidation, polar groups, such as–OH,-COO, H_2N-C=O were incorporated onto chitosanmembrane surface. The optimal plasma condition for chitosan surface treatment was100Wand60s. Higher plasma power or longer exposure time will bring about surface etching. Noevident morphology change was observed, but surface roughness increased slightly afterplasma treatment. Nitrogen plasma can also improve the surface hydrophilicity of chitosanmembrane. But the effect was not so evident compared with oxygen plasma. The surfacecontact angle decreased to about65°when the plasma power was100W and exposure timewas60s. Some kind of nitrogen can be incorporated onto chitosan membrane surface with theform of O=C-NH_2. The surface oxidation of chitosan membrane after exposing to air was still the main mechanism of surface activitation, which induced the improvement of surfacehydrophilicity and surface free energy.
For the ageing effect of plasma treatment, the incorporated polar groups on chitosanmembrane surface will migrated into the membrane bulk gradually. This kind of redistributionof polar groups induced the recovery of surface hydrophobicity. The aging effect of chitosanmembrane after plasma treatment lasted for about10days before the surface contact anglereaching the value of untreated sample.
Ar plasma induced surface graft of PEG onto chitosan membrane surface was carried out inthe study. The thickness of the grafted PEG layer was in the order of10nm indicated by XPSresults. The surface graft ratio depends on the monomer concentration and argon plasmacondition. The optimal monomer concentration was10%under the plasma power of100Wand plasma exposure time of60s. Methoxy polyethylene Glycol Monomethacrylate (PEGMA)was more favorable for surface graft of chitosan membrane than Polyethylene GlycolDimethacrylate (PEGDA) for its higher graft ratio under the same condition. The surfacehydrophilicity and surface free energy of chitosan membrane were improved by the graft ofPEGMA on membrane surface. The surface roughness was increased by the grafted PEGchain. The surface modified chitosan membrane has a good protein adsorption property. Andthe human corneal epithelial cell (HCEC) cell adhesion on chitosan membrane surface wasstrengthened.
Oxygen, nitrogen and argon plasma were applied in the study for RGP lens surfacetreatment respectively. They all could improve the surface hydrophilicity of RGP lens. Themain reason for the improvement of surface hydrophilicity was the incorporation ofoxygen-containing groups on RGP lens surface. The chemical state of Si was changed fromsilixane (Si-CH3) into hydrophilic Si-O through the broken of Si-C bond. The formation of theinorganic Si-O was the main reason for the surface hydrophilicity improvement. Some C-Obond on RGP surface was also broken by plasma treatment, and transformed into O-C=O,which was another reason for the surface hydrophilicity improvement. Nitrogen plasma canintroduce O=C-NH_2group onto RGP lens surface. The reaction between the radicalsgenerated by plasma treatment and air oxygen was the main mechanism for surface oxidation.And the introduced oxygen was mainly in the state of Si-O. The effect of plasma treatment on surface morphology depends on the plasma power. Compared with oxygen and nitrogenplasma, argon plasma has a more strong influence on surface morphology by surface etching.The optimal condition for plasma treatment is100W and1_20s. Ar plasma with higher powerwill induced the increase of surface roughness by surface etching.
PEGMA can be grafted successfully onto RGP surface through Ar plasma induced graftpolymerization. The thickness of the grafted PEG layer is in the order of several nanometers.The surface graft ratio depends on the monomer concentration. And the graft ratio reaches themaximum when the monomer concentration is10%with water as the solvent. After the graftof PEGMA, the surface hydrophilicity of RGP was improved, and the surface adsorption ofprotein on RGP surface was decreased greatly compared with the untreated sample. Thesurface modified RGP lens showed good cell compatibility.
引文
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