PEG水凝胶接枝活性肽及其表面细胞粘附和抗菌性的研究

发布时间：2018-04-15 15:50

本文选题：抗菌材料 + PEG水凝胶　；参考：《河南大学》2012年硕士论文

【摘要】：本论文针对目前生物水凝胶存在的表面功能化问题，对聚乙二醇（PEG）水凝胶材料表面进行光聚合接枝生物活性肽，，提高其表面细胞相容性和抗菌性。分别接枝细胞粘附因子RGDS（精氨酸-甘氨酸-天冬氨酸-丝氨酸）、抗菌性杆菌肽(bacitracin)，并对表面功能化的PEG（聚乙二醇）水凝胶进行小鼠胚胎成纤维细胞相容性、金黄色葡萄球菌抗菌性测试。本论文所做的工作主要有： (1)自行分离得到昆明白小鼠的成纤维细胞和神经细胞，并且能够对分离所得到的细胞进行培养、传代、冻存、复苏。为之后PEG水凝胶表面改性细胞粘附实验提供细胞源。 (2)通过管碟法用本实验室提供的四种革兰氏阳性细菌对活性蛋白杆菌肽的敏感性进行初步检测，并从中选出敏感性显著、人体表面创口易感染的菌株金黄色葡萄球菌作为杆菌肽接枝PEG水凝胶薄膜上后的检测菌株。 (3)使用不同分子量的分子两端有双键的PEG大单体聚乙二醇双丙烯酸酯（PEGDA）为原料，通过紫外光聚合技术制备出一系列物理性能不同的PEG水凝胶薄膜，并测定了PEG水凝胶薄膜的吸水溶胀等物理参数 (4)根据实验要求设计制备了转移试剂ACRL-PEG-RGDS，并利用此转移试剂和二次光聚合技术把多肽RGDS接枝到PEG水凝胶薄膜上，使改性后的PEG水凝胶薄膜具有细胞粘附能力。通过MTT(3-(4，5-二甲基噻唑-2)-2，5-二苯基四氮唑溴盐)法检测了改性后PEG水凝胶薄膜上的细胞活性。 (5)根据实验要求设计制造了转移试剂ACRL-PEG-bacitracin，并利用此转移试剂和二次光聚合技术把杆菌肽(bacitracin)接枝到PEG水凝胶薄膜上，使改性后的PEG水凝胶薄膜具有抗菌能力。通过MTT法检测了改性后PEG水凝胶薄膜上的细菌活性。 (6)利用荧光胺与多肽的伯氨基能够发生荧光显色反应的原理对PEG水凝胶薄膜表面接枝的多肽RGDS和杆菌肽进行荧光显色检查。通过荧光显微镜观察，验证了PEG水凝胶薄膜表面成功地接枝上了多肽RGDS和杆菌肽。 (7)利用转移试剂和二次光聚合技术把多肽RGDS和杆菌肽混合接枝到PEG水凝胶薄膜上，通过MTT法检测了改性后PEG水凝胶薄膜上的细胞和细菌活性。改性后的PEG水凝胶薄膜兼有细胞黏附和抗细菌感染的能力。
[Abstract]:In order to improve the surface compatibility and antimicrobial activity of PEG hydrogels, the surface of PEG hydrogels was grafted with bioactive peptides by photopolymerization.The graft of RGDS (arginine glycine aspartate serine) and bacitracinine (Bacillus sp.) were used to study the compatibility of mouse embryonic fibroblasts with PEG (polyethylene glycol) hydrogel.Antibacterial test of Staphylococcus aureus.The main work of this paper is as follows:1) the fibroblasts and nerve cells of Kunming white mice were isolated and cultured, subcultured, frozen and resuscitated.It provides a cell source for PEG hydrogel surface modified cell adhesion test.(2) the sensitivity of four Gram-positive bacteria to active protein bacilli was preliminarily tested by tube disc method, and the sensitivity was significant.Staphylococcus aureus, which is susceptible to infection on human surface, is used as a detection strain of bacitracin grafted onto PEG hydrogel film.A series of PEG hydrogel films with different physical properties were prepared by ultraviolet photopolymerization using poly (ethylene glycol) diacrylate (PEGDAA), a macromonomer with double bonds at both ends of the molecules with different molecular weights, as the raw material, and a series of PEG hydrogel films with different physical properties were prepared by ultraviolet photopolymerization.The physical parameters such as water absorption and swelling of PEG hydrogel film were determined.The transfer reagent ACRL-PEG-RGDS was designed and prepared according to the experimental requirements. The polypeptide RGDS was grafted onto the PEG hydrogel film by using the transfer reagent and secondary photopolymerization technique. The modified PEG hydrogel film had cell adhesion ability.The cell activity of the modified PEG hydrogel film was determined by MTT- 3-trimethythiazolium-5-dimethylthiazolium-2-diphenyl tetrazolium bromine salt (MTT-3-dimethylthiazolium-2-diphenyl tetrazolium bromide) method.The transfer reagent ACRL-PEG-bacitracin was designed and manufactured according to the experimental requirements. The modified PEG hydrogel film was grafted onto the PEG hydrogel film by using the transfer reagent and secondary photopolymerization technique. The modified PEG hydrogel film had antibacterial activity.The bacterial activity of modified PEG hydrogel film was determined by MTT method.The fluorescence color reaction of RGDS and bacitracin on the surface of PEG hydrogel film was studied by using the principle that fluorescein can react with the primary amino group of polypeptide.The surface of PEG hydrogel film was successfully grafted with polypeptide RGDS and bacitracin by fluorescence microscope.The polypeptide RGDS and bacitracin were grafted onto PEG hydrogel film by transfer reagent and secondary photopolymerization. The cellular and bacterial activity of modified PEG hydrogel film was determined by MTT method.The modified PEG hydrogel film has the ability of cell adhesion and anti-bacterial infection.
【学位授予单位】：河南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.08

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