本文選題：抗菌材料 + PEG水凝膠　； 參考：《河南大學(xué)》2012年碩士論文

【摘要】：本論文針對(duì)目前生物水凝膠存在的表面功能化問(wèn)題，對(duì)聚乙二醇（PEG）水凝膠材料表面進(jìn)行光聚合接枝生物活性肽，，提高其表面細(xì)胞相容性和抗菌性。分別接枝細(xì)胞粘附因子RGDS（精氨酸-甘氨酸-天冬氨酸-絲氨酸）、抗菌性桿菌肽(bacitracin)，并對(duì)表面功能化的PEG（聚乙二醇）水凝膠進(jìn)行小鼠胚胎成纖維細(xì)胞相容性、金黃色葡萄球菌抗菌性測(cè)試。本論文所做的工作主要有： (1)自行分離得到昆明白小鼠的成纖維細(xì)胞和神經(jīng)細(xì)胞，并且能夠?qū)Ψ蛛x所得到的細(xì)胞進(jìn)行培養(yǎng)、傳代、凍存、復(fù)蘇。為之后PEG水凝膠表面改性細(xì)胞粘附實(shí)驗(yàn)提供細(xì)胞源。 (2)通過(guò)管碟法用本實(shí)驗(yàn)室提供的四種革蘭氏陽(yáng)性細(xì)菌對(duì)活性蛋白桿菌肽的敏感性進(jìn)行初步檢測(cè)，并從中選出敏感性顯著、人體表面創(chuàng)口易感染的菌株金黃色葡萄球菌作為桿菌肽接枝PEG水凝膠薄膜上后的檢測(cè)菌株。 (3)使用不同分子量的分子兩端有雙鍵的PEG大單體聚乙二醇雙丙烯酸酯（PEGDA）為原料，通過(guò)紫外光聚合技術(shù)制備出一系列物理性能不同的PEG水凝膠薄膜，并測(cè)定了PEG水凝膠薄膜的吸水溶脹等物理參數(shù) (4)根據(jù)實(shí)驗(yàn)要求設(shè)計(jì)制備了轉(zhuǎn)移試劑ACRL-PEG-RGDS，并利用此轉(zhuǎn)移試劑和二次光聚合技術(shù)把多肽RGDS接枝到PEG水凝膠薄膜上，使改性后的PEG水凝膠薄膜具有細(xì)胞粘附能力。通過(guò)MTT(3-(4，5-二甲基噻唑-2)-2，5-二苯基四氮唑溴鹽)法檢測(cè)了改性后PEG水凝膠薄膜上的細(xì)胞活性。 (5)根據(jù)實(shí)驗(yàn)要求設(shè)計(jì)制造了轉(zhuǎn)移試劑ACRL-PEG-bacitracin，并利用此轉(zhuǎn)移試劑和二次光聚合技術(shù)把桿菌肽(bacitracin)接枝到PEG水凝膠薄膜上，使改性后的PEG水凝膠薄膜具有抗菌能力。通過(guò)MTT法檢測(cè)了改性后PEG水凝膠薄膜上的細(xì)菌活性。 (6)利用熒光胺與多肽的伯氨基能夠發(fā)生熒光顯色反應(yīng)的原理對(duì)PEG水凝膠薄膜表面接枝的多肽RGDS和桿菌肽進(jìn)行熒光顯色檢查。通過(guò)熒光顯微鏡觀察，驗(yàn)證了PEG水凝膠薄膜表面成功地接枝上了多肽RGDS和桿菌肽。 (7)利用轉(zhuǎn)移試劑和二次光聚合技術(shù)把多肽RGDS和桿菌肽混合接枝到PEG水凝膠薄膜上，通過(guò)MTT法檢測(cè)了改性后PEG水凝膠薄膜上的細(xì)胞和細(xì)菌活性。改性后的PEG水凝膠薄膜兼有細(xì)胞黏附和抗細(xì)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.
【學(xué)位授予單位】：河南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R318.08

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