胆红素血液净化吸附剂的制备与吸附性能评价
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摘要
胆红素是一种胆色素,体内胆红素水平过高,可以导致高胆红素血症,对于症状较轻的患者,特别是新生儿高胆红素血症患者,临床上一般采用药物和光线疗法进行治疗,而对于病情比较严重的患者,则要借助血液净化疗法。血液灌流技术作为血液净化疗法中的一种,在治疗高胆红素血症方面逐渐显示出其优势作用,并且越来越受到人们的重视。血液灌流技术的关键点是血液净化吸附剂的研发,国外已经有特异性吸附胆红素的吸附剂上市,并广泛应用于临床。我国对吸附剂的研究起步比较晚,虽然也有相应的产品出现,但是其安全性和有效性均有待于进一步提高。本文目的在于研制出适合临床应用的、具有高度的特异性、较高的吸附容量和生物相容性好的胆红素吸附剂。全文共分为五个部分:
第一部分为绪论,主要介绍了胆红素及高胆红素血症,高胆红素血症的危害,形成原因以及治疗方法;综述了血液净化疗法及血液净化吸附剂,并主要介绍了高胆红素血症血液灌流吸附剂的研究及临床应用情况;最后提出本论文进行的工作。
第二部分研究了以纤维素为载体的吸附剂,首先制备了纤维素微球,经环氧氯丙烷活化,然后固定8种胺化试剂作为功能基得到相应的吸附剂,并对其进行了吸附性能测试。结果显示,以1,6-己二胺为功能基的吸附剂吸附效果最佳,对胆红素水溶液的吸附率最高可以达到82%。
第三部分制备了以壳聚糖为载体的吸附剂,在最佳活化条件下分别固定8种多烯多胺功能基,并对相应得到的吸附剂进行了吸附性能测试,结果显示,以1,6己二胺为功能基的吸附剂吸附效果最佳,对胆红素水溶液的吸附率最高可以达到98%,吸附量为1.5mg/ml,和空白壳聚糖微球相比,吸附率提高了60%,表明该类吸附剂对胆红素具有特异性。而且与纤维素载体吸附剂相比,该类载体吸附剂机械强度和吸附性能都有了整体的改善,也表明壳聚糖是胆红素吸附剂的较好载体。
第四部分研究了以聚乙烯醇微球为载体的胆红素吸附剂,首先用悬浮聚合的方法制备聚乙烯醇微球载体,并对其进行了物理性能表征,表明微球具有较好的粒径分布,较大的孔径和孔隙率,适合作为吸附剂的载体。然后用环氧氯丙烷活化,环氧基最高含量为175umol/ml。活化后的微球固定一系列胺化试剂作为功能配基形成吸附剂,并对其进行了系统的体外吸附性能评价。体外静态吸附性能测试表明以三甲胺,三乙胺和1,6己二胺为配基的吸附剂对胆红素具有较好的吸附性能,三种吸附剂对胆红素模拟血清液的的吸附率均达到80%以上,最大吸附量达4.3mg/ml。通过对吸附剂的吸附动力学研究表明该类吸附剂0.5h基本达到吸附平衡,达到最大吸附率的80%,该类吸附剂具有很快的吸附速率及较大的吸附容量。吸附剂的体外动态吸附性能评价表明10倍胆红素模拟血清液用量时,上述三种功能基的吸附剂对胆红素的吸附量分别为1.577 mg/ml,1.584mg/ml,1.784 mg/ml,吸附率均接近100%。本文还考察了三类吸附剂对高胆红素病人血浆的吸附性能,在5倍体积的高胆红素血浆情况下,以己二胺为功能基的吸附剂最大吸附率可以达到50%,而对其他成分没有明显的吸附。表明该类吸附剂具有较高吸附效率,较大的吸附容量和较好选择性和血液相容性,具有一定的临床应用前景。
最后一部分是对全文的总结,概况了整篇论文所进行的工作和取得的成果及以后工作方向。
Bilirubin is a yellow-orange bile pigment.High level of bilirubin can result in Hyperbilirubinemia,which is harmful to health.In clinical,medicine and phototherapy are usually applied for mild cases,especially newborn infants.But for severe ones,blood purification technology must be adopted to save their lives. Hemoperfusion,a kind of blood purification methods,is demonstrating its advantages in the treating hyperbilirubinemia and attracts more and more people's attentions. And key factor of hemoperfusion is the adsorbent.Though some products have been adopted in clinical,security and efficacy both need to be elevated.The objective of my dissertation is to prepare kinds of adsorbents,which are suitable in clinical application for bilirubin removal and possessing high specificity,large adsorptive capacity,and good biocompatibility.There are five parts in this dissertation.
The first part is an introduction about basic knowledge of bilirubin, hyperbilirubinemia,the harmfulness and reasons of hyperbilirubinemia and methods for treatment.It also summarizes the blood purification technology and adsorbents utilized in processes of hemoperfusion.And it also chiefly gives an overview of the researches and applications of adsorbents for hyperbilirubinemia.At last,I propose the relative work in this research.
The second part is about adsorbents with cellulose as support.First we prepared cellulose microspheres,and then immobilized 8 kinds of amination agents to obtain corresponding adsorbents and carried out vitro test.Results show that it is cel-HDA has the best adsorption performance,and adsorption percentage is 82%to bilirubin solution.
The third part is about adsorbents with chitosan as carrier.Under the optimum active condition,we prepared adsorbents immobilized 8 kinds of amination agents and carried out vitro tests on them.Results still show that it is CS-HDA has the best adsorption performance.Meanwhile chitosan adsorbents' performances improve significantly compared with cellulose ones,which means chitosan is a good support for bilirubin adsorbents.
The fourth part is about adsorbents with polyvinyl alcohol(PVA)as support.First PVA microspheres are produced as support by suspension polymerization in aqueous medium.Then we research its physical characterizations,and all of them indicate PVA particles are carrier matrix.Second we obtained the max amino group content, 175umol/ml.Under the optimum active condition,we prepared corresponding adsorbents with a series of amination agents and carry out vitro tests on them.Results show adsorbents of PVA-TMA,PVA-TEA and PVA-HDA have the good adsorption performances,with adsorption percentages higher than 80%and maximum capacity up to 4.3mg/ml.Adsorption dynamics curves show this kind of adsorbents have high adsorption velocity and large capacities for they can achieve equilibrium in 0.5h and get 80%of the maximum percentages.In vitro adsorption performance evaluations show their adsorption capacities are respectively 1.577mg/ml,1.584mg/ml and 1.784mg/ml,while volume ratio of adsorbent to bilirubin simulating serum solution is 10:1.We also inspected their performances to human plasma.PVA-HDA's maximum adsorption percentage is 50%without obvious adsorption to other components,while volume ratio is 5:1.All results indicate adsorbents have good performances,large adsorption capacities,good selectivity and biocompatibility.There will be a prospect of clinical application.
The last part is the summary of the whole dissertation.It summarizes all the work and achievements,points out the new ideas and fingers out the work in next step.
第一部分为绪论,主要介绍了胆红素及高胆红素血症,高胆红素血症的危害,形成原因以及治疗方法;综述了血液净化疗法及血液净化吸附剂,并主要介绍了高胆红素血症血液灌流吸附剂的研究及临床应用情况;最后提出本论文进行的工作。
第二部分研究了以纤维素为载体的吸附剂,首先制备了纤维素微球,经环氧氯丙烷活化,然后固定8种胺化试剂作为功能基得到相应的吸附剂,并对其进行了吸附性能测试。结果显示,以1,6-己二胺为功能基的吸附剂吸附效果最佳,对胆红素水溶液的吸附率最高可以达到82%。
第三部分制备了以壳聚糖为载体的吸附剂,在最佳活化条件下分别固定8种多烯多胺功能基,并对相应得到的吸附剂进行了吸附性能测试,结果显示,以1,6己二胺为功能基的吸附剂吸附效果最佳,对胆红素水溶液的吸附率最高可以达到98%,吸附量为1.5mg/ml,和空白壳聚糖微球相比,吸附率提高了60%,表明该类吸附剂对胆红素具有特异性。而且与纤维素载体吸附剂相比,该类载体吸附剂机械强度和吸附性能都有了整体的改善,也表明壳聚糖是胆红素吸附剂的较好载体。
第四部分研究了以聚乙烯醇微球为载体的胆红素吸附剂,首先用悬浮聚合的方法制备聚乙烯醇微球载体,并对其进行了物理性能表征,表明微球具有较好的粒径分布,较大的孔径和孔隙率,适合作为吸附剂的载体。然后用环氧氯丙烷活化,环氧基最高含量为175umol/ml。活化后的微球固定一系列胺化试剂作为功能配基形成吸附剂,并对其进行了系统的体外吸附性能评价。体外静态吸附性能测试表明以三甲胺,三乙胺和1,6己二胺为配基的吸附剂对胆红素具有较好的吸附性能,三种吸附剂对胆红素模拟血清液的的吸附率均达到80%以上,最大吸附量达4.3mg/ml。通过对吸附剂的吸附动力学研究表明该类吸附剂0.5h基本达到吸附平衡,达到最大吸附率的80%,该类吸附剂具有很快的吸附速率及较大的吸附容量。吸附剂的体外动态吸附性能评价表明10倍胆红素模拟血清液用量时,上述三种功能基的吸附剂对胆红素的吸附量分别为1.577 mg/ml,1.584mg/ml,1.784 mg/ml,吸附率均接近100%。本文还考察了三类吸附剂对高胆红素病人血浆的吸附性能,在5倍体积的高胆红素血浆情况下,以己二胺为功能基的吸附剂最大吸附率可以达到50%,而对其他成分没有明显的吸附。表明该类吸附剂具有较高吸附效率,较大的吸附容量和较好选择性和血液相容性,具有一定的临床应用前景。
最后一部分是对全文的总结,概况了整篇论文所进行的工作和取得的成果及以后工作方向。
Bilirubin is a yellow-orange bile pigment.High level of bilirubin can result in Hyperbilirubinemia,which is harmful to health.In clinical,medicine and phototherapy are usually applied for mild cases,especially newborn infants.But for severe ones,blood purification technology must be adopted to save their lives. Hemoperfusion,a kind of blood purification methods,is demonstrating its advantages in the treating hyperbilirubinemia and attracts more and more people's attentions. And key factor of hemoperfusion is the adsorbent.Though some products have been adopted in clinical,security and efficacy both need to be elevated.The objective of my dissertation is to prepare kinds of adsorbents,which are suitable in clinical application for bilirubin removal and possessing high specificity,large adsorptive capacity,and good biocompatibility.There are five parts in this dissertation.
The first part is an introduction about basic knowledge of bilirubin, hyperbilirubinemia,the harmfulness and reasons of hyperbilirubinemia and methods for treatment.It also summarizes the blood purification technology and adsorbents utilized in processes of hemoperfusion.And it also chiefly gives an overview of the researches and applications of adsorbents for hyperbilirubinemia.At last,I propose the relative work in this research.
The second part is about adsorbents with cellulose as support.First we prepared cellulose microspheres,and then immobilized 8 kinds of amination agents to obtain corresponding adsorbents and carried out vitro test.Results show that it is cel-HDA has the best adsorption performance,and adsorption percentage is 82%to bilirubin solution.
The third part is about adsorbents with chitosan as carrier.Under the optimum active condition,we prepared adsorbents immobilized 8 kinds of amination agents and carried out vitro tests on them.Results still show that it is CS-HDA has the best adsorption performance.Meanwhile chitosan adsorbents' performances improve significantly compared with cellulose ones,which means chitosan is a good support for bilirubin adsorbents.
The fourth part is about adsorbents with polyvinyl alcohol(PVA)as support.First PVA microspheres are produced as support by suspension polymerization in aqueous medium.Then we research its physical characterizations,and all of them indicate PVA particles are carrier matrix.Second we obtained the max amino group content, 175umol/ml.Under the optimum active condition,we prepared corresponding adsorbents with a series of amination agents and carry out vitro tests on them.Results show adsorbents of PVA-TMA,PVA-TEA and PVA-HDA have the good adsorption performances,with adsorption percentages higher than 80%and maximum capacity up to 4.3mg/ml.Adsorption dynamics curves show this kind of adsorbents have high adsorption velocity and large capacities for they can achieve equilibrium in 0.5h and get 80%of the maximum percentages.In vitro adsorption performance evaluations show their adsorption capacities are respectively 1.577mg/ml,1.584mg/ml and 1.784mg/ml,while volume ratio of adsorbent to bilirubin simulating serum solution is 10:1.We also inspected their performances to human plasma.PVA-HDA's maximum adsorption percentage is 50%without obvious adsorption to other components,while volume ratio is 5:1.All results indicate adsorbents have good performances,large adsorption capacities,good selectivity and biocompatibility.There will be a prospect of clinical application.
The last part is the summary of the whole dissertation.It summarizes all the work and achievements,points out the new ideas and fingers out the work in next step.
引文
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