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Master's Dissertation
DOI
https://doi.org/10.11606/D.85.2013.tde-19112013-145148
Document
Author
Full name
Caroline Cristina Ferraz
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Lugão, Ademar Benévolo (President)
Consiglieri, Vladi Olga
Silva, Classius Ferreira da
Title in Portuguese
Desenvolvimento de uma membrana nanoestruturada à base de poliacrilamida para veiculação de proteínas
Keywords in Portuguese
adesão celular
BSA
citotoxicidade
hidrogel
Papaína
poliacrilamida
radiação ionizante
Abstract in Portuguese
Hidrogéis são membranas formadas pela reticulação de cadeias poliméricas, empregados na área farmacêutica como produtos biomédicos. Dentre os principais polímeros selecionados para a síntese de hidrogéis, destaca-se a poliacrilamida (PAAM) devido às suas propriedades como hidrofilicidade e alto grau de intumescimento. Proteínas terapêuticas e enzimas são veiculadas em hidrogéis como carreadores de fármaco ou como dispositivos para tratamento de feridas e escaras na pele. Este trabalho teve como objetivo a síntese de uma membrana à base de PAAM favorável para veiculação de proteínas. As proteínas empregadas foram papaína e albumina de soro bovino (BSA) e as etapas do processo englobaram síntese da membrana, adição das proteínas no sistema, irradiação em condições específicas e caracterização da membrana. Ao utilizar temperaturas criogênicas na síntese e na irradiação das amostras, houve predomínio de reticulação da cadeia polimérica, fato que não ocorria em temperatura ambiente. As membranas foram obtidas com incorporação dos ativos na concentração de 0,2 a 1% (p/p), obtendo-se concentração de PAAM entre 4% a 10% (p/p), as quais receberam irradiação com raios gama provenientes de uma fonte 60Co, na dose de 25 kGy. Nas condições realizadas, as membranas não apresentaram citotoxicidade nem adesão celular, o perfil de liberação das proteínas foi adequado, a papaína manteve sua bioatividade preservada apesar do decaimento biológico e, segundo estudos de carga das moléculas, a membrana possui maior afinidade com a papaína, liberando-a mais lentamente. Desta forma, o método proposto e as membranas obtidas foram apropriados para a obtenção de um biomaterial.
Title in English
Radio-synthesized polyacrylamide nanostructured hydrogels for proteins release
Keywords in English
BSA
cell adhesion
cytotoxicity
hydrogels
ionizing radiation
Papain
polyacrylamide
Abstract in English
The use of hydrogels for biomedical purposes has been extensively investigated. Polyacrylamide (PAAM) is widely used due to properties such as hydrophilicity and swelling degree. Pharmaceutical proteins correspond to highly active substances which may be applied for distinct purposes. This work concerns the development of radio-synthesized hydrogel for protein release using papain and bovine serum albumin (BSA) as model proteins. The polymer was solubilized (1% w/v) in water and lyophilized. The proteins were incorporated into the lyophilized polymer and the hydrogels were produced by simultaneous crosslinking and sterilization using gamma radiation at 25 kGy under frozen conditions. The produced systems were characterized in terms of swelling degree, gel fraction, crosslinking density, fluid handling capacity, determination pH at point of polymer zero charge and evaluated according to protein release, bioactivity, cytotoxicity and cell adhesion. The hydrogels developed presented different properties as a function of polymer concentration and the optimized results were found for the samples containing 4-10% polyacrylamide. Protein release was controlled by the electrostatic affinity of acrylic moieties of polymer and proteins. This selection was based on the release of the proteins during the experiment period (up to 50 hours), maintenance of enzyme activity and the nanostructure developed. The system was suitable for protein loading and release and according to the cytotoxic assay and cell adhesion it was also adequate for biomedical purposes and this method was able to generate a matrix to protein release.
 
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Publishing Date
2013-12-04
 
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