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用于整形矫正的阴离子胶原蛋白:鼠李聚糖复合物注射凝胶:制备、表征及流变学性质

Injectable gels of anionic collagen:rhamsan composites for plastic correction: preparation, characterization, and rheological properties.

作者信息

de Paula Márcio, Goissis Gilberto, Martins Virgínia C A, da Silva Trindade José Carlos

机构信息

Departamento de Química e Física Molecular do Instituto de Química de São Carlos, Universidade de São Paulo (IQSC/USP).

出版信息

J Biomed Mater Res B Appl Biomater. 2005 Nov;75(2):393-9. doi: 10.1002/jbm.b.30326.

Abstract

The present article describes the preparation and characterization of anionic collagen gels obtained from porcine intestinal submucosa after 72 h of alkaline treatment and in the form of rhamsan composites to develop injectable biomaterials for plastic reconstruction. All materials were characterized by SDS/polyacrylamide gel electrophoresis, infrared spectroscopy, thermal stability, potentiometric titration, rheological properties, and fluidity tests. Biocompatibility was appraised after the injection of anionic collagen: rhamsan composites at 2.5% in 60 North Folk rabbits. Independently of processing, the collagen's secondary structure was preserved in all cases, and after 72 h of hydrolysis the collagen was characterized by a carboxyl group content of 346+/-9, which, at physiological pH, corresponds to an increase of 106+/-17 negative charges, in comparison to native collagen, due to the selective hydrolysis of asparagine and glutamine carboxyamide side chain. Rheological studies of composites at pH 7.4 in concentrations of 2, 4, and 6% (in proportions of 75:1 and 50:1) showed a viscoelastic behavior dependent on the frequency, which is independent of concentration and proportion. In both, the concentration of the storage modulus always predominated over the loss modulus (G'>G'' and delta<45 degrees ). The results from creep experiments confirmed this behavior and showed that anionic collagen:rhamsan composites at pH 7.4 in the proportion of 50:1 are less elastic and more susceptible to deformation in comparison to gels in the proportion of 75:1, independent of concentration. This was further confirmed by flow experiments, indicating that the necessary force for the extrusion of anionic collagen:rhamsan composites, in comparison to anionic collagen, was significantly smaller and with a smooth flow. Biocompatibility studies showed that the tissue reaction of anionic collagen:rhamsan composites at 2.5% in the proportion of 75:1 was compatible with the application of these gels in plastic reconstruction. These results suggest that the association of collagen with rhamsan may be a good alternative in the replacement of glutaraldehyde to stabilize the microfibril assembly of commercial collagen gel preparations.

摘要

本文描述了经72小时碱性处理后从猪小肠黏膜下层获得的阴离子胶原蛋白凝胶的制备与表征,以及其作为鼠李聚糖复合材料的形式,用于开发用于整形重建的可注射生物材料。所有材料均通过SDS/聚丙烯酰胺凝胶电泳、红外光谱、热稳定性、电位滴定、流变学性质和流动性测试进行表征。在60只北方民间兔中注射2.5%的阴离子胶原蛋白:鼠李聚糖复合材料后评估其生物相容性。无论加工方式如何,在所有情况下胶原蛋白的二级结构均得以保留,水解72小时后,胶原蛋白的羧基含量为346±9,在生理pH值下,与天然胶原蛋白相比,由于天冬酰胺和谷氨酰胺羧酰胺侧链的选择性水解,负电荷增加了106±17。对pH值为7.4、浓度为2%、4%和6%(比例为75:1和50:1)的复合材料进行流变学研究表明,其粘弹性行为取决于频率,与浓度和比例无关。在这两种情况下,储能模量的浓度始终高于损耗模量(G'>G''且δ<45度)。蠕变实验结果证实了这种行为,并表明pH值为7.4、比例为50:1的阴离子胶原蛋白:鼠李聚糖复合材料与比例为75:1的凝胶相比,弹性较小且更容易变形,与浓度无关。流动实验进一步证实了这一点,表明与阴离子胶原蛋白相比,挤出阴离子胶原蛋白:鼠李聚糖复合材料所需的力明显更小且流动顺畅。生物相容性研究表明,2.5%、比例为75:1的阴离子胶原蛋白:鼠李聚糖复合材料的组织反应与这些凝胶在整形重建中的应用相容。这些结果表明,胶原蛋白与鼠李聚糖的结合可能是替代戊二醛稳定商业胶原蛋白凝胶制剂微原纤维组装的良好选择。

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