Santos T C, Marques A P, Silva S S, Oliveira J M, Mano J F, Castro A G, Reis R L
3B's Research Group - Biomaterials, Biodegradables and Biomimetics, Department of Polymer Engineering, Campus de Gualtar, 4710-057 Braga, Portugal.
J Biotechnol. 2007 Oct 31;132(2):218-26. doi: 10.1016/j.jbiotec.2007.07.497. Epub 2007 Jul 12.
Several novel biodegradable materials have been proposed for wound healing applications in the past few years. Taking into consideration the biocompatibility of chitosan-based biomaterials, and that they promote adequate cell adhesion, this work aims at investigating the effect of chitosan-based membranes, over the activation of human polymorphonuclear neutrophils (PMNs). The recruitment and activation of polymorphonuclear neutrophils (PMNs) reflects a primary reaction to foreign bodies. Activation of neutrophils results in the production of reactive oxygen species (ROS) such as O(2)(-) and HO(-) and the release of hydrolytic enzymes which are determinant factors in the inflammatory process, playing an essential role in the healing mechanisms. PMNs isolated from human peripheral blood of healthy volunteers were cultured in the presence of chitosan or chitosan/soy newly developed membranes. The effect of the biomaterials on the activation of PMNs was assessed by the quantification of lysozyme and ROS. The results showed that PMNs, in the presence of the chitosan-based membranes secrete similar lysozyme amounts, as compared to controls (PMNs without materials) and also showed that the materials do not stimulate the production of either O(2)(-) or HO(-). Moreover, PMNs incubated with the biomaterials when stimulated with phorbol 12-myristate 13-acetate (PMA) or formyl-methionyl-leucyl-phenylalanine (fMLP) showed a chemiluminescence profile with a slightly lower intensity, to that observed for positive controls (cells without materials and stimulated with PMA), which reflects the maintenance of their stimulation capacity. Our data suggests that the new biomaterials studied herein do not elicit activation of PMNs, as assessed by the low lysozyme activity and by the minor detection of ROS by chemiluminescence. These findings reinforce previous statements supporting the suitability of chitosan-based materials for wound healing applications.
在过去几年中，已经提出了几种新型可生物降解材料用于伤口愈合。考虑到壳聚糖基生物材料的生物相容性以及它们能促进细胞充分黏附，这项工作旨在研究壳聚糖基膜对人多形核中性粒细胞（PMN）激活的影响。多形核中性粒细胞（PMN）的募集和激活反映了对异物的主要反应。中性粒细胞的激活导致活性氧（ROS）如超氧阴离子（O₂⁻）和羟基自由基（HO⁻）的产生以及水解酶的释放，这些是炎症过程中的决定性因素，在愈合机制中起重要作用。从健康志愿者外周血中分离出的PMN在壳聚糖或新开发的壳聚糖/大豆膜存在下进行培养。通过溶菌酶和ROS的定量评估生物材料对PMN激活的影响。结果表明，与对照组（无材料的PMN）相比，在壳聚糖基膜存在下的PMN分泌的溶菌酶量相似，并且还表明这些材料不会刺激超氧阴离子（O₂⁻）或羟基自由基（HO⁻）的产生。此外，当用佛波酯12 - 肉豆蔻酸酯13 - 乙酸酯（PMA）或甲酰甲硫氨酰亮氨酰苯丙氨酸（fMLP）刺激时，与生物材料一起孵育的PMN显示出化学发光强度略低于阳性对照组（无材料且用PMA刺激的细胞），这反映了它们刺激能力的维持。我们的数据表明，通过低溶菌酶活性和化学发光法对ROS的少量检测评估，本文研究的新型生物材料不会引发PMN的激活。这些发现强化了先前支持壳聚糖基材料适用于伤口愈合应用的说法。
