Laboratório de Controle de Qualidade Farmacêutico/Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Ipiranga, Porto Alegre, Brazil.
Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy.
Eur J Pharm Biopharm. 2019 Sep;142:247-257. doi: 10.1016/j.ejpb.2019.06.030. Epub 2019 Jun 29.
In healthy individuals, wound healing is a highly efficient process. However, interruptions of normal healing give rise to chronic wounds, characterized by inflammation with impaired angiogenesis and re-epithelialization. The aim of this work was the design and the development of electrospun nanofibrous scaffolds based on sodium alginate (SA) and pullulan (PUL) and loaded with human platelet lysate (PL) intended for skin reparation, to take the advantage of nanofibrous scaffolds (with improved physical structure) and of SA as biopolymer. Two preparation approaches have been used to load PL in the scaffolds: as component of the PUL/SA matrix, to be electrospun, or as coating component, to cover the previously prepared electrospun PUL based membranes. A preformulation study to assess pullulan entanglement concentration and alginate or citric acid critical concentration, to obtain electrospun nanofibers, has been performed. The preparation process allowed to obtain insoluble systems starting from aqueous solutions and these were able to act as scaffolds for tissue engineering with suitable mechanical properties and PL release. PL loading in PUL/SA matrix nanofibers did not substantially modify the nanofiber morphology before crosslinking, while the crosslinking process, in presence of PL, determined less sharp nanofibers probably due to an increase in hydrophilicity caused by PL proteins. On the contrary, the coated nanofibers showed an increase in diameters due to PL loading. The two different approaches affected the fiber dimension and scaffold elasticity, especially for PL loaded systems. Anyhow, these differences were not crucial for fibroblast adhesion and proliferation which were mainly influenced by PL loading. In particular, fibroblasts presented different conformation and orientation mainly due to the presence of PL. This caused a cell random orientation compatible to a fibroblast-to-myofibroblast transition that could enhance wound healing.
在健康个体中,伤口愈合是一个高效的过程。然而,正常愈合过程的中断会导致慢性伤口,其特征为伴有受损血管生成和再上皮化的炎症。本工作的目的是设计和开发基于海藻酸钠(SA)和普鲁兰(PUL)的电纺纳米纤维支架,并负载人血小板裂解物(PL),用于皮肤修复,以利用纳米纤维支架(改善物理结构)和 SA 作为生物聚合物。采用两种方法将 PL 负载到支架中:作为 PUL/SA 基质的组成部分进行电纺,或作为涂层成分覆盖先前制备的基于 PUL 的电纺膜。进行了预配方研究以评估普鲁兰缠结浓度和海藻酸钠或柠檬酸的临界浓度,以获得电纺纳米纤维。制备过程允许从水溶液中获得不溶性体系,这些体系可作为组织工程的支架,具有合适的机械性能和 PL 释放。PL 负载在 PUL/SA 基质纳米纤维中不会在交联前实质上改变纳米纤维形态,而交联过程在存在 PL 的情况下会导致纳米纤维变得不那么锋利,这可能是由于 PL 蛋白引起的亲水性增加所致。相反,负载 PL 的涂层纳米纤维的直径增加。这两种不同的方法影响纤维的尺寸和支架的弹性,特别是对于 PL 负载的系统。无论如何,这些差异对于成纤维细胞的粘附和增殖并不关键,而成纤维细胞的粘附和增殖主要受 PL 负载的影响。特别是,由于 PL 的存在,成纤维细胞呈现出不同的构象和取向。这导致细胞随机取向,与成纤维细胞向肌成纤维细胞转化兼容,从而促进伤口愈合。