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In vitro and in vivo testing of nanofibrous membranes doped with alaptide and L-arginine for wound treatment
Biomedical Materials ( IF 3.9 ) Pub Date : 2020-11-26 , DOI: 10.1088/1748-605x/ab950f
Petr Mikeš 1 , Antonín Brož 2, 3 , Alla Sinica 4, 5 , Nikifor Asatiani 1 , Lucie Bačáková 2
Affiliation  

We have prepared a candidate biocompatible construct for skin wound healing based on electrospun polycaprolactone (PCL) nanofibrous membranes. The membrane material was loaded either with L-arginine or with alaptide, or with a mixture of both bioactive components. Alaptide is a spirocyclic synthetic dipeptide, an analogue of melanocyte-stimulating hormone release-inhibiting factor. L-arginine is an amino acid with a basic guanidine side chain. It is a direct precursor of nitric oxide, which plays a pivotal role in skin repair. The presence and the distribution of the additives were proved with high-performance liquid chromatography, Fourier-transform infrared spectroscopy and Raman spectroscopy. The influence of L-arginine and alaptide on the morphology of the membrane was characterized using scanning electron microscopy. No statistically significant correlation between fiber diameter and drug concentration was observed. The membranes were then tested in vitro for their cytotoxicity, using primary human dermal fibroblasts, in order to obtain the optimal concentrations of the additives for in vivo tests in a rat model. The membranes with the highest concentration of L-arginine (10 wt. %) proved to be cytotoxic. The membranes with alaptide in concentrations from 0.1 to 2.5 wt.%, and with the other L-arginine concentrations (1 and 5 wt.%), did not show high toxicity. In addition, there was no observed improvement in cell proliferation on the membranes. The in vivo experiments revealed that membranes with 1.5 wt.% of alaptide or with 1.5 wt.% of alaptide in combination with 5 wt.% of L-arginine markedly accelerated the healing of skin incisions, and particularly the healing of skin burns, i.e. wounds of relatively large extent. These results indicate that our newly-developed nanofibrous membranes are promising for treating wounds with large damaged areas, where a supporting material is needed.



中文翻译:

掺杂阿拉肽和L-精氨酸的纳米纤维膜用于伤口治疗的体外和体内测试

我们已经制备了一种基于电纺聚己内酯(PCL)纳米纤维膜的用于皮肤伤口愈合的候选生物相容性结构。膜材料负载有L-精氨酸或阿拉肽,或者负载有两种生物活性成分的混合物。Alaptide 是一种螺环合成二肽,是黑素细胞刺激激素释放抑制因子的类似物。L-精氨酸是一种具有碱性胍侧链的氨基酸。它是一氧化氮的直接前体,一氧化氮在皮肤修复中发挥着关键作用。通过高效液相色谱、傅里叶变换红外光谱和拉曼光谱证实了添加剂的存在和分布。使用扫描电子显微镜表征L-精氨酸和阿拉肽对膜形态的影响。未观察到纤维直径和药物浓度之间具有统计学上显着的相关性。然后使用原代人真皮成纤维细胞在体外测试膜的细胞毒性,以获得用于大鼠模型体内测试的添加剂的最佳浓度。具有最高浓度 L-精氨酸(10 wt.%)的膜被证明具有细胞毒性。含有浓度为 0.1 至 2.5 wt.% 的阿拉肽以及含有其他 L-精氨酸浓度(1 和 5 wt.%)的膜没有表现出高毒性。此外,没有观察到膜上细胞增殖的改善。体内实验表明,含有1.5wt.%的alaptide或含有1.5wt.%的alaptide与5wt.%的L-精氨酸组合的膜显着加速了皮肤切口的愈合,特别是皮肤烧伤的愈合,即比较大面积的伤口。这些结果表明,我们新开发的纳米纤维膜有望用于治疗需要支撑材料的大面积受损伤口。

更新日期:2020-11-26
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