Pinilla Yudi T, Patarroyo Manuel A, Velandia Myriam L, Segura Nidya A, Bello Felio J
Medical and Forensic Entomology Research Group, School of Medicine and Health Sciences, Universidad del Rosario, Bogotá, Colombia.
Basic Sciences Department, School of Medicine and Health Sciences, Universidad del Rosario, Bogotá, Colombia; Molecular Biology and Immunology Department, Fundación Instituto de Inmunología de Colombia, Bogotá, Colombia.
Acta Trop. 2015 Feb;142:26-33. doi: 10.1016/j.actatropica.2014.11.003. Epub 2014 Nov 8.
Sarconesiopsis magellanica is a necrophagous blowfly which is relevant in both forensic and medical sciences. Previous studies regarding this species have led to understanding life-cycle, population and reproduction parameters, as well as identifying and characterising proteolytic enzymes derived from larval excretions and secretions (ES). As other studies have shown that ES proteolytic activity plays a significant role in wound healing and fibroblasts play a relevant role in granulation tissue formation during such healing, the present study was aimed at analysing the biological effect of S. magellanica larval ES on fibroblasts. ES were obtained from third-instar larvae and added to fibroblast cells at three concentrations (10, 5 and 1 μg/mL) to evaluate their behaviour. MTT assays were used for analysing cell proliferation and viability, whilst cell adhesion was measured by optical density with 10% SDS. Fibroblast migration and morphology was recorded by microscopic observation. ES did not affect fibroblast viability and induced an increase in cell proliferation; cell adhesion became reduced, whilst cell migration through extracellular matrix increased. ES also induced a decreased cell surface and morphological alterations. Changes in all the above-mentioned parameters were reduced when ES were incubated at 60 °C, probably due to protease denaturation. These results suggested that the proteases contained in S. magellanica larval ES contributed towards granulation tissue formation, increased cell migration and promoted cell proliferation. All these data support carrying out further experiments aimed at validating S. magellanica usefulness in larval therapy.
麦哲伦拟肉蝇是一种食尸性丽蝇,在法医学和医学领域都具有重要意义。此前针对该物种的研究有助于了解其生命周期、种群和繁殖参数,以及鉴定和表征源自幼虫排泄物和分泌物(ES)的蛋白水解酶。由于其他研究表明,ES的蛋白水解活性在伤口愈合中发挥着重要作用,而成纤维细胞在这种愈合过程中的肉芽组织形成中起着相关作用,因此本研究旨在分析麦哲伦拟肉蝇幼虫ES对成纤维细胞的生物学效应。从三龄幼虫中获取ES,并以三种浓度(10、5和1μg/mL)添加到成纤维细胞中,以评估其行为。采用MTT法分析细胞增殖和活力,同时用10%十二烷基硫酸钠通过光密度测量细胞粘附。通过显微镜观察记录成纤维细胞的迁移和形态。ES不影响成纤维细胞的活力,并诱导细胞增殖增加;细胞粘附减少,而细胞通过细胞外基质的迁移增加。ES还诱导细胞表面减少和形态改变。当ES在60℃孵育时,上述所有参数的变化均减少,这可能是由于蛋白酶变性所致。这些结果表明,麦哲伦拟肉蝇幼虫ES中含有的蛋白酶有助于肉芽组织形成、增加细胞迁移并促进细胞增殖。所有这些数据支持开展进一步实验,以验证麦哲伦拟肉蝇在幼虫疗法中的效用。