Jayatilake J A M S, Samaranayake Y H, Samaranayake L P
Microbiology, Department of Oral Medicine and Periodontology, Faculty of Dental Sciences, University of Peradeniya, Peradeniya, Sri Lanka.
Mycopathologia. 2008 Jun;165(6):373-80. doi: 10.1007/s11046-008-9096-1. Epub 2008 Mar 5.
The purpose of this study is to compare the light and scanning electron microscopic (SEM) features of tissue invasion by three Candida species (C. albicans, C. tropicalis, and C. dubliniensis) in two different tissue culture models: rabbit tongue mucosal explants (RTME) and reconstituted human oral epithelium (RHOE). Tongue mucosal biopsies of healthy New Zealand rabbits were maintained in explant culture using a transwell system. RHOE was obtained from Skinethic Laboratory (Nice, France). RTME and RHOE were inoculated with C. albicans, C. tropicalis, and C. dubliniensis separately and incubated at 37 degrees C, 5% CO(2), and 100% humidity up to 48 h. Light microscopic and SEM examinations of uninfected (controls) and infected tissues were performed at 24 and 48 h. C. albicans produced characteristic hallmarks of pathological tissue invasion in both tissue models over a period of 48 h. Hyphae penetrated through epithelial cells and intercellular gaps latter resembling thigmotropism. SEM showed cavitations on the epithelial cell surfaces particularly pronounced at sites of hyphal invasion. Some hyphae on RTME showed several clusters of blastospores attached in regular arrangements resembling "appareil sporifere". C. tropicalis and C. dubliniensis produced few hyphae mainly on RTME but they did not penetrate either model. Our findings indicate that multiple host-fungal interactions such as cavitations, thigmotropism, and morphogenesis take place during candidal tissue invasion. RTME described here appears to be useful in investigations of such pathogenic processes of Candida active at the epithelial front.
本研究的目的是比较三种念珠菌(白色念珠菌、热带念珠菌和都柏林念珠菌)在两种不同组织培养模型中组织侵袭的光学显微镜和扫描电子显微镜(SEM)特征:兔舌黏膜外植体(RTME)和重组人口腔上皮(RHOE)。健康新西兰兔的舌黏膜活检组织使用Transwell系统维持在外植体培养中。RHOE购自法国尼斯的Skinethic实验室。将RTME和RHOE分别接种白色念珠菌、热带念珠菌和都柏林念珠菌,并在37℃、5%二氧化碳和100%湿度下孵育长达48小时。在24小时和48小时对未感染(对照)和感染组织进行光学显微镜和SEM检查。在48小时内,白色念珠菌在两种组织模型中均产生了病理性组织侵袭的特征性标志。菌丝穿透上皮细胞和细胞间隙,后者类似于向触性。SEM显示上皮细胞表面有空洞,在菌丝侵袭部位尤为明显。RTME上的一些菌丝显示有几簇芽生孢子规则排列附着,类似于“孢子形成装置”。热带念珠菌和都柏林念珠菌主要在RTME上产生少量菌丝,但它们未穿透任何一种模型。我们的研究结果表明,在念珠菌组织侵袭过程中发生了多种宿主 - 真菌相互作用,如空洞形成、向触性和形态发生。本文描述的RTME似乎有助于研究念珠菌在上皮前沿活跃的此类致病过程。