title: Study on the Structure of Candida Albicans-Staphylococcus Epidermidis Mixed Species Biofilm on Polyvinyl Chloride Biomaterial.
abstract: The objective of the study was to establish an in vitro model of Candida albicans-Staphylococcus epidermidis mixed species biofilm (BF) on polyvinyl chloride (PVC) material, and to investigate the formation and the structure of mixed species BF formation using a combined approach of confocal laser scanning microscope (CLSM), scanning electron microscope (SEM), and 3D image reconstruction technique. Mixed species BF is achieved by co-incubating Staphylococcus epidermidis bacteria (ATCC35984) and Candida albicans fungal (ATCC10231) with PVC pieces in Tris-buffered saline. BF formation was examined at 2, 6, 12, 24, 48, and 72 h of co-culture. Thickness of these BFs and the number, and percentage of viable cells in BFs were measured. CT scan images of BFs were obtained using CLSM and SEM and reconstructed 3D images of mixed species BF were acquired, in an effort to examine structure of the BF. Staphylococcus epidermidis attached to various forms of candida albicans (spores, pseudohyphae, and hyphae), formed complex and dense mesh arrays. The BF is constituted of a large number of viable and dead pathogens, the surface of mixed species BF is uneven, with living pathogens predominating protrusive portions and dead pathogens aggregating in concaves. Mixed species BF formation on the surface of PVC material was found to be a dynamic process, with rapid growth being at 24 h of co-culture, maximal thickness peaked at 48 h. These mixed species BF matured at 48-72 h. Significant differences were observed in the proportion of viable cells between interior, middle, and outer layers of BFs (p < 0.05). Mixed species BF Candida albicans-Staphylococcus epidermidis is sophisticated in structure. The combined approach involving CLSM, SEM, and 3D image reconstruction technique is ideal for the investigation of mixed species BF on PVC material.
authors.affiliation: Department of Thoracic Surgery, Key Laboratory of Lung Cancer Research of Yunnan Province, The Third Affiliated Hospital of Kunming Medical University, Tumor Hospital of Yunnan Province, Kunming, 650118, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Thoracic Surgery, Key Laboratory of Lung Cancer Research of Yunnan Province, The Third Affiliated Hospital of Kunming Medical University, Tumor Hospital of Yunnan Province, Kunming, 650118, China.
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Wang
authors.affiliation: Department of Thoracic Surgery, Key Laboratory of Lung Cancer Research of Yunnan Province, The Third Affiliated Hospital of Kunming Medical University, Tumor Hospital of Yunnan Province, Kunming, 650118, China. huangych2001@163.com.
authors.forename: Yun-Chao
authors.initials: YC
authors.lastname: Huang
authors.affiliation: Department of Thoracic Surgery, Key Laboratory of Lung Cancer Research of Yunnan Province, The Third Affiliated Hospital of Kunming Medical University, Tumor Hospital of Yunnan Province, Kunming, 650118, China.
authors.forename: Guang-Qiang
authors.initials: GQ
authors.lastname: Zhao
authors.affiliation: Department of Thoracic Surgery, Key Laboratory of Lung Cancer Research of Yunnan Province, The Third Affiliated Hospital of Kunming Medical University, Tumor Hospital of Yunnan Province, Kunming, 650118, China.
authors.forename: Yu-Jie
authors.initials: YJ
authors.lastname: Lei
authors.affiliation: Department of Thoracic Surgery, Key Laboratory of Lung Cancer Research of Yunnan Province, The Third Affiliated Hospital of Kunming Medical University, Tumor Hospital of Yunnan Province, Kunming, 650118, China.
authors.forename: Lian-Hua
authors.initials: LH
authors.lastname: Ye
authors.affiliation: Department of Thoracic Surgery, Key Laboratory of Lung Cancer Research of Yunnan Province, The Third Affiliated Hospital of Kunming Medical University, Tumor Hospital of Yunnan Province, Kunming, 650118, China.
authors.forename: Qiu-Bo
authors.initials: QB
authors.lastname: Huang
authors.affiliation: Department of Thoracic Surgery, Key Laboratory of Lung Cancer Research of Yunnan Province, The Third Affiliated Hospital of Kunming Medical University, Tumor Hospital of Yunnan Province, Kunming, 650118, China.
authors.forename: Wan-Shi
authors.initials: WS
authors.lastname: Duan
mesh_terms: D001672:Biocompatible Materials / Q000737:chemistry / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects* / Q000254:growth & development; D002176:Candida albicans / Q000502:physiology*; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D008954:Models, Biological; D011143:Polyvinyl Chloride / Q000737:chemistry / Q000494:pharmacology*; D013212:Staphylococcus epidermidis / Q000502:physiology*
chemical_list: D001672:Biocompatible Materials; D011143:Polyvinyl Chloride
keywords: 3D image reconstruction; Candida albicans; Confocal laser scanning microscope (CLSM); Mixed species biofilm; Polyvinyl chloride (PVC); Scanning electron microscope (SEM); Staphylococcus epidermidis