Three/four-dimensional (3D/4D) microscopic imaging and processing in clinical dental research

BMC Oral Health

Research Article

Mojgan Houshmandi¹  Ping Ye²  Hong Yu³ 

[1] Institute of Dental Research, Oral Health, Westmead Hospital, Westmead, Australia;Institute of Dental Research, Oral Health, Westmead Hospital, Westmead, Australia;Affiliation of Faculty of Dentistry, the University of Sydney, Sydney, Australia;Microscopy Laboratory, Westmead Institute for Medical Research, Westmead, Australia;

关键词: 3D reconstruction;    4D imaging;    Live cell imaging;    Carious lesion;    Periodontitis;    Porphyromonas gingivalis;   

DOI  :  10.1186/s12903-016-0282-0

 received in 2016-04-26, accepted in 2016-08-20,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundConfocal laser scanning microscope (CLSM) has been widely employed in our laboratory for structural and functional analysis of clinical dental specimens and live cell imaging of cultured oral epithelial cells.MethodsIn this vitro study, a Fluoview 1000 (Olympus) confocal system was utilised to study thick sections of carious lesions (40–100 μm) and periodontal disease tissue samples (20–40 μm) by 2D Z stacking imaging and 3-dimentional (3D) reconstruction. Four-dimensional (4D) imaging when including time or position points was used for live cells to assess penetration/localisation/co-localization of oral pathogen proteins and therapeutic drugs.ResultsThree-dimensional (3D) reconstruction revealed latent features of carious hard tissues (strongly expressed amelogenin proteins in dentin tubules), and soft tissues (increased glial markers GFAP and S100B in pulp components). We also found the oral microbial specific pathogens, Porphyromonas gingivalis to be widely localised inside the periodontal pocket epithelial tissues as detected by 3D reconstruction from a series of 2D sections from periodontal disease tissue samples. 4D live cell imaging showed the diffusion patterns of fluorescent molecules in response to a bacterial virulence factor, the pathogen (gingipain haemagglutinin) domain that attacked epithelial integrity. This technology also showed uptake of a novel porphyrin-linked metronidazole antibiotic into epithelial cells to kill intracellular oral pathogen, P. gingivalis.ConclusionsThree/four-dimensional (3D/4D) imaging and processing in confocal microscopy is of great interest and benefit to clinical dental researchers.

【 授权许可】

CC BY   
© The Author(s). 2016