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Atomic Force Microscopy: A three-dimensional reconstructive tool of oral microbiota in gingivitis and periodontitis

AIM: This study aims to ascertain the advantages of Atomic Force Microscopy (AFM) in the morphologic study of microorganisms and their interactions within the subgingival biofilm in patients with gingivitis and periodontitis. SETTINGS AND DESIGN: Conducted a study on twenty patients, ten patients wi...

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Autores principales: Salavadhi, Shyam Sunder, Chintalapani, Srikanth, Ramachandran, Radhika, Nagubandi, Kirankumar, Ramisetti, Arpita, Boyapati, Ramanarayana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Medknow Publications & Media Pvt Ltd 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5813339/
https://www.ncbi.nlm.nih.gov/pubmed/29456299
http://dx.doi.org/10.4103/jisp.jisp_209_17
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author Salavadhi, Shyam Sunder
Chintalapani, Srikanth
Ramachandran, Radhika
Nagubandi, Kirankumar
Ramisetti, Arpita
Boyapati, Ramanarayana
author_facet Salavadhi, Shyam Sunder
Chintalapani, Srikanth
Ramachandran, Radhika
Nagubandi, Kirankumar
Ramisetti, Arpita
Boyapati, Ramanarayana
author_sort Salavadhi, Shyam Sunder
collection PubMed
description AIM: This study aims to ascertain the advantages of Atomic Force Microscopy (AFM) in the morphologic study of microorganisms and their interactions within the subgingival biofilm in patients with gingivitis and periodontitis. SETTINGS AND DESIGN: Conducted a study on twenty patients, ten patients with severe periodontitis with probing the pocket depth of ≥8 mm, with a clinical attachment loss (CAL) of ≥6 mm CAL and ten patients with gingivitis: ≥5 mm pocket depth, and no attachment loss, was selected for the study. MATERIALS AND METHODS: Bacterial biofilms were collected and slide preparation done. Morphological study was done using AFM. AFM consists of a cantilever-mounted tip, a piezoelectric scanner, a photodetector diode, a laser diode, and a feedback control. The laser beam is reflected from back of the cantilever into the quadrant of the photodetector. AFM works on the principle of interaction between the tip and the sample which causes the cantilever to deflect, thereby changing the position of laser onto the photodetector. Methodology used for studying the bacteria through AFM includes the following: (1) Probe type: Platinum coated silicon nitrate tip. (2) Probe force: 0.11 N/m. (3) Probe geometry: Triangular shaped tip. (4) Probe frequency: 22 KHz. (5) Probe immobilization: Used in Contact mode. AFM Solver Pro-M (NT-MDT) equipped with ETALON probe was used to take images in Nova software. RESULTS: The investigation showed various morphological features, such as shape, size, and secretory product-like vesicles of the bacterial species involved in gingivitis and periodontitis. More bacterial surface details were studied by reproducing a three-dimensional reconstruction using AFM. CONCLUSIONS: The morphological variations of bacteria of different sizes, and shapes, cell wall structures, secretory product-like vesicles flagellated and filamentous microorganisms, polymorphonuclear leukocytes, and bacterial coaggregation analysis were done by AFM. Results of the present study conclude that AFM is a quite a reliable method for studying morphology of bacterial species involving periodontal diseases and is also used to study microbial interactions in biofilm.
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spelling pubmed-58133392018-02-16 Atomic Force Microscopy: A three-dimensional reconstructive tool of oral microbiota in gingivitis and periodontitis Salavadhi, Shyam Sunder Chintalapani, Srikanth Ramachandran, Radhika Nagubandi, Kirankumar Ramisetti, Arpita Boyapati, Ramanarayana J Indian Soc Periodontol Original Article AIM: This study aims to ascertain the advantages of Atomic Force Microscopy (AFM) in the morphologic study of microorganisms and their interactions within the subgingival biofilm in patients with gingivitis and periodontitis. SETTINGS AND DESIGN: Conducted a study on twenty patients, ten patients with severe periodontitis with probing the pocket depth of ≥8 mm, with a clinical attachment loss (CAL) of ≥6 mm CAL and ten patients with gingivitis: ≥5 mm pocket depth, and no attachment loss, was selected for the study. MATERIALS AND METHODS: Bacterial biofilms were collected and slide preparation done. Morphological study was done using AFM. AFM consists of a cantilever-mounted tip, a piezoelectric scanner, a photodetector diode, a laser diode, and a feedback control. The laser beam is reflected from back of the cantilever into the quadrant of the photodetector. AFM works on the principle of interaction between the tip and the sample which causes the cantilever to deflect, thereby changing the position of laser onto the photodetector. Methodology used for studying the bacteria through AFM includes the following: (1) Probe type: Platinum coated silicon nitrate tip. (2) Probe force: 0.11 N/m. (3) Probe geometry: Triangular shaped tip. (4) Probe frequency: 22 KHz. (5) Probe immobilization: Used in Contact mode. AFM Solver Pro-M (NT-MDT) equipped with ETALON probe was used to take images in Nova software. RESULTS: The investigation showed various morphological features, such as shape, size, and secretory product-like vesicles of the bacterial species involved in gingivitis and periodontitis. More bacterial surface details were studied by reproducing a three-dimensional reconstruction using AFM. CONCLUSIONS: The morphological variations of bacteria of different sizes, and shapes, cell wall structures, secretory product-like vesicles flagellated and filamentous microorganisms, polymorphonuclear leukocytes, and bacterial coaggregation analysis were done by AFM. Results of the present study conclude that AFM is a quite a reliable method for studying morphology of bacterial species involving periodontal diseases and is also used to study microbial interactions in biofilm. Medknow Publications & Media Pvt Ltd 2017 /pmc/articles/PMC5813339/ /pubmed/29456299 http://dx.doi.org/10.4103/jisp.jisp_209_17 Text en Copyright: © 2018 Indian Society of Periodontology http://creativecommons.org/licenses/by-nc-sa/3.0 This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.
spellingShingle Original Article
Salavadhi, Shyam Sunder
Chintalapani, Srikanth
Ramachandran, Radhika
Nagubandi, Kirankumar
Ramisetti, Arpita
Boyapati, Ramanarayana
Atomic Force Microscopy: A three-dimensional reconstructive tool of oral microbiota in gingivitis and periodontitis
title Atomic Force Microscopy: A three-dimensional reconstructive tool of oral microbiota in gingivitis and periodontitis
title_full Atomic Force Microscopy: A three-dimensional reconstructive tool of oral microbiota in gingivitis and periodontitis
title_fullStr Atomic Force Microscopy: A three-dimensional reconstructive tool of oral microbiota in gingivitis and periodontitis
title_full_unstemmed Atomic Force Microscopy: A three-dimensional reconstructive tool of oral microbiota in gingivitis and periodontitis
title_short Atomic Force Microscopy: A three-dimensional reconstructive tool of oral microbiota in gingivitis and periodontitis
title_sort atomic force microscopy: a three-dimensional reconstructive tool of oral microbiota in gingivitis and periodontitis
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5813339/
https://www.ncbi.nlm.nih.gov/pubmed/29456299
http://dx.doi.org/10.4103/jisp.jisp_209_17
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