Salivary miRNA as a potential biomarker of susceptibility to periodontitis / Sharul Nisha Ali

Introduction: Salivary transcriptome is increasingly recognised as a highly potential and futuristic diagnostic fluid since it is readily available, non-invasive and may be used to elucidate the disease susceptibility, including periodontitis. Acknowledging this, salivary microRNA (miRNA), part of t...

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Main Author:	Ali, Sharul Nisha
Format:	Thesis
Language:	English
Published:	2021
Subjects:	Oral and dental medicine Pathology Diseases
Online Access:	https://ir.uitm.edu.my/id/eprint/60910/1/60910.pdf
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spelling	my-uitm-ir.609102022-06-02T08:25:17Z Salivary miRNA as a potential biomarker of susceptibility to periodontitis / Sharul Nisha Ali 2021-11 Ali, Sharul Nisha Oral and dental medicine. Pathology. Diseases Introduction: Salivary transcriptome is increasingly recognised as a highly potential and futuristic diagnostic fluid since it is readily available, non-invasive and may be used to elucidate the disease susceptibility, including periodontitis. Acknowledging this, salivary microRNA (miRNA), part of the transcriptomic biomarkers, may present as the source of a potential biomarker in periodontitis patients. Objective: The general aim of this study is to elucidate the expression of miRNA from the saliva of periodontitis and healthy participants. Specifically, this study aimed to ascertain the feasibility of salivary miRNA as a biomarker in participants susceptible to periodontitis Stage III and IV and compare the expression of novel miRNA in periodontitis and healthy participants. Materials and Methods: A total of 21 research participants (thirteen localised or generalised periodontitis Stage III and IV and eight healthy participants) participated in this study. Whole saliva samples were collected using the unstimulated saliva collection technique. Total RNA, including small RNA, was extracted and purified from the whole saliva using miRNeasy Micro kit from Qiagen. Based on the minimal concentration for library preparation, nine out of 21 purified miRNA samples were converted into complementary DNA (cDNA) from the single-stranded RNA using the reverse transcription-polymerase chain reaction (RT-PCR). Library preparation was prepared, and each sample was given an index for identification before sequencing. Only two samples from healthy (H6) and periodontitis (D12) met the criteria for sequencing with the NextSeq 500/550 from Illumina. Known and novel miRNA expression from the NGS were later subjected to small RNA sequencing analysis. Result: After sequencing with the NGS, 814 novel and 338 known miRNAs identified from D12, while 1035 novel and 365 known miRNAs identified from H6. Based on small RNA sequencing analysis, 16 novel miRNAs were upregulated in the periodontitis group, while 15 novel miRNAs were upregulated in the healthy group. novelMiR_48 was expressed in both groups wherein D12 was upregulated but downregulated in H6. MITF, one of the targeted genes of novelMiR_48, is speculated to be responsible for osteoclast differentiation via the RANK-RANKL-Calcium signaling pathway, hence initiate alveolar bone loss, which is the hallmark for periodontitis. Conclusion: From this study, it can be concluded that novel miRNA expression differs between periodontitis and healthy participants. In addition, salivary miRNA is feasible to be use as a biomarker in patients susceptible to periodontitis Stage III and IV. 2021-11 Thesis https://ir.uitm.edu.my/id/eprint/60910/ https://ir.uitm.edu.my/id/eprint/60910/1/60910.pdf text en public phd doctoral Universiti Teknologi MARA Faculty of Dentistry Hidayat, Mohd Faizal Hafez (Dr.)
institution	Universiti Teknologi MARA
collection	UiTM Institutional Repository
language	English
advisor	Hidayat, Mohd Faizal Hafez (Dr.)
topic	Oral and dental medicine Pathology Diseases
spellingShingle	Oral and dental medicine Pathology Diseases Ali, Sharul Nisha Salivary miRNA as a potential biomarker of susceptibility to periodontitis / Sharul Nisha Ali
description	Introduction: Salivary transcriptome is increasingly recognised as a highly potential and futuristic diagnostic fluid since it is readily available, non-invasive and may be used to elucidate the disease susceptibility, including periodontitis. Acknowledging this, salivary microRNA (miRNA), part of the transcriptomic biomarkers, may present as the source of a potential biomarker in periodontitis patients. Objective: The general aim of this study is to elucidate the expression of miRNA from the saliva of periodontitis and healthy participants. Specifically, this study aimed to ascertain the feasibility of salivary miRNA as a biomarker in participants susceptible to periodontitis Stage III and IV and compare the expression of novel miRNA in periodontitis and healthy participants. Materials and Methods: A total of 21 research participants (thirteen localised or generalised periodontitis Stage III and IV and eight healthy participants) participated in this study. Whole saliva samples were collected using the unstimulated saliva collection technique. Total RNA, including small RNA, was extracted and purified from the whole saliva using miRNeasy Micro kit from Qiagen. Based on the minimal concentration for library preparation, nine out of 21 purified miRNA samples were converted into complementary DNA (cDNA) from the single-stranded RNA using the reverse transcription-polymerase chain reaction (RT-PCR). Library preparation was prepared, and each sample was given an index for identification before sequencing. Only two samples from healthy (H6) and periodontitis (D12) met the criteria for sequencing with the NextSeq 500/550 from Illumina. Known and novel miRNA expression from the NGS were later subjected to small RNA sequencing analysis. Result: After sequencing with the NGS, 814 novel and 338 known miRNAs identified from D12, while 1035 novel and 365 known miRNAs identified from H6. Based on small RNA sequencing analysis, 16 novel miRNAs were upregulated in the periodontitis group, while 15 novel miRNAs were upregulated in the healthy group. novelMiR_48 was expressed in both groups wherein D12 was upregulated but downregulated in H6. MITF, one of the targeted genes of novelMiR_48, is speculated to be responsible for osteoclast differentiation via the RANK-RANKL-Calcium signaling pathway, hence initiate alveolar bone loss, which is the hallmark for periodontitis. Conclusion: From this study, it can be concluded that novel miRNA expression differs between periodontitis and healthy participants. In addition, salivary miRNA is feasible to be use as a biomarker in patients susceptible to periodontitis Stage III and IV.
format	Thesis
qualification_name	Doctor of Philosophy (PhD.)
qualification_level	Doctorate
author	Ali, Sharul Nisha
author_facet	Ali, Sharul Nisha
author_sort	Ali, Sharul Nisha
title	Salivary miRNA as a potential biomarker of susceptibility to periodontitis / Sharul Nisha Ali
title_short	Salivary miRNA as a potential biomarker of susceptibility to periodontitis / Sharul Nisha Ali
title_full	Salivary miRNA as a potential biomarker of susceptibility to periodontitis / Sharul Nisha Ali
title_fullStr	Salivary miRNA as a potential biomarker of susceptibility to periodontitis / Sharul Nisha Ali
title_full_unstemmed	Salivary miRNA as a potential biomarker of susceptibility to periodontitis / Sharul Nisha Ali
title_sort	salivary mirna as a potential biomarker of susceptibility to periodontitis / sharul nisha ali
granting_institution	Universiti Teknologi MARA
granting_department	Faculty of Dentistry
publishDate	2021
url	https://ir.uitm.edu.my/id/eprint/60910/1/60910.pdf
_version_	1783735183234564096

Salivary miRNA as a potential biomarker of susceptibility to periodontitis / Sharul Nisha Ali

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