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Title Can the Gingival Crevicular Fluid Transcriptome Predict Healing After Dental Trauma?
URL
Publication Date
Degree MS
Discipline/Department Dentistry
Degree Level masters
University/Publisher The Ohio State University
Abstract Can the Gingival Crevicular Fluid Transcriptome Predict Healing After Dental Trauma?, Vollmar C, McTigue D, Kumar P, Subramanian K, Kumar A, (Nationwide Children’s Hospital and The Ohio State University, Columbus, OH)Purpose: Determine if gingival crevicular fluid transcriptome is altered after dental trauma and if these changes persist over time.Methods: Gingival crevicular fluid (GCF) samples were obtained from patients following permanent anterior tooth dental trauma who presented to Nationwide Children’s Hospital Emergency Department or Emergency Dental Clinic. Samples were obtained from the gingival sulcus using PerioPaper strip, placed in a sterile 10ml vial containing RNAlater, and stored at minus twenty degrees Fahrenheit. GCF sampling was repeated at routine follow-up encounters at 2-4 weeks, 6 weeks, 2 months, and 4-6 months when possible. Samples that were analyzed were divided into one of four categories of trauma: subluxation, uncomplicated crown fracture, extrusion, and avulsion. Four patients were selected from each group that demonstrated the most homogenous characteristics for sequencing and data analysis. RNA was isolated using epicenter baseline-ZERO DNase and Ribo-Zero Magnetic Kit. mRNA was enriched using ice-cold ethanol. cDNA conversion was performed using the Scriptseq kit from the stabilized mRNA. Pooled cDNA libraries were clustered on the HiSeq and 250bp paired-end sequencing was performed. The filtered sequences were uploaded to the Kallisto program, which was used to align and quantify abundances of transcripts from the filtered sequences. Sequences were aligned to GRCh38 and estimated gene counts were obtained and uploaded in PANTHER (Protein Annotation Through Evolutionary Relationships) gene analysis platform for analysis. Statistical over-representation test with Bonferroni correction was carried out to obtain the difference in fold enrichment of genes and functions between the groups. Significantly different pathways were visualized using PANTHER mapping systems.Results: At the initial encounter, the avulsion group shows the least number of gene transcript families present, whereas the uncomplicated crown fracture and extrusion groups had the most. By the final encounter, the avulsion group had the most transcript families expressed, while the crown fracture and extrusion groups showed the least. The subluxation group had an intermediate number of transcript families present immediately after dental trauma that slightly increased at the final visit. The quantity and quality of gene expression varied between groups at both the initial and final encounter. Looking at the PANTHER pathway maps, specific pathways were up regulated in different groups.Conclusion: GCF Transcriptome displays changes in gene expression immediately after dental trauma representing a rapid response to injury at the level of the genome. The transcriptome remains elevated overtime in all groups, however, some types of trauma show more of a prolonged response while others return closer to baseline.
Subjects/Keywords Dentistry; Dental Care; dental trauma; dental injury; GCF; human transcriptomics
Contributors McTigue, Dennis (Advisor)
Language en
Rights unrestricted ; This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
Country of Publication us
Format application/pdf
Record ID oai:etd.ohiolink.edu:osu1435011386
Repository ohiolink
Date Indexed 2021-01-29
Grantor The Ohio State University

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…31 viii List of Tables Table 1: Sample characteristics and demographic data .................................................... 15 ix List of Figures Figure 1: Number of gene families differing from GRCh38………………...…………..16 Figure 2: GCF

…Transcriptome immediately following dental trauma…………………...18 Figure 3: GCF Transcriptome at final encounter………………………………………...19 Figure 4: Inflammation mediated by chemokine and cytokine signaling pathway……...23 Figure 5: Toll-like receptor signaling pathway…

…radiographic or clinical 4 signs have manifested. Gingival crevicular fluid (GCF) is a biological substance secreted within the gingival sulcus of a tooth, and it can exist as either a serum transudate or perhaps more commonly in a disease state as…

…periodontal ligament during the healing process 20. Gingival crevicular fluid has been used in numerous studies of periodontal disease and orthodontic root resorption 20-22. GCF is readily available within the periodontal sulcus and its collection is simple…

…cells and biochemical molecules. This inflammatory response could provide valuable insight into healing after dental trauma. Previous studies of the GCF after dental trauma and the differences in cytokine profiles have been conducted at Nationwide…

…thus resulting in better clinical outcomes. Hypothesis: The GCF transcriptome will be altered immediately following dental injury and these changes overtime will correlate with the healing process. 9 Chapter 2: Methods A. Sample Collection Research…

…were individually wiped with gauze to remove supragingival dental plaque, saliva or blood, so as not to contaminate the sample. The teeth were isolated with cotton rolls and gingival crevicular fluid (GCF) samples were taken individually with…

…extrusion first (EF), extrusion last (EL), subluxation first (SF), subluxation last (SL), crown fracture first (CFF), crown fracture last (CFL). When the transcriptome of the GCF from our samples…

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