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Can the Gingival Crevicular Fluid Transcriptome Predict Healing After Dental Trauma?

Vollmar, Christine
http://rave.ohiolink.edu/etdc/view?acc_num=osu1435011386

Abstract Details

2015, Master of Science, Ohio State University, Dentistry.
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.
Dennis McTigue, DDS, MS (Advisor)
Purnima Kumar, BDS, MS, PhD (Committee Member)
Kumar Subramanian, DDS, MSD (Committee Member)
Ashok Kumar, DDS, MS (Committee Member)
44 p.
Dental Care; Dentistry
dental trauma; dental injury; GCF; human transcriptomics

Recommended Citations

Citations

  • Vollmar, C. (2015). Can the Gingival Crevicular Fluid Transcriptome Predict Healing After Dental Trauma? [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1435011386

    APA Style (7th edition)

  • Vollmar, Christine. Can the Gingival Crevicular Fluid Transcriptome Predict Healing After Dental Trauma? 2015. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1435011386.

    MLA Style (8th edition)

  • Vollmar, Christine. "Can the Gingival Crevicular Fluid Transcriptome Predict Healing After Dental Trauma?" Master's thesis, Ohio State University, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=osu1435011386

    Chicago Manual of Style (17th edition)

osu1435011386
501
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This open access ETD is published by The Ohio State University and OhioLINK.