Thesis
The effect of non-demineralized dentin matrix components and induced inflammation on dental pulp stem cells
University of Iowa
Master of Science (MS), University of Iowa
Summer 2023
DOI: 10.25820/etd.006938
Abstract
Introduction: During the caries process, the secretion of acid metabolites by the microbiome of carious lesions results in the demineralization of dentin. This process liberates dentin matrix components that influence the inflammatory and reparative processes of the dental pulp in response to caries.
Methods: Dentin matrix components were extracted from sound, non-carious permanent third molars, purified, and concentrated. Undigested components and components digested by a cocktail of matrix metalloproteinases were used to stimulate third passage dental pulp stem cells alone and in multiple combinations with and without lipopolysaccharide and/or lipoteichoic acid for 24 hours. Cytokine production and gene expression changes were evaluated using LEGENDplex and RNA-sequencing, respectively. Parametric and nonparametric statistical analysis was performed as appropriate to compare differences among multiple groups (α = 0.05).
Results: The cytokines CXCL10 (IP-10), CCL2 (MCP-1), IL-6, and CXCL8 (IL-8) were produced by dental pulp stem cells. No differentially expressed genes were detected in pairwise comparisons not involving a bacterial byproduct or matrix metalloproteinases. Variation between dental pulp stem cell donors was significant for differential gene expression.
Conclusion: Dental pulp stem cells respond to induced inflammation, matrix metalloproteinases, and dentin matrix components by modifying cytokine production and gene expression. Differences between individuals are a significant factor for variations in gene expression. The modulation of matrix metalloproteinase activity may be valuable for repair of the pulp-dentin complex.
Details
- Title: Subtitle
- The effect of non-demineralized dentin matrix components and induced inflammation on dental pulp stem cells
- Creators
- Christopher T Hogden
- Contributors
- Cristina De Mattos Pimenta Vidal (Advisor)Carolina Cucco (Committee Member)Fabricio Teixeira (Committee Member)Sukirth M Ganesan (Committee Member)
- Resource Type
- Thesis
- Degree Awarded
- Master of Science (MS), University of Iowa
- Degree in
- Oral Science
- Date degree season
- Summer 2023
- DOI
- 10.25820/etd.006938
- Publisher
- University of Iowa
- Number of pages
- xii, 122 pages
- Copyright
- Copyright 2023 Christopher T Hogden
- Language
- English
- Date submitted
- 07/25/2023
- Description illustrations
- tables, graphs
- Description bibliographic
- Includes bibliographical references (pages 108-122).
- Public Abstract (ETD)
- Carious lesions (i.e., cavities) result in the breakdown of tooth structure. Underneath tooth enamel resides a tissue known as dentin. This mineralized tissue is made by specialized cells known as odontoblasts. During development, odontoblasts secrete proteins essential for dentin formation. When a cavity reaches dentin, these proteins are released, which influences the level of inflammation in the dental pulp. The dentin proteins can be modified by enzymes known as proteases, which can digest other proteins. Protease modifications can change the behavior of dentin proteins favoring repair over continued inflammation. To investigate the effects of these proteins with and without protease modification, we cultured these proteins with stem cells from the dental pulp and bacterial byproducts in various combinations. After 24 hours, we evaluated the signaling proteins (cytokines and chemokines) released by the stem cells using a technique known as flow cytometry. This method detects special dyes attached to the signaling proteins using several lasers. Additionally, we removed RNA from the stem cells and determined its sequence using a technique known as RNA-sequencing. This reveals which genes are expressed following each of the treatments. We found dental pulp stem cells respond to induced inflammation, proteases, and dentin proteins by modifying their cytokine production and gene expression. Differences between individuals may be a significant factor in the response to bacterial byproducts or the proteins from dentin. This information might assist in the development of new techniques for the treatment of cavities.
- Academic Unit
- Oral Pathology, Radiology and Medicine
- Record Identifier
- 9984454318802771
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