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Complete Thesis _ Angelika Dzieza5.70 MB
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Thesis
Assessing the biological effect of a novel non-thermal plasma (NTP) gas on periodontal disease
University of Iowa
Master of Science (MS), University of Iowa
Spring 2025
DOI: 10.25820/etd.007941
Abstract
Introduction:
Periodontitis is a prevalent chronic inflammatory disease affecting tooth-supporting structures associated with an exaggerated host immune response to plaque dysbiotic bacteria. Affecting 42% of U.S. adults over 30, it leads to tooth loss, if not managed properly, and increases systemic inflammatory burden. Conventional treatments, such as scaling and root planing (SRP), often struggle to remove biofilms hiding in the deep periodontal pockets, necessitating adjunctive therapies. Non-thermal plasma (NTP) gas, composed of reactive oxygen and nitrogen species (RONS), has emerged as a novel therapeutic approach due to its antimicrobial and tissue-regenerative properties. Preclinical studies indicate that NTP disrupts bacterial biofilms, modulates immune responses, and enhances periodontal tissue repair.
The purpose of this study is to evaluate NTP’s effects on inflammatory gene expression in gingival epithelial cells and its impact on alveolar bone loss in an experimentally induced murine periodontitis model. The data will provide evidence to evaluate the potential application of NTP in clinic as a minimally invasive adjunctive therapy for improving periodontitis treatment outcomes.
Materials and Methods:
A ligature-induced periodontitis (LIP) model was established in C57BL/6 mice (n=9), with NTP treatment applied at 6- and 48-hours post-ligation. Mice were euthanized on day 10 for micro-CT imaging to assess bone loss and RNA-sequencing (RNA-seq) analysis for gene expression. Histological analyses, including tartrate-resistant acid phosphatase (TRAP), hematoxylin and eosin (H&E), and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, were performed to evaluate osteoclast activity and tissue necrosis. In iv vitro, primary human gingival keratinocytes (TIGK cells) were exposed to NTP under various conditions, with or without E. coli lipopolysaccharide (LPS) co-stimulation, with RT-qPCR used to quantify inflammatory gene expression. Statistical analysis included unpaired Student’s t-test (bone loss) and one-way ANOVA (RT-qPCR), while RNA-seq data were analyzed using DESeq2 with gene ontology (GO) enrichment.
Results:
NTP treatment significantly reduced alveolar bone loss in LIP mice, as evidenced by micro-CT analysis (0.58 ± 0.16 mm vs. 0.84 ± 0.07 mm, p = 0.032). Histological analysis confirmed tissue integrity upon NTP treatment. TUNEL staining showed no increase in apoptosis, while TRAP staining revealed reduced osteoclast activity by NTP treatment, suggesting inhibition of bone resorption. RNA-seq analysis identified significant transcriptional changes in genes related to epithelial integrity, connective tissue repair, and immune modulation. GO analysis highlighted enrichment in pathways involved in keratin filament organization, leukocyte activation, and VEGF production. KEGG pathway analysis revealed enrichment in inflammatory signaling and metabolic pathways. In vitro, NTP exposure upregulated inflammatory (CXCL8/IL8, CCL20, ICAM), microbial defensive (DEFB4B), and epithelial repair genes (EPCAM), particularly in the presence of oxygen, suggesting NTP enhances mucosal immune responses and barrier function.
Conclusion:
NTP demonstrates promising therapeutic potential in periodontitis by modulating host responses and preserving tissue integrity. In the murine model, NTP reduced alveolar bone loss, maintained epithelial homeostasis, and decreased osteoclast activity, indicating its protective role against bone resorption. In vitro, NTP enhanced mucosal immune responses and epithelial v barrier function by upregulating inflammatory and repair-associated genes. These findings highlight NTP’s dual capability of mitigating periodontal tissue destruction and potentially enhancing epithelial regeneration. With further research and clinical evaluation, NTP could emerge as a valuable adjunctive therapy to improve periodontal treatment outcomes and overall oral health.
Details
- Title: Subtitle
- Assessing the biological effect of a novel non-thermal plasma (NTP) gas on periodontal disease
- Creators
- Angelika Dzieza
- Contributors
- Shaoping Zhang (Advisor)Erliang Zeng (Committee Member)Sukirth Ganesan (Committee Member)Sivaraman Prakasam (Committee Member)
- Resource Type
- Thesis
- Degree Awarded
- Master of Science (MS), University of Iowa
- Degree in
- Oral Science
- Date degree season
- Spring 2025
- DOI
- 10.25820/etd.007941
- Publisher
- University of Iowa
- Number of pages
- xi, 64 pages
- Copyright
- Copyright 2025 Angelika Dzieza
- Language
- English
- Date submitted
- 04/16/2025
- Description illustrations
- Illustrations, tables, graphs, charts
- Description bibliographic
- Includes bibliographical references (pages 58-64).
- Public Abstract (ETD)
Periodontitis is a common inflammatory disease affecting the gums and tooth-supporting structures, leading to tooth loss and systemic health complications. It affects approximately 42% of U.S. adults over 30 and is initiated by bacterial biofilms and an exaggerated immune response. Conventional treatments like scaling and root planing (SRP) often fail to eliminate biofilms in deep periodontal pockets, necessitating alternative therapies. Non-thermal plasma (NTP) gas, composed of reactive oxygen and nitrogen species (RONS), has emerged as a novel treatment due to its antimicrobial, host-modulatory, and tissue-regenerative properties. This study aims to evaluate NTP’s effects on inflammatory gene expression in gingival epithelial cells and its impact on alveolar bone loss in a murine periodontitis model.
In the pre-clinical animal periodontitis model, NTP treatment significantly reduced bone loss compared to untreated controls. Micro-CT analysis showed decreased alveolar bone resorption, while histological analysis confirmed preserved tissue integrity and reduced osteoclast activity, suggesting NTP’s protective effect against bone degradation. Additionally, TUNEL staining indicated no increase in apoptosis, confirming its safety.
In vitro, primary human gingival epithelial cells treated with NTP demonstrated increased expression of genes associated with immune response, epithelial integrity, and tissue repair, particularly when oxygen and argon plasma gas were present. These findings suggest that NTP enhances immune modulation and epithelial barrier function, further supporting its therapeutic potential. Together, these results indicate that NTP could serve as a minimally invasive adjunctive therapy for periodontitis by reducing inflammation, mitigating bone loss, and promoting tissue regeneration. Further research and clinical trials may establish NTP as an innovative tool for improving periodontal treatment outcomes.
- Academic Unit
- Oral Pathology, Radiology and Medicine
- Record Identifier
- 9984831231002771
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