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Apoptosis Regulation in Dental Pulp Cells and PD-1/PD-L1 Expression Dynamics Under Ozone Exposure – A Pilot Approach

Archivum Immunologiae et Therapiae Experimentalis
Volume 72 (2024): Issue 1 (January 2024)
By:
Open Access
|Sep 2024

Figures & Tables

Fig 1.

An example of the flow cytometry analysis of antigens expression on T and B lymphocytes. (A) Gating strategy for lymphocytes as well as their subpopulations, B lymphocytes (CD19+) and T lymphocytes (CD3+); (B) Gating strategy for Tc (CD8+) and Th (CD4+) lymphocytes among T cell subpopulations (CD3+); (C) Strategy for gating PD-1 expression among the Th cell population; (D) Gating strategy for PD-1 expression among the Tc cell population; (E) Gating strategy for PD-1 expression among the B cell population. PD-1, programmed cell death protein 1.
An example of the flow cytometry analysis of antigens expression on T and B lymphocytes. (A) Gating strategy for lymphocytes as well as their subpopulations, B lymphocytes (CD19+) and T lymphocytes (CD3+); (B) Gating strategy for Tc (CD8+) and Th (CD4+) lymphocytes among T cell subpopulations (CD3+); (C) Strategy for gating PD-1 expression among the Th cell population; (D) Gating strategy for PD-1 expression among the Tc cell population; (E) Gating strategy for PD-1 expression among the B cell population. PD-1, programmed cell death protein 1.

Fig 2.

Average relative expression of Bcl-2 and Bax genes (logRQ) in dental pulp cells cultures depending on the ozonation time compared to cells without ozone treatment. The cells not subjected to ozonation (ozone dose 0 s.) or exposed to ozone for 6 s or 12 s were analyzed for the expression of Bax and Bcl-2. Data are representative of six independent experiments, each repeated in duplicate ± SD; *Indicates significant difference between two indicated groups (p < 0.05, ANOVA test).
Average relative expression of Bcl-2 and Bax genes (logRQ) in dental pulp cells cultures depending on the ozonation time compared to cells without ozone treatment. The cells not subjected to ozonation (ozone dose 0 s.) or exposed to ozone for 6 s or 12 s were analyzed for the expression of Bax and Bcl-2. Data are representative of six independent experiments, each repeated in duplicate ± SD; *Indicates significant difference between two indicated groups (p < 0.05, ANOVA test).

Fig 3.

Differences in B lymphocyte measurements before and after ozonation. The percentages of CD19+ B lymphocytes (A) expressing PD-1 (B) and PD-L1 (C) molecules in the studied cell populations isolated from dental pulp of healthy donors (n=15) before and after ozonation. CD19+CD200 expression (D) and CD19+CD200R expression in B lymphocytes (E) before and after ozonation. Dots represent the cases (n = 15), (—) median, (×) mean, whiskers indicate quartiles. PD-1, programmed cell death protein 1; PD-L1, programmed death-ligand 1.
Differences in B lymphocyte measurements before and after ozonation. The percentages of CD19+ B lymphocytes (A) expressing PD-1 (B) and PD-L1 (C) molecules in the studied cell populations isolated from dental pulp of healthy donors (n=15) before and after ozonation. CD19+CD200 expression (D) and CD19+CD200R expression in B lymphocytes (E) before and after ozonation. Dots represent the cases (n = 15), (—) median, (×) mean, whiskers indicate quartiles. PD-1, programmed cell death protein 1; PD-L1, programmed death-ligand 1.

Fig 4.

Differences in T lymphocyte measurements before and after ozonation. The percentages of CD4+ (A) and CD8+ (B) T lymphocytes expressing PD-1 (B, E) and PD-L1 (C, F) molecules in the studied cell populations isolated from dental pulp of healthy donors (n = 15) before and after ozonation. Dots represent the cases (n = 15), (—) median, (×) mean, whiskers indicate quartiles. PD-1, programmed cell death protein 1; PD-L1, programmed death-ligand 1.
Differences in T lymphocyte measurements before and after ozonation. The percentages of CD4+ (A) and CD8+ (B) T lymphocytes expressing PD-1 (B, E) and PD-L1 (C, F) molecules in the studied cell populations isolated from dental pulp of healthy donors (n = 15) before and after ozonation. Dots represent the cases (n = 15), (—) median, (×) mean, whiskers indicate quartiles. PD-1, programmed cell death protein 1; PD-L1, programmed death-ligand 1.

Bax/Bcl-2 expression ratio in response to ozonation of dental pulp cells

Ozonation time (s)Bax mean (logRQ)Bcl-2 mean (logRQ)Bax/Bcl-2 ratiop-value
00.00480.057300.199
60.1883−0.0903−20.0011*
120.16110.30.50.0005*,**

Medium conductivity (mS) depending on time of ozonation and time after ozonation

Ozonation time (s)Conductivity measurement after ozonation (mS) Average of 5 measurements, deviation ± 1 mS
Immediately after ozonationAfter 24 h of keeping medium in 4°CAfter 48 h of keeping medium in 4°C
612119
12151210
DOI: https://doi.org/10.2478/aite-2024-0019 | Journal eISSN: 1661-4917
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Language: English
Submitted on: Apr 3, 2024
Accepted on: Aug 12, 2024
Published on: Sep 14, 2024
Published by: Hirszfeld Institute of Immunology and Experimental Therapy
In partnership with: Paradigm Publishing Services
Publication frequency: 1 times per year
Keywords:
Ozone,
Apoptosis,
Dental pulp cells,
CD19 B lymphocyte,
CD4 T lymphocyte,
CD8 T lymphocyte
Related subjects:
Medicine,
Basic medical science,
Biochemistry,
Immunology,
Clinical medicine,
Clinical medicine, other,
Clinical chemistry

© 2024 Maja Ptasiewicz, Mirosław Orłowski, Agnieszka Magryś, Janusz Kocki, Krzysztof Gosik, Piotr Stachurski, Renata Chałas, published by Hirszfeld Institute of Immunology and Experimental Therapy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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