MMP-8 in Periodontal Sites of Postpartum and without-Any-Pregnancy Women
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Design
2.2. Sample Selection
2.3. Data Collection
2.4. Periodontal Assessment
- Probing pocket depth (PPD): measured with a millimeter periodontal probe, determined as the distance from the gingival margin to the bottom of the sulcus or periodontal pocket.
- Bleeding on probing (BOP): the occurrence of probing bleeding was recorded 10 s after removing the millimeter probe inside the groove or periodontal pocket [19].
- Clinical attachment level (CAL): distance in millimeters between the cementoenamel junction and the bottom of the groove or periodontal pocket [27].
2.5. Gingival Fluid Collection
2.6. Sample Preparation
2.7. ELISA Test
2.7.1. Plate Preparation
- The capture antibody was diluted to the working concentration in PBS without carrier protein. Each well was coated with 100 μL of the capture antibody. The plate was sealed and incubated overnight at room temperature.
- The next day, the wells were aspirated and washed with buffer solution, repeating the process twice for a total of three washes. Each well was washed with wash solution (400 μL) using a squirt bottle, manifold dispenser, or automatic washer. Complete removal of liquid at each step was essential for good performance. After the last wash, it was necessary to remove any remaining wash buffer by vacuuming or inverting the plate and rubbing it against paper towels.
- Plates were blocked by adding 300 μL of reagent diluent to each well. These were incubated at room temperature for at least 1 h.
- Aspiration/washing was repeated as in step 2. Then, the plates were ready for sample addition.
2.7.2. Assay Procedure
- A total of 100 μL of sample or standard was added to the reagent diluent or an appropriate diluent per well. It was covered with adhesive tape and incubated for 2 h at room temperature.
- Aspiration/washing was repeated as in step 2 of the plates’ preparation.
- A total of 100 μL of detection antibody, diluted in NGS diluent reagent, was added to each well. This was covered with new adhesive tape and incubated for 2 h at room temperature.
- Aspiration/washing was repeated as in step 2 of plate preparation.
- In total, 100 μL of the streptavidin–HRP working dilution was added to each well. The plate was covered and incubated for 20 min at room temperature. Placing the plate in direct light was avoided.
- Vacuuming/washing was repeated as in step 2.
- A total of 100 μL of substrate solution was added to each well. These were incubated for 20 min at room temperature, and we avoided placing the plate in direct light.
- In total, 50 μL of stop solution was added to each well. We gently tapped the plate to ensure thorough mixing.
- The optical density of each well was determined immediately using a microplate reader set at 450 nm.
2.8. Statistical Analysis
3. Results
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Group | ||||
---|---|---|---|---|
Postpartum Women | No Pregnancy | p * | Total | |
Behavioral habits | 20 (100%) | 20 (100%) | 40 (100%) | |
Brushing after meals | ||||
Yes | 16 (80.00%) | 20 (100.00%) | p = 0.1060 | 36 (90.00%) |
No | 4 (20.00%) | 0 | 4 (10.00%) | |
Daily brushing frequency | ||||
Two or fewer times | 12 (55.00%) | 6 (30.00%) | p = 0.056 * | 18 (45.00%) |
Three or more times | 8 (35.00%) | 14 (70.00%) | 22 (55.00%) | |
Flossing | ||||
Yes | 8 (40.00%) | 17 (85.00%) | p = 0.0079 * | 25 (62.50%) |
No | 12 (60.00%) | 3 (15.00%) | 15 (37.50%) | |
Visit to the dentist | ||||
Yes | 4 (20.00%) | 12 (60.00%) | p = 0.0128 * | 14 (40.00%) |
No | 16 (80.00%) | 8 (40.00%) | 24 (60.00%) | |
History of alcohol | ||||
Yes | 7 (35.00%) | 18 (90.00%) | p = 0.0008 | 25 (62.50%) |
No | 13 (65.00%) | 2 (10.00%) | 15 (37.50%) | |
Past smoking history | ||||
Yes | 2 (10.00%) | 2 (10.00%) | p = 1.0000 | 4 (10.00%) |
No | 18 (90.00%) | 18 (90.00%) | 36 (90.00%) |
Oral Parameters | Group | p-Value * | |
---|---|---|---|
Postpartum Women (n = 20) | No Pregnancy (n = 20) | ||
Number of bleeding sites | 29.10 ± 18.74 | 19.85 ± 10.71 | 0.0632 |
Bleeding on probing (POB) | 26.80 ± 16.78 | 17.10 ± 9.67 | 0.0339 * |
Number of teeth | 26.50 ± 1.64 | 27.70 ± 0.93 | 0.0024 * |
Summary Measurements of MMP-8 Level in pg/mL | Groups | |||
---|---|---|---|---|
Postpartum Women | No Pregnancy | |||
Place | Handbag | Place | Handbag | |
Minimum | 104 | 145.3 | 30.25 | 120.3 |
Maximum | 4280 | 4898 | 1742 | 4895 |
Variance | 4176 | 4753 | 1711 | 4775 |
Average | 2560 | 2653 | 540.3 | 2735 |
Standard deviation | 1315 | 1652 | 449.6 | 1596 |
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Lima, K.M.; Calado, K.A.A.; Pereira, A.d.F.V.; Silva, M.C.P.d.; Lopes, F.F. MMP-8 in Periodontal Sites of Postpartum and without-Any-Pregnancy Women. Int. J. Environ. Res. Public Health 2024, 21, 739. https://doi.org/10.3390/ijerph21060739
Lima KM, Calado KAA, Pereira AdFV, Silva MCPd, Lopes FF. MMP-8 in Periodontal Sites of Postpartum and without-Any-Pregnancy Women. International Journal of Environmental Research and Public Health. 2024; 21(6):739. https://doi.org/10.3390/ijerph21060739
Chicago/Turabian StyleLima, Karyne Martins, Keiko Aramaki Abreu Calado, Adriana de Fátima Vasconcelos Pereira, Mayara Cristina Pinto da Silva, and Fernanda Ferreira Lopes. 2024. "MMP-8 in Periodontal Sites of Postpartum and without-Any-Pregnancy Women" International Journal of Environmental Research and Public Health 21, no. 6: 739. https://doi.org/10.3390/ijerph21060739