Flow cytometry for assessing pulp viability in pediatric dentistry: An exploratory study
Josué Zuriel Ortiz García1, Ismael Secundino1, José Luis Ayala Herrera1
1Research Department, Faculty of Dentistry, La Salle Bajío University, León, Guanajuato, Mexico.
Summary
Flow cytometry effectively measured pediatric dental pulp viability, showing minimal differences between healthy, reversible, and irreversible pulpitis. Protein profiling and matrix metalloproteinase-2 activity offer potential for improved diagnostics in pediatric endodontics.
Area of Science:
- Pediatric Dentistry
- Biomaterials Science
- Cellular Biology
Background:
- Dental pulp (DP) is crucial for pediatric tooth health, but diagnosing pulpal pathologies is challenging due to limited patient cooperation and lack of reliable viability tests.
- Current diagnostic methods are often invasive, with histopathology as the gold standard.
- Flow cytometry offers a precise method for cellular analysis applicable to assessing pulp viability.
Purpose of the Study:
- To evaluate flow cytometry as a tool for assessing pulp viability in pediatric dental patients.
- To explore protein profiles and proteolytic activity as potential diagnostic markers for pulpal conditions.
- To correlate cellular viability and molecular markers with different stages of pulpal disease.
Main Methods:
- Collected 44 pediatric pulp samples (ages 2-7) classified as healthy, reversible pulpitis, irreversible pulpitis, or pulp necrosis.
- Utilized 7-aminoactinomycin D (7-AAD) staining with flow cytometry to quantify cell viability.
- Performed SDS-PAGE for protein profiling and gelatin zymography to assess matrix metalloproteinase (MMP) activity.
Main Results:
- Flow cytometry accurately quantified cell viability, showing minimal differences between healthy, reversible, and irreversible pulpitis, aligning with diagnostic challenges.
- Protein profiling indicated a progressive decrease in protein bands with advancing disease severity.
- Proteolytic activity, associated with MMP-2, increased with disease progression, notably higher in pulp necrosis.
Conclusions:
- Flow cytometry is a valuable method for quantifying pediatric pulp cell viability, underscoring the difficulty in clinically differentiating early pulpal conditions.
- Protein profiling shows promise for identifying diagnostic biomarkers to support less invasive diagnostic strategies.
- While MMP-2 activity increased with disease, it was not confirmed as a specific biomarker for disease progression.
