RNA preservation in human dental pulp for transcriptomic profiling: a comparative multi-parameter study
Raksha Bhat1, Preethesh Shetty1, Shishir Shetty1
1Department of Conservative Dentistry and Endodontics, Nitte (Deemed to be University), AB Shetty Memorial Institute of Dental Sciences (ABSMIDS), Mangalore, Karnataka, India.
Frontiers in Dental Medicine
|November 10, 2025
Summary
RNAlater solution is the optimal method for preserving dental pulp tissue for RNA analysis, significantly improving RNA yield, purity, and integrity compared to snap freezing or RNAiso Plus. This finding supports standardizing methods for dental transcriptomics.
Area of Science:
- Molecular Biology
- Dental Research
- Biotechnology
Background:
- Dental pulp tissue presents unique challenges for RNA analysis due to its fibrous nature, high RNase activity, and susceptibility to degradation.
- Maintaining RNA integrity and quality is crucial for accurate transcriptomic studies and clinical applications.
Purpose of the Study:
- To systematically evaluate and compare three distinct preservation methodologies for human dental pulp tissue.
- To determine the optimal approach for preserving RNA integrity and quality in dental pulp samples.
Main Methods:
- Dental pulp samples from 36 patients with irreversible pulpitis were preserved using three methods: snap freezing, RNAiso Plus, and RNAlater solution.
- RNA quality was assessed using Nanodrop spectrophotometry, Qubit fluorometry, and Bioanalyzer capillary electrophoresis, evaluating yield, purity, and integrity.
Main Results:
- RNAlater storage demonstrated statistically superior performance across all parameters.
- Yield was significantly higher with RNAlater (11.5-fold over snap freezing) and RNAiso Plus (1.8-fold).
- RNAlater samples showed significantly better RNA integrity (mean RIN 6.0 vs. 3.34 for snap freezing) and achieved optimal quality in 75% of cases.
Conclusions:
- RNAlater storage is the optimal preservation method for dental pulp RNA investigations, enhancing yield, purity, and integrity.
- These findings support the standardization of preservation methodologies in dental transcriptomics and clinical settings.
