Comparative analysis of two colorimetric assays in dental pulp cell density.
OBJECTIVE
To compare and contrast two colorimetric assays used for the measurement of proliferation using two dental pulp cell types: dental pulp stem cells (DPSC) and human dental pulp fibroblasts (HDPF).


METHODS
Dental pulp stem cells or HDPF were seeded at 0.25×10(4) cells per well in 96-well plates. Cell proliferation was evaluated after 24-72h. At the end of the experimental period, the sulforhodamine B (SRB) assay or a water-soluble tetrazolium salt (WST-1) assay was performed. Optical densities were determined in a microplate reader (Genius; TECAN). Data were analysed by Student's t-test (comparison between cell types) and one-way anova followed by Tukey test (time-point intervals). Pearson' correlation tests were performed to compare the two assays for each cell line.


RESULTS
Both assays showed that DPSC had higher proliferation rates than HDPF. A positive significant correlation between the two colorimetric assays tested for both cell types DPSC (Pearson's correlation coefficient=0.847; P<0.05) and HDPF (Pearson's correlation coefficient=0.775; P<0.05).


CONCLUSIONS
Both tests demonstrated similar trends of cell proliferation, and thus are both appropriate for the evaluation of DPSC and HDPF. The choice of assay is therefore one of the practical applications. SRB stained plates can be dried and stored so may have utility in laboratories where data may require review or when access to analytical equipment is limited. WST-1 assays have the benefit of both ease and speed and may have utility in laboratories requiring either high throughput or rapid analyses.
Nedel
F
F
School of Dentistry Center of Biotechnology, Federal University of Pelotas, Pelotas, RS, Brazil.
Soki
F N
FN
Conde
M C M
MC
Zeitlin
B D
BD
Tarquinio
S B C
SB
Nör
J E
JE
Seixas
F K
FK
Demarco
F F
FF
D053687:Adult Stem Cells / Q000166:cytology*; D000704:Analysis of Variance; D049109:Cell Proliferation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay / Q000379:methods; D003124:Colorimetry / Q000379:methods*; D004396:Coloring Agents / Q000378:metabolism; D003782:Dental Pulp / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D012235:Rhodamines / Q000378:metabolism; D018709:Statistics, Nonparametric; D013778:Tetrazolium Salts / Q000378:metabolism
D004396:Coloring Agents; D012235:Rhodamines; D013778:Tetrazolium Salts; C022027:lissamine rhodamine B