Background:
Proteomic is capable of elucidating complex biological systems through protein expression, function, and interaction under a particular condition.
Objective:
This study was aimed to determine the potential of ascorbic acid alone in inducing differentially expressed osteoblast-related proteins in dental stem cells via liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) approach.
Methods:
The cells were isolated from deciduous (SHED) and permanent teeth (DPSC) and induced with 10 µg/mL ascorbic acid. Bone mineralisation and osteoblast gene expression were determined using von Kossa staining and reverse transcriptase-polymerase chain reaction, respectively. The label-free protein samples were harvested on days 7 and 21 followed by protein identification and quantification using LC-MS/MS. Based on the similar protein expressed throughout treatment and controls for both SHED and DPSC, overall biological processes followed by osteoblast-related protein abundance were determined using the PANTHER database. STRING database was performed to determine differentially expressed proteins as candidates for SHED and DPSC during osteoblast development.
Results:
Both cells indicated brownish mineral stain and expression of osteoblast-related genes at day 21. Overall, a total of 700 proteins were similar among all treatments on both days 7 and 21 with 482 proteins appearing in the PANTHER database. Osteoblast-related protein abundance indicated 31 and 14 proteins that related to SHED and DPSC, respectively. Further analysis by the STRING database identified only 22 and 11 proteins involved from the respective group. Differential expressed analysis of similar proteins from these two groups revealed ACTN4 and ACTN1 as proteins that are involved in both SHED and DPSC. In addition, three (PSMD11/RPN11, PLS3, and CLIC1) and one (SYNCRIP) proteins were differentially expressed specifically for SHED and DPSC, respectively.
Conclusion:
Proteome differential expression showed that ascorbic acid alone can induce osteoblast-related proteins in both SHED and DPSC and generates specific differentially expressed protein markers.