| Summary: | In vitro stem cell differentiation into osteoblast involved a combination of ascorbic acid and β-glycerophosphate. However, β-glycerophosphate reduced cell viability in the culture. Therefore, this study was conducted to determine the potential of ascorbic acid to induce osteoblast differentiation by identifying differentially expressed osteoblast-related secretome. Stem cells isolated from permanent (DPSC) tooth pulp was induced with 10 µg/mL ascorbic acid for 21 days, and osteoblast gene expression was determined via reverse transcriptase-polymerase chain reaction (RT-PCR). After 21 days of induction, cells were incubated in a serum-free basal medium for 12 h and analyzed via liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) approach. The biological processes and differentially expressed osteoblast-related secretome were determined using UniProt and PANTHER based on similar proteins expressed in ascorbic acid induction and positive control. Interaction between secretome was determined using STRING for DPSC differentiation pathways determination. As a result, the BSP gene was expressed in ascorbic acid induction. A total of 57 secretome proteins were identified in ascorbic acid induction and control, where three differentially expressed osteoblast-related secretome were determined in DPSC. For secretome interaction, only two secretomes were involved in three pathways of DPSC after STRING analysis. In conclusion, ascorbic acid alone without β-glycerophosphate successfully induced osteoblast-related secretome. Inhibin beta A chain and fibrillin-2 were suggested as secretome markers for DPSC during osteoblast differentiation. © 2022 Penerbit Universiti Kebangsaan Malaysia. All rights reserved.
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