Journal
SAINS MALAYSIANA
Volume 51, Issue 7, Pages 2173-2186Publisher
UNIV KEBANGSAAN MALAYSIA
DOI: 10.17576/jsm-2022-5107-19
Keywords
Ascorbic acid; osteoblast differentiation; secretome; stem cells from human permanent tooth pulp (DPSC)
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This study aimed to determine the potential of ascorbic acid to induce osteoblast differentiation by identifying differentially expressed osteoblast-related secretome. The results showed that ascorbic acid alone without beta-glycerophosphate successfully induced osteoblast-related secretome. Inhibin beta A chain and fibrillin-2 were suggested as secretome markers for DPSC during osteoblast differentiation.
In vitro stem cell differentiation into osteoblast involved a combination of ascorbic acid andfl-glycerophosphate. However, fl-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 mu g/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 beta-glycerophosphate successfully induced osteoblast-related secretome. Inhibin beta A chain and fibrillin-2 were suggested as secretome markers for DPSC during osteoblast differentiation.
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