Abstract
Hyperglycemia, a major characteristic of diabetes, is considered to play a vital role in diabetic complications. High glucose levels have been found to inhibit the mineralization of dental pulp cells. However, gene expression associated with this phenomenon has not yet been reported. This is important for future dental therapeutic application. Objective Our study aimed to investigate the effect of high glucose levels on mineralization of human dental pulp-derived cells (hDPCs) and identify the genes involved.Methodology hDPCs were cultured in mineralizing medium containing 25 or 5.5 mM D-glucose. On days 1 and 14, RNA was extracted and expression microarray performed. Then, differentially expressed genes (DEGs) were selected for further validation using the reverse transcription quantitative polymerase chain reaction (RT-qPCR) method. Cells were fixed and stained with alizarin red on day 21 to detect the formation of mineralized nodules, which was further quantified by acetic acid extraction.Results Comparisons between high-glucose and low-glucose conditions showed that on day 1, there were 72 significantly up-regulated and 75 down-regulated genes in the high-glucose condition. Moreover, 115 significantly up- and 292 down-regulated genes were identified in the high-glucose condition on day 14. DEGs were enriched in different GO terms and pathways, such as biological and cellular processes, metabolic pathways, cytokine–cytokine receptor interaction and AGE-RAGE signaling pathways. RT-qPCR results confirmed the significant expression of pyruvate dehydrogenase kinase 3 (PDK3), cyclin-dependent kinase 8 (CDK8), activating transcription factor 3 (ATF3), fibulin-7 (Fbln-7), hyaluronan synthase 1 (HAS1), interleukin 4 receptor (IL-4R) and apolipoprotein C1 (ApoC1).Conclusions The high-glucose condition significantly inhibited the mineralization of hDPCs. DEGs were identified, and interestingly, HAS1 and Fbln-7 genes may be involved in the glucose inhibitory effect on hDPC mineralization.
Highlights
Diabetes mellitus constitutes a group of metabolic disorders related to hyperglycemia
Our study aimed to investigate the effect of high glucose levels on mineralization of human dental pulp-derived cells and identify the genes involved
reverse transcription quantitative polymerase chain reaction (RT-qPCR) results confirmed the significant expression of pyruvate dehydrogenase kinase 3 (PDK3), cyclin-dependent kinase 8 (CDK8), activating transcription factor 3 (ATF3), fibulin-7 (Fbln-7), hyaluronan synthase 1 (HAS1), interleukin 4 receptor (IL4R) and apolipoprotein C1 (ApoC1)
Summary
Diabetes mellitus constitutes a group of metabolic disorders related to hyperglycemia. Studies have reported alteration in dental pulp structure, increased inflammation and impaired pulpal healing in the dental pulp of people with diabetes.. Hyperglycemia has been considered to be a major factor involved in the pathogenesis of diabetes, as it induces alterations at the cellular level of the affected tissue. High levels of glucose have been reported to inhibit the osteogenic differentiation and mineral tissue formation.. Research shows that hyperglycemia exerts a negative effect on pulpal healing by inhibiting dentin bridge formation and increasing pulpal inflammation.. Treating dental pulp cells with high glucose levels was found to inhibit mineralization.. Even though a high level of glucose was reported to inhibit the mineralization of pulp cells, gene expression associated with the influence of glucose has never been explored and may be important as basis for future studies to improve the dental therapeutic application in diabetic patient Research shows that hyperglycemia exerts a negative effect on pulpal healing by inhibiting dentin bridge formation and increasing pulpal inflammation. treating dental pulp cells with high glucose levels was found to inhibit mineralization. Even though a high level of glucose was reported to inhibit the mineralization of pulp cells, gene expression associated with the influence of glucose has never been explored and may be important as basis for future studies to improve the dental therapeutic application in diabetic patient
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