Effect of Membrane Potential on Osteogenic Differentiation of Human Dental Follicle Stem Cells and Its Electrophysiological Mechanisms

doi:10.13701/j.cnki.kqyxyj.2025.05.010

Abstract

Abstract: Objective: To explore the effect of membrane potential on osteogenic differentiation of human dental follicle stem cells (hDFCs) and its electrophysiological mechanisms. Methods: hDFCs were isolated and cultured from human dental follicle tissue by enzymatic digestion methods. To evaluate the effects of Kir2.1 channels on the osteogenic differentiation capacity and membrane potentials of hDFCs, osteogenic differentiation induction, Alizarin red staining, real-time reverse transcriptase-polymerase chain reaction (RT-qPCR), and whole cell patch clamp were conducted by using lentiviral particles carrying human Kir2.1 channels specific short hairpin RNA (shRNA) and Kir2.1 channels specific inhibitor (ML133). Ca²⁺ imaging was performed to identify the store-operated Ca²⁺ channels (SOCs) mediated Ca²⁺ influx in hDFCs by using La³⁺ (a SOCs inhibitor) and Thapsigargin (an endoplasmic reticulum Ca²⁺ pump inhibitor), and to evaluate the effect of membrane potential hyperpolarization caused by decreased extracellular potassium level (1 mmol/L) on intracellular Ca²⁺ concentration. Results: Alizarin red staining, RT-qPCR, and patch clamp results showed that blocking the function of Kir2.1 channels or inhibition of Kir2.1 channels expression could inhibit the osteogenic differentiation ability of hDFCs and reverse the membrane potential hyperpolarization associated with osteogenic differentiation. Ca²⁺ imaging results showed that: (1) SOCs channels mediated Ca²⁺ influx in hDFCs. (2) Promotion of membrane hyperpolarization increased the intracellular Ca²⁺ levels in hDFCs, which could be inhibited by blocking the function of Kir2.1 channels or inhibition of Kir2.1 channels expression; by removal of extracellular Ca²⁺; or by blocking the function of SOCs channels. Conclusion: Kir2.1-linked membrane hyperpolarization controls osteogenic differentiation of hDFCs by increasing intracellular Ca²⁺ levels and SOCs channels-mediated Ca²⁺ influx plays an important role in this process.

Key words: human dental follicle cells, osteogenic differentiation, Kir2.1 channel, membrane hyperpolarization, calcium

GU Yujie, YANG Yidan, LIAO Siqi, WANG Heyi, ZHOU Rui, LAN Xiaorong, XU Xiaomei, ZUO Dongchuan, ZENG Jin. Effect of Membrane Potential on Osteogenic Differentiation of Human Dental Follicle Stem Cells and Its Electrophysiological Mechanisms[J]. Journal of Oral Science Research, 2025, 41(5): 413-419.

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