Effects of ascorbic acid on dental pulp stem cells from human deciduous (SHED) and permanent teeth (DPSC)

Cytotoxicity: Concentration screened were 10, 30, 50, 80, 100, 150, 250, 500 ug/mL ascorbic acid (AA) on SHED and DPSC. Based on IC50 values, SHED can withstand higher AA concentration compared to DPSC. Both cells survived in 10-150 ug/mL but died in 250 and 500 ug/mL AA. <br>Osteoblast differentiation:We performed viability (MTT assay), ALP specific enzyme activity, von Kossa stain and RT-PCR for bone molecular markers. Ascorbic acid could differentiate SHED and DPSC to osteoblast. Its effects were similar to the positive control of the study (AA+BGP)

细胞毒性实验：本研究以浓度分别为10、30、50、80、100、150、250、500 μg/mL的抗坏血酸（ascorbic acid，AA）处理人脱落乳牙干细胞（Stem Cells from Human Exfoliated Deciduous Teeth, SHED）与牙髓干细胞（Dental Pulp Stem Cells, DPSC）。基于半数抑制浓度（half maximal inhibitory concentration, IC50）的分析结果显示，SHED可耐受的AA浓度显著高于DPSC；两类细胞在10~150 μg/mL的AA环境中均可存活，但在250 μg/mL与500 μg/mL浓度下均会死亡。<br>成骨分化实验：本研究通过MTT比色法（MTT assay）检测细胞活性、碱性磷酸酶（alkaline phosphatase, ALP）特异性酶活性、Von Kossa染色法（von Kossa stain）以及逆转录聚合酶链式反应（reverse transcription-polymerase chain reaction, RT-PCR）以检测骨相关分子标志物的表达情况。结果表明，抗坏血酸可将SHED与DPSC诱导分化为成骨细胞，其诱导效果与本研究的阳性对照组（AA+BGP）相近。