Effects of low level of calcium on the biological behavior of rat primary ameloblasts cultured in vitro

Yonggang Wang; Jianping Ruan; Jing Zhou; Jiangang Tian; Ruizhe Huang; Jianghong Gao

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Effects of low level of calcium on the biological behavior of rat primary ameloblasts cultured in vitro

VernacularTitle:低钙对SD大鼠原代成釉细胞生物学特性影响的体外实验研究
Author: Yonggang WANG ^{1
,

2
,

3} ; Jianping RUAN ^{1
,

2} ; Jing ZHOU ^{1
,

2} ; Jiangang TIAN ^{1
,

2} ; Ruizhe HUANG ^{1
,

2} ; Jianghong GAO ^{1
,

2}
Author Information

1. Clinical Research Center of Shaanxi Province for Dental and Maxillofacial Diseases
2. Department of Preventive Dentistry, College of Stomatology, Xi’an Jiaotong University, Xi’an 710004
3. Department of Stomatology, Dongying People’s Hospital, Dongying 257091, China
Publication Type:Journal Article
Keywords: ameloblast; proliferation; apoptosis; differentiation; calcium
From: Journal of Xi'an Jiaotong University(Medical Sciences) 2021;42(2):257-261,266
CountryChina
Language:Chinese
Abstract: 【Objective】 To explore the effect of low levels of calcium on the biological characteristics of ameloblasts. 【Methods】 Rat primary ameloblasts were cultured in standard DMEM medium. After five days they were identified by RT-PCR and immunohistochemistry. Then 0, 0.6 and 1.2 mmoL/L Ca²⁺ and 100 mL/L fetal bovine serum were added into DMEM medium without calcium. After 48 hours, the cell morphology was observed by inverted microscope. The proliferation and apoptosis of cells were separately examined by MTT and AnnexinV-PI. Real-time quantitative PCR was used to detect the expression levels of amelogenin and KLK4 mRNA. 【Results】 After Five days in standard DMEM medium, the cells were shaped like the paving pattern. RT-PCR showed that both amelogenin and KLK4 were expressed in the cells. Immunohistochemical staining showed that most cells had positive staining for amelogenin. After 48 hours of calcium intervention, some cells in 1.2 mmoL/L Ca²⁺ group had higher nuclear density and poor light transmittance, and more high columnar cells could be observed in 1.2 mmoL/L Ca²⁺ group than those in 0 and 0.6 mmoL/L Ca²⁺ groups. With the decrease in calcium concentration in the medium, MTT showed that the proliferation activity of ameloblasts reduced (P<0.01). Annexin V-PI showed that the percentage of apoptotic cells decreased, and there was a significant difference between 1.2 mmoL/L and 0 mmoL/L Ca²⁺ groups (P<0.05). Real time-PCR showed that the expressions of amelogenin and KLK4 mRNA reduced (P<0.01). 【Conclusion】 Low-level calcium may inhibit the differentiation of ameloblasts, thereby affecting the formation of enamel mineralization.