Boron Induces Early Matrix Mineralization via Calcium Deposition and Elevation of Alkaline Phosphatase Activity in Differentiated Rat Bone Marrow Mesenchymal Stem Cells

Bent-Al-Hoda Movahedi Najafabadi, M. H. Abnosi
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引用次数: 33

Abstract

Objective Boron (B) is essential for plant development and might be an essential micronutrient for animals and humans. This study was conducted to characterize the impact of boric acid (BA) on the cellular and molecular nature of differentiated rat bone marrow mesenchymal stem cells (BMSCs). Materials and Methods In this experimental study, BMSCs were extracted and expanded to the 3rdpassage, then cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) complemented with osteogenic media as well as 6 ng/ml and 6 µg/ml of BA. After 5, 10, 15 and 21 days the viability and the level of mineralization was determined using MTT assay and alizarin red respectively. In addition, the morphology, nuclear diameter and cytoplasmic area of the cells were studied with the help of fluorescent dye. The concentration of calcium, activity of alanine transaminase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) as well as sodium and potassium levels were also evaluated using commercial kits and a flame photometer respectively. Results Although 6 µg/ml of BA was found to be toxic, a concentration of 6 ng/ml increased the osteogenic ability of the cell significantly throughout the treatment. In addition it was observed that B treatment caused the early induction of matrix mineralization compared to controls. Conclusion Although more investigation is required, we suggest the prescription of a very low concentration of B in the form of BA or foods containing BA, in groups at high risk of osteoporosis or in the case of bone fracture.
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硼通过钙沉积和碱性磷酸酶活性升高诱导分化大鼠骨髓间充质干细胞早期基质矿化
目的硼(B)是植物发育所必需的微量元素,可能是动物和人类必需的微量元素。本研究旨在探讨硼酸(BA)对分化大鼠骨髓间充质干细胞(BMSCs)细胞和分子性质的影响。材料与方法本实验将骨髓间充质干细胞提取并扩增至第3代,在Dulbecco 's Modified Eagle 's Medium (DMEM)中添加成骨培养基以及6 ng/ml和6µg/ml BA进行培养。培养5、10、15、21 d后,分别用MTT法和茜素红法测定细胞活力和矿化水平。此外,利用荧光染料对细胞的形态、核直径和细胞质面积进行了研究。用商品化试剂盒和火焰光度计分别测定钙浓度、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(LDH)和碱性磷酸酶(ALP)活性以及钠和钾水平。结果虽然6µg/ml BA具有毒性,但在整个治疗过程中,浓度为6 ng/ml的BA显著提高了细胞的成骨能力。此外,与对照组相比,B处理引起基质矿化的早期诱导。结论虽然还需要进一步的研究,但我们建议骨质疏松或骨折高危人群使用极低浓度的BA或含BA的食物。
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