Center for Research on Dental Implants, Federal University of Santa Catarina, Delfino Conti Street, Florianópolis, 88040-900, Brazil.
Laboratory of Applied Virology, Federal University of Santa Catarina, Henrique da Silva Fontes Avenue, Florianópolis, 88040-900, Brazil.
J Mater Sci Mater Med. 2021 Jan 19;32(1):1. doi: 10.1007/s10856-020-06475-6.
In in vitro culture systems, dexamethasone (DEX) has been applied with ascorbic acid (ASC) and β-glycerophosphate (βGLY) as culture media supplementation to induce osteogenic differentiation of mesenchymal stem cells. However, there are some inconsistencies regarding the role of DEX as osteogenic media supplementation. Therefore, this study verified the influence of DEX culture media supplementation on the osteogenic differentiation, especially the capacity to mineralize the extracellular matrix of stem cells from human exfoliated deciduous teeth (SHED). Five groups were established: G1-SHED + Dulbecco's Modified Eagles' Medium (DMEM) + fetal bovine serum (FBS); G2-SHED + DMEM + FBS + DEX; G3-SHED + DMEM + FBS + ASC + βGLY; G4-SHED + DMEM + FBS + ASC + βGLY + DEX; G5-MC3T3-E1 + α Minimal Essential Medium (MEM) + FBS + ASC + βGLY. DNA content, alkaline phosphatase (ALP) activity, free calcium quantification in the extracellular medium, and extracellular matrix mineralization quantification through staining with von Kossa, alizarin red, and tetracycline were performed on days 7 and 21. Osteogenic media supplemented with ASC and β-GLY demonstrated similar effects on SHED in the presence or absence of DEX for DNA content (day 21) and capacity to mineralize the extracellular matrix according to alizarin red and tetracycline quantifications (day 21). In addition, the presence of DEX in the osteogenic medium promoted less ALP activity (day 7) and extracellular matrix mineralization according to the von Kossa assay (day 21), and more free calcium quantification at extracellular medium (day 21). In summary, the presence of DEX in the osteogenic media supplementation did not interfere with SHED commitment into mineral matrix depositor cells. We suggest that DEX may be omitted from culture media supplementation for SHED osteogenic differentiation in vitro studies.
在体外培养系统中，地塞米松（DEX）与抗坏血酸（ASC）和β-甘油磷酸（βGLY）一起作为培养基补充剂，以诱导间充质干细胞的成骨分化。然而，DEX 作为成骨培养基补充剂的作用存在一些不一致之处。因此，本研究验证了 DEX 培养基补充剂对成骨分化的影响，特别是对人脱落乳牙（SHED）干细胞细胞外基质矿化能力的影响。建立了五组：G1-SHED+DMEM+胎牛血清（FBS）；G2-SHED+DMEM+FBS+DEX；G3-SHED+DMEM+FBS+ASC+βGLY；G4-SHED+DMEM+FBS+ASC+βGLY+DEX；G5-MC3T3-E1+α最低必需培养基（MEM）+FBS+ASC+βGLY。在第 7 天和第 21 天，通过 DNA 含量、碱性磷酸酶（ALP）活性、细胞外基质中游离钙的定量分析以及von Kossa、茜素红和四环素染色定量分析细胞外基质矿化，对 SHED 进行了检测。在存在或不存在 DEX 的情况下，补充 ASC 和β-GLY 的成骨培养基对 SHED 的 DNA 含量（第 21 天）和根据茜素红和四环素定量分析的细胞外基质矿化能力（第 21 天）具有相似的作用。此外，成骨培养基中 DEX 的存在促进了较少的 ALP 活性（第 7 天）和根据 von Kossa 测定的细胞外基质矿化（第 21 天），以及更多的细胞外基质中游离钙定量分析（第 21 天）。综上所述，DEX 存在于成骨培养基补充剂中并不干扰 SHED 向矿化基质沉积细胞的分化。我们建议在体外研究 SHED 成骨分化时，可以从培养基补充剂中省略 DEX。
