Grafting biomaterials associated to topical glucocorticoid: effects on pre-osteoblastic cells (MC3T3-E1)

Silva, Alice de Araújo Ferreira; Pereira, Carolina Nemesio de Barros; Dias, Danilo Rocha; Lages, Frederico Santos; Maltos, K. L. M.; Moreira, Allyson Nogueira; Zenóbio, Élton Gonçalves; Diniz, Ivana Márcia Alves

doi:10.1590/1807-3107bor-2022.vol36.0090

Cited by 2 publications

(1 citation statement)

References 21 publications

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“…In the current literature, there is a variety of Asc/βGP combinations used for MC3T3-E1 subclone 4 cells, e.g., 100 µM/2 mM [28], 50 µg.mL −1 /5 mM [50], 200 µM/10 mM [51], 25 µg.mL −1 /5 mM [52], and 50 µg.mL −1 /4 mM [53], while inorganic phosphate is also used in other forms such as NaH 2 PO 4 [54] or KH 2 PO 4 [55]. Consequently, cells respond differently in diverse osteogenic conditions, which produces various levels of mineralisation and specificity in ARS staining across publications.…”

Section: Discussionmentioning

confidence: 99%

Defining the Most Potent Osteoinductive Culture Conditions for MC3T3-E1 Cells Reveals No Implication of Oxidative Stress or Energy Metabolism

Semicheva,

Ersoy,

Vasilaki

et al. 2024

IJMS

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The MC3T3-E1 preosteoblastic cell line is widely utilised as a reliable in vitro system to assess bone formation. However, the experimental growth conditions for these cells hugely diverge, and, particularly, the osteogenic medium (OSM)’s composition varies in research studies. Therefore, we aimed to define the ideal culture conditions for MC3T3-E1 subclone 4 cells with regard to their mineralization capacity and explore if oxidative stress or the cellular metabolism processes are implicated. Cells were treated with nine different combinations of long-lasting ascorbate (Asc) and β-glycerophosphate (βGP), and osteogenesis/calcification was evaluated at three different time-points by qPCR, Western blotting, and bone nodule staining. Key molecules of the oxidative and metabolic pathways were also assessed. It was found that sufficient mineral deposition was achieved only in the 150 μg.mL−1/2 mM Asc/βGP combination on day 21 in OSM, and this was supported by Runx2, Alpl, Bglap, and Col1a1 expression level increases. NOX2 and SOD2 as well as PGC1α and Tfam were also monitored as indicators of redox and metabolic processes, respectively, where no differences were observed. Elevation in OCN protein levels and ALP activity showed that mineralisation comes as a result of these differences. This work defines the most appropriate culture conditions for MC3T3-E1 cells and could be used by other research laboratories in this field.

show abstract

Section: Discussionmentioning

confidence: 99%

Defining the Most Potent Osteoinductive Culture Conditions for MC3T3-E1 Cells Reveals No Implication of Oxidative Stress or Energy Metabolism

Semicheva,

Ersoy,

Vasilaki

et al. 2024

IJMS

View full text Add to dashboard Cite

show abstract

Defining the most potent osteoinductive culture conditions for MC3T3-E1 cells reveals no implication of oxidative stress or energy metabolism

Semicheva,

Ersoy,

Vasilaki

et al. 2024

Preprint

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MC3T3-E1 preosteoblastic cell line is widely utilised as a reliable in vitro system to assess bone formation. However, the experimental growth conditions for these cells hugely diverge and, particularly, the osteogenic medium (OSM) composition varies in research studies. Therefore, we aimed to define the ideal culture conditions for MC3T3-E1 subclone 4 cells with regards to mineralization capacity and explore if oxidative stress or cellular metabolism processes are implicated. Cells were treated with 9 different combinations of long-lasting ascorbate (Asc) and beta-glycerophosphate (betaGP) and osteogenesis/calcification were evaluated at 3 different time-points by qPCR, Western blotting and bone nodule staining. Key molecules of oxidative and metabolic pathways were also assessed. It was found that sufficient mineral deposition was achieved only in 150 ug.mL-1/2mM Asc/betaGP combination at 21d in OSM and this was supported by Runx2, Alpl, Bglap and Col1a1 expression levels increase. NOX2 and SOD2 as well as PGC1alpha and Tfam were also monitored as indicators of redox and metabolic processes, respectively, where no differences were observed. Elevation in OCN protein levels and ALP activity showed that mineralisation comes as a result of these differences. This work defines the most appropriate culture conditions for MC3T3-E1 cells and could be used by other research laboratories in this field.

show abstract

Grafting biomaterials associated to topical glucocorticoid: effects on pre-osteoblastic cells (MC3T3-E1)

Cited by 2 publications

References 21 publications

Defining the Most Potent Osteoinductive Culture Conditions for MC3T3-E1 Cells Reveals No Implication of Oxidative Stress or Energy Metabolism

Defining the Most Potent Osteoinductive Culture Conditions for MC3T3-E1 Cells Reveals No Implication of Oxidative Stress or Energy Metabolism

Defining the most potent osteoinductive culture conditions for MC3T3-E1 cells reveals no implication of oxidative stress or energy metabolism

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