Insulin‑like growth factor 2‑enhanced osteogenic differentiation of stem cell spheroids by regulation of Runx2 and Col1 expression

Min,Sae Kyung; Kim,Minji; Park,Jun-Beom

doi:10.3892/etm.2021.9814

Insulin‑like growth factor 2‑enhanced osteogenic differentiation of stem cell spheroids by regulation of Runx2 and Col1 expression

Authors:
- Sae Kyung Min
- Minji Kim
- Jun-Beom Park
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Affiliations: Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea, College of Dentistry, Chosun University, Gwangju 61452, Republic of Korea
Published online on: February 22, 2021 https://doi.org/10.3892/etm.2021.9814
Article Number: 383
Copyright: © Min et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Insulin‑like growth factor 2 (IGF‑2) is a growth factor that is involved in various functions of cells, including stem cells. The effects of IGF‑2 on the cellular viability and osteogenic differentiation of stem cell spheroids were investigated in the present study. Stem cell spheroids were formed using concave microwells in the presence of IGF‑2 at final concentrations of 0, 10 and 100 ng/ml. Cellular viability was measured qualitatively using a microscope and quantitatively using an assay kit based on water‑soluble tetrazolium salt. The level of alkaline phosphatase activity, and an anthraquinone dye assay for calcium deposit evaluation, were used to assess osteogenic differentiation. A quantitative PCR analysis was conducted to evaluate the expression of Runx2 and Col1. Spheroid formation was noticed on day 1 in the microwells, and the spheroidal shape was maintained up to day 7. The cell viability assay values for IGF‑2 at 0, 10 and 100 ng/ml at day 1 were 0.193±0.002, 0.191±0.002 and 0.201±0.006, respectively (P>0.05). The absorbance values at 405 nm for the alkaline phosphatase activity assays on day 21 were 0.221±0.006, 0.375±0.010 and 0.280±0.015 for IGF‑2 at 0, 10 and 100 ng/ml, respectively. There were significantly higher values for IGF‑2 in the 10 and 100 ng/ml groups when compared with the control (P<0.05). Significantly higher Alizarin red staining was noted for IGF‑2 in the 10 ng/ml group when compared with the unloaded control at day 21 (P<0.05). Quantitative PCR revealed that mRNA levels of Runx2 and Col1 were significantly higher at 100 ng/ml on day 7. Conclusively, the present study demonstrated that the application of IGF‑2 increased alkaline phosphatase activity, Alizarin red staining, and Runx2 and Col1 expression of stem cell spheroids.

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