J Dent Rehabil Appl Sci.  2024 Feb;40(1):13-23. 10.14368/jdras.2024.40.1.13.

A study of growth factors, chondrogenic differentiation of mesenchymal stem cells and cell response by needle size differences in vitro

• Park J
• Hwang YJ
• Choi JJ
• Chon JY
• Lee SW

• KMID: 2553627
• DOI: http://doi.org/10.14368/jdras.2024.40.1.13

Abstract

Purpose
This aim of this study was to demonstrate growth factors that differentiate human mesenchymal stem cells into chondrocytes and to evaluate cell proliferation enhancement by needle size differences.
Materials and Methods
Human mesenchymal stem cells were cultured in chondrogenic medium supplemented with BMP-2, BMP-4, BMP-6, BMP-7, BMP-13, FGF-2, FGF-18, IGF-1, TGF-β1, TGF-β2, TGF-β3 and without growth factors for 14, 21, and 28 days. Then, the expression levels of SOX-5, SOX-6, SOX-9 and FOXO1A were comparatively analyzed. Human mesenchymal stem cells were inoculated into culture dishes using 18, 21, and 26 gauge (G) needles, and cell proliferation was measured after 24, 48, and 72 hours, respectively.
Results
In addition to the previously known FGF, IGF-1, and TGFβ1,the BMP family growth factors such as BMP-2, BMP-4, BMP-6, and BMP-7 increased the expression of chondrocyte differentiation genes SOX-5, SOX-6, SOX-9, and FOXO1A. At 48 hours, the 26G group, the smallest needle, showed significant cell proliferation improvement compared to the control group and the 18G group. At 72 hours, the 26G group, the smallest needle, showed significant increase in cell proliferation compared to the control group.
Conclusion
Through this study, growth factors with the ability to induce chondrocyte differentiation of human mesenchymal stem cells were investigated, and cell proliferation changes by needle size differences were determined.

Keyword

human mesenchymal stem cells; chondrocytes; growth factors; needles; cell proliferation

Figure

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  Fig. 1 Human mesenchymal stem cells were cultured in a chondrogenic medium supplemented with BMP-2, BMP-4, BMP-6, BMP-7, BMP-13, FGF-2, FGF-18, IGF-1, TGF-β1, TGF-β2, TGF-β3.

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  Fig. 2 Human mesenchymal stem cells were inoculated into mesenchymal stem cell growth medium using 18, 21, and 26 gauge (G) needles.

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  Fig. 3 Effect of growth factors (BMP-2, BMP-4, BMP-6, BMP-7, BMP-13, FGF-2, FGF-18, IGF-1, TGFβ1. TGFβ2, TGFβ3) on SOX-5, SOX-6, Sox9 and β-actin DNA expression after 14, 21, 28 days of mesenchymal stem cell culture.

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  Fig. 4 Result from the proliferation assay using ethidium bromide incorporation after 24, 48 and 72 h of mesenchymal stem cell culture. Note that only significant differences between the highest OD levels and other OD levels are presented. Oneway ANOVA (n = 5). **: significant difference (P < 0.01). Note that only significant differences between the highest gene expression levels and other gene expression levels are presented.

Reference

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