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. 2020 Jul 19;10(7):116.
doi: 10.3390/life10070116.

Anti-Inflammatory Effects of Adenine Enhance Osteogenesis in the Osteoblast-Like MG-63 Cells

Yu-Pin Chen  1 , Yo-Lun Chu  1 , Yang-Hwei Tsuang  2   3 , Yueh Wu  1 , Cheng-Yi Kuo  4 , Yi-Jie Kuo  1   3
Affiliations

Anti-Inflammatory Effects of Adenine Enhance Osteogenesis in the Osteoblast-Like MG-63 Cells

Yu-Pin Chen et al. Life (Basel). .

Abstract

Background: Adenine is a purine with a role in cellular respiration and protein synthesis. It is considered for its pharmacological potential. We investigated whether anti-inflammatory effect of adenine benefits on the proliferation and maturation of osteoblastic cells.

Methods: Human osteoblast-like cells (MG-63) were cultured with adenine under control conditions or pre-treated with 10ng/mL of tumor necrosis factor-α (TNF-α) followed by adenine treatment. Cell viability was examined using dimethylthiazol diphenyltetrazolium bromide (MTT) assay. Expression of cytokines and osteogenic markers were analyzed using quantitative PCR (qPCR) and ELISA. Enzyme activity of alkaline phosphatase (ALP) and collagen content were measured.

Results: TNF-α exposure led to a decreased viability of osteoblastic cells. Treatment with adenine suppressed TNF-α-induced elevation in IL-6 expression and nitrite oxide production in MG-63 cells. Adenine induced the osteoblast differentiation with increased transcript levels of collage and increased ALP enzyme activity.

Conclusions: Adenine exerts anti-inflammatory activity in an inflammatory cell model. Adenine benefits osteoblast differentiation in normal and inflammatory experimental settings. Adenine has a potential for the use to treat inflammatory bone condition such as osteoporosis.

Keywords: IL-6; adenine; inflammatory bone disease; osteoblast; osteogenesis.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effects of adenine on cell proliferation of MG-63 cells. (A) MG-63 cells were exposed to serial concentrations of adenine (0–250 µM/mL) for 24 h. (B) MG-63 cells were pre-cultured with TNF-α (10 ng/mL) and treated with serial concentrations of adenine (0–250 µM) for 24 h. Cell viability was determined using MTT assay. Data were expressed as mean ± SD.
Figure 2
Figure 2
Effect of adenine on pro-inflammatory response in MG-63 cells. Cells were treated with or without 10ng/mL of TNF-α and cultured with serial concentrations of adenine (0–250 µM) for 24 h. (A,B) IL-6 production were determined by ELISA. (C,D) mRNA expression of iNOS upon TNF-α stimulation was examined using qPCR. (E,F) Nitrite production in culture medium of MG-63 stimulated with or without TNF-α was evaluated. Data were expressed as mean ± SD. ‡, p < 0.05 as comparing to 0 µg/mL; †, p < 0.05 as comparing to control (Con).
Figure 3
Figure 3
Adenine induced alkaline phosphatase (ALP) activity in MG-63 cells treated without (A) or with (B) stimulation of TNF-α. Cells were treated with serial concentrations of adenine (0–250 µM) for 24 h. Data were expressed as mean ± SD. ‡, p < 0.05 as comparing to 0 µg/mL; †, p < 0.05 as comparing to control (Con).
Figure 4
Figure 4
Adenine increased ALP mRNA expression in MG-63 cells treated without (A) or with (B) TNF-α. Cells were treated with serial concentrations of adenine (0–250 µM) for 24 h and mRNA expressions were determined by qPCR. Data were expressed as mean ± SD. ‡, p < 0.05 as comparing to 0 µg/mL; †, p < 0.05 as comparing to control (Con).
Figure 5
Figure 5
Effect of adenine on collagen synthesis in MG-63 cells treated without (A) or with (B) stimulation of TNF-α. Cells were treated with serial concentrations of adenine (0–250 µM) for 24 h and collagen synthesis in MG-63 cells was determined by ELISA. Data were expressed as mean ± SD. ‡, p < 0.05 as comparing to 0 µg/mL; †, p < 0.05 as comparing to control (Con).
Figure 6
Figure 6
Adenine induced collagen type I mRNA expression in MG-63 cells treated without (A) or with (B) stimulation of TNF-α. Cells were treated with serial concentrations of adenine (0–250 µM) for 24 h and mRNA expressions were determined by qPCR. Data were expressed as mean ± SD. ‡, p < 0.05 as comparing to 0 µg/mL; †, p < 0.05 as comparing to control (Con).
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