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. 2018 Jun 23;23(7):1517. doi: 10.3390/molecules23071517

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Osteogenic gene expression in calvarial primary osteoblasts. Calvariae from 8-week-old GF and SPF mice were harvested, and primary osteoblasts were isolated and cultured in osteoblast differentiation medium. After culture for the indicated times, total RNA was isolated, and real-time PCR analysis was performed to assess osteogenic gene expression. Significant differences in expression of the osteoprotegerin (Opg) gene (a), receptor activator of nuclear factor-kappa B ligand (Rankl) (b) gene, the Opg/Rankl ratio (c), osteocalcin (Ocn) (d) gene, and alkaline phosphatase (Alp) (e) gene were observed between GF and SPF osteoblasts. At 5 weeks of culture, no significant differences were observed in expression of the collagen type I alpha (Col1a) 1 (f) and Col1a2 (g) genes between GF and SPF osteoblasts. Expression of the insulin-like growth factor (Igf-1 (h) and Igf-2 (i) genes was significantly higher in SPF osteoblasts than GF osteoblasts at 5 weeks. * p < 0.05, t-test (n = 4/group).