Boron Accelerates Cultured Osteoblastic Cell Activity through Calcium Flux

Abstract

A low concentration of boron (B) accelerates the proliferation and differentiation of mammalian osteoblasts. The aim of this study was to investigate the effects of 0.1 mM of B on the membrane function of osteoblastic cells in vitro. Genes involved in cell activity were investigated using gene expression microarray analyses. The Ca influx and efflux were evaluated to demonstrate the activation of L-type Ca channel for the Ca influx, and that of Na/K-ATPase for the Ca efflux. A real-time PCR analysis revealed that the messenger RNA (mRNA) expression of four mineralization-related genes was clearly increased after 3 days of culture with a B-supplemented culture medium. Using microarray analyses, five genes involved in cell proliferation and differentiation were upregulated compared to the control group. Regarding the Ca influx, in the nifedipine-pretreated group, the relative fluorescence intensity for 1 min after adding B solution did not increase compared with that for 1 min before addition. In the control group, the relative fluorescence intensity was significantly increased compared with the experimental group (P < 0.05). Regarding the Ca efflux, in the experimental group cultured in 0.1 mM of B-supplemented medium, the relative fluorescence intensity for 10 min after ouabain treatment revealed a significantly lower slope value compared with the control group (P < 0.01). This is the first study to demonstrate the acceleration of Ca flux by B supplementation in osteoblastic cells. Cell membrane stability is related to the mechanism by which a very low concentration of B promotes the proliferation and differentiation of mammalian osteoblastic cells in vitro.

Fish Collagen Promotes the Expression of Genes Related to Osteoblastic Activity

“…The primers are indicated in Table 1. The analyses of RT-PCR were carried out according to the similar methods to our recent study [6], except the following two steps. The first was that the protocol of amplification and quantification was 40 cycles (58-59 ∘ C, 1 min).…”

“…MC3T3-E1 cells were seeded in a 60 mm culture dish covered with or without TAC gel at a density of 1.5 × 10 6 cells. After 3-day culture, total RNA from cells was isolated using TRIzol reagent (1 mL/10 cm 2 ) (Invitrogen, Carlsbad, CA, USA) and purified in accordance with the manufacturer's instructions [6]. [6].…”

“…After 3-day culture, total RNA from cells was isolated using TRIzol reagent (1 mL/10 cm 2 ) (Invitrogen, Carlsbad, CA, USA) and purified in accordance with the manufacturer's instructions [6]. [6]. The cRNA was amplified and hybridized to an Affinity Mouse Genome 430 2.0 array (for 39,000 transcriptional products), scanned by an Affymetrix scanner, and calculated using the Affymetrix Expression Console6.…”

“…The cRNA was amplified and hybridized to an Affinity Mouse Genome 430 2.0 array (for 39,000 transcriptional products), scanned by an Affymetrix scanner, and calculated using the Affymetrix Expression Console6. [6]. We then established the criteria for upregulated genes ( score > 3.0, ratio > 1.5fold).…”

Effects of Nutritional Deficiency of Boron on the Bones of the Appendicular Skeleton of Mice

“…Recently, Capati et al [72] demonstrated the acceleration of the Ca 2+ influx and efflux by 0.1 mM B supplementation in cultured NOS-1 osteoblastic cells. Results indicated that cell membrane stability may be related to the mechanism by which a very low concentration of B promotes the proliferation and differentiation in mammalian osteoblastic cells in vitro [72].…”