primary rat osteoblast cells  (Cell Applications Inc)

Journal: Experimental Cell Research

Article Title: Allopurinol and oxypurinol promote osteoblast differentiation and increase bone formation

doi: 10.1016/j.yexcr.2016.03.004

Figure Lengend Snippet: Allopurinol and oxypurinol inhibit osteoblast XO activity. (A) The level of XO mRNA expression is the same in differentiating cells (day 7 of culture) and mature, bone-forming osteoblasts (day 14). (B) XO activity was 20% lower in mature osteoblasts compared to differentiating cells. Allopurinol (≥0.1 µM) inhibits XO activity by 30% and 20% at day 7 and day 14, respectively. (C) Oxypurinol (≥0.1 µM) reduced XO activity by up to 32%. Values are means±SEM (n=6), ***/###=p<0.001, **=p<0.01, *=p<0.05.

Article Snippet: Primary rat osteoblast cells were obtained from 2-day-old neonatal Sprague-Dawley rats euthanised by cervical dislocation, as described previously , .

Techniques: Activity Assay, Expressing

Journal: Experimental Cell Research

Article Title: Allopurinol and oxypurinol promote osteoblast differentiation and increase bone formation

doi: 10.1016/j.yexcr.2016.03.004

Figure Lengend Snippet: Allopurinol and oxypurinol increase bone formation by osteoblasts. (A) Allopurinol (≥1 nM) stimulates bone formation by up to 4-fold. No effect was seen with 10 µM allopurinol. The (B) number and (C) size of the mineralised nodules were also increased by allopurinol treatment (≥10 nM). (D) Oxypurinol (≥1 nM) increased bone formation by up to 3-fold; the (E) number and (F) size of the mineralised nodules were also increased. (G) Allopurinol, oxypurinol and BMP2 promoted bone formation to a similar extent (~2-fold). Febuxostat induced the largest increase in bone formation (3-fold). Values are means±SEM (n=6), ***=p<0.001, **=p<0.01, *=p<0.05. (H) Representative whole well scans (unstained) and phase contrast microscopy images (alizarin red stained) show the increased bone formation seen with allopurinol and oxypurinol. Scale bars: whole well=0.5 cm, phase contrast images=500 µm.

Article Snippet: Primary rat osteoblast cells were obtained from 2-day-old neonatal Sprague-Dawley rats euthanised by cervical dislocation, as described previously , .

Techniques: Microscopy, Staining

Journal: Experimental Cell Research

Article Title: Allopurinol and oxypurinol promote osteoblast differentiation and increase bone formation

doi: 10.1016/j.yexcr.2016.03.004

Figure Lengend Snippet: No effect of allopurinol or oxypurinol on cell number, viability or ATP release. Cell number was measured after 7 or 14 days of treatment with 1 nM–10 µM allopurinol or oxypurinol; ATP release and viability were assessed in mature osteoblasts. At all concentrations tested, (A-B) allopurinol and (C-D) oxypurinol had no effect on cell number, controlled ATP release (solid bars, primary y axis) or viability (lines, secondary y axis). Values are means±SEM (n=6–12).

Article Snippet: Primary rat osteoblast cells were obtained from 2-day-old neonatal Sprague-Dawley rats euthanised by cervical dislocation, as described previously , .

Techniques:

Journal: Experimental Cell Research

Article Title: Allopurinol and oxypurinol promote osteoblast differentiation and increase bone formation

doi: 10.1016/j.yexcr.2016.03.004

Figure Lengend Snippet: Increased TNAP activity in osteoblasts treated with allopurinol or oxypurinol. TNAP activity was measured in differentiating (day 7) and mature (day 14) osteoblasts treated with 10 nM–0.1 µM allopurinol or oxypurinol. Basal TNAP activity was ~6-fold higher in mature, bone-forming osteoblasts. (A) Allopurinol (≥10 nM) increased TNAP activity by up to 50% and 65% at day 7 and day 14, respectively. (B) Oxypurinol (≥10 nM) stimulated TNAP activity by ≤60% in differentiating cells and mature osteoblasts. (C) Allopurinol and (D) oxypurinol have no effect on soluble collagen levels. Values are means±SEM (n=6), ***/###=p<0.001, **=p<0.01.

Article Snippet: Primary rat osteoblast cells were obtained from 2-day-old neonatal Sprague-Dawley rats euthanised by cervical dislocation, as described previously , .

Techniques: Activity Assay

Journal: Experimental Cell Research

Article Title: Allopurinol and oxypurinol promote osteoblast differentiation and increase bone formation

doi: 10.1016/j.yexcr.2016.03.004

Figure Lengend Snippet: Allopurinol and oxypurinol influence the expression of key osteoblast genes. The effect of allopurinol and oxypurinol (0.1 μM) on the expression of collagen type I (Col1α1), osteocalcin (Ocn), TNAP, XO and ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1) was in investigated in differentiating and mature osteoblasts. (A) Allopurinol decreased Npp1 expression ~5-fold in differentiating osteoblasts. (B) In mature osteoblasts, allopurinol increased TNAP and Ocn expression 2-fold and 5-fold, respectively. (C and D) Oxypurinol reduced Npp1 expression 5-fold in differentiating and mature osteoblasts. Levels of TNAP and Ocn expression were increased 2-fold and 5-fold, respectively in mature osteoblasts. Values are means±SEM (n=4), *=p<0.05.

Article Snippet: Primary rat osteoblast cells were obtained from 2-day-old neonatal Sprague-Dawley rats euthanised by cervical dislocation, as described previously , .

Techniques: Expressing