Table 5.
Femur (trabecular) static/dynamic histomorphometry: Wild type & Mutant Mice.
| Absolute Values | ||||||||
|---|---|---|---|---|---|---|---|---|
| Femur (Cancellous-bone) | WT (n=9) | MN (n=6) | MNAt (n=6) | HYP (n=9) | ||||
| * | § | ρ | # | |||||
| BV/TV (%) | 20.7 ± 1.5 | §# | 26.7 ± 1.9 | *ρ# | 21.4 ± 1.4 | §# | 2.76 ± 0.49 | *§ρ |
| Tb. Th (um) | 51.3 ± 4.1 | § | 62.3 ± 2.4 | *ρ# | 46.0 ± 1.6 | § | 47.5 ± 3.9 | § |
| Tb. Sp (um) | 209.4 ± 2.5 | §# | 163.5 ± 4.5 | *ρ# | 200.3 ± 6.6 | §# | 782.7 ± 245.10 | *§ρ |
| Tb.N | 3.8 ± 0.07 | §# | 4.6 ± 0.12 | *ρ# | 3.9 ± 0.11 | §# | 1.0 ± 0.20 | *§ρ |
| Oc.S/BS (%) | 14.9 ± 3.4 | # | 20.9 ± 3.2 | 22.5 ± 4.8 | 29.2 ± 6.7 | * | ||
| Oc.N/BS (#/mm) | 2.5 ± 0.5 | # | 2.9 ± 0.32 | # | 2.3 ± 0.48 | # | 5.49 ± 0.29 | *§ρ |
| O.Wi (μm) | 1.94 ± 0.29 | ρ# | 1.95 ± 0.32 | ρ# | 3.1 ± 0.48 | *§# | 6.1 ± 0.32 | *§ρ |
| OV/BV (%) | 0.05 ± 0.03 | ρ# | 0.08 ± 0.03 | ρ# | 0.36 ± 0.07 | *§# | 227.10 ± 28.90 | *§ρ |
| Ob.S (mm) | 1.5 ± 0.07 | § | 2.6 ± 0.28 | *ρ | 1.4 ± 0.22 | § | ND | — |
| Ob.S/BS (%) | 14.8 ± 1.6 | § | 22.1 ± 2.18 | *ρ | 15.4 ± 1.82 | § | ND | — |
| Ob.N/BS (#/mm) | 11.1 ± 1.8 | § | 16.7 ± 0.9 | *ρ | 11.2 ± 0.96 | § | ND | — |
| BFR/TV (per/day)*1000 | 5.4 ± 0.4 | § | 7.8 ± 0.66 | *ρ | 5.8 ± 0.48 | § | ND | — |
| MAR (μm/day) | 1.39 ± 0.03 | § | 1.57 ± 0.05 | *ρ | 1.27 ± 0.05 | § | ND | — |
Bone (femur, epiphyseal cancellous bone) histomorphological analysis of histology stained slides (see methods) for wild type mice (WT), MEPE−/− mice (MN), MEPE−/−/ASARM-transgenic mice (MNAt) and X-linked hypophosphatemic mice (HYP): Male mice (age 5.4 months; N=6) fasted for 16 hours and housed singly in metabolic cages (water ad libitum) were sacrificed and femurs removed for histology and μCT analyses. Columns show absolute values with standard error of the means (SEM). Statistical differences between columns were measured by one-way analysis of variance (ANOVA) followed by Tukey's post multi comparison test. Changes that are statistically significant different (p<0.01) are denoted by the following superscript codes:
statistically different to wild type mice (WT)
statistically different to MEPE−/− mice (MN)
statistically different to MEPE−/−/ASARM transgenic mice (MNAt) and
statistically different to X-linked hypophosphatemic rickets mice (HYP). Explora-Nova histomorphometric software (F17011 LaRochelle Cedex 1France) was used to analyze the microscopically imaged samples and the guidelines as described by us previously and discussed in methods (14,16,33). The methods and guidelines of Ott 2008 (44) were used to calculate histomorphometric measurements of bone turnover, mineralization and volume. Bone histomorphometric nomenclature, symbols and units were standardized as described by Parfitt et al 1987 (43) with guidelines for assessment from Bouxsein et al 2010 (42).
Index: BV/TV (%) = Bone-volume/total-volume(%); Tb.Th (μm) = trabecular thickness (μm); Tb.Sp(μm) = trabecular spacing(μm); Tb.N = trabecular number (mm−1); Oc.S/BS (%) = osteoclast-surface/bone-surface (%); Oc.N/BS (#/mm) = osteoclast-number/bone-surface (#/mm); O.Wi (μm) = Osteoid Width (%); OV/BV (%) = osteoid-volume/bone-volume (%); Ob.S (mm) = osteoblast-surface (mm); Ob.S/BS (%) = osteoblast-surface/bone-surface (%); Ob.N/BS (#/mm) = osteoblast-number/bone-surface (#/mm); BFR/TV (per/day)*1000 = bone formation rate with tissue volume referent in per day units (μm/day*1000); MAR (μm/day) = mineral apposition rate (μm/day).