Introduction
Materials and Methods
Animals and Diets
Experimental Design
Structure of Tibias (In Vivo) and Femurs and Lumbar Vertebrae (Ex Vivo)
Post-Scanning Image Processing and Analysis
Bone Mineral Density (BMD) of Tibias, Femurs, and LV1-3
Peak Load of Tibias and Femurs
Statistical Analysis
Results
Food Intake and Body Weight
In Vivo Trabecular and Cortical BMD and Structure at the Proximal and Midpoint Tibia
Outcomes | CON | HSP + NAR |
p value | ||||||
---|---|---|---|---|---|---|---|---|---|
Age (months) | |||||||||
2 | 4 | 6 | 2 | 4 | 6 | Age | Diet | Age × diet | |
Trabecular structure | |||||||||
BMD (g/cm3) | 0.200 ± 0.008 | 0.193 ± 0.006 | 0.194 ± 0.008 | 0.216 ± 0.006 | 0.219 ± 0.011 | 0.199 ± 0.011 | 0.082 | 0.143 | 0.051 |
TV (mm3) | 1.496 ± 0.410a
| 1.163 ± 0.042b
| 1.134 ± 0.049b
| 1.324 ± 0.034*a
| 1.126 ± 0.030b
| 1.020 ± 0.039c
|
p < 0.001 | 0.114 | 0.028 |
BV (mm3) | 0.101 ± 0.014a
| 0.066 ± 0.007b
| 0.066 ± 0.009ab
| 0.100 ± 0.011ab
| 0.098 ± 0.014*a
| 0.064 ± 0.013b
| 0.005 | 0.485 | 0.006 |
BV/TV (%) | 6.66 ± 0.770 | 5.58 ± 0.49 | 5.73 ± 0.68 | 7.46 ± 0.67ab
| 8.65 ± 1.15*a
| 6.02 ± 1.09b
| 0.059 | 0.164 | 0.010 |
Tb.Th (mm) | 0.057 ± 0.001c
| 0.063 ± 0.001b
| 0.070 ± 0.001a
| 0.059 ± 0.001b
| 0.068 ± 0.001*a
| 0.072 ± 0.002a
|
p < 0.001 | 0.033 | 0.299 |
Tb.N (1/mm) | 1.151 ± 0.113 | 0.885 ± 0.077 | 0.824 ± 0.100 | 1.264 ± 0.098a
| 1.285 ± 0.173*a
| 0.858 ± 0.165b
| 0.002 | 0.230 | 0.020 |
Tb.Sp (mm) | 0.324 ± 0.013 | 0.359 ± 0.014 | 0.381 ± 0.016 | 0.309 ± 0.0149b
| 0.330 ± 0.026ab
| 0.382 ± 0.029a
| 0.002 | 0.522 | 0.366 |
DA (no unit) | 2.544 ± 0.103 | 2.396 ± 0.197 | 2.196 ± 0.216 | 2.568 ± 0.154 | 2.044 ± 0.147 | 2.518 ± 0.279 | 0.051 | 0.993 | 0.140 |
Conn.D (1/mm3) | 46.4 ± 3.7a
| 21.3 ± 2.4b
| 22.8 ± 3.8b
| 44.2 ± 4.89a
| 36.9 ± 6.6*ab
| 24.5 ± 2.4b
|
p < 0.001 | 0.240 | 0.075 |
Cortical structure | |||||||||
TMD (g/cm3) | 0.716 ± 0.007c
| 0.882 ± 0.008b
| 0.919 ± 0.011a
| 0.749 ± 0.008c
| 0.892 ± 0.011b
| 0.935 ± 0.019a
|
p < 0.001 | 0.098 | 0.299 |
Ct.Ar (mm2) | 1.091 ± 0.053c
| 1.222 ± 0.035b
| 1.323 ± 0.041a
| 1.284 ± 0.051*b
| 1.372 ± 0.063*b
| 1.507 ± 0.059*a
|
p < 0.001 | 0.014 | 0.633 |
Tt.Ar (mm2) | 4.376 ± 0.100a
| 3.748 ± 0.108b
| 3.802 ± 0.121b
| 4.179 ± 0.094a
| 3.811 ± 0.101b
| 3.812 ± 0.107b
|
p < 0.001 | 0.784 | 0.007 |
Ct.Ar/Tt.Ar (%) | 24.8 ± 0.8c
| 32.7 ± 0.6b
| 34.9 ± 1.0a
| 30.7 ± 0.8*c
| 36.0 ± 1.1*b
| 39.6 ± 1.2*a
|
p < 0.001 | p < 0.001 | 0.165 |
Ct.Th (mm) | 0.088 ± 0.004c
| 0.139 ± 0.002b
| 0.150 ± 0.004a
| 0.109 ± 0.004*c
| 0.152 ± 0.005*b
| 0.169 ± 0.007*a
|
p < 0.001 | 0.003 | 0.162 |
Ps.Pm (mm) | 8.578 ± 0.072a
| 8.174 ± 0.116b
| 8.292 ± 0.121b
| 8.397 ± 0.116a
| 8.165 ± 0.148b
| 8.311 ± 0.133ab
|
p < 0.001 | 0.044 | 0.095 |
Ec.Pm (mm) | 7.125 ± 0.099a
| 6.349 ± 0.114b
| 6.406 ± 0.127b
| 6.760 ± 0.124a
| 6.198 ± 0.088b
| 6.134 ± 0.152b
|
p < 0.001 | 0.110 | 0.189 |
Ma.Ar (mm2) | 3.285 ± 0.062a
| 2.526 ± 0.082b
| 2.479 ± 0.104b
| 2.895 ± 0.067*a
| 2.439 ± 0.071b
| 2.306 ± 0.084c
|
p < 0.001 | 0.058 | 0.005 |
Ecc (no unit) | 0.394 ± 0.026b
| 0.464 ± 0.020ab
| 0.475 ± 0.014a
| 0.388 ± 0.017c
| 0.436 ± 0.031b
| 0.451 ± 0.037a
| 0.026 | 0.505 | 0.852 |
Outcomes | CON | HSP + NAR |
p value | ||||||
---|---|---|---|---|---|---|---|---|---|
Age (months) | |||||||||
2 | 4 | 6 | 2 | 4 | 6 | Age | Diet | Age × diet | |
Cortical structure | |||||||||
TMD (g/cm3) | 1.043 ± 0.003c
| 1.135 ± 0.006b
| 1.168 ± 0.005a
| 1.070 ± 0.006*c
| 1.145 ± 0.008b
| 1.184 ± 0.008a
|
p < 0.001 | 0.020 | 0.144 |
Ct.Ar (mm2) | 0.829 ± 0.039c
| 0.977 ± 0.041b
| 1.080 ± 0.039a
| 0.879 ± 0.015b
| 1.091 ± 0.045a
| 1.121 ± 0.045a
|
p < 0.001 | 0.186 | 0.131 |
Tt.Ar (mm2) | 1.469 ± 0.072b
| 1.538 ± 0.079ab
| 1.640 ± 0.073a
| 1.450 ± 0.028b
| 1.661 ± 0.041a
| 1.642 ± 0.047a
|
p < 0.001 | 0.661 | 0.062 |
Ct.Ar/Tt.Ar (%) | 56.2 ± 0.6c
| 63.9 ± 0.9b
| 66.2 ± 1.0a
| 60.6 ± 1.2c
| 65.6 ± 1.4b
| 68.2 ± 1.0a
|
p < 0.001 | 0.066 | 0.130 |
Ct.Th (mm) | 0.200 ± 0.005c
| 0.239 ± 0.004b
| 0.256 ± 0.005a
| 0.220 ± 0.005*c
| 0.255 ± 0.010b
| 0.273 ± 0.009a
|
p < 0.001 | 0.044 | 0.910 |
Ps.Pm (mm) | 4.872 ± 0.146b
| 5.096 ± 0.179ab
| 5.336 ± 0.157a
| 4.851 ± 0.065b
| 5.403 ± 0.100a
| 5.241 ± 0.100a
|
p < 0.001 | 0.718 | 0.069 |
Ec.Pm (mm) | 3.384 ± 0.129a
| 3.100 ± 0.143b
| 3.092 ± 0.136b
| 3.154 ± 0.051a
| 3.163 ± 0.069a
| 2.956 ± 0.048b
|
p < 0.001 | 0.510 | 0.016 |
Ma.Ar (mm2) | 0.640 ± 0.035a
| 0.560 ± 0.041b
| 0.559 ± 0.038b
| 0.574 ± 0.026ab
| 0.570 ± 0.021a
| 0.520 ± 0.014b
| 0.001 | 0.468 | 0.026 |
Ecc (no unit) | 0.705 ± 0.010 | 0.711 ± 0.017 | 0.704 ± 0.013 | 0.712 ± 0.013 | 0.732 ± 0.011 | 0.712 ± 0.013 | 0.125 | 0.502 | 0.541 |
Ex Vivo BMD, Trabecular, and Cortical Bone Structure at the Femur and Lumbar Vertebrae
CON | HSP + NAR |
p value | |
---|---|---|---|
Femur BMD (g/cm2) | 0.089 ± 0.003 | 0.091 ± 0.003 | 0.540 |
Femur BMC (g) | 0.044 ± 0.002 | 0.044 ± 0.002 | 0.997 |
Femur Area (cm2) | 0.497 ± 0.011 | 0.482 ± 0.008 | 0.287 |
Femur neck trabecular structure | |||
TV (mm3) | 0.540 ± 0.046 | 0.504 ± 0.032 | 0.533 |
BV (mm3) | 0.205 ± 0.020 | 0.211 ± 0.010 | 0.782 |
BV/TV (%) | 37.9 ± 1.7 | 42.7 ± 2.5 | 0.128 |
Tb.Th (mm) | 0.098 ± 0.002 | 0.102 ± 0.004 | 0.453 |
Tb.N (1/mm) | 3.854 ± 0.158 | 4.190 ± 0.175 | 0.176 |
Tb.Sp (mm) | 0.189 ± 0.009 | 0.166 ± 0.006* | 0.049 |
Femur midpoint cortical structure | |||
Ct.Ar (mm2) | 1.401 ± 0.055 | 1.486 ± 0.082 | 0.408 |
Tt.Ar (mm2) | 2.823 ± 0.139 | 2.736 ± 0.093 | 0.087 |
Ct.Ar/Tt.Ar (%) | 50.0 ± 1.6 | 54.2 ± 5.3 | 0.112 |
Ct.Th (mm) | 0.245 ± 0.008 | 0.267 ± 0.011 | 0.119 |
Distal femur trabecular structure | |||
BV/TV (%) | 11.5 ± 1.2 | 13.4 ± 0.981 | 0.232 |
Tb.Th (mm) | 0.073 ± 0.001 | 0.076 ± 0.002 | 0.203 |
Tb.N (1/mm) | 1.594 ± 0.166 | 1.778 ± 0.139 | 0.411 |
Tb.Sp (mm) | 0.381 ± 0.050 | 0.339 ± 0.027 | 0.465 |
CON | HSP + NAR |
p value | |
---|---|---|---|
LV1-3 BMD (g/cm2) | 0.077 ± 0.002 | 0.079 ± 0.002 | 0.500 |
LV1-3 BMC (g) | 0.032 ± 0.001 | 0.034 ± 0.001 | 0.370 |
LV1-3 Area (cm2) | 0.413 ± 0.013 | 0.422 ± 0.013 | 0.630 |
LV2 trabecular structure | |||
TV (mm3) | 1.071 ± 0.081 | 1.435 ± 0.035* | 0.002 |
BV (mm3) | 0.250 ± 0.024 | 0.364 ± 0.035* | 0.016 |
BV/TV (%) | 22.9 ± 1.04 | 26.0 ± 2.2 | 0.337 |
Tb.Th (mm) | 0.070 ± 0.001 | 0.072 ± 0.001 | 0.331 |
Tb.N (1/mm) | 3.270 ± 0.138 | 3.512 ± 0.262 | 0.397 |
Tb.Sp (mm) | 0.235 ± 0.008 | 0.232 ± 0.012 | 0.978 |
LV2 cortical structure | |||
Ct.Ar (mm2) | 0.268 ± 0.008 | 0.266 ± 0.017 | 0.918 |
Ct.Th (mm) | 0.086 ± 0.003 | 0.081 ± 0.004 | 0.290 |