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Histomorphometric changes in the tibia of ovariectomized aged wistar rats treated with calcitonin
Abstracts The 7th International
146s
Congress
Bone Vol. 19, No. 3, Supplement September 1996: 129S- 169s
of Bone Morphometry
69
67 LONGITUDINAL TERM
AND
SKELETAL
z,
H.
Department
CROSS-SU-TIONAL
EFFECTS
OF AGING
CHARAClZRIZATION AND
OF LONG-
ORCHIDECTOMY
Qi. D.T. Crawford. CM. Pie, H.A. of Metabolic Diseases, Pfizer Centi
IN THE
MALE
Simmons, D.D. Thompson Research, Gmton.
CT (U.S.A.)
We rat is widely used as a” animal model for skeletal research. The effects of aging and sex hormone deficiency on skeletal metabolism have been studied in great detail in female mts. However, only a few imd incomplete studies have bee” pufo”ncd
in male
rats.
and sex
Therefae.
OUT potpose was to detail the long-term
hormone deficiency on skeletal metabolivn In the longilodinal ray absqtiometry Body wcighr
(ORX.
(DXA)
were shan~qexated
(SHAM.
was per-fom~ed at 3.4.6.8,
x-
10, 12. and 20 months of age.
tolal body bone area, bone mineral content and density wcz w
study. Spmgoe-Hawley nwnlhs
male rau
n=lO) at 3 months of age. Whole body dual-energy
Fat and lean body mass were also &te”“ined SHAM
of aging
rats and eight
male rats were SHAM
or ORX
at 3 months of age. Eight 3,4.6,8.10.12,15,
femoral bone awa, bone mineral
density were determined by DXA histomorphomcuic
al 20 months of age. In the cross-sectional
ORX rats were serially sacrificed at
of age. Tolal
ex
viva.
Static
and 23
conten& and bone mineral
and dynamic
canceUcus bone
analyses were performed on proximal tibial metaphyses.
The age-related changes in male ra~$ can be stannwized
as following:
(a) Bcdy weight
increased gradually from 520 g at 3 months of age to 913 g at 15 months of age ml thereafter maintained this level. (b) Bone mass rapidly increased until 6 months of age, and the” slightly increased behvcen 6 to 10 months of age. &crease in bone mass was found between 12 to 23 nwxbs The long-term following:
effects of
(a) Signifzantly
ORX
at 3 month
weight was maintained lower than SHAM less weight than SHAM).
(b) Significantly and nuinlained
the slwiy (approximately inc%wd
ostwclastic
Tiwwdtw
a” aged-r&cd
of age.
old male rats ca” be summarized
as
inhibited weight gain 2 months post-surgery and the tody
mass at 1 month post-ORX
20%
throughout the study (appmximaIely
20%
inhibited lhe age-related incwseinbone at lower levels thmughout the remainder d
less peak bone mass than SHAM).
bone resorption
during
the fmt
3
(c) Significantly
months
post-ORX.
ad
significantly decreased osteoblastic bone formation beyond 3 nmnths when compared 10 shvn connols. (d) The lower bone mass in ORX rats WBSdoe to the lack of age&a&d bone gain rather than the net loss from basal contmls (3 months of age). (e) Lean tcdy mass was signiIica”tly
lowered (-22%)
L. Khaldi, G. Skarantavos, G. Lyritis Laboratory for the Research of Musculoskeletal KAT Hospital, Athens (Greece)
System,
in male rats. Two studies were pzfcaned.
study. Sprague-Da&y
“=lO) OTorchidccbnnized
effeas
HISTOMORPHOMETRIC CHANGES IN THE TIBIA OF OVARIECTOMIZED AGED WISTAR RATS TREATED WITH CALCITONIN
in ORX than in SHAM
Ow data indicated thahat (a) peak boot mass axaned
at 20 months of age.
Calcitonin (CT) is known to restrain bone loss in early postmenopausal period, but its effect in aged bone is obscure. The aim of this study is to investigate the histological and morphometrical changes in the tibias of aged wistar rats treated with CT. 4 groups of 9 aged rats (ZO-22months) were used. which underwent ovarisctomy at the age of 3-4 months. Group A was killed at the beginning of the experiment. Groups B.C were treated respectively with 6 and 20 IUlkg of CT subcutaneously, daily for 2 months. Group D was placebo treated Qroups B.C and D were killed after the two months period and the upper part of the tibias was formalin fixed, decalcified and paraffin embedded, histological remarks were registered were done using the following and morphometrical observations parameters: trabscular perimeter (TP), trabecular width m. bone area, TBV. % calcified length, % fibrosis, % active resorbtion. % inactive resorbtion. % total resorbtion, no. of osteoclastes (cl), no. of fields. ANOVA statistical analysis was performed. Our results were listed as followed: TP {AA. B>D. C>A p=O.Ol}. TBV {B>A. B>D. C>A p=O 01). % cal. cart leng. {DcA. DB p=O.O013. A>C p=O.O085. BC p=O.O061. B>C p=O.O015. BzD p=O.O64. CC p=O.O047. B>C p=O.O036. CB p=O.O49. BD p=O.O065. C>D p=O.O45}. It is concluded that CT depresses the no. of osteoclasts and increases the TBV of the upper tibia1 metaphyisis. Low CT doses seems to be efficient for this effect.
at 10 months of age, and age-reLated
decrease in bone mass occurred at 12 months of age in male rats. and (b) sex hormone is a” important factor for the accumulation of peak bone and muscle mars in male rats.
70
68 OBSERVATION OF MORPHOMETRICAL GROWTH PLATE OF ADULT MALE WISTAR TREATED WITH SALMON CALCITONIN. L. Kha@i, G. Lenis, Th. Karachalios, G. Lyritis Laboratory for the Research of Musculoskeletal KAT Hospital, Athens (Greece)
THE RATS
System,
Calcitonin is used widely as an antiosteoclastic drug, but their is no information of its effect on the epiphysial plate after the long term treatment. Aim of this study is to signify histomorphometricaly action of salmon Calcitonin on the growth plate. We used 18 male wlstar rats 3 month old. Srats were treated with subcutaneous injection of 5 IU/kg of calcitonin for 12 weeks, the other 9 received a placebo dose for the same period. The rats were killed at the end of the experiment. The distal femoral’s epiphysises were formalin fixed, decalcified and paraffin embedded, stained with H/E, Alcian blue. We measured the number of chondrocytes per column of the proliferative and hypertrophic zone. Our results were estimated with per student t-test. A significant increase (p=O.O012) of condrocytes column were found in the calcitonin treated group. It is concluded that calcitonin has promotive effect on the epiphysial plate function.
Mean No. Of chondrocytes /column rats with SCT rats with placebo
6.4 4.9
p=o.o012
DIRECT INCORPORATION OF 3-DIMENSIONAL MICROTOMOGRAPHY DATA INTO AN ELASTIC MODEL OF HUMAN TRABECULAR BONE. J.H. Kinney, A.J.C. Ladd, D.L. Haupt, S. Majumdar, Lawrence Livermore National Laboratory, Livermore, CA 94551 and U.C. San Francisco, San Francisco, CA 94143. We have developed a lattice spring model that can calculate the elastic properties of materials having very complex microstructures. This model is much faster than traditional finite element methods. Complex microstlvctures with up to 10’ nodes can be analyzed in a few hours on a standard workstation. Each volume element (voxel) of a 3D x-ray computed tomography image is treated like an elastic element with nodes at the eight vertices. The nodes are connected by springs in the [lo01 and [I101 crystallographic directions. A suitable choice of spring constants for the different directions leads to elastic isotropy on the voxel scale. The elastic model of the bulk structure is obtained by combining the elastic elements. External displacements are then applied to the surfaces. Equilibrium displacements and stress fields are found by minimizing the strain energy. In this study, 3D microtomography was used to image the structure of fully hydrated human cancellous bone in the distal radius. These images were directly incorporated into the model, and the Young’s moduli in compression were calculated along the proximal-distal and mediallateral directions. The samples were then tested in compression. The experimental Young’s moduli ranged from 450 MPa to 614 MPa, indicating that the tissue modulus varied between 11-15 GPa. The ratios of the moduli between prox/dist and med/lat directions ranged from 2.13-3.8 (experiment) and 2.1-3.6 (calculated). The results of this study indicate that a lattice spring model, incorporating detailed 3d bone morphology, accurately predicts elastic properties of cancellous bone. An extension of this model that incorporates nonlinear and fracture behavior is in progress. Work performed under the auspices
of
the US Department
of Energy,
contract
W-7405-Eng-48.
146s
Congress
Bone Vol. 19, No. 3, Supplement September 1996: 129S- 169s
of Bone Morphometry
69
67 LONGITUDINAL TERM
AND
SKELETAL
z,
H.
Department
CROSS-SU-TIONAL
EFFECTS
OF AGING
CHARAClZRIZATION AND
OF LONG-
ORCHIDECTOMY
Qi. D.T. Crawford. CM. Pie, H.A. of Metabolic Diseases, Pfizer Centi
IN THE
MALE
Simmons, D.D. Thompson Research, Gmton.
CT (U.S.A.)
We rat is widely used as a” animal model for skeletal research. The effects of aging and sex hormone deficiency on skeletal metabolism have been studied in great detail in female mts. However, only a few imd incomplete studies have bee” pufo”ncd
in male
rats.
and sex
Therefae.
OUT potpose was to detail the long-term
hormone deficiency on skeletal metabolivn In the longilodinal ray absqtiometry Body wcighr
(ORX.
(DXA)
were shan~qexated
(SHAM.
was per-fom~ed at 3.4.6.8,
x-
10, 12. and 20 months of age.
tolal body bone area, bone mineral content and density wcz w
study. Spmgoe-Hawley nwnlhs
male rau
n=lO) at 3 months of age. Whole body dual-energy
Fat and lean body mass were also &te”“ined SHAM
of aging
rats and eight
male rats were SHAM
or ORX
at 3 months of age. Eight 3,4.6,8.10.12,15,
femoral bone awa, bone mineral
density were determined by DXA histomorphomcuic
al 20 months of age. In the cross-sectional
ORX rats were serially sacrificed at
of age. Tolal
ex
viva.
Static
and 23
conten& and bone mineral
and dynamic
canceUcus bone
analyses were performed on proximal tibial metaphyses.
The age-related changes in male ra~$ can be stannwized
as following:
(a) Bcdy weight
increased gradually from 520 g at 3 months of age to 913 g at 15 months of age ml thereafter maintained this level. (b) Bone mass rapidly increased until 6 months of age, and the” slightly increased behvcen 6 to 10 months of age. &crease in bone mass was found between 12 to 23 nwxbs The long-term following:
effects of
(a) Signifzantly
ORX
at 3 month
weight was maintained lower than SHAM less weight than SHAM).
(b) Significantly and nuinlained
the slwiy (approximately inc%wd
ostwclastic
Tiwwdtw
a” aged-r&cd
of age.
old male rats ca” be summarized
as
inhibited weight gain 2 months post-surgery and the tody
mass at 1 month post-ORX
20%
throughout the study (appmximaIely
20%
inhibited lhe age-related incwseinbone at lower levels thmughout the remainder d
less peak bone mass than SHAM).
bone resorption
during
the fmt
3
(c) Significantly
months
post-ORX.
ad
significantly decreased osteoblastic bone formation beyond 3 nmnths when compared 10 shvn connols. (d) The lower bone mass in ORX rats WBSdoe to the lack of age&a&d bone gain rather than the net loss from basal contmls (3 months of age). (e) Lean tcdy mass was signiIica”tly
lowered (-22%)
L. Khaldi, G. Skarantavos, G. Lyritis Laboratory for the Research of Musculoskeletal KAT Hospital, Athens (Greece)
System,
in male rats. Two studies were pzfcaned.
study. Sprague-Da&y
“=lO) OTorchidccbnnized
effeas
HISTOMORPHOMETRIC CHANGES IN THE TIBIA OF OVARIECTOMIZED AGED WISTAR RATS TREATED WITH CALCITONIN
in ORX than in SHAM
Ow data indicated thahat (a) peak boot mass axaned
at 20 months of age.
Calcitonin (CT) is known to restrain bone loss in early postmenopausal period, but its effect in aged bone is obscure. The aim of this study is to investigate the histological and morphometrical changes in the tibias of aged wistar rats treated with CT. 4 groups of 9 aged rats (ZO-22months) were used. which underwent ovarisctomy at the age of 3-4 months. Group A was killed at the beginning of the experiment. Groups B.C were treated respectively with 6 and 20 IUlkg of CT subcutaneously, daily for 2 months. Group D was placebo treated Qroups B.C and D were killed after the two months period and the upper part of the tibias was formalin fixed, decalcified and paraffin embedded, histological remarks were registered were done using the following and morphometrical observations parameters: trabscular perimeter (TP), trabecular width m. bone area, TBV. % calcified length, % fibrosis, % active resorbtion. % inactive resorbtion. % total resorbtion, no. of osteoclastes (cl), no. of fields. ANOVA statistical analysis was performed. Our results were listed as followed: TP {A
at 10 months of age, and age-reLated
decrease in bone mass occurred at 12 months of age in male rats. and (b) sex hormone is a” important factor for the accumulation of peak bone and muscle mars in male rats.
70
68 OBSERVATION OF MORPHOMETRICAL GROWTH PLATE OF ADULT MALE WISTAR TREATED WITH SALMON CALCITONIN. L. Kha@i, G. Lenis, Th. Karachalios, G. Lyritis Laboratory for the Research of Musculoskeletal KAT Hospital, Athens (Greece)
THE RATS
System,
Calcitonin is used widely as an antiosteoclastic drug, but their is no information of its effect on the epiphysial plate after the long term treatment. Aim of this study is to signify histomorphometricaly action of salmon Calcitonin on the growth plate. We used 18 male wlstar rats 3 month old. Srats were treated with subcutaneous injection of 5 IU/kg of calcitonin for 12 weeks, the other 9 received a placebo dose for the same period. The rats were killed at the end of the experiment. The distal femoral’s epiphysises were formalin fixed, decalcified and paraffin embedded, stained with H/E, Alcian blue. We measured the number of chondrocytes per column of the proliferative and hypertrophic zone. Our results were estimated with per student t-test. A significant increase (p=O.O012) of condrocytes column were found in the calcitonin treated group. It is concluded that calcitonin has promotive effect on the epiphysial plate function.
Mean No. Of chondrocytes /column rats with SCT rats with placebo
6.4 4.9
p=o.o012
DIRECT INCORPORATION OF 3-DIMENSIONAL MICROTOMOGRAPHY DATA INTO AN ELASTIC MODEL OF HUMAN TRABECULAR BONE. J.H. Kinney, A.J.C. Ladd, D.L. Haupt, S. Majumdar, Lawrence Livermore National Laboratory, Livermore, CA 94551 and U.C. San Francisco, San Francisco, CA 94143. We have developed a lattice spring model that can calculate the elastic properties of materials having very complex microstructures. This model is much faster than traditional finite element methods. Complex microstlvctures with up to 10’ nodes can be analyzed in a few hours on a standard workstation. Each volume element (voxel) of a 3D x-ray computed tomography image is treated like an elastic element with nodes at the eight vertices. The nodes are connected by springs in the [lo01 and [I101 crystallographic directions. A suitable choice of spring constants for the different directions leads to elastic isotropy on the voxel scale. The elastic model of the bulk structure is obtained by combining the elastic elements. External displacements are then applied to the surfaces. Equilibrium displacements and stress fields are found by minimizing the strain energy. In this study, 3D microtomography was used to image the structure of fully hydrated human cancellous bone in the distal radius. These images were directly incorporated into the model, and the Young’s moduli in compression were calculated along the proximal-distal and mediallateral directions. The samples were then tested in compression. The experimental Young’s moduli ranged from 450 MPa to 614 MPa, indicating that the tissue modulus varied between 11-15 GPa. The ratios of the moduli between prox/dist and med/lat directions ranged from 2.13-3.8 (experiment) and 2.1-3.6 (calculated). The results of this study indicate that a lattice spring model, incorporating detailed 3d bone morphology, accurately predicts elastic properties of cancellous bone. An extension of this model that incorporates nonlinear and fracture behavior is in progress. Work performed under the auspices
of
the US Department
of Energy,
contract
W-7405-Eng-48.