Ca45 incorporation into the bones of the limbs, particularly in the area of the muscle attachment increased in dogs as a result of 6 month hypokinesia. There were no phenomena of osteoporosis in the cortical layer of the diaphyses; however, changes in the form of osteons, an increase in the number of anastomoses between the channels and the thinning of the subperiosteal layer pointed to disturbances of the bone tissue reconstruction. Mineral saturation of the bone microstructures of the experimental dogs had a tendency to rise. No changes in the mechanical properties of the long bones occurred as a result of hypokinesia in dogs

Korzhenyants, V. A.

Muradov, I. S.

Pavlova, M. P.

Stupakov, G. P.

Volozhin, A. I.

English

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NASA TECHNICAL MZHORANDUl1 NASA TM-76168 
EFFECT OF SIX-MONTll HYPOKINESIA IN DOGS ON MINERAL 
COMPONENT, RECONSTRUCTION AND MECHANICAL PROPERTIES OF BONE TISSUE 
A. I. Volozhin, M. P. Pavlova, I. Sh. Muradov, 
G. P. Stupakov, V. A. Korzhen'yants 
Translation of "Vliyaniye. 6-mesyachnoy kipokinezii u sobak na 
mineral 'nyy komponent, perestroyku i mekhanicheskiye svovstva 
kostnoy tkani", Patologicheskaya l?hiologiya i Ekspedmenta1'naya 
Terapiya, No.6, Nov-Dec 1976, pp 34-38 
(NASA-XM-7b160) EFFECT OF SIX-MONTH \ 
HYPOKiNESIA IN DOGS ON MINEBAL COMPONENT, 
RECONSTRUCTION AND MECHANICAL ~BOP~RTIES OF 
BONE TISSUE (National Aeronaut~cs and spa~e. G5/51 
Administration) 10 p HC Au2/MF A01 CSCL Que 
HBl-2069B 
Unclas 
41871 
NATIONAL AERONAUTICS AND SPACE ADMINISTRATION 
WASHIN{~JON, D. c. 20546 ~.Ay 1980 
llAC.A. Sdonti.fic :,l.l1d 'i'f.)..!~mlcal Il1i'orma tion ,I"acilitl' '. 
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/ -----~----------~------------------------r_----------------------~ I \, Rtp."I'\O. _____ ..JI'L..,: '_,_Ci_O_,,_._rn_m.n. A .. uHlon N., i 3. lhcipi .. n'" C.IGI.o tho NASA TM-76l63 _ 
.1, T.lt. Gnd $111011,1. EFFECT OF SIX-MONTH HYPOKINESIA I t .s'~yOl880 
DOCS ON MINERAL COMPONENT, RECONSTRUCTION AND I 
! ~mCHANICAL PROPERTIES Oll' BONE TISSUE ' 6, P.t/.,mlno Olo~nl .. ItCln Cod. 
I 1, Alllh\\r( I) lAo I. Volozhin, M. P. Pavlova, I. She Mllradov, 
\0, WOII. Uni' No. I 
I--~-
I G. P. Stullakov, V. A. Korzhen'yanta 
: 15. S\l,.pl.l'II.nl~y Nol •• 
l 'franslation of: "Vliyuniye 6-mesyachnoy kipokinezii U sobak na mineral 'nyy 
f komponent, perestroykll :l mekhanicheskiye svoystva kostnoy tkan:l", 
. Patologicheskaya Fiziologiya i Eksperimental' nayn Terapiya, No.6, 
I Nov-Dec. 1976, pp 34 .. 38 
I 
!';. 4'5,'IGCI' ~ 
Ca incl'1rporation into the bones of the limbs, particularly in the area 01 
• the muscle a ttachment increased il\ dogs as a result of 6-lIlonth hypokinesia 
i There. were no phenomena of osteoporosis in the cortical layer of the dia-
physes; however, chang(;..I:J in the form of osteons, an increase in the number 
\ of anastOmoses between the channels and the thinning of th~ subperiosteal 
, layer pointed to distu);bances of the bone tissue );econstrllction. Mineral 
I satu:r.ation of the bone microstructures of the experimental dogs had a ten-
I dency to rise. 
No changes in tbe mechanical p);operties of the long bones occurred as a 
1 result of hypokinesia in dogs. ) 
. . ~ 
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THIS COPYRIGU'rED SOVIET WORK IS REPRO-
DUCED AND SOLD BY NTIS UWOER LICENSE 
FROM V AAP, THE S(WtET COPYRIGHT AGENCY. 
~O FURTHER COPYING IS PERHITTED WITH-
OUT PERMISSION FROM VAAP 
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uno 612.766.2-08.612.751.1.015.31 
EFFEOT OF SIX-MONTH HYFOKmESJA IN DOGS ON M:rnERAL 
COw. ·.JNENT, REOONSTRUOTION AND MEOHANIOAL PROPERTIES 
OF~<jNE TISSUE 
By 
A. I. Volozhin, M. P. Pavlova, I. She Muradov, G. P. 
stupakov, V. A. Korzhen'yants*' 
As a consequonce of the prolonged xeduct1.on in the functional load on the 
suppoxt-motox apparatus the bone tissue is exposed to atxophic changes. Da-
minex8,lization of the bones and a negative calcium balance wete found in pa:r:alyzed 
patients [11, 14J, as well as in pXactically healthy human volunteers undex con-
ditions of hypokinesia for several weeks and even months [4,10, 15]. In expexi-
ments on rats and rabbits restricted movement induced changes in the calCium, 
phosphorous and protein metabolism L1, 5,7J, as well as a slowing down of the 
xenewal of calcium salts L2,3J. There ro1e no published data on too state of 
minaxal saturation of the bone tissue micros'bructures and the mechanioal ;pro-
pexties of the bone in. relation to the peoulia:r:ities of its reoonstruotion with 
xestricted motor activity. 
Technique 
E~ximents wera set up on 16 dogs of both sexes in age li-2 yaaxs w~ighing 
8-12 kg. Hypokinesia lasting 6 months was created in 8 animals bit· :placing' them 
in spac~_ally designed metal cages Whose volume roughly corresponded to the size of 
the dog. Eight dogs were tha controls. During the entixe pexiod of hypokinesia 
the dogs wexe in the lying position, and thorough sanitary and hygienic ca:r:e was 
* Depaxtrnent of Pathological pr.ysiology of the N. A. Semashko Moscow Medical 
Stomatological Institute and the Laboratoxy of Biophysics of the N. N. Pxiorov 
Oentxal ~.Jlstitute of Txawnatology and Orthopedics • 
. If* 
Numbers in ma:r:gin indicate pagination in oxiginal fr . ...:eign text. 
1 
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taken of them. The calcium metabolism in the mineralized tissues was studied in 
6 experimental and 6 control dogs with the help of Ca4S that in too form of a 
chloride salt diluted in an isotonic solution of NaCl was administered intraveno'~ly 
in a dose ot' 100 mOi per 10 Kg of animal weight on the 180th day of the experioont, 
Within 24 h after administration of Ca4S the dogs were killed by etoor vapors and 
chloroform vapors, the maxillaxy, humeral, femoral and tibial bones were removed 
from the right side, they were cleaned of the soft tissues and fixed in 10% neutral 
formalin. The inCisors, premolaxs and molaxs were extrac'~ed from the jaw. With the 
help of 13. fine metal cutter fragments were sawed off the middle of the diaphysB's /35 
of the long bones, heads of the humerus and femur, trochanter major of the femur 
and tubercle major of the humer us. The bone fragments were dried to a constant 
weight, weighed, incinerated in a muffle furnace, the ash was again weighed and 
the content of ash rasldue in percentages of dry weight of the tissue was calculated. 
After pulverization in mo:rtax standard suspensions were made of the ash of 20 mg 
each that were placed in a targe'c, radiometry was conducted on the ,unit "Tesla" with 
the help of a window counter T-2S-BFL. The t:adioactivity' was computed in impulses 
~er 1 mg of ash. 
~, In the cortical bone measurements were made of the average mineral saturation 
by a direct method--by determination of the volume of diaphysis fragment (by weighing 
on torsion 'Scales in air and in "Tater) with its subsequent incineration. From the 
microsections the mineral saturation was determined in the microstructures of the 
subperiosteal, intern~dial and subendosteal zones of the cortical layer in the bone 
diaphyses with the help of the method of quantitative microroentgenography [6,12J. 
The. results of the analysis of average mineral saturation and in the bone micro-
struc~ures ware expressed in gra~s of mineral salts per 1 cm3 of tissue. 
For histological study the fragments of the bone diaphyses were decalcinated 
in 10% nitric acid, and poured into celloidin. Sections 10 !lm thick were stained 
with hematoxylin-eosin. From the histological preparations on the cross sections 
of the diaphyses the width afthe osteons and the haversian canals was determined. 
The average dimensions and frequency of distribution of these structures with 
respect to width were determined. 
The left femoral and humeral bones of the dogs were fixed in a 1% solution of 
neutral formalin and x-rayed in the sagittal and frontal projections without a 
2 
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--~- ~------
--'-"'<~-
magnification screen. From the x-ray photographs with the help of a MBS-l micro-
scope in the middle third of the bone diaphysis the external diameter, width of 
the ~dull~ canal and the thickness of the cortical layer were determined. No 
later than in 48 h after the extraction the mechanical ~roperties of the whole 
femur and humerus waxe studied in 4 dogs by testing them for static bending on 
a standard machine with maximum force SOO kg-f. The error in oomputing the 
indices of the strength-measuring device is about ~ of the measured load. The 
load conditions: di~tance between the supports 6.5 mm, axis of the bending knife 
coincided with the middle of this distance and with the middle of the length of 
the bone~ the rate of loading was 10 mm/min. The tensile strength was computed 
M· 
according to the formula: ~, where M --bending moment, Wx--moment of resistanoe. 
"x x The magnitude of the moment of inertia necessary to calculate the moment of 
resistance was determined with the assumption that in the cross section the external 
shape of the bone and the shape of the medull~ oanal represent a regular ellipse. 
The numerical data were statistically prooessed according to Fishe~-Student. 
Results and Discussion 
.,. - - .. 
After hypokinesia the inclusion of Ca4.5 (see table) was increased in the 
diaphysis of the femur and trochanter major, while it was reduced in the diaphysis 
of the humerus. The trend towards the increase in inclusion of Ca4.5 was revealed 
in the head of the humerus and feml~, and a considerable increase--in the tu-
bercle major of the humerus. Ca4S assimilation in the lower jaw of the experimental 
animals was redUced, and in the upper jaw was not altered. Inclusion of the 
calcium isotope was considerably reduced in the lower premolars and molars, and in 
the other teeth corres]Onded to the control. No significant changes wexe revealed 
in the content of the ash residue in all the mineralized tissues of the experimental 
animals as compared to the control. 
Hypokinesia in the clogs did not affect the ratio of the thickness of the 
cortical layer, as well as the medullar canal to the external width of the bone 
diaphyses. 
As a result of the morphometric study it was shown that the average width 
of the osteons in the diaphyses of different long bones in the control dogs was 
191-264~m, and of the haversian canals--38.9-.5.5.0 ~m. In dogs after hypokinesia 
3 
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the ave~age width of the osteons oo~~esponded to the oont~ol, while the ava~age 
diamete~ of the have~sian canals rose insignifioantly. 
About .50% of the have~sian canals in the diaphyses of the long bones in 
the cont~ol dogs have a dia.mete~ up to 40 J.Lm, .5O-55%--up to 80 11m (fig. 1). The 
number of wider canals is 5-8.%, which apparently reflects the process of reoon-
struction of the osteons. Uncl.~J' the influonce of hypokinesj.a in dogs the number 
of vascular oanals over 80 Ilm wide is incr~ased and with width 30-40 Ilm--decreased. 
The distribution of the d~aphyses in the bones of the aont~ol and experimental 
dogs according to width of the osteons was the same. 
The average mineral saturation of the fragment of t~ humeral diaphysis in 
the control dogs was 1.11J,tO.021 g/cm3, and the femoral--l.16~0.017 g/om3• After 
hypokinesia practically the same amounts were obtained, respeotively 1.118+0.014 
and 1.15~~0.019 g/cm3• 
Figure 1. Frequency of Distribution of Haversian Canals (I) and Osteons (~I) 
according to Width in Diaphyses of Bones of Dogs 
A--humerusl B--femur, l--control, 2--hy:pokinesia 
The microroentgenographic study of the long bones in the control dogs re'leale,d 
the nonunif'ormity in the mineral saturation of different zones of the di!1Physis: 
the intermedial zon~ was the most minerali~ed (1,39tl.50 g/om3) as compared to the 
subperiosteal and subendosteal (1.22-1.39 g/cm), r<Ci.05). 
During hy:pokin~sia an increase was found in the indices of mineral saturation 
of the microstructures jf all the zones of the diaphysiS of the humerus to l.~, 
1.52 and 1.43 g/cm) (in the control 1.28, 1.50 and 1.24 g/cm) respectively for the 
subperiosteal, intermedi al and subendosteal zones). In the femur of the ex;peri-
mental animals the mineral saturation was increased in all the zones, except the 
4 
136 
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EFFECT OF SIX-MONTH HYFOKlNESIA IN DOGS ON INCLUSIOn OF CA 4,5 IN MmERAL~ED TISSUES 
("'m) 
Groul> Tissue • Radioactivity 1 % ~f I Tissue 
of impulses ontrol. 
Radioactivity 
of impuloes 
par 1 .mg of as . ----~~T-ee ......... -o"~~~·,c,~"'-.--" ...... ,.",~ 130ne eli ... per 1 mg of ~h 
Oontrol 
upper jaw; aphysE'SI 
incisors 43i8r2.6 femur 
Exper1me~t 
Oontrol tpremolars 
~ 
Experiment' 
o.antrol i,molars 
Experimont 
iTeeth of 
~ lower J'awl 
I Oontrol ~ incisors 
Experiment; 
Oontrol ~premolars 
Experiment; 
Oontrl:)l lmola.:rs 
Experiment ~ 
Control ~lower jaw 
Experiment i 
Control ~~r jaw 
Experiment 
.)(. 
1'-;:0. OS 
44.§4.9 
39.J!2.3 
49.2i-6.4 
J 41~ ~2.,5 
';;4.~,6 
101.6 
I 
112,5.2 
i 
, 106.S 
I 
1103.7 
! 176.9)(-
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i 82 • .5* 
tubercle 
major 
I humerus 
* Haads of 
bones: 
14,5 • .5.!:.7.2 
93.5±9.2 
74. ?!.3. 7 
femur JfO,5t26.0 
, 4,51+23.0 
Ihumerus 1,519!37.6 
! 1,57?!.29.2 ltrochanter~ 298!.21.9 
imajor of ' 
!femur 
~ 
~u'bercle 
(major of 
lhumerus 
I 
, % of 
control 
111.5.4 
I 
! 
i 
I 128.,5* ! i 79.9 
110.4 
, 111.2 
-118.5 
142.8* 
. 
subperiosteal. In contrast to tho humerus and femur in the tubercle major the 
corresponding indices in all zones were somewhat reduced. 
On the h1stological preparations the thickness of the layer of the general 
plates of long bones in the experimental animals was smaller than in the control, 
especially iil the subperiosteal zone of the femur. The osteons weI'S characterized 
by a less regular form; among them anastomoses were observed more often than in the 
control, and the area occupied by the intercalative plates was increased (fig. 2). 
In t' ~ control dogs the magnitUde of tertsile st:rength of the femur fluctUated 
from lS.OJ to 18./+,5 Itg-f/mm2, and the hu.merus--from 16.34 to 22.6,5 kg-f/mm2• The 
fluctuations in the tensile st:rengthof the bones of the dogs in the expe:rimental 
group were :roughly the same, 
,5 
Fi ur 2. Dia hy is of F mur 
a--con rol l b--af 6- nth hypokin sias anastomos s ar visl'lJl b tw n t 
hav rs1an c als 1n th cortical la~ r , irr gular sha of t ost ons . Staini 
with toxylin- 0 M . x 120 . 
grou 
Totalli th t nsil str h of h f Inur and hu rus in dogs of tho control 
w~. 17. 56-0 . 76 - Imm2, and aft r 6-rnonth hypok1n s1a--17. 37-0 . 55 _f/mm2, 
no alt r d . 
137 
Thus , a r sult of 6-month hypokinesia in ogs a ch occurs in th" calcium 
met olis in th bon s of th xtr miti s consisting of an intensification of Ca45 
assimilation in t zon of muscl attac nt . It is ssible that this fact 
ment in t indlcat d r 
liz tissues of the tooth-jaw syst m incor 
ions of ost oporosis . In the min ra-
ratton of Ca 5 pr1marily is reduced , 
which can e lain d hy its ro istri ution tw n different sections of th skele-
ton un er con itions of h;rpokinosia. 
n ogs as a result of th restrict d movement no int nsified resorption of 
bon issue occurs and the av r 
are not significantly alt r 
dimensions of the osteons and haversian ~anals 
However the data of the histological study make 
it ssible to consi er that during hypokinesia c rtain disorders develop in the 
6 
, t G AL PAGE t 
\0' P )( )R QU Al m ' 
. ( 
"- ~----. ,.-,--~ p u 
process of bone tissue r&construotion. Here the mineral s&~uration of the mioro-
structures haa a tendency to inc;..:ease almost in all zones. Based on our studies 
one oan draw the oonclusion that under conditions of hypokinesia demineralization 
is intens1f~.ed in the oompact substance of the dog bones. 
As a result of hypokinesia no decrease ooours ).n the strength of the long 
/?IS 
bones of adult dogs, which agrees with the published data that the mechanioal pro-
perties of the bone depend not on the dimensions of the haversian canals but are 
linked to the width of the osteons [9, 13, 17J and mainly with the mineral saturation 
of the bone tissue [9, 16J. The main factors on which the strength of th6 bones 
depends are not Significantly changed as a result of hypokinesia. 
1. Volozhin, A. I. 'at. fiziob,., No.6 (1971), p. 65. 
2. Voloshin, A. I.., Vasil' yev, l? V. I Uglova, N. N., et al., Kosm1c~J3jt.&~ l:l~21" 
No. ~ (1972), p. 10. 
3. Volozhin, A.I. I Shashkov, V. Sol Smitriyev, B. S., et al.,p'~.f £iziol,., No.2 
(1974), p. 42. 
~. 1{rasnykh, I. G. in ~~ble~ ~.£hes~QX 'll1Ql-~, ["Problems of Space Biology" J, 
Vol. 1;, MOSCOW, 1;09. 
5. Lobanchik, V. I. N~ot9ml'?, ~oronx Jtal'tsix~~,ogo ~ . .!L~{rys,,:V~ l;lsloviya!tlL 
d~ii~'J.K~!lj)Y ~..PQ~itl(3Z.,U., L"Certain Aswcts of Calcium Metabolism in Rats under 
Conditions of 'Prolonged HytlOkinesia" J, Author's abstract of candidate disser-
tation, l-loscow, 1970. 
6. Polyakov, A. N •. Ort.~. ~Eavmatol., No. 3 (1970), p. 41. 
7. Prokhonchukov, A. D., Kovalanko, Ya. A.; Kolesnik, A. G.; at all §~q1Jl8-to~l!,. 
No.4 (1970), 1'. 1. 
8. Currey, J. D. L ~, Vol. 93 (1959), p. 8'l. 
9. Curr~y, J. D •• J., .B,~,Q£halJ4 Vol. 2 (1969), ,P. 1. 
10. Deitrick, J. E., Whedon, G. D.; an.a Shorr, E. ,Am,.!. J. Mad., Vol. 4 (15*8), 1'. 3. 
11. Dunning, M. F.; and l'lum, F •. A:l;,cili,Wiern. ~, Vol. 99 (1957), p. 716-731. 
12. Engstrom, A. ~ ~~iol (Stockh.), 1946, Supple 63. 
13. Evans, F. G. h£EL~. (Basel), Vol. 35 (1958), p. 285. 
14. Frueta, J. J and Geiser, H. 1:. Bone ~ Surg., Vol. 40B(1958), ,P. 282. 
7 
, 
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i 
1 
., 
I 
1.5. Iesokutz, B •• Blizzand, J. J .. Birkhead, N. C. et al., b Appl. ~ysiol!" 
Vol. 21 (1966) t p. 101:3. 
16. V o ISO , G. P., and Kuba.la., A. L. l:!l.W. Biol., Vol. :31 (19.59), p. 261. 
17. Voso, G. P. }n!i-t~.~oc.~ Vol. 144 (1962), 1'. 31. 
1 
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Effect of six-month hypokinesia in dogs on mineral component, reconstruction and mechanical properties of bone tissue

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