Method of estimating state of bone tissue in case of immobilisation osteoporosis

FIELD: medicine.

SUBSTANCE: invention relates to field of medicine, namely to orthopedics. In order to estimate state of bone tissue in case of immobilisation osteoporosis in a laboratory animal examined are homogenates: bone, muscular, bone marrow of any extremity and peripheral blood. Biochemical and integral parametres are determined. Five factor variables F1-F5 are calculated using values of biochemical and integral parametres, constant values of factor coefficients of biochemical parametres and free coefficients. After that calculated is the value of discriminant function, whose value is used to estimate bone state as normal or conclusion about presence of immobilisation osteoporosis is made.

EFFECT: method increases accuracy and efficiency, has high stability of immobilisation osteoporosis recognition.

1 ex, 3 tbl

 

The invention relates to medicine, namely to orthopedics, and can be used to identify immobilization osteoporosis in laboratory animals.

In clinical practice to examine the state of bone tissue metabolism using the definition of many of the parameters in the serum, including activity markers of osteoblasts and osteoclasts, for example teramobile fraction of alkaline phosphatase and tetrameristaceae acid phosphatase, and daily urinary concentrations of total hydrooxyproline, calcium, inorganic phosphate and other(1, 2, 3, 4). However, these parameters are not sensitive enough to immobilization osteoporosis, as a feature of immobilization osteoporosis is the lack of load on the limb and therefore the decrease in bone mineral density (BMD) differs from the change in BMD in systemic osteoporosis.

The known method for diagnosing osteoporosis in laboratory animals (U.S. Pat. RF 2325107), based on identifying the relationship of the diameter of the diaphysis of the tibia bone to the mass, and increase of this index concludes the presence of osteoporosis in animals.

However, the known biometric indicator is not sensitive and reliable for evaluating the status of bone tissue in immobilization osteoporosis, as immobilization osteoporosis are the two which is secondary and mechanism of formation differs from primary systemic osteoporosis.

The objective of the invention: improving the accuracy and sensitivity of the method with immobilization osteoporosis, is solved as follows.

To assess the status of bone tissue in immobilization osteoporosis in laboratory animal research homogenates of bone and muscle tissue, bone marrow and peripheral blood, determine biochemical parameters, including integral (X1-X44table 1), count five factor variables (F1-F5and the value of the discriminant functions (canonical root - Y) by the formula:

where a0-44b0-44c0-44d0-44, g0-44- constant values of factor coefficients biochemical parameters and available ratios (table 2),

where F1F2F3F4F5values of factor variables with rounded numerical coefficients

and when Y>0 determine the presence of immobilization osteoporosis.

The choice of biochemical parameters, including integral, due to experimental research. In exp is Rimante participated 136 laboratory animals: 32 - intact, 64 immobilization osteoporosis. Modeling of immobilization osteoporosis was performed by resection of the bones of the lower leg right hind limb at the level of its proximal part. Experimental conditions limit the motor function of rats due to the amputation of the lower leg caused the phenomenon immobilization osteoporosis, which was clearly expressed to 90 days after surgery (according to morphology).

All animals were identified 80 laboratory parameters, including the integral in various biological material: bone homogenate, muscle homogenate, bone marrow, peripheral blood. All the indicators are standardized methods for the Specific analyzers basic Stat Fax 1904, Gell Dyn 1300. From the entire dataset selected the most significant indicators - 44 (table 1). The values of these parameters by means of dispersion, correlation, discriminant, canonical and factor analysis processed (5), and identified five independent variables that determine the contribution of biochemical parameters on biological processes in the bone tissue, allowing you to more accurately assess the condition of the bone tissue of the laboratory animal, and to determine the metrics you can use the biological material of any limb of an animal.

Independent factors:

F1 factor determined by calcium content in bone,

F2factor associated with the concentration of magnesium in bone,

F3factor associated with calcium homeostasis in the body of a laboratory animal,

F4factor associated with the dynamics of the concentration of inorganic phosphate in bone,

F5factor associated with hemopoesis blood.

Factor coefficients biochemical parameters are presented in table 2 with an accuracy of up to four digits after the decimal point.

Using independent variables (F1-F5)calculate the linear discriminant function (canonical root-Y), the value of which articulates the animals in groups: intact (Y<0) and immobilization osteoporosis (Y>0). Sensitivity - 93-95%. Thus, a more accurate assessment of the status of bone tissue allows to identify immobilization osteoporosis.

The method is as follows.

In laboratory animal (use any limb) determine quantitative values of 44 indicators, calculated variables, and the value of the discriminant functions (formulas 1-6) and on the value of the discriminant functions to determine the presence of the animal immobilization osteoporosis.

Example. In table 3 before the taulani values of biochemical parameters of two laboratory rats (with normal bone and in the presence of immobilization osteoporosis) and the values of factor coefficients.

Free coefficients in formulas 1-5 equal to: a01,257; bo-0,041; c04,396; d0-0,495; go-2,511.

Substitute the values of rats "Faith" and factor coefficients from table 3 in the formula (1-5) and calculated values of factors:

F1=and1X1+...and44X44-a0=(-0,02512)×5,1+...0,001022×4,5-1,257=-1,0109

F2=b1X1+...b44X44-b0=(-0,01584)×5,1+... 0,005399×4,5-(-0,041)=-0,8867

F3=C1X1+...44X44-C0=0,06755×5,1+...0,04365×4,5-4,396=0,1615

F4=d1X1+...d44X44-d0=(0,01372)×5,1+...(-0,05111)×4,5-(-0,495)=-0,1084

F5=g1X1+...g44X44-g0=(0,06803)×5,1+...0,1966×4,5-(-2,511)=1,6728

Calculated for rats "Faith" is the discriminant functions (canonical root-Y) by the formula (6):

Y=0,01003×(-1,0109)+0,5207×(-0,8867)-1,0369×0,1615+0,7815×(-0,1084)-0,4994×1,6728= -1,558

The value of the discriminant function is less than zero (Y<0), so the condition of the bone tissue of the rat "Faith" is estimated normal.

Calculated values of factors of the rat "Orion" (values from table 3) by the formulas (1-5):

F1=and1X1+...and44X44-a0=(-0,02512)×5,7+...0,001022×5,0-1,257=-0,9742

F2=b1X1+...b44X44-b0=(-0,01584)×5,7+...0,005399×5,0-(-0,041)=0,1287

F3=C1X1+...44 X44-C0=0,06755×5,7+...0,04365×5,0-4,396=-0,8754

F4=d1X1+...d44X44-d0=(0,01372)×5,7+...(-0,05111)×5,0-(-0,495)=0,4576

F5=g1X1+...g44X44-g0=(0,06803)×5,7+...0,1966×5,0-(-2,511)=1,9427

Determine the value of the discriminant functions (canonical root-Y) by the formula (6):

Y=0,01003×(-0,9742)+0,5207×0,1287-1,0369×(-0,8754)+0,7815×0,4576-0,4994×1,9427=0,3526

The value of the discriminant function is greater than zero (Y>0), so the rats Orion determine the presence of immobilization osteoporosis.

Thus, the proposed method is effective, has a high resistance recognition immobilization osteoporosis, which can be used in the research on optimization of bone regeneration after surgical correction of necromany bones.

Literature used

1. Vladislav Dolgov, Ermakova I.P. Laboratory diagnosis metabolism of bone tissue. //Osteoporosis and osteopathy, 2000 - No. 4. - P.29-39.

2. The cake A.A., Dedukh N.V., Shevchenko S.D., Filipenko, VA and other Diagnosis and conservative treatment of diseases and injuries of musculoskeletal system. // Book 1: Osteoporosis. - Kharkov. Is "basis. - 1995. - 52 S.

3. TIC N. Encyclopedia of clinical laboratory tests. M, Labelform, 1997, 960 S.

4. Franke J., Runge, Osteoporosis. Translation from German. - M.: Medicine, 1995, pp. 27, 30.

5. Draper N.R., Smith, of Priklad the th regression analysis. - M.: Dialectics. 2007. - 911 S.

The method of evaluating the status of bone tissue in immobilization osteoporosis by bone tissue, characterized in that in laboratory animal examined bone homogenate, muscle homogenate, bone marrow any limbs, and peripheral blood, determine the biochemical and integral parameters X1-X44in table 1, are presented in the description; count five factor variables F1-F5and the value of the discriminant function (Y) by the formula: F1=a1X1+...a44X44-a0; F2=b1X1+...b44X44-b0; F3=c1X1+...c44X44-c0; F4=d1X1+...d44X44-d0; F5=g1X1+...g44X44-g0where X1-X44values of biochemical and integral parameters; and0-44b0-44with0-44d0-44, g0-44- constant values of factor coefficients biochemical parameters and available coefficients presented in table 2 contained in the description, Y=0,01003F1+0,5207F2-1,00369F3+0,7815F4-0,4994F5where F1F2F3F4F5values of factor variables with rounded numerical coefficients, and Y>0, the condition of the bone is estimated normal, and when EIT is the situation discriminant function is greater than zero Y> 0 determine the presence of immobilization osteoporosis.



 

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