**2. Materials and methods**

The study involved 104 patients (mean age 54.03 ± 9.56 years) who were divided into three groups. Group 1 included 39 healthy women, group 2 – 30 patients with AH and osteopenia, and group 3 – 35 women with AH and OP. The study protocol was approved by the Ethics Committee of Tyumen Cardiologic Research Center, Tomsk National Research Medical Center of the Russian Academy of Sciences, Tomsk, Russia. Before enrollment, each study participant gave a written informed consent to use the study results for scientific purposes.

Exclusion criteria included: presence of acute cerebrovascular accident less than 6 months ago, coronary artery disease, type 2 diabetes mellitus, chronic heart failure of functional class (FC) III-IV (according to the New York Heart Association classification–NYHA), cancer, and mental illness. AH was diagnosed according to the current recommendations of the European Society of Cardiology and Russian Society of Cardiology. The scope of diagnostic measures included: clinical examination, laboratory, and instrumental methods to evaluate the cardiovascular and skeletal systems. The study of the parameters of 24-hour ambulatory blood pressure monitoring (ABPM) was carried out for all examined patients according to the standard scheme, using the oscillometric method, on the equipment of BPLAB LLC "Petr Telegin" (Russian Federation), with the study of standard parameters.

The study of the elastic properties of the vascular walls was carried out using a sphygmograph Vasera VS-1000 Series (Fukuda Denishi, Japan), with the assessment of the following parameters: pulse wave velocity for the elastic-type arteries on the right or left (PWV-R/L) and ankle-brachial index (ABI-R, ABI-L) as a parameter of peripheral vascular blood flow and a screening parameter for the presence of AS of the vessels of the lower extremities. Osteodensitometry was performed using the Siemens Somatom

Emotion spiral computed tomograph. Calcium content CA-HA and the standard deviation of the T-Score peak were assessed (standard values: from 2.0 to −1.0, normal values: from −1.0 to −2.5, and osteopenia: from −2.5 and lower - OP).

Ultrasound scanning of the brachiocephalic arteries was performed; the parameters of intima-media thickness (IMT) of the carotid artery, state of the vascular wall, and the presence of atherosclerotic plaques were taken into account. IMT was determined at a distance of 2 cm from the bifurcation of the common carotid artery on the posterior wall (normal – less than 0.8 mm, the upper limit of normal was 0.9 mm, the thickening was more than 0.9 mm). Atherosclerotic plaque was a local thickening of the arterial wall exceeding 50% or more of the thickness of the adjacent unchanged IMT, protruding into the lumen of the vessel and having different structure compared to unchanged arterial wall and/or thickening of the IMT of more than 1.3 mm [13]. Fasting venous blood was collected into the Vacuette disposable tubes (Japan); the blood was centrifuged for 15 min at 2500 rpm in the Sigma centrifuge (Germany). Patients' blood serum was aliquoted for further freezing (at −70°C).

The parameters of lipid metabolism were studied using the Cobas Integra 400 plus automatic biochemical analyzer (Switzerland). Total cholesterol and triglycerides (TG) in blood serum were determined using the enzymatic colorimetric method; high-density lipoprotein (HDL) and low-density lipoprotein (LDL) were determined using the direct enzymatic colorimetric method; concentrations of apolipoprotein A-I (Apo A-I), apolipoprotein B (Apo-B), and lipoprotein a (Lp(a)) were obtained using immunoturbidimetry with the analytical kits and control materials by Roche Diagnostics Gmb (Germany).

The following biochemical markers of inflammation were determined: highsensitivity C-reactive protein (hs-CRP) by immunoturbidimetric method using the "C-reactive protein hs" analytical kit (BioSystem, Spain) on the Clima MC-15 semiautomatic, open type analyzer (Spain); interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α) –"sandwich" and homocysteine (HYC) by competitive methods (solid-phase chemiluminescence enzyme immunoassay) using the following analytical kits: IL-1β, IL-6, IL-8, TNF-α, HYC, respectively. The level of sex hormones (estradiol, progesterone, testosterone) and parathyroid hormone were determined using a competitive, solid-phase, chemiluminescent ELISA method with the Siemens Diagnostics reagents; myeloperoxidase was investigated by the sandwich ELISA method using the eBioscience reagents.

Carbohydrate metabolism was assessed based on glucose and glycated hemoglobin (HbA1c) concentrations. Blood glucose was determined by the hexokinase method using the Cobas Integra 400 plus biochemical analyzer. Glycated hemoglobin was determined by chromatography using the Bio-Rad D10 analyzer, USA. The atherogenic coefficient (AC) was calculated as = Apo B/Apo A-I. The parameters of functional activity of endothelium in blood serum: nitrite levels were determined using the Humalyzer 2000 Human biochemical analyzer (Germany) and endothelin-1-21 using the Dynatech semi-automated immunoassay analyzer (Germany).

Statistical data processing was carried out using the Statistica software package (SPSS Inc., ver 11.5). Parameter distribution was tested using the Kolmogorov-Smirnov test. To determine the statistical significance of the differences in continuous values depending on the distribution parameters, one-factor analysis of variance with the Holm-Sidak correction for multiple comparisons or the Kruskal−Wallis criterion with the Bonferroni correction for multiple comparisons were used. Continuous variables represented as M ± SD (mean ± standard deviation) or Me [Q25; Q75] (median and interquartile range). To assess the differences in qualitative variables,

*Characteristics of Pathogenetic Links in Vascular Remodeling and Bone Tissue Destruction… DOI: http://dx.doi.org/10.5772/intechopen.107444*

the chi-square criterion and the Fisher exact criterion were used. Spearman and Pearson linear correlation coefficient, logistic regression method, and discriminant analysis were used to identify the relationship between the variables.
