RU2270586C2 - Food calcium-containing supplement and functional foodstuff containing the same - Google Patents

Abstract

FIELD: food processing industry, in particular production of calcium-containing supplements and functional fermented milky foodstuffs.

SUBSTANCE: invention relates to food supplement containing calcium source, vitamin D2 and vitamin E. As calcium source homogenized fish protein concentrate, containing calcium, magnesium and phosphorus in ratio of 1:0.4:0.4 is used. Said concentrate is obtained from bone chine with flesh lump of oceanic salmonic fishes by aqueous fermentation at 75-85°C followed by drying at 20°C or less. Functional fermented foodstuff containing milky protein and calcium-containing supplement also is disclosed. As milky protein kefir is used. Claimed product contains (g/100 g of product): fish protein concentrate 3.0±1.0; vitamin D2 300±10 IU, vitamin E 10±0.1 IU, and balance: kefir.

EFFECT: supplement and product having high calcium digestibility useful in osteoporosis prophylaxis.

3 cl, 10 tbl, 3 dwg

Description

The invention relates to the food industry, namely the production of calcium-containing food additives and dairy products for functional purposes, and can be used in the dairy industry, as well as medicine as a prophylactic.

Currently, the food industry, including the dairy, supports all types of human nutrition: traditional, preventive, therapeutic and prophylactic, medical, specialized, functional and non-traditional.

Functional foods include foods that, in addition to traditional nutritional effects, have a positive effect on a particular function of a macroorganism (TP Pilat, AA Ivanov, Biologically active food additives (theory, production, use). - M. Avvallon , 2002. - S.221-223).

Several physiological functions and conditions of a person have been identified, the positive effect on which would allow us to attribute food products to one or another category of functional nutrition products. Among these conditions, one of the most common today is a metabolic change in the skeletal system associated with a lack of calcium in the body, leading to skeletal disease - osteoporosis (V.A. Tutelyan, V.A. Knyazhev Implementation of the concept of the state policy of healthy nutrition of the Russian population: scientific support // Nutrition Issues. 2000. Volume 69, No. 3, C.4-7).

In recent years, significant progress has been made in the understanding of the pathophysiology and treatment of osteoporosis, including the proof that the prevention and treatment of osteoporosis is a real challenge (E.I. Marova Osteoporosis // Russian Medical Journal. 2001, v. 9, No. 9).

It is known that the most important and most common cause of osteoporosis is a deficiency of calcium and vitamin D in the body, which is primarily associated with insufficient intake of these ingredients with food. Currently, an adequate intake of calcium and vitamin D is considered to be the consumption of 1200 mg of elemental calcium and 800 IU of vitamin D per day. Such a dose is recommended for all persons over 60 years of age and those at risk of developing osteoporosis (E.L. Nasonov Osteoporosis in the practice of a doctor // Russian Medical Journal, 2002, Volume 10, No. 6, P.288-293; L.I. Benevolenskaya General principles for the prevention and treatment of osteoporosis // Consilium. 2000. Volume 2, No. 6. S.240-244).

Nutritional prevention of the appearance and development of osteoporosis involves a number of measures and recommendations for people at risk of this disease, one of which is the appropriate diet.

It is known that most calcium is found in dairy products (cheese, cottage cheese), leafy greens, spinach, cabbage, lettuce, carrots, fish bones, figs and apricots.

It is also known that in food products, calcium is in the form of poorly soluble or completely water insoluble compounds and that calcium is a difficult to assimilate element and its digestibility depends not only on the calcium content in the products, but also on its ratio with other micro and macronutrients, primarily phosphorus, magnesium, proteins and vitamins. At the same time, the absorption of calcium in the gastrointestinal tract decreases both with a high amount of fat in the diet and with a diet low in fat (milk fats, egg yolk, fish liver), i.e., unfortunately, just those products , which contains vitamin D, necessary for the absorption of calcium (Pilat T.A., Ivanov A.A. Biologically active food additives (theory, production, use). M: Avvallon. 2002. P.221-223).

It is known that the ratio of calcium and magnesium in products that is optimal for the assimilation of calcium is 1: 0.6, and that of calcium and phosphorus is 1: 1.5 (according to some sources, 1: 1). However, in nature there are no natural foods in which such ratios are observed. In bread, meat and potatoes, the ratio of calcium and magnesium is 1: 2, in milk and kefir - 1: 0.1; in many vegetables and fruits - 1: 4,5. (Pilat T.A., Ivanov A.A. Biologically active food additives (theory, production, use). M: Avvallon. 2002. S.221-223).

Calcium found in dairy products is easily digestible, but its amount in them is clearly insufficient. The daily norm of calcium for people with osteoporosis is contained in 2 liters of milk or lactic acid products. Eating so many dairy products daily for most people is not realistic. The process of simply adding calcium ions to dairy products is difficult because of the physical properties of inorganic calcium salts (their insignificant solubility), and because of the interaction of calcium with calcium-sensitive components of milk, such as proteins. These interactions can lead to coagulation of protein components during heat treatment, a decrease in the nutritional value of milk proteins, and the appearance of milk sugar salts - lactates. Ultimately, all this reduces the absorption of dairy products in the digestive tract.

Thus, although there is an urgent need to obtain functional foods enriched with calcium, the assortment of such products, which have both the necessary degree of absorption of calcium and sufficient nutritional and biological value, is very small.

A known method of obtaining a dry dietary dietary milk protein enriched with calcium, by means of thermal calcium enrichment using calcium alginate and as a coagulant of calcium lactate (Z. RF No. 2000126558, publ. 10.11.2003).

It is known that calcium-enriched milk, in which calcium hydrolyzed pectin, calcium citrate, a metastable lactate-citrate calcium complex or a balanced mixture of calcium lactate and calcium carbonate are used, and food grade polyphosphate, for example sodium hexametaphosphate (c. RF No. 2001131719), is used as an additional source of calcium. , published on September 20, 2003).

Known liquid food product based on milk protein containing a metastable calcium complex obtained by the interaction of an alkaline source of calcium (calcium hydroxide, calcium oxide, calcium carbonate) and citric and lactic acid (c. RF No. 2000127754, publ. 09/20/2003).

However, all these products are characterized by the lack of a ratio of mineral composition and protein necessary for nutrition and assimilation of calcium, an insufficient amount of vitamins is contained, and difficult-to-digest components appear in food products, such as calcium lactate, which is hydrolyzed in the human intestinal tract by the enzyme lactase (β-glycosidase), the level of which in adults is often reduced, and therefore an increase in the amount of lactate in the product can lead to digestion (Food chemistry / echaev AP Traubenberg SE, AA Kochetkov et al, Ed A.P.Nechaeva -... SPb .: GIORD, 2001. - S.288-289 (259 p.)).

At the same time, the range of calcium-fortified food additives is quite large. Known, for example, is a mineral fortifier for baby and preventive nutrition, obtained from meat bones (as.with. No. 1593605, p. RF 2212821), however, their use is limited to the age of the person, there are contraindications for children less than 12 years old.

Closest to the claimed is a food supplement for the prevention of calcium deficiency and the optimization of calcium metabolism, in which an artificial mixture of chemical compounds containing calcite of biological origin (egg shells, shells of mollusks) and various chemical compounds of calcium and an additional mineral-organic complex are used as an enrichment consisting of organic and / or inorganic compounds of magnesium, zinc, copper, potassium, an organic compound covalently with yazannym iodine nonhormonal nature, lysine, arginine, in free and / or bound and / or tryptophan in free and / or bound state, citric and succinic acids and / or their salts, vitamin D _3, thiosulfate and / or polyuronic acid (s. RF 2001135433, published on July 20, 2003).

However, the known food additive and the food product containing it are not a natural product, they are an artificially created mixture and do not have the necessary ratio between calcium, magnesium and protein.

The main disadvantage of all the above-mentioned food mineral fortifiers is the non-observance of the optimal physiological ratio between calcium, magnesium and phosphorus, which means that the digestibility of both the product and the calcium mineral supplement is reduced, their biological and nutritional value, as well as their importance as preventive agents for prevent osteoporosis.

Closest to the claimed is a calcium-fortified food product containing milk protein and an enriching amount of the calcium complex obtained by the interaction of a calcium source and a negatively charged emulsifier in the presence or absence of an organic or inorganic acid or their salts (Cl. RF No. 2218800). At the same time, calcium hydroxide, calcium carbonate, calcium chloride, calcium phosphate, calcium sulfate, calcium nitrate, calcium lactate, calcium fumarate, calcium citrate, calcium acetate, calcium glycerophosphate or calcium oxide are used as a source of calcium, and a negatively charged emulsifier is complex ester of citric acid and monoglycerides, stearoyl lactylate in the form of sodium, calcium or acid salts, enzyme-modified lecithin, stearyl citrate, fatty acids and their salts, or esters of diacetyl tartaric acid and monogl tseridov.

However, although the known product contains an increased amount of calcium, non-compliance with the optimal physiological ratio between calcium, magnesium and phosphorus reduces its biological value and its value as a prophylactic to prevent osteoporosis. The production of such a product requires careful technological control, since clogging by-product synthetic components can occur at different stages of the process flow, the presence of which always occurs in chemically synthesized compounds, and the additional introduction of an emulsifier into the mixture leads to a rise in the cost of the food product.

The objective of this invention is to develop a functional food product, as well as a calcium supplement for the prevention of osteoporosis, which, in addition to food and high biological values, has a high digestibility of calcium and the ability to correct metabolic disorders in bone tissue, manifested by a disease such as osteoporosis, by creating in the claimed products of a physiologically justified ratio between the mineral and protein components that make up their composition.

The problem is achieved by a functional food product containing milk protein in the form of kefir and additional vitamins D ₂ and E, and as a source of calcium, a homogenized fish protein concentrate (RBC) obtained from bone ridges with meat cuts of fish of the oceanic salmon species (genus Oncorhynchus) by water fermentation at a temperature of 75-85 ° C followed by drying at a temperature not exceeding 20 ° C and characterized by a ratio between the content of calcium, magnesium and phosphorus equal to 1: 0.4: 0.4, in the following ratio comrade, per 100 g of product:

RBC - 3.0 ± 0.1 g;

Ergocalciferol (vitamin D ₂ ) - 300 ± 10 IU;

α-tocopherol acetate (vitamin E) - 10 ± 0.1 ME;

Kefir - the rest.

The task is also achieved by a calcium-containing food additive with a ratio between the content of calcium, magnesium and phosphorus equal to 1: 0.4: 0.4, obtained from bone ridges with cuts of fish meat of oceanic salmon species (genus Oncorhynchus) by fermentation at a temperature of 75 -85 ° C and subsequent drying at a temperature not exceeding 20 ° C, and additionally containing vitamins D ₂ and E in a physiologically acceptable amount.

The inventive functional food product (hereinafter the product) is a natural product containing the necessary components in physiologically justified proportions, has the necessary nutritional, energy value, as well as prophylactic activity in case of osteoporosis. 200 g of the claimed functional product (one glass, 200 ml) contains the daily norm of biologically significant nutrients for the prevention of osteoporosis. Given the composition of kefir in the claimed functional product, the ratio between the content of calcium, magnesium and phosphorus is 1: 0.6: 1, which is optimal for the absorption of these components and, above all, calcium in the gastrointestinal tract. Introduced vitamins (mainly vitamin D ₂ ) accelerate the absorption of calcium ions in the intestinal mucosa and further intensify the metabolism of this element in the body. The inventive product is a full-fledged natural nutritious product, because it contains easily digestible muscle proteins of fish with a high percentage of essential amino acids. The presence of animal fats containing lipotropic factors (phospholipids) enhances anabolic processes in the body and therefore favorably affects the state of bone tissue. The nutritional and energy value of the product (in 100 g) in cases of using kefir as a base with varying degrees of fat content is given in table 1.

Table 1 product name Content, in%, not less than: Calories, kcal / 100 g squirrel fat carbohydrates Kefir 1% supplemented 6.0 1,2 4.05 51.0 Kefir 2.5% with additive 6.0 2.7 4.05 64.5

Our experimental studies have shown high efficiency in the use of the claimed product and additives according to the schemes of the experimental groups of the first and second. Against the background of experimental glucocorticoid osteoporosis, the claimed food supplement reduced bone porosity, increased calcium in serum and bones, and decreased urinary calcium excretion.

For the preparation of a food-grade calcium-containing supplement (hereinafter supplements) used for the prevention of osteoporosis, for the first time we proposed using bone ridges with meat cuts as waste from the processing of salmon species of ocean fish (Oncorhynchus genus) as an economically and biomedically-based raw material. The population of these fish is at a good level, catches are stable. After processing the fish into loin and waste for the manufacture of additives used waste - spines with meat cuts.

We experimentally discovered for the first time that it is in the bone residues of fish ridges with cuts of meat from the salmon family (genus Oncorhynchus) that the ratio between calcium, magnesium, and phosphorus that is closest to physiologically justified for calcium absorption is observed.

Studies of the biological value of waste in the form of bone ridges with meat cuts (in total) showed that they have a well-balanced protein composition, low content of polar lipids and, which is especially important, are balanced by mineral micronutrients, especially Ca, Mg, P (table 2). Studies of the mineral composition of raw materials under various conditions of the technological process of its processing: autoclaving with subsequent degreasing with alcohol, autoclaving without degreasing with alcohol, changing cooking conditions, drying and homogenization, showed that for such basic micronutrients as Ca, Mg and P, the total protein lipid and moisture, the food additive proposed by the applicant, characterized by the claimed ratio between calcium, magnesium and phosphorus and obtained by aqueous fermentation at a temperature of 75-85 ° C followed by drying at a temperature of no higher than 20 ° C, is a stable, biologically valuable product with prophylactic activity in case of osteoporosis.

To increase the biological and nutritional value, as well as to increase the digestibility of calcium and increase the shelf life, vitamins D ₂ and E are additionally added to the supplement in a physiologically acceptable ratio.

table 2 Cooking mode Sa Mg P Lipids Protein Moisture Water fermentation for 30 minutes at a temperature of 80 ° C, freeze drying 20.7 5.2 5.7 6.8 54.1 7.1

To speed up the drying process while maintaining the necessary properties of the additive, it is advisable to carry out freeze drying at a pressure of 0.015-0.02 mbar and a temperature of 12-17 ° C.

To obtain additives, use the raw materials obtained after processing commercial fish of salmon breeds - spines with meat cuts. Fins, heads and caudal fins are not used. Processing with organic solvents to remove fat is not performed, since the lipid content in the feedstock is low (below 10%). After water fermentation, the raw materials are dried using, for example, freeze drying for typically five hours at a temperature of 15 ± 2 ° C and a pressure of 0.015-0.02 mbar. Dehydration can be carried out at normal pressure, but the drying time increases to 72 hours. The dried material is homogenized to a fine powder with granules of no more than 0.5 mm, obtaining a fish protein concentrate, to which vitamins D ₂ (P. 71.893.5) and E (P. 001153 / 01-2002) are immediately added in physiological ratio. The finished food supplement is packaged, labeled and sent for storage (storage for 1 month at a temperature of 0 to -5 ° C).

To obtain a functional food product suitable for the prevention of osteoporosis, a fish protein concentrate with the claimed properties and vitamins D ₂ and E in a predetermined ratio are added to the prepared standard kefir (OST 4929-84) of varying degrees of fat content or the claimed food supplement is added.

The quality of the raw materials and auxiliary materials used for the preparation of kefir with RBC meets the requirements of the current regulatory documentation and the Hygienic requirements for the quality and safety of food raw materials and food products (SanPiN 2.3.2.1078-01).

Characterization of food quality indicators.

Sampling and preparation for testing was carried out according to GOST 26809, GOST 26929.

Organoleptic characteristics of the product correspond to GOST 28283 (Table 3).

According to physico-chemical parameters, the claimed product meets the requirements specified in table 4. The mass fraction of solids was determined according to GOST 3626; determination of the mass fraction of fat - according to GOST 5867; determination of the mass fraction of protein - according to GOST 23327; determination of the mass fraction of total sugar - according to GOST 3628;

Table 4 Name of indicator Indicator value kefir 1% with the addition kefir 2.5% supplemented Mass fraction of solids,%, not less 9.0 Mass fraction of protein,%, not less 6.0 Mass fraction of fat,%, not less than 1,2 2.7 Mass fraction of total sugar (by sucrose) in terms of dry matter,%, not less 4.05 Acidity, T ° 70-110 Temperature at the outlet from the enterprise, ° С, not higher 6

According to the content of toxic elements, mycotoxins, antibiotics, pesticides and radionuclides, the food product meets the requirements of SanPin 2.3.2.1078-01 (index 1.2.1.), The data are shown in table 5. Their determination was carried out according to GOST 26927; GOST 26930; GOST 26932; GOST 26933.

Table 5 Name of substance (element) Permissible level of its content, mg / kg (for radionuclides - Bq / kg), no more Toxic elements Lead 0.1 Arsenic 0.05 Cadmium 0,03 Mercury 0.005 Mycotoxins (aflatoxin M ₁ ) 0,0005 Antibiotics (chloramphenicol, tetracycline group, streptomycin, penicillin) Not allowed Organochlorine Pesticides Hexachlorocyclohexane (α, β, γ - isomers) 0.05 DDT and its metabolites 0.05 Radionuclides Cesium - 137 one hundred Strontium - 90 25

According to microbiological parameters, the product meets the requirements of SanPiN 2.3.2.1078-01 (index 1.2.1.6.) Indicated in table 6. Sampling and preparation of samples for microbiological analyzes was carried out according to GOST 26668, GOST 26669, GOST 26670. The determination of microbiological parameters was carried out according to GOST R 50474; GOST 30347; GOST R 50480.

Table 6 Name of indicator Indicator value The mass of the product (cm ³ ) in which it is not allowed: BGKP (coliforms) 0.01 Pathogenic (including salmonella) 25 S.aureus 1,0

Experimental evaluation of the functional effectiveness of a food supplement for the prevention of osteoporosis.

Objects and materials of experimental research:

White rats of the Wistar strain with an initial weight of 150-200 were used as experimental material. The animals were kept in a vivarium in accordance with the sanitary rules for the design, equipment and maintenance of experimental biological clinics dated April 6, 1973. Feeding was carried out in accordance with the standards approved by order of the Ministry of Health of the USSR No. 163 of 03/10/1966, additional food additives that increase the calcium content in the diet of animals were not introduced.

Experimental model of osteoporosis:

Osteoporosis was modeled by the method of Ziganshina et al. (Ziganshina L.E., Burnashova Z.A., Valeeva I.Kh., Galyautdinova A.Yu., Samoilova N.S. Comparative study of the effectiveness of dimephosphone and xidiphon in steroid osteoporosis in rats // Experimental and Clinical Pharmacology. 2000. T 63. No. 6. P.39-42.)

According to the above method, steroid glucocorticoid osteoporosis was modeled by intragastric administration of 5 mg of prednisolone per kg of animal weight daily for 20 days. Every 10 days, the state of calcium-phosphorus metabolism was evaluated. On the 20th day, animals were decapitated under light ether anesthesia, blood serum was obtained, and femurs and lumbar vertebrae were isolated for further biochemical, histomorphological studies and biomechanical tests.

To study the effectiveness of the additive as a prophylaxis of experimental osteoporosis, animals were divided into 4 groups. The group of intact animals included rats not receiving any drugs. The control group included animals that were injected with prednisolone according to the above procedure for 20 days. Animals of the two experimental groups daily consumed food supplement containing RBC with vitamins in a dose of 0.1 g per 100 g of rat weight. The daily dose contained Ca - 0.03 g, vitamin D ₂ - 0.0002 mg, vitamin E - 0.00001 mg. Rats of the first experimental group used the supplement for 20 days simultaneously with the introduction of prednisone (the first experimental group). Rats of the second experimental group consumed the supplement for 40 days: within 20 days before the start of prednisolone administration and then for 20 days simultaneously with the administration of prednisolone (second experimental group).

Experimental research methods:

The determination of calcium-phosphorus metabolism was evaluated by the level of serum total calcium and inorganic phosphorus concentration, as well as by the level of urine excretion per day.

Calcium and phosphorus were determined spectrophotometrically using the methods and reagent kits Calcium-Novo and Phosphorus-Novo (Novosibirsk), respectively. The measurements were carried out on a Shimadzu UV-1601 spectrophotometer. For measurements, quartz cuvettes 1 cm thick were used. Calcium was determined at a wavelength of 650 nm, phosphorus was determined at 340 nm.

For analysis of calcium and magnesium in the bones, dry burning was used. The amount of Ca ²⁺ and Mg ²⁺ in the bones was determined by complexometric titration. Calcium content was determined by titration of ethylenediaminetetraacetic acid with indicator murexide at pH 12. The total content of calcium and magnesium was determined by titrating a sample of ethylenediaminetetraacetic acid with indicator eriochrome black at pH 10. The magnesium content was determined by the difference between these titrations in milliequivalents per 100 g of dry matter.

Bone density was calculated as the ratio of bone mass (g) to the volume of a given bone (cm ³ ). Bone volume was determined by the pycnometric method according to the volume of displaced water.

To compare the various characteristics of the state of the bone tissue, the mechanical loading of whole bones is currently preferred in accordance with the most probable load application in real conditions. Resistance to fracture during compression was characterized by the value of the bearing capacity of the sample, numerically equal to the maximum value of the load, which entails the destruction of the bone. Bearing capacity was expressed in kg. The determination of the maximum axial load on the bone was carried out in the laboratory of the Far Eastern State Technical University. To determine the maximum axial load on the joint, the universal educational SM-1 complex was used to determine the resistance of materials.

A histomorphological study of bone material was performed on the lumbar and femur of rats. After rat slaughter, the bones were removed and fixed in 10% neutral formalin. Then, the samples were dehydrated in alcohols of increasing density, and after decalcification with Evans reagent, they were poured into paraffin. Survey sections were stained with hematoxylin and eosin. After that, a complex analysis of the morphofunctional state of the structures was carried out.

Statistical processing of the results of biochemical and biomechanical studies included an assessment of the scatter of the analysis results and the spread of the error from the average value.

Statistical data processing was performed in Excel 6.0.

The results of an experimental study:

Indicators of calcium-phosphorus metabolism in serum, urine and bones of experimental animals.

As you know, in the pathogenesis of the development of glucocorticoid osteoporosis, a certain role is played by disturbances in calcium-phosphorus metabolism, especially those related to bone tissue and blood serum. Our studies showed that in the control group of animals receiving glucocorticoids for 20 days, there was a significant decrease in serum calcium levels while increasing calcium excretion in daily urine (tables 7 and 8).

Table 7
The content of calcium and phosphorus (in mmol / l) in the blood serum of experimental animals. Groups of animals Intact The control Experience 1 Experience 2 Calcium 2.55 ± 0.16 1.78 ± 0.16 ^# 2.67 ± 0.15 * 3.01 ± 0.16 * Phosphorus 2.43 ± 0.06 2.47 ± 0.08 2.22 ± 0.09 2.47 ± 0.09 Note: ^{# - the} difference with indicators of intact rats is significant, * - the difference with indicators of control rats is significant. Table 8
The content of calcium and phosphorus (in mmol / l) in the daily urine of animals. Groups of animals Intact The control Experience 1 Experience 2 Calcium 2.12 3.0 2.6 1,6 Phosphorus 0.49 0.365 0.277 0.16

When consuming the inventive product containing a significant amount of easily digestible calcium and vitamins, a distinct normalizing effect was revealed. The serum calcium level in animals that took the supplement for 20 days (first group) and 40 days (second group) remained at the level of intact rats. The state of calcium in serum was characterized as normal and corresponded to that in intact animals (table 7).

The results of a study of calcium and magnesium in bone tissue are presented in table 9. It should be noted the presence of positive dynamics of an increase in calcium and magnesium in the bones of experimental rats, which is especially characteristic of a group of animals that took the product for 40 days.

Table 9.
The content of calcium and magnesium (in meq / 100 ml) in the femur of experimental animals. Groups of animals Intact The control First group Second group Calcium 17.81 ± 1.74 17.16 ± 0.44 17.44 ± 1.07 19.3 ± 0.95 Magnesium 1.63 ± 0.26 1.83 ± 0.1 2.0 ± 0.24 1.93 ± 0.26

Studies of bone density and biomechanical indicators of the femur (bearing capacity) of experimental animals revealed a number of changes in control animals and experimental rats (table 10). After administration of glucocorticoids, the bone density of the control rats decreased, while the simultaneous administration of the product normalized this indicator in experimental animals. The carrying capacity of the bone during the entire experiment remained unchanged.

Table 10.
Indicators of bone density (in g / cm ³ ) and bearing capacity of the femur (in kg) in experimental animals. Groups of animals Intact The control Experience 1 Experience 2 Bone density 1,236 ± 0,049 1,135 ± 0,074 1.237 ± 0.062 1.269 ± 0.017 Bone bearing capacity 16.33 ± 1.21 17.60 ± 2.00 15.13 ± 1.23 18.48 ± 1.19

Pathomorphological examination of bone tissue;

Pathological examination of the bone tissue of animals with experimental osteoporosis and taking the supplement showed a number of changes. In animals of the intact group, the bone tissue had a normal structure (Fig. 1). The bone tissue components were well expressed: lamellar bone tissue, which forms the compact substance of the diaphysis and the outer thin layer of the pineal gland pineal gland, as well as the spongy bone tissue, which forms the inner part of the pineal gland. The periosteum (periosteum) covered a compact layer with a thin layer. In the inner fibrous layer of the periosteum and in the endoste lining the bone from the side of the bone marrow, a layer of osteoblasts was weakly expressed. The bone cuff is clearly visible. Signs of violation of osteogenesis are not identified.

In animals of the control group with experimental osteoporosis, morphological changes characteristic of osteoporosis were detected (Fig. 2). In the periosteum there were reactive changes in the form of thickening and sagging, osteoblasts were not visualized. The cortical bone, which forms the surface layer of the diaphysis of the femur, is of uneven thickness and is used under the periosteum; throughout the entire length, the diaphyseal part of the bone is thinned, compared with intact animals, with varicose veins. The perichondral and enchondral bone did not have uniform staining with the presence of basophilic foci, which indicated a violation of the mineralization of the bone. The bone cuff in some areas had a cribriform structure and was uneven in thickness. A distinct fatty degeneration of bone marrow cells was determined and, as a result, vacuolization was observed. Haversian canals were few in number. From the endosteum, osteoclasts were encountered in the Gauship lacunae; due to the expansion of the Gauship lacunae, the endost surface had a scalloped pattern.

In animals of the 2nd experimental group receiving the supplement according to the scheme for 20 days before the administration of glucocorticoids and then 20 days at the same time, the picture of osteoporosis was revealed, however, its degree was less pronounced than in animals in the control (Fig. 3) . Compared with the last animals, the activation of periosteum cells was visible in rats of this group, the layer of osteoblasts adjacent to the inner surface of the periosteum was better expressed. Gross bone osteoporosis was not found in two of the five animals examined.

In animals of the 1st experimental group receiving the supplement according to the scheme for 20 days simultaneously with the administration of glucocorticoids, pathological changes in the bone tissue were significant and comparable with those in the control (osteoporosis without taking the supplement). There was an uneven thinning of the tubular bone and an overdose of its surface, probably as a result of an increase in the activity of osteoclasts. As in the control, there was a noticeable increase in the number of Haversian canals, also associated with increased intraosseous resorption. As a result, in the metaphysical part, the cortical layer had a structure resembling a trabecular bone. In addition, there was an expansion of the cavities of the bone marrow canal, while the endosteal part of the bone was separated from the bone marrow cells by a wide space. Focal vacuolization of bone marrow due to fatty degeneration was noted. Osteoblasts both in the periosteum and in the endosteum were few. The periosteum with areas of thickening due to its fibrous layer.

Thus, the results of an experimental study indicate that the use of the claimed nutritional supplement and functional product has a significant effect in experimental glucocorticoid osteoporosis, reducing its manifestations. It should be noted that the positive effect of the product is more obvious with its prolonged use and increased dose (second experimental group). This effect is due to metabolic patterns characteristic of the macroorganism as a whole and bone tissue in particular. This result was also confirmed by studies in the practice of prevention of osteoporosis when taking the claimed fermented milk product in a hospital in Vladivostok. Apparently, the use of a product containing easily assimilated calcium should last a long time (at least two months).

Claims (3)

1. A functional food product containing milk protein and a calcium source, characterized in that the product additionally contains vitamins D ₂ and E, kefir is used as milk protein, and a homogenized fish protein concentrate (RBC) with a ratio between calcium content is used as a calcium source. , magnesium and phosphorus equal to 1: 0.4: 0.4 obtained from bone ridges with meat cuts of fish of oceanic salmon species (genus Oncorhynchus) by fermenting them at a temperature of 75-85 ° С followed by drying at a temperature not higher 20 ° C, in the following ratio of components per 100 g of product: RBC 3.0 ± 0.1 g Ergocalciferol (Vitamin D ₂ ) 300 ± 10 IU α-Tocopherol Acetate (Vitamin E) 10 ± 0.1 ME Kefir Rest 2. The functional food product according to claim 1, characterized in that the RBC is dried at a temperature of 12-17 ° C and a pressure of 0.015-0.02 mbar. 3. A calcium-containing food supplement containing a source of calcium and vitamin D, characterized in that a homogenized fish protein concentrate with a ratio between calcium, magnesium and phosphorus equal to 1: 0.4: 0.4 obtained from bone ridges is used as a calcium source with meat cuts of fish of oceanic salmon species (genus Oncorhynchus) by water fermentation at a temperature of 75-85 ° C and subsequent drying at a temperature of no higher than 20 ° C, and additionally containing vitamin E, and as vitamin D - vitamin D ₂ physiologically acceptable amount.

Images