Pisarev A.A., Shaimardanova L.R., Soboleva A.A.

Crimea State Medical University named after S.I, Georgienvskyj

MORPHOLOGICAL CHANGES OF METAPHYSEAL CARTILAGE AS A CRITERION OF BIOLOGICAL EFFECT OF PARENTERAL INJECTED CEREBROSPINAL FLUID

Cerebrospinal fluid - it is a humoral surrounding of the brain that performs multiple functions. Physiological properties of cerebrospinal fluid are caused by the presence of a wide range of biologically active substances - it's the hormones of the hypothalamus, pituitary gland, pineal, neuropeptides, neurotransmitters, endogenous opioids, biogenic amines and other active metabolites. The protein content in CSF is low, the parenteral administration does not cause immunopathological reactions. Several authors established a variety of effects of liquor after parenteral administration and indicated possible prospect of using it as a basis of drugs. Many authors have applied for the correction of pathological states xenogeneic cerebrospinal fluid obtained from cattle, as the closest in composition to the cerebrospinal fluid of human. The experiments proved the absence of teratogenic, embryotoxic properties of xenogenic cerebrospinal fluid, as well as immunopathological reactions after its introduction. The aim of our study was to study the effect of xenogenic cerebrospinal fluid on the structure of the proximal epiphyseal cartilage of femur. Cartilaginous tissue (textus cartilagineus) - a kind of connective tissue consisting of cartilage cells (chondrocytes) and dense intercellular substance. Performs the support function. Chondroblasts (from the greek - Chondros - cartilage, blastos - bud) - a young flattened cells capable of proliferation and synthesis of intercellular substance of cartilage (proteoglycans). The cytoplasm of chondroblasts has a well developed granular and agranular endoplasmic reticulum, the Golgi apparatus. With the participation of chondroblasts peripheral growth cartilage is commited. These cellsin a process of development of cartilage become chondrocytes. Chondrocytes can be oval, round or polygonal shapes - depending on the degree of differentiation. Chondrocytes locate in special cavities (lacunae) in the intercellular substance singly or in groups. Epiphyseal cartilage (cartilago epiphysialis) - connects the epiphysis and diaphysis plate cartilage, which is localized on the growth zone of the bone. In the course of human life metaepifizal cartilage experience a significant static and kinetic pressure, because they are the kind of strut seal between the rigid surfaces of bones. In our study, we used morphofunctional classification by Koveshnikov V.G. , according to the epiphyseal cartilage secrete 5 areas: 1) zone of the indifferent cartilage adjacent to the pineal gland; signs of the proliferation of chondrocytes in this zone are absent ;2) the zone of cartilage cell proliferation represented by a large number of actively growing cells, stacked and form a "coin columns", 3) zone difinitive cartilage - contains a large, cube-shaped chondrocytes, also forming a column, and these cells at the final stage of maturation and are not able to proliferate; 4) the zone of destruction - the cells are irregular in shape, their size is larger than the cells of all zones, with many missing the kernel in the zone of destruction is the destruction of cells begins to calcified matrix; 5) zone of primary bone formation - most deepest lied sites of epiphyseal cartilage, places ingrowth of blood vessels and osteogenic cells that form bone beams primary spongiosis. Tendency of increasing in diseases of the sceletal apparatus under the influence of various factors, both internal and external environment, and social significance, due to frequent injuries of middle working age and possible disabilities, torpid and recurrent determine the relevance of further study of theoretical and practical aspects of these pathologies . The interaction of various factors in the development of the disease complicates the choice of treatment methods and reduce their effectiveness. A large amount of literature contain information that characterizes the role of cartilage in the development of degenerative joint disease. State of the articular cartilage largely determines the normal function of the joint. As donor animals used lactating cows, xenogen cerebrospinal fluid which contains a wide range of BAS биологич акт вва.. To gathering material we punctured of a large tank of the brain during its processing of the biological product - a seamless, taken in vivo, cryopreserved, purified membrane filtration systems "Millipore" cerebrospinal fluid, which was used for further experimental studies. For the experiment, we selected 96 Wistar rats of both sexes of four age categories: newborn, juvenile, mature and old. Cerebrospinal fluid newborn animals were administered parenterally once; biopsy specimens was performed at 7 and 30 days. The other three groups of cerebrospinal fluid was injected three times and ten times - the fence material was carried out on 7 and 30 days. Control group was injected NaCl solution in the same dosage and in the same way. For histological examination to isolate the thighs, locking in 10% neutral formalin, decalcified in 5% formic acid, dehydrated in alcohols and embedded in paraffin blocks. Preparing histological sections 5-10 microns thick, which were stained with hematoxylin Weigert - eosin. Number of chondrocytes was counted in the column, respectively, measure the width of the place of epiphyseal cartilage. Measured by the usual ratio of cellular elements and the intercellular substance epiphyseal cartilage and volume fraction of primary spongiosis in the area of bone formation. In animals, early postnatal age of a single injection of xenogenic cerebrospinal fluid led to a narrowing of the epiphyseal cartilage by 14,7% compared with control. Indifferent zone of cartilage has not significantly changed, but the width of the zone of proliferative cartilage was significantly decreased by 14.7%. Mitotic figures are rarer, the proliferative zone chondrocytes occupy less orderly situation, "coin columns" are collected on 4-5 cells, in contrast to the control group, where the chondrocytes arranged in columns of 5-8 cells. Partitioning between the posts, well-defined in the control of virtually non-existent. Band width of the definitive cartilage also decreased by 25%. Definitive zone contains cells at different stages of maturation and are also organized in columns of 5-6 cells each, in contrast to controls, where the number of cells in a column of this zone was 6-8. At 30 day experiment, the trend continued. The width of epiphyseal cartilage is even more behind the corresponding figure in control of 20%. The width of the zone of proliferative cartilage and definitive decrease in the experimental group by 20% and 35% respectively, relative to the width of the same bands in control. The number of cells in the column zone of proliferative cartilage still lagged behind the controls, and is 4-5 cells in the bars. Cells of the definitive zone of the cartilage, the number of 4-5, differed from the control series of eccentrically located nucleus, a greater ordering in the columns. In areas of cartilage destruction, both on day 7 and 30 day experiment, significant differences from control were found. In the experimental group, the primary trabeculae spongiosis have branched structure at all stages of the experiment. Thus, after a single injection of xenogenic cerebrospinal fluid newborn animals develop persistent changes in epiphyseal cartilage, manifested in the reduction of its proliferative activity by reducing the number of proliferating cells. These results indicate a slowing growth rate of bones in animals early postnatal period in response to the introduction of xenogenic cerebrospinal fluid. Despite numerous literature data on the effect of growth stimulation xenogeneic cerebrospinal fluid in different age groups, newborn animals are the only category of recipients, showing a pronounced inverse effect, which may be due to hormonal interference with the physiological processes of adaptation to extrauterine existence. Further studies of cartilage tissue in other age groups will help shed light on chondrogenesis in the introduction growth stimulation xenogenic cerebrospinal fluid, which in the long term may be used for hondrodisplastic states, as well as cartilage injuries in sports medicine.