Comparative analysis of the morphological changes of the joints when using the sodium salt of 4- (3-methyl-2-oxo-2h-[1,2,4] triazino [2,3-c] quinazolin-6-yl) butyric acid (compound DSK-38 ) and diclofenac sodium on the adjuvant arthritis model
Introduction. The presence of adverse events in modern NSAID encourages the search for new chemicals, suitable for the creation of more effective and safer antiphlogistics.
Objective: on the basis of pathological studies provide a comparative assessment of the therapeutic effect of the compound DSK-38 and diclofenac in the model of adjuvant arthritis (AA).
Materials and methods. The study was conducted on 35 nonlinear male rats weighing 140-150g, divided into 5 groups of 7 animals each:
1 - intact rats; 2 - AA rats without treatment (control pathology, euthanasia on the 14 day); 3 - AA rats without treatment (control pathology, euthanasia on the 28 day); 4 - rats with AA treated with compound DSK-38 (2 mg / kg i/p); 5 - AA rats treated with diclofenac (4 mg / kg / i/p). Doses of both substances were equal to their ED50. Arthritis is modeled by introducing under the planter aponeurosis of rats 0.1 ml of complete Freund’s adjuvant. Treatment was carried out with 14 to 28 day experiment. The degree of treatment effect was assessed at 28 day.
For evaluation of the morphological changes of the synovium their ankle joints were fixed in 10% neutral formalin solution and subjected to decalcification. Histological sections (5-7mkm) were stained with hematoxylin and eosin, Van Gison pikrofuksine main brown for Shubich, a combination of basic brown and strong green dye, PAS - reaction with alcian blue.
Microscopy and photography of histological preparations was performed using a light microscope BX OLIMPUS 41 at a magnification of 40, 100, 200 and 400 times. Morphometry and statistical analysis was performed using the program "Quick PHOTO MICRO 2.3."
Results. On day 28 of the experiment in untreated animals in the intermediate and deep areas of the synovial membrane of affected joints revealed the broad field of granulation tissue of varying degrees of maturity. They consist mainly of capillaries, fibroblasts, lymphocytes and histiocytes. In the central regions of these infiltrations were degenerative changes of the basic substance, surrounded by connective tissue fibers.
The treatment DSK-38, as well as diclofenac, the most pronounced changes were observed in the superficial parts of the articular cartilage (surface and intermediate zones), which grows in the granulation tissue pannus and damage while articular cartilage. Unlike the untreated animals full invasion into the cartilage synovium were observed.
Applications DSK-38 caused a decrease or absence of inflammation in the joint, and in periarticular tissues. This cartilage was Gorny, with clearly defined areas and constant presence of chondrocytes. Thus bone destruction caused by inflammation, accompanied by reduction in bone tissue and restoration of function focal synovial and cartilage tissue, as evidenced by the appearance of acidic glycosaminoglycans in dissociated chondrocytes.
Regeneration of bone tissue was realized by proliferation of connective tissue and its transformation into the cartilage. Only in some places synovium remained moderate signs of swelling and slight focal lymphocytic infiltration. On the background of DSK-38 there has been a significant decrease in the number of fibroblasts compared with untreated rats and treatment with Diclofenac.
A compound DSK-38 reduces inflammation of the synovial membrane ankle and performance is not inferior to comparator diclofenac sodium, as evidenced by the presence of the compound antiproliferative properties.
Along with antiproliferative properties unlike diclofenac sodium in a new compound DSK-38 also observed antysklerohennyy effect, as evidenced by the low level of fibroblastic reaction (1350 ± 152,93, p <0,05).