ABSTRACT
The aim of the work was a comprehensive assessment of the system of cytokines and peripheral blood osteocalcin with the establishment of features of their interconnections in children with congenital cleft lip and palate (CCLP) in comparison with the corresponding controls at different age periods. In the peripheral blood of 80 children (0-12 months, 1-3 years, 4-9 years, 10-15 years) with osteocalcin and IL17, IL4, congenital cleft lip and palate and age-appropriate control of healthy individuals (40 people) IL6, IL1ß, IFNγ by enzyme immunoassay. An analysis of the data obtained with CCLP revealed significant differences in the dynamics of the relationship between pro-inflammatory (IL1ß, IL6, IL17), regulatory (IFNγ), anti-inflammatory (IL4) cytokines and osteocalcin compared with controls. The patterns of changes and effects of IL17, IFNγ, IL4 and osteocalcin in healthy children and in children with congenital cleft lip and palate in postnatal ontogenesis were established. Cytokine dysregulation of immunosteogenesis in CCLP, leading to a significant deficit of osteocalcin in the first year of life due to an imbalance of the cytokine profile: discordant IL17, IFNγ and IL4 were detected. Thus, the data obtained are undoubtedly important in the future for developing new strategies for targeted therapy aimed at normalizing osteocalcin levels at different age periods in children with congenital cleft lip and palate.
Subject(s)
Cleft Lip , Cleft Palate , Child , Cytokines , Humans , Osteocalcin , OsteogenesisABSTRACT
It was found that the chemiluminescence intensity in native and recellularized tissues of rat muscular organs as well as in their decellularized scaffolds can serve as an express criterion that, along with ultrastructural analysis, makes it possible to perform quantitative assessment of the viability of cellular structures in biological samples of the diaphragm.
Subject(s)
Diaphragm/chemistry , Luminescent Measurements , Tissue Engineering , Tissue Scaffolds/chemistry , Animals , RatsABSTRACT
Automated image analysis methods are highly important for biotechnology research. The authors developed and tested a program for the morphometric analysis of photomicrographs of the sections processed using the standard immunohistochemical examination protocols. The color deconvolution method used in the algorithm was proven to be effective in mapping the distribution of DAB chromogen in the sample containing multiple dyes. The experiment demonstrated that the level of extracellular matrix proteins could be comparatively quantified in different groups of samples. The effective methods for the quantitative analysis of the Ki-67 labelling index were also tested using the same algorithms. The developed program was published under free GPL 3.0.
Subject(s)
Antigens, Nuclear/isolation & purification , Extracellular Matrix/ultrastructure , Image Processing, Computer-Assisted/methods , Immunohistochemistry/methods , Algorithms , Antigens, Nuclear/ultrastructure , Coloring Agents/chemistry , Coloring Agents/isolation & purification , Humans , SoftwareABSTRACT
Experimental skin ischemia in rats was induced by suturing a skin fold on the back with a silk thread. Combined pretreatment with superoxide dismutase (intraperitoneally) and Reamberin (intravenously) in doses of 0.01 and 6.25 mg/kg (by succinate concentration), respectively, produced a strong protective effect on the skin. The index of cytolysis decreased by 39%. The more pronounced antinecrotic effect of combined treatment with superoxide dismutase and Reamberin compared to the effect of Reamberin alone was related to a sharp increase in the reserve capacity of the antioxidant system. After combined therapy, activity of antioxidant defense enzymes not only increased, but even exceeded the normal level. The increase in activity of endogenous superoxide dismutase under the influence of combined therapy was accompanied by suppression of superoxide anion production.
Subject(s)
Antioxidants/therapeutic use , Ischemia/drug therapy , Skin/blood supply , Skin/drug effects , Superoxide Dismutase/pharmacology , Adenosine Diphosphate/analysis , Adenosine Triphosphate/analysis , Animals , Antioxidants/administration & dosage , Antioxidants/pharmacology , Dose-Response Relationship, Drug , Drug Therapy, Combination , Injections, Intraperitoneal , Ischemia/enzymology , Ischemia/etiology , Ischemia/pathology , Male , Necrosis/drug therapy , Random Allocation , Rats , Rats, Inbred Strains , Skin/enzymology , Superoxide Dismutase/administration & dosage , Superoxide Dismutase/metabolism , Superoxides/antagonists & inhibitorsABSTRACT
Antinecrotic activity of SOD was studied in rats with experimental skin ischemia. Treatment with SOD increased activity of endogenous SOD in skin homogenates (by 70 and 26% compared to the ischemic and intact skin, respectively). However, the rate of superoxide anion generation remained unchanged after SOD treatment. Creatine phosphate content and NAD/NADH redox potential increased by 16 and 21%, respectively, on day 3 after SOD administration. The increase in functional activity of the energy supply system and rise in the reserve capacity of the antioxidant protection system contribute to inhibition of lactate dehydrogenase and creatine phosphokinase and decrease in the cytolysis index under the influence of SOD. Our results indicate that SOD produces the antinecrotic effect and holds much promise for the therapy of skin ischemia.
