[New bioplastic materials for reconstructive surgery].

Results of creation and clinical application of bioplastic materials using specimens from the tissue bank of the Institute of Traumatology and Orthopedics are presented. The use of materials of the new generation, such as perfoost, alomatrix-implant, and osteomatrix, is illustrated by examples of the treatment of various bone diseases. High biocompatibility and efficiency, osteoconductive and osteoinductive properties of these materials are reported. Technical developments in modern bioimplantology are analysed with reference to their status in the Institute's tissue bank.

[Identification of genes induced to chondrocytes by nitric oxide]. / Poisk i identifikatsiia genov, indutsiruemykh v khondrotsitakh pod deistviem okisi azota.

Nitric oxide (NO) acts as a short-lived paracrine factor and selectively activates transcription of certain genes. The spectrum of inducible genes was studied in primary chondrocytes. A cDNA library was obtained by subtraction hybridization with RNAs isolated from rabbit chondrocytes before and after treatment with nitrosoglutathione, an NO-generating agent. Some of the cloned cDNAs were homologous to known mammalian genes and human EST. NO-dependent transcriptional activation was demonstrated for the stromelysin 1 and cyclooxygenase 2 genes and, for the first time, for mcl1 coding for an apoptosis suppressor.

[Pathogenetic aspects of chlamydia-associated urogenic arthritis: feasibility of microorganism reproduction in cells of articular cartilage]. / O patogeneticheskikh aspektakh urogennykh artritov, assotsiirovannykh s khlamidiiami: vozmozhnost' mikroorganizma razmnozhat'sia v kletkakh systavnogo khriashcha.

AIM: The study of feasibility of Chlamydia trachomatis infection and reproduction. This microorganism is an essential etiologic factor in urogenic arthritis, in chondrocytes and fibroblasts of human skin. MATERIALS AND METHODS: Infection of human skin chondrocytes and fibroblasts was made with chlamydia CP-1 strain isolated from joint fluid of the patient and serially passaged in the hen's embryo yolksacs. The inoculation results were assessed by direct staining with the use of monoclonal and fluorescent antibodies and hematoxiline. RESULTS: Chlamydial infection of human skin connective tissue, chondrocytes of the auricular cartilage and fibroblasts in particular, is possible. CONCLUSION: The findings confirm the ability of Chlamydia trachomatis to reproduce in the cartilage tissue.

[Effect of free radicals on proliferation and metabolism of chondrocytes and on sensitivity of a cellular monolayer to proteases]. / Vliianie svobodnykh radikalov na proliferatsiiu i obmen khondrotsitov i na chuvstvitel'nost' kletochnogo monosloia k deistviiu proteaz.

Oxygen radicals (OR) were demonstrated to suppress the proliferation of chondrocytes and to decrease the synthesis of collagen by these cells. The concomitant action of OR and proteases (collagenase or trypsin) sharply increased the detachment of cells and destruction of the intercellular matrix at very low enzymatic concentrations (10-100-fold dilution of routine doses for cultural techniques) as compared with the control.

[Significance of estradiol in the development of fibrosis in systemic scleroderma (bases of sexual predisposition to the disease)]. / Znachenie éstradiola v razvitii fibroza pri sistemnoi sklerodermii (osnovy polovoi predraspolozhennosti k zabolevaniiu).

The influence of estradiol (E2) was studied on the production of type III procollagen by cultivated fibroblasts from the skin of healthy donors and SSD patients. It has been found that when in the norm E2 (concentration 10(-8) and 10(-7) M) inhibits the production of type III procollagen. In SSD the hormone's effect is not found. Here the quantity of E2 receptors in cells from SSD patients is 1.96 time lower than the norm. The resulted data testify to the fact that the decrease of E2 receptors in fibroblasts from SSD patients may be one of the factors contributing to the increase in the functional activity of cells and to the fibrosis progression.

[The effect of cyclic nucleotides on the specific binding of estradiol by skin fibroblasts in the normal state and in systemic scleroderma]. / Vliianie tsiklicheskikh nukleotidov na spetsificheskoe sviazyvanie éstradiola fibroblastami kozhi v norme i pri sistemnoi sklerodermii.

Specific binding of estradiol E2 was studied in homogenates of cultivated skin fibroblasts from healthy persons and patients with systemic sclerodermia as well as effect of cAMP and cGMP on the hormone binding was evaluated. Binding of estradiol E2 was decreased 4-fold in systemic sclerodermia. cAMP and cGMP were of importance in activation of the estradiol specific binding. Under conditions of normal state cAMP 10(-6) elevated 2-fold the estradiol specific binding, while the stimulation was not observed in systemic sclerodermia. However, cGMP 10(-6) M did not affect distinctly the estradiol E2 binding by fibroblasts of healthy volunteers but stimulated the hormone binding in systemic sclerodermia as a result of which the binding values were increased practically up to the basal level of normal state.

[Phospholipids and glycosphingolipids in cultured skin fibroblasts from healthy donors and patients with systemic scleroderma]. / Fosfolipidy i glikosfingolipidy kul'tiviruemykh fibroblastov kozhi zdorovykh donorov i bol'nykh sistemnoi sklerodermiei.

A comparative study of phospho- and glycosphingolipids of cultured skin fibroblast from healthy donors and from patients with systemic sclerodermia (SSD) was carried out. It was shown that the total phospholipid content in SSD fibroblasts is elevated. No significant changes in the concentration of neutral glycosphingolipids were observed. The ganglioside composition of SSD cell cultures differs significantly from that of healthy donor cells. The concentration of the gangliosides, GM3 and GM1, is decreased; no ganglioside GD1a was found in SSD fibroblasts. The data obtained are suggestive of changes in the properties of fibroblast surface which can be manifested both in the impaired reception of matrix proteins and in the impairment of basic properties of the membrane. These changes are well correlated with the results of previous studies on the AMP cyclase system.

[The regulation of DNA synthesis by fibronectin and its proteolytic products in the skin fibroblasts of healthy donors and patients with systemic scleroderma and rheumatoid arthritis]. / Reguliatsiia sinteza DNK fibronektinom i produktami ego proteoliza v fibroblastakh kozhi zdorovykh donorov i bol'nykh sistemnoi sklerodermiei i revmatoidnym artritom.

To study the effect of fibronectin isolated from plasma and culture media and the effect of its tryptic hydrolyzates on DNA synthesis, cultured skin fibroblasts of healthy donors and these of patients with systemic scleroderma (SSD) and rheumatoid arthritis (RA) were employed. It was shown that both fibronectin and total products of its proteolysis markedly stimulated DNA synthesis only in skin fibroblasts of patients with SSD. Fibronectin fragments inhibited DNA synthesis in all fibroblast strains studied. The effect of fibronectin and all its Gel fragments on the DNA synthesis in skin fibroblasts of patients with SSD was dose-dependent. The activity of total fibronectin tryptate, Gel-fragment-free tryptate, and Gel fragments themselves depended on the duration of fibronectin proteolysis, i. e. on the size of the fragments obtained. Culture media collected after treatment of fibroblast monolayer with trypsin and subsequent removal of fibronectin Gel fragments had mitogenic effect on skin fibroblasts, especially on those of patients with SSD and RA. It is supposed that fibronectin Gel fragments are inhibitors of growth factors produced by fibroblasts. The results suggest that fibronectin and its fragments have an important regulatory role in fibroblast proliferation.

[Experimental secondary cataract induced by immune complexes]. / Eksperimental'naia vtorichnaia katarakta, indutsirovannaia immunnymi kompleksami.

The aim of this experimental study was to elucidate the contribution of the immune complexes to the formation of a secondary cataract. The immune complexes are formed after the lens extraction in case specific antibodies to the lens and its capsule and to the lens mass are present in the circulation. The immune complexes, fixed on the posterior capsule, induce antibody-dependent fibrosis, in other words, a secondary cataract.

[Monoclonal antibodies cross-reacting with fibroblasts of interstitial connective tissue of the myocardium and cell wall protein antigens of group A Streptococcus]. / Monoklonal'nye antitela, perekrestno reagiruiushchie s fibroblastami interstitsial'noi soedinitel'noi tkani miokarda i belkovymi antigenami kletochnoi stenki streptokokka gruppy A.

Monoclonal antibodies (MCA) B6/5 and C5/3 were obtained after immunization of BALB/c mice with the protein non-type-specific antigens (NTSA) of streptococcal group A cell wall. MCA B6/5 in the indirect immunofluorescence react with human and animal interstitial connective tissue (ICT) of the myocardium and human fibroblast culture cells. MCA C5/3 react with the bands of muscle fibers of the myocardium. MCA B6/5 and C5/3 are autoantibodies. It was revealed that these MCA are directed to two streptococcal cross-reacting antigens (CRA). Production of B6/5 and C5/3, apparently, does not depend on the possibility of some streptococcal antigens to bind fibrinogen. Bound immunoglobulins were not revealed in the ICT and in the muscle fibres by the cultivation of the C5/3 monoclone. Firstly it was stated that, MCA B6/5, reacting with fibroblasts and with streptococcal CRA, are capable to fix in the ICT of myocardium, what is typical for the phenomenon described in rheumatic fever.

[Clinico-immunologic aspects of development of cataracts in patients with myopia]. / Izuchenie kliniko-immunologicheskikh aspektov obrazovaniia katarakty u bol'nykh s miopiei.

The present study has been aimed at the detection of specific antibodies to the lens and its capsule in the circulating blood of patients with complicated myopic cataracts. The presence of these antibodies results in the development of the posterior capsule fibrosis, this leading to the migration of the cells from the equatorial area to the site of the capsule injury. The detection of the specific antibodies in the blood will help single out the risk group among the patients for whom implantation of a negative intraocular lens is indicated for the correction of high myopia. The presence of these antibodies may also help predict the development of a cataract in the postoperative period; besides, it may be used to create an immunological certificate, informing on the presence of antibodies specific for the lens or its capsule in the patient's circulating blood.

[Interferon in rheumatoid arthritis]. / Interferon pri revmatoidnom artrite.

The paper is concerned with the role played by interferon (IF) in rheumatoid arthritis (RA). The content of IF was measured in the patients' blood serum and the possibility of the use of the preparations of exogenous interferon for the treatment of RA patients was demonstrated. Assay of serum IF in RA patients showed that its content did not exceed normal. The results of the treatment with IF preparations has demonstrated their beneficial effect on certain parameters. Further study of the role played by IF in the management of rheumatic diseases is required in order to develop the drug doses and schedules for the treatment.

[Metabolism of glycoproteins and fibronectin in skin fibroblasts of patients with rheumatoid arthritis]. / Metabolizm glikoproteinov i fibronektina v fibroblastakh kozhi bol'nykh revmatoidnym artritom.

The distribution in the cellular monolayer of the de novo synthetized pre-labeled glycoproteins and fibronectin upon culturing of fibroblasts in the medium with low serum content was analyzed. It was found that in rheumatoid arthritis (RA) the amount of total glycoproteins on the surface and within fibroblasts is higher and in the extracellular matrix is lower than in skin fibroblasts of healthy donors (HD). However, the amount of pre-labeled fibronectin on the surface of skin fibroblasts from patients with RA was considerably lower than in those from HD This finding as well as a rapid decrease in the amount of pre-labeled fibronectin in the extracellular matrix of RA fibroblasts is indicative of a more rapid metabolism of this protein in RA. In the skin fibroblasts from HD there was a practically uniform decrease in the amount of pre-labeled fibronectin in the cellular monolayer. The presence of caseinolytic activity in the culture medium even upon the first day of cell culturing in the serum-free medium, as well as the effect of various proteinase inhibitors on glycoprotein content in the cellular monolayer provide evidence that the rate of glycoprotein and fibronectin metabolism, especially in connective tissue cells of patients with RA, might possibly be determined not only by the level of their synthesis but also by the level of proteolytic activity in the connective tissue cells.

[Study of the interaction of low-density lipoproteins with immobilized fibrinogen and fibronectin using an immunoenzyme assay]. / Izuchenie vzaimodeistviia lipoproteidov nizkoi plotnosti s immobilizovannymi fibrinogenom i fibronektinom metodom immunofermentnogo analiza.

The interaction of serum low density lipoproteins (LDL) with fibrinogen, fibronectin and fibrinogen-fibronectin complex immobilized on polystyrene was studied. LDL binding by these proteins was assessed by enzyme-linked immunosorbent assay. It was shown that LDL interacts with both immobilized fibronectin and immobilized fibrinogen. The fibrinogen-fibronectin complex bound a greater amount of LDL than either component alone, in the same concentration as in the complex. The binding of LDL with immobilized fibronectin or fibrinogen increased in the presence of low concentrations of diluted fibrinogen or fibronectin, respectively, which suggests the formation of the three-component complex on the surface. The formation of fibrinogen-fibronectin-LDL complexes during homeostasis may promote the accumulation of LDL in the vascular wall.

[Effect of a medium with a low serum content on protein synthesis and secretion and RNA and DNA synthesis in a skin fibroblast culture from patients with rheumatoid arthritis and systemic scleroderma]. / Vliianie sredy s nizkim soderzhaniem syvorotki na sintez i sekretsiiu belka, sintez RNK i DNK v kul'ture fibroblastov kozhi bol'nykh revmatoidnym artritom i sistemnoi sklerodermiei.

Synthesis and secretion of protein, as well as synthesis of RNA and DNA by skin fibroblasts of patients with systemic sclerodermia (SSD) and rheumatoid arthritis (RA) upon prolonged culturing of fibroblasts in the medium with low (0.5-1%) serum content differ markedly in their direction and intensity from protein, RNA and DNA synthesis by skin fibroblasts of healthy donors (HD) and by fetal fibroblasts. It has been found that skin fibroblasts of patients with RA and SSD, as well as those of HD, secrete 75-80% of protein synthesized by fibroblasts de novo upon their culturing in DMEM medium with 1% human serum. Under the same conditions, on days 2-5 of culturing, RNA synthesis in the fibroblasts of patients with RA and SSD was increased 3-4-fold, while DNA synthesis was increased 2-3-fold. Collagenolytic and caseinolytic activity in the culture medium of skin fibroblasts from HD and patients with RA reached maximal levels on days 3-5. High protein secretion was observed in DMEM serum-free medium in the presence of vitamin mixture upon culturing skin fibroblasts of patients with SSD. The results obtained show that skin fibroblasts from HD differ in their functional activity from those of patients with rheumatic disorders. It might be suggested, therefore, that the mechanism of protein secretion plays an important role in the maintenance of constant intracellular protein levels in resting cells.

[Protein synthesis and secretion and RNA and DNA synthesis in human skin fibroblasts in a medium with a low serum content]. / Sintez i sekretsiia belka, sintez RNK i DNK u fibroblastov kozhi cheloveka v srede s nizkim soderzhaniem syvorotki.

Human skin fibroblasts, both postnatal and embryonic, were cultured in the stationary phase of growth for 6-10 days in the DMEM with bovine serum (BS), 0.1-0.5% fetal calf serum (FCS) or 1% human serum (HS). On the day 4 of culturing, a considerable increase was observed in the synthesis and secretion of protein by postnatal fibroblasts in the Eagle medium with 0.1-0.5% FCS, or with 0.5% BS, and in medium 199 with 0.1-0.5 BS, or with 0.1 FCS. Maximum synthesis and secretion of 14C-proline labeled protein was observed on day 2 of culturing of cells in the DMEM medium with 1% HS. In the DMEM medium with low serum content, protein synthesis being virtually unchanged, 75-80% of protein was secreted by cells into the culture medium with BS on days 2-4; in the medium with FCS such a high secretion of protein was observed only on day 4. High synthesis of protein by fetal fibroblasts in the DMEM medium with 0.1% BS and high protein secretion in all the media with 0.1% BS or 0.5% FCS were observed. The maximum level of secretion of protein by fibroblasts coincided with a considerable increase in both RNA and DNA syntheses. The data obtained suggest that cells in deep resting state actively react to the composition of the medium as well as to the quality and quantity of the serum. It may also be suggested that the mechanism of protein secretion has an important role in maintenance of the constant level of intracellular proteins in resting cells.