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I V Voronkina

Publications and source records attributed to I V Voronkina.

11 recordsLinked to original sources

[Isolation of human basal keratinocytes by selective adhesion to extracellular matrix proteins].

Epidermal human cells (keratinocytes) differently interact with extracellular matrix proteins of the skin basal membrane depending on the stages of their differentiation. The pool of basal keratinocytes commonly includes stem cells and transient amplifying cells. They directly attach to the skin basal membrane. Keratinocytes change their adhesive properties during differentiation, lose direct interaction with the basal membrane and move to suprabasal epidermal strata. From this, it is suggested that basal and primarily stem cells can be isolated from a heterogenous keratinocyte population due to their selective adhesion to the extracellular matrix proteins. In the current study, we analysed the specificity of interaction between primary keratinocytes and extracellular matrix proteins (collagens of I and IV types, laminin-2/4, fibronectin and matrigel). We have demonstrated that the basal keratinocytes extracted from the skin have different adhesive abilities. The rapidly spreading cells usually interacted with collagen and fibronectin rather that with laminin-2/4 or matrigel. The majority of these cells being represented by basal keratinocytes. Our data demonstrate that the applied method of keratinocyte selection may be directed for precise isolation of skin stem from a common cell population.

Cell Adhesion↗

[Changes in human burn fluid biological activity during normal burn healing].

The main goal of this work was monitoring the changes occurring in human burn fluid biological activity during normal burn healing. The fluid available in the burn until healing makes a good material for controlling biochemical microenvironment of burn cells. This environment involves factors, such as extracellular matrix proteins and matrix metalloproteinases. In this work our previous studies of the influence of wound and burn fluids on the functional activity of cells were extended to include the effect of burn fluid on fibroblasts and keratinocytes, i. e. human skin cells present in the wound and involved in wound healing. It was shown that human burn fluid biological activity depends on the time that passed after burning, and on the correctness of healing. Migration of human fibroblasts becomes more intensive under the influence of such a fluid independently on the time of fluid sampling. Unlike, keratinocyte migration was inhibited by burn fluid sampled 1-3 days after burning but was enhanced by fluids sampled 6 days following burning. The obtained data are to be necessarily taken into consideration at burn treatment and also at transplantation of cells for healing of wounds of different nature.

Biological Factors↗

[Complex evaluation of wound healing process by a deep wound rat model with implanted polychlorvinyl camera].

A simultaneous study of wound proteolytic activity and morphological picture of the first stages of wound healing on rat deep wound model has been shown. The process of wound healing can be evaluated by dynamics of matrix metalloproteinase activities in wound fluid. Changes in activities of different matrix metalloproteinases correlate with different stages of healing. Implantation of polychlorvinyl camera in the wound makes it possible to obtain the volume of wound fluid sufficient for a complex evaluation of healing at the initial stages of wound process.

Animals↗

[Effect of burn exudate on functional activity of Balb/3T3 cell line and the role of matrix metalloproteinases in this process].

The influence of burn fluid and its matrix metalloproteinases (MMPs) on Balb/3T3 cells was studied. The influence of burn fluid was assessed by morphology and specific functional activities of cells characteristic of the healing process--proliferation, monolayer contraction and migration of cells in wound model. The presence of burn fluid in cultivating medium accelerated cell proliferation by 2.5 times compared to normal conditions, promoted fibroblast monolayer contraction, and accelerated cell migration on the wound surface, thus stimulating cell functions necessary for successful heating. This effect is partly due to MMPs. The burn fluid contains, presumably, some additional factors not inhibited by specific MMP inhibitors EDTA and 1,10-phenantrolin. These factors may stimulate migration and proliferation of cells. The presence of 1-2% burn fluid is sufficient for enhancing cell proliferation.

3T3 Cells↗

[Matrix metalloproteinases MMP-2 and MMP-9 of wound and burn exudates and their effect on extracellular matrix proteins].

The dynamics of matrix metalloproteinases (MMP), as well as of fibronectin concentration in wound and burn fluids was traced. The wound fluid proteolytic activity was studied by gelatin zymography method. The data on degradation of fibronectin and various laminin isoforms by wound fluid proteases show that laminin-1, laminin-2/4 and fibronectin were degraded by wound fluid into small fragments. Remodelling of extracellular matrix proteins occurs. Dynamics of MMP-2 and MMP-9 content in wound or burn fluids as well as that of adhesive protein fibronectin content could be used as a base for development of method of controlling the extracellular matrix remodelling process.

Burns↗

[The "air pouch" model in mice and a study of the wound fluid proteolytic activity].

The experimental model of a cutaneous wound in mice has been offered, aimed to study the proteolytic activity of the wound fluid produced at early stages of wound healing, and to examine the continuous inflammatory state. The presence of metalloproteinases MMP-2 and MMP-9 in the wound fluid matrix was found to correlate with the existence of neutrophils and macrophages in the tissue.

Animals↗

[Various effects of laminin-1 and laminin-2/4 on adhesion and migration of cultured human keratinocytes].

The cell-matrix interaction is one of the factors defining the cell behavior in normal and wounded tissues. To determine the function of laminin-2/4, one of components of the skin basement membrane in the process of reepithelization, we studied its interaction with human keratinocytes. The adhesive properties of laminin-2/4 and its effect on keratinocytes migration in vitro were analysed. For comparison with our present investigation, we used the earlier studied laminin-1 from EHS mouse sarcoma. Laminin-2/4 appeared to be a good substrate for human keratinocytes, and this correlates with a greater number of cell surface receptors compared with laminin-1. Laminin-2/4 alone does not stimulate keratinocyte migration, but, in contrast to laminin-1, supports EGF-mediated migration. The obtained results give an insight into the function of laminin-2/4 in normal skin and during wound healing.

Animals↗

[Cell interaction with extracellular proteins during two-phase polymer system formation].

A modified method of investigation of surface properties of cells and proteins with the help of a two-phase polymer system dextran-500/polyethylenglycol-6000 was used. This method is based on changing the kinetics of two-phase system partitioning into phase on adding cells or macromolecules. These changes were registered by measuring the top phase optical density during the system partitioning at 500 nm. Cell lines L (NCTC clone 929), LS and A431, human keratinocytes and platelets, collagen I, laminin-1, laminin-2/4, and fibronectin were studied. The interaction between collagen and all cell types with the formation of complexes takes place during co-partitioning of cells and proteins in the two-phase system. Laminins differ in surface properties and in interaction with cells. Laminin-1 makes preferable complexes with cells of monolayer subline L (NCTC clone 929), but not with cells of suspension subline LS. No interaction of laminin-2/4 with L cells was detected, but, in contrast to laminin-1, this protein has the affinity to A431 cells. No interaction of L cells with fibronectin were detected.

Animals↗

Laminin-2/4 from human placenta is a better adhesion agent for primary keratinocytes than laminin-1 from EHS sarcoma.

A comparison of the adhesion of human primary keratinocytes to laminin-1 from murine EHS sarcoma and laminin-2/4 from human placenta was carried out using two methods, cell adhesion to substrates covered with the laminin isoforms, and interaction of keratinocytes from suspension with latex beads coated with the proteins. Laminin-2/4 was considerably more potent as a promoter of attachment of primary human keratinocytes than laminin-1 (and fibronectin), with increased attachment of cells correlating well with the number of latex bead binding sites. Only small cells of diameter of less than 20 microm bound more than 5 beads. Staining of keratinocytes with involucrin antibodies confirmed the existence of an inverse relationship between laminin-2/4-coated bead binding and differentiation.

Animals↗

[Features of formation of phases of dual-phase polymeric system in the presence of collagen I, laminin I, and their mixtures].

A study was made of the phase formation kinetics of a two-phase aqueous polymer system, consisting of 7.8% (w/w) dextran-500 and 4.6% (w/w) polyethylene glycol-6000, in the presence of various concentrations of collagen (0.07-0.20 mg/ml), laminin I (0.01-0.03 mg/ml) and their mixture. The phase formation was evaluated by registration of its optical density on a spectrophotometer. The obtained two-phase polymer system optical density curves and also the partitioning coefficients for the studied objects depend on surface properties of these objects. It has been shown that the surface properties of collagen I and laminin I differ according to differences in their molecules conformations, and that the phase formation kinetics points to their interaction during a mutual partitioning of these proteins in the system. The authors made a conclusion that collagen I and laminin I in the two-phase polymer system conditions could make complexes.

Biopolymers↗