Blinova Mi

Enhancement of the blood growth promoting activity after exposure of volunteers to visible and infrared polarized light. Part I: stimulation of human keratinocyte proliferation in vitro

The systemic mechanisms of the wound healing effect of low intensity lasers remain largely uninvestigated. The goal of this randomized, placebo controlled, double blind study is to prove that irradiation of a small area of the human body with visible and infrared polarized (VIP) light (400-3400 nm, 95% polarization, 40 mW cm(2), 12 J cm(2)) leads to an increase of the growth promoting (GP) activity of the entire circulating blood for primary cultures of human keratinocytes (KCs). Thirty minutes after the VIP-irradiation of a sacral area of volunteers, the GP activity of circulating blood was seen to increase through the elevation of the number of KCs cultured with the isolated plasma by 20 +/- 3%, p < 0.001. A similar increase in GP activity was seen in plasma derived from the in vitro irradiated blood of each volunteer, and from the mixture of irradiated and non-irradiated autologous blood 1:10. Enhanced GP activity was also recorded at 24 h after the 1st and 4-9th daily phototherapeutic sessions. Hence, exposure of volunteers to VIP light leads to a fast increase in the GP activity of the entire circulating blood for human KCs in vitro, which is a consequence of the transcutaneous photomodification of blood and its effect on the rest of the circulating blood volume.

Two‐stage implantation of the skin‐ and bone‐integrated pylon seeded with autologous fibroblasts induced into osteoblast differentiation for direct skeletal attachment of limb prostheses

Angio- and osteogenesis following the two-stage (TS) implantation of the skin- and bone-integrated pylon seeded with autologous fibroblasts was evaluated. Two consecutive animal substudies were undertaken: intramedullary subcutaneous implantation (15 rabbits) and a TS transcutaneous implantation (12 rabbits). We observed enhanced osseointegrative properties of the intramedullary porous component seeded with fibroblasts induced into osteoblast differentiation, as compared to the untreated porous titanium pylon. The three-phase scintigraphy and subsequent histological analysis showed that the level of osteogenesis was 1.5-fold higher than in the control group, and significantly so (p < 0.05). The biocompatibility was further proved by the absence of inflammatory response or encapsulation and sequestration on the histology assay. Treatment of the transcutaneous component with autologous fibroblasts was associated with nearly a 2-fold decrease in the period required for the ingrowth of dermal and subdermal soft tissues into the implant surface, as compared to the untreated porous titanium component. Direct dermal attachment to the transcutaneous implant prevented superficial and deep periprosthetic infections in rabbits in vivo.

Effect of melanins from black yeast fungi on proliferation and differentiation of cultivated human keratinocytes and fibroblasts

The effects of melanin preparations from black yeast fungi (BYF) on the proliferation and differentiation of normal cultivated human skin keratinocytes and embryonic pulmonary fibroblasts have been investigated. Melanin preparations in the range of 5–0.1 μg/ml were optimally active, with a more pronounced effect on keratinocyte than on fibroblast proliferation. Of 17 dihydroxynaphthalene (DHN) natural melanin preparations and two commercial dihydroxyphenylalanine (DOPA) melanin preparations, only one preparation—DOPA melanin (of animal origin) significantly stimulated proliferation of keratinocytes at 5 μg/ml; four preparations (DHN melanin from BYF) significantly inhibited proliferation of these cells at 5 or 1 μg/ml. The remaining preparations had no significant effect. Similarly, of the 17 preparations of DHN melanin from BYF, one preparation significantly stimulated fibroblast proliferation, and four significantly inhibited proliferation at 5 μg/ml, one at all the concentrations, and three from 1 down to 0.1 μg/ml. These melanin preparations were also shown to affect the in vitro differentiation of keratinocytes.

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 20μm 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.

Experimental bladder regeneration using a poly-l-lactide/silk fibroin scaffold seeded with nanoparticle-labeled allogenic bone marrow stromal cells

In the present study, a poly-l-lactide/silk fibroin (PL-SF) bilayer scaffold seeded with allogenic bone marrow stromal cells (BMSCs) was investigated as a potential approach for bladder tissue engineering in a model of partial bladder wall cystectomy in rabbits. The inner porous layer of the scaffold produced from silk fibroin was designed to promote cell proliferation and the outer layer produced from poly-l-lactic acid to serve as a waterproof barrier. To compare the feasibility and efficacy of BMSC application in the reconstruction of bladder defects, 12 adult male rabbits were divided into experimental and control groups (six animals each) that received a scaffold seeded with BMSCs or an acellular one, respectively. For BMSC tracking in the graft in in vivo studies using magnetic resonance imaging, cells were labeled with superparamagnetic iron oxide nanoparticles. In vitro studies demonstrated high intracellular incorporation of nanoparticles and the absence of a toxic influence on BMSC viability and proliferation. Following implantation of the graft with BMSCs into the bladder, we observed integration of the scaffold with surrounding bladder tissues (as detected by magnetic resonance imaging). During the follow-up period of 12 weeks, labeled BMSCs resided in the implanted scaffold. The functional activity of the reconstructed bladder was confirmed by electromyography. Subsequent histological assay demonstrated enhanced biointegrative properties of the PL-SF scaffold with cells in comparison to the control graft, as related to complete regeneration of the smooth muscle and urothelium tissues in the implant. Confocal microscopy studies confirmed the presence of the superparamagnetic iron oxide nanoparticle-labeled BMSCs in newly formed bladder layers, thus indicating the role of stem cells in bladder regeneration. The results of this study demonstrate that application of a PL-SF scaffold seeded with allogenic BMSCs can enhance biointegration of the graft in vivo and support bladder tissue regeneration and function.

Cell cultivation on porous titanium implants with various structures

The paper presents data on the cultivation of human dermal fibroblasts and rabbit mesenchymal stromal cells on two types of porous titanium implants, i.e., those with irregular pores formed by pressed titanium particles and those with regular pores formed by the cohesion of one-size titanium particles inside the implant. The goal of this study was to determine what type of titanium implant porosity ensured its strongest interaction with cells. Cells were cultivated on implants for 7 days. During this period, they formed a confluent monolayer on the implant surface. Cells grown on titanium implants were monitored by scanning electron microscopy. Fibroblasts interaction with implants depended on the implant porosity structure. On implants with irregular pores cells were more spread. On implants with regular pores fibroblasts enveloped particles and were only occasionally bound with neighboring particles by small outgrowths. There was no tight interaction of particles inside the implant. In implants formed by pressed particles, cells grow not only on surface, but also in the depth of the implant. Thus, we suppose that a tighter interaction of cells with the titanium implant and, supposedly, tissues with the implant in the organism will take place in the variant when the implant structure is formed by pressed titanium particles, i.e., cellular interaction was observed inside the implant. In implants with irregular pores, cells grew both on the surface and in the depth. Thus, cells exhibited more adequate interactions with irregular pore titanium implants in vitro and hopefully the same interaction will be true in tissues after the implantation of the prosthesis into the organism.

Ultrastructure of coelomic epithelium and coelomocytes of the starfish Asterias rubens L. in norm and after wounding

Samples of coelomic epithelium (CE) and coelomocyte suspension of intact and wounded starfish Asterias rubens L. were studied by electron microscopy. The CE was shown to be composed of three types of cells: flagellar (approximately 60%), secretory (approximately 3%), and myoepithelial (approximately 37%); flagellar and secretory cells form the CE apical surface. Secretory cells are represented by two subtypes, i.e., granular and mucous secretory cells. Myoepithelial cells are located in the basal zone of the epithelium. In 4–5% of cases, adjacent flagellar cells are separated by various sizes of intercellular gaps. These gaps seem to be lacunae left by the flagellar cells after their release into the coelomic cavity. The morphological pattern of the conversion of CE flagellar cells into coelomocytes was characterized. After a moderate wounding used in the present study, no significant structural alterations in the CE organization were revealed. In coelomocyte suspension, small rounded young coelomocytes (approximately 3%) and the larger mature coelomocytes (approximately 97%) were found. On the surface of one of the young coelomocytes, a flagellum was revealed. Surface of the mature coelomocytes forms processes of various size and structure; their cytoplasm contains lysosomes and phagocytic vacuoles of different size. After wounding, a coelomocyte activation was found that consisted of a sharp rise in the number and length of filopodia on their surface, as well as the formation of multicellular aggregates. The complex of ultrastructural data allows it to be suggested that the histogenesis of coelomocytes from CE flagellar cells is a process of cell transdifferentiation.

Culture of mussel ( Mytiuls edulis L.) mantle cells

To date, cell lines derived from marine invertebrates have not been available. Hence primary cell cultures serve as model systems for various experiments. In the present study, we established a primary culture of mussel Mytilus edulis L. mantle cells. Cells were isolated by means of explant culture and enzymatic dissociation of mantle tissue. They maintained viability up to 22 months regardless of the initiation method. In course of culturing, cells that were transferred onto new plates successfully attached to a new surface. The physiological activity of cultured cells was also confirmed by formation of crystals, which appeared after 4–6 months. After a continuous time of culturing, mantle cells may be successfully cryopreserved. We used 5% DMSO with postfreezing survival up to 50%. These results demonstrate that M. edulis mantle cells can maintain viability and physiological activity for an exceptionally long time and can be cryopreserved for further examination.

Matrigel increases the rate of split wound healing and promotes keratinocyte `take' in deep wounds in rats

The influence of matrigel, a mixture of the components of thebasement membrane, on the wound healing was studied in a modelof experimental wounds in rats. Matrigel was found to increasethe rate of epithelization of split-thickness wounds. The modelof deep wound was developed in which the host animal could notprovide enough migrating and proliferating keratinocytes tocover the wound area. The model is relevant to severe burns andinjuries in humans. When rat keratinocyte suspension wastransplanted into deep wounds, cell retention in the wound bedwas only observed if matrigel was added together with the cells.Increasing matrigel concentration in the wound was seen toenhance the rate of wound area coverage by the cells. Althoughthe process of healing seemed macroscopically normal, afterhistological screening of the biopsies cell in the wouldappeared as amorphous aggregates and tubules rather thenstratified epidermis.

A comparative evaluation of antimicrobial eye drops cytotoxicity

In addition to the spectrum of antibacterial activity of antimicrobial medicines and their pharmacokinetic and pharmacodynamic properties, their safety is also an important issue. Currently, there is no consensus on the fluoroquinolone toxicity. The purpose of this study was to compare in vitro the overall cytotoxic effect of six antibacterial fluoroquinolone eye drops: 1. Cipromed™ (ciprofloxacin 0.3 %; Sentiss Pharma Pvt. Ltd., India); 2. Floxal™ (ofloxacin 0.3 %; Dr. Gerhard Mann, Chem.-Pharm. Fabrik GmbH, Germany); 3. Oftaquix™ (levofloxacin 0.5 %; Santen Oy, Finland); 4. Signicef® (levofloxacin 0.5 %; Sentiss Pharma Pvt. Ltd., India); 5. Vigamox® (moxifloxacin 0.5 %; Alcon Laboratories, Inc., USA); 6. Zymar® (gatifloxacin 0.3 %; Allergan Sales LLC, USA). The study showed the possibility of using cultured cells for comparative evaluation of cytotoxic effects of various ophthalmic preparations. We found that tested antimicrobial medicines may have a cytostatic effect in vitro and differ in their cytotoxic potential.