Minimally Manipulated Bone Marrow-Derived Cells Can Be Used for Tissue Engineering In Situ and Simultaneous Formation of Personalized Tissue Models.

Red bone marrow and autologous bone tissue (bone fragments and bone chips) of the donor were harvested intraoperatively during autoplasty of talus bone defect. Titanium chips were obtained by grinding a fragment of a microporous titanium-coated hip arthroplasty (Zimmer). Bone marrow mononuclear cells were isolated in the operating room, and bone and titanium fragments were incubated with a suspension of mononuclear cells. The quality of revitalization was assessed by fluorescence microscopy and histological examination after culturing of adherent cells on the bone and titanium fragments. During culturing on bone chips, bone marrow mononuclear fraction cells demonstrated significantly higher metabolic activity than bone marrow cells (p=0.04). Mononuclear fraction cells were also capable of stable colonization of titanium fragments with the formation of composite tissue model.

[The Possibilities of Cell Technologies in the Treatment of Cicatricial Lesions of the Vocal Folds].

The article is a brief review of publications devoted to the problem of persistent dysphonia. The main cause of voice disorders is the scarring of the vocal folds resulting from trauma, surgical manipulation, inflammatory process. Treatment of cicatricial lesions of the vocal folds remains a challenge, as far as existing methods do not ensure the recovery of the ultrastructure of the vocal folds. The authors present modern data on the structure of the vocal folds at the cellular level. Considered pathologic processes occur in different stages of scarring. Applied technologies of phonosurgery and conservative treatment, their effectiveness and shortcomings are covered. Analysis of experimental research conducted in the world demonstrates the promise of using the methods of tissue engineering to treat scarring of the vocal folds and to restore the microstructure of the latter. Identified current issues remain unresolved, which leads to the need for further experimental and clinical studies in the treatment of this pathology.

[PLGA mesh-collagen hybrid scaffold and tissue-engineered product in substitution urethroplasty: experimental validation].

Urethral strictures are a pressing issue in modern medicine. Substitution urethroplasty is considered one of the most effective treatment methods. However, despite the surgery showing good results, many problems remain unresolved, one being substitute material deficiency in extensive or recurrent strictures, as well as in cases requiring multistage surgeries, including those used to treat hypospadias. Graft removal also leaves the donor area prone to diseases and increases the length of surgery leading to a higher risk of intra- and postoperative complications. Tissue engineering (namely tissue-engineered products comprised of scaffolds and cells) may be a useful tool in dealing with these issues. The authors assessed the characteristics of a novel hybrid scaffold created from "reconstructed" collagen and a poly(lactic-co-glycolic acid) mesh. The resulting composite product showed good mechanical properties and functional performance. The hybrid scaffold was non-cytotoxic and provided an adequate base for cell adhesion and proliferation. Biodegradation resulted in the scaffold being replaced by urothelium and urethral mucosa. The newly formed tissues possessed adequate structural and functional properties. Only one rabbit out of 12 developed urethral stricture at the site of scaffold implantation. The above-mentioned facts suggest that the novel hybrid scaffold is a promising tissue-engineered product with potential implication in substitution urethroplasty.

[Correction of cronic liver failure by transplantation of liver cells suspension and cell-engineering designs (experimental investigation)].

On an experimental model of chronic fibrotic liver damage (male rats Wistar (n-60), damage of CCl4, the duration of the experiment 90 days) it was studied the effectiveness of cell therapy for the correction of chronic liver failure. These rats were divided into 3 experimental groups: in the Ist-group (control, n=10) isotonic saline (650 mkl.) was injected; in the IInd-group (n=20) suspension of liver cells was applicated in a dose 8 - l0 x 10(6) cells; in the IIIrd-group (n=30) suspension of liver cells and bone marrow cells (mesenchymal stromal cells) in ratio 5:1 were used as cell associates on microparticles intjectable heterogeneous biopolymer hydrogel "SpheroGEL" (cell-engineering design) in common dose 8 - l0 x 10(6) It was ascertained that in the 2nd and in the 3rd groups the accelerated normalization of disturbed liver functional indices (ALT, AST, ALP) took place - to 30 days, but in the control group only to 90 days. The reliable differences in rats ofnormalization offunctional indices were absent between the IInd and the IIIrd groups. But in 90 days by using special histological dyeing it was found out that defibrotic processes in liver tissue were more expressed in the IIIrd group in comparison with the IIIrd group. Received results were consequence of prolonged vital activity of cells (liver cells and mesenchymal stromal bone marrow cells) into cell-engineering designs, which were transplanted in the IIIrd group. The obtained effect can be explained by that the developed cell-engineering designs provide adequate conditions for prolonged vital activity of the transplanted cells.

[The influence of multipotent mesenchymal stromal cells of bone marrow on process of local radiation injury in rats after local beta-irradiation].

The influence of multipotent mesenchymal stromal cells (MMSC), got from allogenic bone marrow, on local radiation injury in rats after beta-irradiation by a source 90Sr/90Y in a dose 140 Gy under various conditions and ways transplantation was studied. It was established, that transplantation MMSC, allocated on biodegradative membranes, which have been carried out for 21 day after an irradiation in conditions minor surgical of ulcers, resulted in reduction of the area of local beam defeats and acceleration of healing of skin, in comparison with the control of an irradiation. The introduction of suspension MMSC subcutaneous around of the center of a defeat at 8 day after an irradiation caused earlier healing of ulcers. It was concluded, that application of MMSC is perspective for treatment of local radiation injury and necessity of development of optimum conditions of their use at cell therapy of radiation injury of skin.

[Use of precultivative multipotent mesenchymal stromal cells isolated from autologic bone marrow in treatment of non-healing autoimmune gastric ulcers].

It is shown that multipotent mesenchymal stromal cells (MMSC) grafted to the zone of ulcer defect contribute to better wound repair in case of chronic non-healing gastric ulcers. It is noted that precultivative MMSC isolated from bone marrow achieve their bioregulatory activity in patient's organism not at once but due to prolonged functioning in the grafting zone (not less than 30 days).

[Effects of the platelet activating factor on the proliferation and IgE secretion by U-266BL human myeloma cells].

Platelet activating factor (PAF) influences the proliferation and IgE secretion by U-266BL human myeloma (HM) cells. At a relatively high PAF concentration (up to 10(-8) M), the rate of proliferation is significantly decreased, while the IgE secretion rate is increased. At low PAF concentrations (10(-11) M), the HM cell proliferation rate increases, whereas their ability to IgE secretion decreases. The specific secretion activity (per cell) is a linear function in the range of PAF concentrations from 10(-11) to 10(-8) M. This behavior can be used for the quantitative determination of PAF in biological media.

[The use of pre-differentiated mesenchymal bone marrow stem cells for treatment of deep burn wounds]. / Ispol'zovanie preddifferentsirovannykh mezenkhimal'nykh stvolovykh kletok kostnogo mozga dlia lecheniia glubokikh ozhogovykh ran.

The opportunity of faster closing deep burn wounds by using fibroblast-like mesenchymal bone marrow stem cells (FMSC) and embryonic fibroblasts (EF) was investigated in rats. It was shown that FMSC as well as EF transplanted onto burn surfaces reduced the expression of cell infiltration but accelerated the formation of vessels de novo and granulation tissue in the wounds. These changes form the conditions for faster closing the burn wounds as compared to the control wounds (without cell transplantation). High rate of wound closing induced by FMSC and EF is thought to be caused by a long period (up to 30 days) of vital activity of the cells grafted on the burn surface. It was also found out that the rate of wound regeneration induced by FMSC was higher that that induced by EF.

[The cultivation of bone marrow cells and cell lines on polymeric films].

The cultivation of multipotent mesenchymal stromal bone marrow cells and cells of A-431, MDCK, Vero, 3T3 and Hep-G2 was performed on polymeric films (PVA) with different hydrophobic fatty acid residues. The cells of different types grew on these films with different intensity, but in the most cases comparable with the cultivation control on usual plastic. The examined films were nontoxic to cells and sufficiently adhesive. They did not changed pH of cultural media, were optically transparent under microscope and comfortable in the experimental work. These films can be used as a model for the artificial organ construction. The covalent binding of different fatty acids to PVA shows possibility of the adaptable changes of films properties (hydrophobity and adhesiveness), and therefore possibility of the creation of optimal conditions for different cell types attachement and growth.

Use of autologous bone marrow mesenchymal stem cells for healing of free full-thickness skin graft in a zone with pronounced hypoperfusion of soft tissues caused by arteriovenous shunting.

A combined graft consisting of a free full-thickness skin flap and cultured autologous fibroblast-like bone marrow mesenchymal stem cells was effectively implanted and healed on the facial soft tissue defect after removal of a pathological vascular conglomeration in a female patient with congenital arteriovenous macrofistulous dysplasia. In order to reduce bloodflow intensity and arteriovenous shunting, repeated endovascular occlusion and transcutaneous ligature of regional vessels from the carotid artery basin feeding the pathological zone was carried out followed by resection of this tumor-like vascular formation.

Use of recombinant AdSV40-BetaGal adenoviral construction for monitoring of transplanted cells.

Original recombinant adenoviral construction carrying E. coli beta-galactosidase LacZ gene designed by the authors is convenient for labeling and monitoring of bone marrow mesenchymal (stromal) progenitor cells and myocardial and skin fetal cells transplanted in damaged rat tissues (in the perinecrotic zone of the myocardium and onto burnt skin surface) for their reparation. This genetic construction after pre-inactivation of endogenous beta-galactosidase allows to detect transplanted cells in the foci of injury; positive effects of transplantation on tissue reparation processes can be attributed to the presence of transplanted cells.

First experience of the use bone marrow mesenchymal stem cells for the treatment of a patient with deep skin burns.

Female patient with extensive skin burn (I-II-IIIAB skin burn, total area 40%, area of IIIB degree 30%) was treated using transplantation of allogenic fibroblast-like bone marrow mesenchymal stem cells onto the surface of deep thermal burn. The study of wound healing dynamics after transplantation of allogenic fibroblast-like mesenchymal stem cells confirmed high tempo of wound regeneration in the presence of active neoangiogenesis. Due to this, autodermoplasty of burn wounds could be carried out with good results as early as on day 4 after transplantation of fibroblast-like mesenchymal stem cells; this led to more rapid healing of donor zones and accelerated rehabilitation of the patient.

Mesenchymal bone marrow stem cells more effectively stimulate regeneration of deep burn wounds than embryonic fibroblasts.

Regeneration of deep burn wounds after transplantation of allogenic and autogenic fibroblast-like bone marrow mesenchymal stem cells and embryonic fibroblasts on burn surface was studied in 40 Wistar rats. Transplantation of allogenic and autogenic fibroblast-like bone marrow mesenchymal stem cells and transplantation of embryonic fibroblasts decreased cell infiltration of the wound and accelerated the formation of new vessels and granulation tissue in the wound in comparison with the control (burn wounds without cell transplantation). Regeneration processes were most active after transplantation of fibroblast-like bone marrow mesenchymal stem cells, in particular, autogenic cells, which was confirmed by more rapid decrease in burn surface area. Wound healing after transplantation of fibroblast-like bone marrow mesenchymal cells and embryonic fibroblasts was associated with long functioning of transplanted cells (as was shown by staining for beta-galactosidase, the cells were transfected with an adenovirus vector carrying the marker gene). It is hypothesized that more rapid regeneration of burn wounds after transplantation of fibroblast-like bone marrow mesenchymal stem cells was due to low differentiation of these cells in comparison with embryonic fibroblasts.

Recovery of contractile function of cryodamaged rat myocardium after transplantation of fetal cardiomyocytes and predifferentiated bone marrow stromal stem cells.

The effect of cell transplantation into cryodamaged rat myocardium was studied on isolated hearts by increasing functional load to the left ventricle. Transplantation of allogeneic fetal cardiomyocytes improved the function of the left ventricle under conditions of considerably increased preload. Transplantation of autologous mesenchymal stem cells repaired left-ventricular function under conditions of increased pre- and afterload.

[In vitro immunization of human lymphocytes with Epstein-Barr virus capsid antigen]. / Immunizatsiia in vitro limfotsitov cheloveka kapsidnymi antigenami virusa Epshteina-Barr.

Conditions for in vitro immunization of human lymphocytes from adult peripheral blood, tonsils and cord blood with Epstein-Barr Virus (EBV) capsid antigens have been studied. Pokeweed mitogen and B cell growth factor from Namalva cell line were shown to induce a significant production of specific antibodies by human lymphocytes stimulated with EBV. This effect made it possible to generate primary immune response in vitro using lymphocytes from EBV seronegative donors.

Differentiation of bone marrow stromal stem cells into cardiomyocyte-like cells in different mammalian species.

We describe the possibility of obtaining cardiomyocyte-like cell cultures from rat, guinea pig, and human bone marrow stromal stem cells. The content of troponin I-positive cells attains 35-45% of the total number of cells in the cultures and persists at this level for up to 4 months under differentiation conditions. Spontaneous contractions of cardiomyocyte-like cells were observed after the formation of cell monolayer under differentiation conditions.