[Use of allogeneic mesenchymal stem cells in the treatment of intestinal inflammatory diseases].

AIM: to determine the whether mesenchymal stem cells (MSC) may be used in the treatment of patients with chrOnic intestinal inflammatory diseases (IID). SUBJECTS AND METHODS: Thirty-nine patients with ulcerative colitis (UC) (Group 1) and 11 with Crohn's disease (CD) (Group 2) were examined. Comparative groups included 30 patients with UC (Group 2) and 10 with CD (Group 4). Two-three days before MSC administration, immunodepressants were discontinued, the dosage of corticosteroids was reduced to 15-20 mg/day, and that of aminosalicylates remained to be 2 g/day. The results were quantified using the mean values of the Rachmilewich clinical activity index, the Crohn's disease activity index and the Mayo and Gebs scales. The patients were followed up for 4-8 months. Humoral immunological indices (cytokines, autologous antibodies) were determined. Bone marrow cells were obtained from the donor sternum or iliac crest. Cultivation at the end of weeks 5-6 provided a population of allogeneic donor MSC in a quantity of (1.5-2) x 10(8) tells required for transplantation to a patient. MSC cultures were once injected intravenously in a dropwise fashion. RESULTS: A statistically significant decrease in the indices of the clinical and morphological activities of an inflammatory process was noted in 39 patients with UC and in 11 patients with CD as compared with the comporison groups after MSC transplantation. Clinicomorphological remission occurred in 40 patients. Inclusion of MSC into the treatment program was ineffective in 8 patients with UC and in 2 patients with CD. The use of MSC made it possible to discontinue corticosteroids in 34 of the 50 patients with the hormone-dependent and hormone-resistant forms of UC and CD and to reduce the dose of prednisolone to 5 mg/day in 7 patients, by administering 5-aminosalicylic acid only. CONCLUSION: The use of MSC may be appreciated as a new strategic direction of therapy for IID. The intravenously administered stem cells exert a potent immunomodulatory effect, reduce the activity of autoimmune inflammation, and stimulate a reparative process in the intestinal mucosa.

[Allogeneic mesenchymal stromal cells in patients with ulcerative colitis: two years of observation].

UNLABELLED: The aim of the study was to determine the efficacy and safety of mesenchymal stromal cells (MSCs) of bone marrow in the treatment of patients with ulcerative colitis (UC). MATERIALS AND METHODS: The study included 44 patients with ulcerative colitis (UC), which was implemented MSC transplantation, 40 patients with UC who received standard therapy with mesalazane (salofalka) 4-6 g/day and corticosteroids (prednisone)--1-2 mg/kg, azathioprine--1.5 mg/kg methotrexate 20-50 mg/m2, and 12 patients who underwent induction and maintenance of infliximab therapy. 2-3 days prior to the induction of MSCs abolish immunosuppressive doses of corticosteroids reduced to 15-20 mg/day dose of aminosalicylates was left at 2.0 g/day. To quantify the results using the average values of indices of Rahmilevich clinical activity, indices of endoscopic and histological activity scales Mayo and Gebs. The patients were observed for 24 months after transplantation. Were studied parameters of the humoral immune status (immunoglobulin A, G, M, autologous antibody), cytokine profile. Bone marrow cells were obtained from the donor's sternum or the iliac crest. By culturing the end of 5 to 6 weeks received a population of allogeneic donor's MSCs in the amount of (1.5-2) x 10(8) cells needed for transplant patient. Culture of MSCs injected in the drip i/v, single dose. RESULTS: In 34 (72.7%) patients with UC after the induction of MSCs was statistically significant compared with the group of patients treated with drugs only 5-aminosalicylic acid and corticosteroids, reducing the clinical and morphological indices of inflammatory activity. In 12 patients with UC include MSCs in the treatment program did not have a therapeutic effect. Application of MSC allowed to cancel corticosteroids in most patients with hormone-dependent and steroid resistance forms of UC, and in 7 to reduce the dose of prednisolone to 5 mg/day, limiting the use of drugs 5-ASA. According to the anti-inflammatory effectiveness of combined therapy with MSCs comparable to infliximab therapy. CONCLUSION: The use of MSCs can be evaluated as a new strategic direction for therapy UC. MSC, introduced in I/O, have powerful immunomodulatory effects, reduce the activity of autoimmune inflammation and stimulate the reparative process in the intestinal mucosa, thereby increasing the duration of remission, reduces risk of recurrence of disease, reduces the frequency of hospitalizations.

[Application of allogeneic mesenchymal stem cells in complex patients treatment with ulcerative colitis].

Systemic transplantation of mesenchymal stem cells (MSC) is known to promote reparative process in a number of tissue damage as well as mucous healing one. It provided the basis for our study which was aimed to the effect of systemic transplantation of allogenic MSC (intravenous transfusion) in complex therapy of patients with Ulcerative Colitis.

["Ischemia/reperfusion" for stem cells of two "critical" cell renewal systems of organism].

In this article is presented the result of the experiments on mice-hybrids F1(CBA x C57B1/6), which indicates the presence of the reaction of "ischemia/reperfusion" for stem cells of two "critical" cell renewal systems of organism (bone marrow and intestinal epithelium) during the irradiation under the conditions of hypoxic radioprotector application. The additional injection of the source of nitric oxide radicals-sodum nitroprusside (SNT) to the mice right after the irradiation under the conditions of hypoxic protection by serotonin, resulted the substantial increase of the survival rate of hematopoietic stem cells (registered by the methods of endogenous and exogenous colony forming in spleen) and stem cells of intestinal epithelium (registered by the method of intestinal "microcolonies"). The similar radioprotective effect was also registered during the test of survival rate of mice under tests of "bone marrow" and "intestinal" forms of radiation lethality that is evidence of the importance of the realization of phenomenon "ischemia/reperfusion" in the reaction of whole organism on the acute radiation injury. As SNP weakens the manifestation apoptosis and necrosis through competition with active forms of oxygen (AFO) during the period of "reperfusion" on basis of the found out phenomenon experimental model for studying mechanisms of stem cells damage in vivo induced by AFO and for the search of the modifiers weakening or strengthening such damage can be developed.

[Radioprotective and antistressful properties of nitric oxide production modulators].

The radioprotective and antistressful activities of L-arginine and the "Pronumol" preparation, in which L-arginine is contained in the complex of proteins with nucleic acids, were studied. In mice repeated peroral intake of L-arginine and "Pronumol" partially prevented radiation-induced and stress-induced lipid peroxidation and DNA degradation in thymus, increased hemopoietic stem cell survival, and prevented an increase in chromosome aberration frequency in bone marrow cells of irradiated mice. When repeatedly administered per os before irradiation, "Pronumol" increased survival of intestinal stem cells in irradiated mice and prevented thymus cell devastation induced by radiation and stress.

[Radioprotective effect of E. coli endotoxin on hematopoietic stem cells is partially suppressed by inhibiting production of nitric oxide by administering N-omega-nitro-L-arginine]. / Radiozashchitnoe deistvie éndotoksina E. coli na stvolovye krovetvornye kletki chastichno podavliaetsia ingibitsieé oksida azota putem vvedeniia N-omega-nitro-L-arginina.

Lipopolysaccharide of E. coli (LPS) injected to mice one day before the total gamma-irradiation caused a substantial increase in the level of endogeneous colonies formed in the spleen. It is known that this type of endotoxin may result in considerable production of nitric oxide from macrophages in different tissues. Therefore it is possible that the activation of hemopoietic stem cells after LPS-treatment was completely or partially stimulated by nitric oxide, which is the most important physiological mediator, or by action some other mediators (cytokines and growth factors) produced by hemopoietic microenvironment elements in response to the action of nitric oxide. This assumption was checked in experience with combined treatment of mice by LPS and a nonspecific inhibitor of nitric oxide production--N omega-nitro-1-arginine (NA). NA used in high dose (250 mg/kg) reduced partially (approximately by 30%) the LPS-increased level of spleen endogeneous colonies. When LPS was injected to mice 15 minutes after gamma-irradiation, this led to a slight increase in level of spleen colonies. In case when LPS was used together with NA after gamma-irradiation, this increase was still found.

[A modification in the radiation reaction of normal and tumor tissues in mice induced by N-methylformamide]. / Modifikatsiia radiatsionnoi reaktsii normal'nykh i opukholevykh tkanei myshei N-metilformamidom.

The effect of potential differentiation-inducing agent N-methylformamide on radiation response of murine normal and tumor cells (Lewis lung carcinoma, hematopoietic tissue and jejunum epithelial stem cells) was studied. The agent reduced or not altered radiation damage of tumor and epithelial cells in mice receiving NMF before irradiation. Sensitization to radiation was observed in endogenous spleen colony forming hemopoietic stem cells. When the agent was injected 15 min before irradiation the sensitizing effect was less pronounced. The highest effect was observed when agent was injected 24 h after irradiation of animals.

[Radiobiological aspects of increased radioresistance of murine epithelial stem cells from patches of Peyer]. / Radiobiologicheskie aspekty povyshennoi radiorezistentnosti stvolovykh kletok épiteliia nonkogo kishechnika myshei v zone Peerovykh bliashek.

The fraction of the survived intestinal crypts associated with patches of Peyer is higher to those non-patch-associated crypts of mice for three inbred strains. The difference in the survival between associated and non-patch-associated crypts increases with dose of gamma-irradiation. This difference for old mice is less than for young mice. Pre-irradiation (5 Gy) of mice one week before the conditioned gamma-irradiation cannot modify the difference in the survival between associated and non-patch-associated crypts. Radioprotection of mice by hypoxic gas mixture (10% O2) cannot modify this difference. Pre-treatment of mice by dextran-sulfate alone or in combination with hypoxic gas mixture decreased the survival of intestinal crypts associated with patches of Peyer to the level of non-path-associated crypts. The difference in the survival between two subpopulation of intestinal stem cells is less after neutron irradiation than after gamma-irradiation.

[Radiosensitizing effect of hyperthermia (41.5C, 30 min) in stem cells of mouse spermatogenic epithelium]. / Radiosensibiliziruiushchii éffect gipertermii (41,5, 30 min) dlia stvolovykh kletok spermatogennogo épiteliia myshei.

Mouse testes preheated up to 41.5 degrees C for 30 min exhibit an increased radiosensitivity of testis epithelium stem cells. High death rate is described by a shift (by approximately 4 Gy) in the "dose-response" curve to the Abscissa left. The DMF value for hyperthermia depends on the cell survival rate, at which calculations are made, and varies from 2.1 to 1.4 within the range under study.

[The effect of gas hypoxia (10% O2) and dextran sulfate on the radiosensitivity of the stem cells of the intestinal epithelium in mice]. / Vliianie gazovoi gipoksii (10% O2) i dekstran-sul'fata na radiochuvstvitel'nost' stvolovykh kletok kishechnogo épiteliia myshei.

Injection of dextran sulphate before irradiation was shown to protect jejunal epithelium stem cells (D0 increased from 1.13 to 1.82 Gy). The protective effect of a combination of dextran sulphate and gas hypoxic mixture (10% O2) did not exceed that of the administration of the gas hypoxic mixture (10% O2) alone.

[The radioprotective effect of a hypoxic respiratory mixture (8% O2) for intestinal epithelial stem cells during single and fractionated irradiation of mice]. / Radiozashchitnoe deistvie gipoksicheskoi dykhatel'noi smesi (8% O2) dlia stvolovykh kletok kishechnogo épiteliia pri odnokratnom i fraktsionirovannom obluchenii myshei.

The method of intestinal "microcolonies" was used to study the radioprotective effect of a gas mixture, containing 8% of O2, on mice subjected to single and fractionated (5 fractions for 30 min) irradiation. The protective effect was indicated by a decreased slope of dose curves of the stem cell injury; the extrapolation number decreased simultaneously. So the values of dose modifying factors (DMF) were higher, when calculated by D0 ratio (where they amounted to 1.76 and 1.39 for single and fractionated exposure respectively), than those determined by equally effective doses (1.19 and 1.26 for single and fractionated effects respectively, which corresponded to LD50/4 when calculated at lg N = 1.9). It is suggested that the radiation response of certain stem cell populations of intestinal epithelium are different: this is attributed to different degrees of hypoxia in cells and to different directions of the hypoxia effects on the injury and the ability of postirradiation repair.

[Radiosensitivity of stem cells in the spermatogenic epithelium of mice of different strains and different ages]. / Radiochuvstvitel'nost' stovolovykh kletok spermatogennogo épiteliia u myshei razlichnykh shtammov i raznogo vozrasta.

In experiments on CBA and BALB/c male mice (3 months of age) and F1(CBA X C57BL/6) hybrids (at the age of 3, 12, and 24 months) a difference was noted in the radiosensitivity of spermatogenic epithelium stem cells displayed by the changes in their colony-forming ability in testicular tubules 42 days following local 60Co-gamma-irradiation. The older the hybrid mice the smaller was the number of spermatogenic epithelium stem cells.

[Radiobiology of stem cells in intestinal epithelium. Effect of single and multiple preliminary sublethal irradiation of mice on the dose dependence of the survival of stem cells in small intestine epithelium]. / Radiobiologiia stvolovykh kletok épiteliia kishechnika. Vliianie odnokratnogo i mnogokratnogo predvaritel'nogo subletal'nogo oblucheniia myshei na dozovuiu zavisimost' vyzhivaemosti stovolovykh kletok épiteliia tonkogo kishechnika.

After a single or three-fold whole body irradiation of mice with a dose of 4 Gy and the time interval for the proliferation to be restored (5 days or 3 weeks) the survival curve for stem cells of small intestine epithelium with regard to radiation dose was the same as that for non-preirradiated mice. This indicated that the proliferative potential of stem cells in these experimental conditions was not reduced.

[Radiosensitizing and damaging action of hyperthermia on different biological systems. Radiosensitizing and damaging action of hyperthermia on mouse hematopoietic stem cells]. / Radiosensibiliziruiushchee i povrezhdaiushchee deistvie gipertermii na razlichnye biologicheskie sistemy. Radiosensibiliziruiushchee i povrezhdaiushchee deistvie gipertermii na stvolovye krovetvornye kletki myshei.

Dose dependences of heat-induced inactivation of CFUs have been described for the temperature range of 41 to 44 degrees C. The energy of activation of the processes of thermal death was 100-120 kcal/M. CFUs were more resistant to heating than the cells of mouse leukaemia. Heating 1 h or immediately before irradiation had a radiosensitizing effect on CFUs. When bone marrow cells were heated 1 h after irradiation the radiosensitizing effect was absent: there was only a simple summation of the effects of the two damaging agents.

[Age-related changes in the radiosensitivity of animals and critical cell systems. Survival of stem cells of the small intestine epithelium 4-5 days after death in mice of various ages]. / Vozrastnye izmeneniia radiochuvstvitel'nosti zhivotnykh i kriticheskikh kletochnyk sistem. Vyzhivaemost' stvolovykh kletok épiteliia tonkogo kishechnika i 4-5 cutochnaia smertnost' myshei raznogo vozrasta posle oblucheniia.

Mice of three strains exhibited similar changes in radiosensitivity tested with a reference to "intestinal" death: juvenile and old animals were more radiosensitive. No age-related changes were detected in radiosensitivity of stem cells of the small intestine epithelium estimated with a reference to average lethal dose D0.