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Altindirek, Didem/0000-0001-5068-2968; Armutak, Elif Ilkay/0000-0002-7359-6568 WOS:000574979000007 PubMed ID: 32871779 Background/Aim: This study was designed to provide further evidence for the interactions between hydrogen sulfide (H2S) and nitric oxide (NO) in ischemia/reperfiision (I/R) injury. Materials and Methods: Rat hearts were studied with the Langendorff technique using the H2S donor sodium hydrosulfide (NaHS, 40 mu M) and the cystathionine gamma-lyase (CTH or CSE) inhibitor DL-propargylglycine (PAG, 1 mM). NO synthase inhibitor L-NG-nitroarginine methyl ester (L-NAME, 30 mg/kg, 7 days) was administered before the isolation. The hearts were homogenized for biochemical and molecular analysis. Results: NaHS reversed I/R-induced cardiac performance impairment, increased tissue nitric oxide production and decreased tissue markers for cardiac injury, while L-NAME inhibited these effects. The expression of CTH was increased with PAG, which was suppressed by L-NAME. Conclusion: H2S and NO increase each other's production suggesting their interaction and cooperation in cardioprotection against I/R injury. Scientific Research Projects Coordination Unit of Istanbul UniversityIstanbul University [31508, 55736, 47109] This work was supported by Scientific Research Projects Coordination Unit of Istanbul University [grant numbers; 31508, 55736 and 47109].

Objects: Benfothiamin is a highly potent form of vitamin B1 protecting endothelial function. On the other hand, vitamin D provides restoration of muscular tissue by inhibition of apoptosis and acceleration of cellular proliferation following muscle injury. We assumed that the administration of these vitamins in ischemia/ reperfusion (I/R) injury, could reduce the damage by alteration of the release of various oxidant and antioxidant mediators leading to cellular damage. Materials and Methods: We assigned 30 Wistar Albino males rats into 5 groups. In the control group (n=6), rats were anaesthetized and total antioxidant capacity (TAS), total oxidant capacity (TOS), malondialdehyde (MDA), superoxide dismutase (SOD) and nitricoxide (NO) level were measured in lower extremity soleus muscle. Benfotiamin and D were given to the groups and the values of these parameters were evaluated in ischemia reperfusion muscle tissue specimens. All tissues were examined histologically. Results: We detected a significant change in groups 3 and 4 for antioxidant NO level after ischemia and reperfusion. Therefore, we observed that the administration of vitamin D and benfothiamin increased NO levels in muscle especially during reperfusion. The level of other oxidants TOS and MDA and antioxidants TAS and SOD were not significant during I/R at given periods. Overall vitamin D and benfothiamin have acute beneficial effects especially in improving I/R injury of lower extremity, even at non-critical periods. Conclusion: Acute term effects of benfothiamin and vitamin D can be useful during where changes due to I/R. The effects can be evaluated during long term I/R.

Stroke is a major cause of death and long-term disability, affecting one in six people worldwide. The only currently available approved pharmacological treatment for ischemic stroke is tissue plasminogen activator; however, relatively few patients are eligible for this therapy. We hypothesized that intravenous (IV) infusion of banked unrelated allogeneic umbilical cord blood (UCB) would improve functional outcomes in patients with ischemic stroke. To investigate this, we conducted a phase I open-label trial to assess the safety and feasibility of a single IV infusion of non-human leukocyte antigen (HLA) matched, ABO matched, unrelated allogeneic UCB into adult stroke patients. Ten participants with acute middle cerebral artery ischemic stroke were enrolled. UCB units were matched for blood group antigens and race but not HLA, and infused 3–9 days post-stroke. The adverse event (AE) profile over a 12 month postinfusion period indicated that the treatment was well-tolerated in these stroke patients, with no serious AEs directly related to the study product. Study participants were also assessed using neurological and functional evaluations, including the modified Rankin Score (mRS) and National Institute of Health Stroke Scale (NIHSS). At 3 months post-treatment, all participants had improved by at least one grade in mRS (mean 2.8 ± 0.9) and by at least 4 points in NIHSS (mean 5.9 ± 1.4), relative to baseline. Together, these data suggest that a single i.v. dose of allogeneic non-HLA matched human UCB cells is safe in adults with ischemic stroke, and support the conduct of a randomized, placebo-controlled phase 2 study.

International audience; Neonatal hypoxic-ischemic encephalopathy (NHIE) is a dramatic perinatal complication, associated with poor neurological prognosis despite neuroprotection by therapeutic hypothermia, in the absence of an available curative therapy. We evaluated and compared ready-to-use human umbilical cord blood cells (HUCBC) and bankable but allogeneic endothelial progenitors (ECFC) as cell therapy candidate for NHIE. We compared benefits of HUCBC and ECFC transplantation 48 hours after injury in male rat NHIE model, based on the Rice-Vannucci approach. Based on behavioral tests, immune-histological assessment and metabolic imaging of brain perfusion using single pho-ton emission computed tomography (SPECT), HUCBC, or ECFC administration provided equally early and sustained functional benefits, up to 8 weeks after injury. These results were associated with total normalization of injured hemisphere cerebral blood flow assessed by SPECT/CT imaging. In conclusion, even if ECFC represent an efficient candidate, HUCBC autologous criteria and easier availability make them the ideal candidate for hypoxic-ischemic cell therapy. STEM CELLS TRANSLATIONAL Neonatal hypoxic ischemic encephalopathy is a dramatic perinatal complication. Neurological and neurosensory sequelae are frequent in survivors, including motor or learning disabilities, cerebral palsy, or epilepsy. Facing the absence of effective curative therapy, many hopes have been credited in cell therapy strategies. Based on behavioral tests, immune-histological assessment and metabolic imaging of brain perfusion using single photon emission computed tomog-raphy, we report in this work that cell transplantation of both human umbilical cord blood cells and endothelial progenitors provided equally early and sustained functional benefits, up to adulthood.

Uncompromised by chronic disease-related comorbidities, human umbilical cord blood (UCB) progenitor cells with high aldehyde dehydrogenase activity (ALDHhi cells) stimulate blood vessel regeneration after intra-muscular transplantation. However, implementation of cellular therapies using UCB ALDHhi cells for critical limb ischemia, the most severe form of severe peripheral artery disease, is limited by the rarity (18-fold over 6-days under serum-free conditions. Consistent with the concept that ALDH-activity is decreased as progenitor cells differentiate, only 15.1% ± 1.3% of progeny maintained high ALDH-activity after culture. However, compared to fresh UCB cells, expansion increased the total number of ALDHhi cells (2.7-fold), CD34+/CD133+ cells (2.8-fold), and hematopoietic colony forming cells (7.7-fold). Remarkably, injection of expanded progeny accelerated recovery of perfusion and improved limb usage in immunodeficient mice with femoral artery ligation-induced limb ischemia. At 7 or 28 days post-transplantation, mice transplanted with expanded ALDHhi cells showed augmented endothelial cell proliferation and increased capillary density compared to controls. Expanded cells maintained pro-angiogenic mRNA expression and secreted angiogenesis-associated growth factors, chemokines, and matrix modifying proteins. Coculture with expanded cells augmented human microvascular endothelial cell survival and tubule formation under serum-starved, growth factor-reduced conditions. Expanded UCB-derived ALDHhi cells represent an alternative to autologous bone marrow as an accessible source of pro-angiogenic hematopoietic progenitor cells for the refinement of vascular regeneration-inductive therapies.

Der Hirntod des Spenders und die konsekutive generalisierte massive IL-6 Produktion und Sezernierung beeinflussen maßgeblich die Immunogenität des Spenderorgans, verstärken den Ischämie-/Reperfusionsschaden und führen in der Folge vermehrt zu akuten Abstoßungen, einer reduzierten initialen Transplantatfunktion und einer beeinträchtigten Langzeitfunktion des Transplantats. In dieser Arbeit wurde der Einfluss einer einmaligen Vorbehandlung des hirntoten Spenders (BD) mit einem monoklonalen anti- IL-6-Rezeptor Antikörper (IL-6R mAb) auf die frühe und späte Immunantwort des Empfängers und den Ischämie/Reperfusionsschaden nach Nierentransplantation (NTx) in einem Rattenmodell untersucht. Es wurden Nieren hirntoter (BD) und lebender (LD) F-344-Spenderratten in Lewis-Empfängerratten transplantiert. Der Hirntod wurde im Spender 6 Stunden (h) vor der Organentnahme und Transplantation induziert. BD und LD Spenderratten wurden 1 h nach Hirntodinduktion mit anti-IL-6R/CD126 mAb behandelt. Empfänger von Transplantaten unbehandelter Spender dienten als Kontrollen. Um den Effekt einer Spendervorbehandlung zu überprüfen, wurden zu den Zeitpunkten 18 h, 7 Tage und 5 Monate nach Transplantation Genexpressionsanalysen (RT-TaqMan-PCR) spezifischer Entzündungsmarker (IL-6, IL-10, IL-4, TNFα, IFNγ, IP-10, CCL19/-20) , histologische und immunhistologische Färbungen (ED1+, MHCII+, CD4+, CD8+) sowie durchflusszytometrische (T-Zellen, Antigen-präsentierende Zellen (APZ), NK-, NKT-Zellen) und ELISA- Untersuchungen (IL-6, TNFα, IFNγ) durchgeführt . Die Langzeittransplantatfunktion wurde zusätzlich durch serielle Messungen der Urin-Proteinkonzentration (Proteinurie) und der Kreatinin-Clearance bewertet. Es zeigte sich, dass durch die Vorbehandlung hirntoter Spender mit einem mAb gegen den IL-6- Rezeptor die Schädigung von Nierentransplantaten in der frühen Phase nach Transplantation deutlich minimiert werden kann. Die Genexpression pro- bzw. anti-inflammatorischer Zytokine (IL-6, TNFα, INFγ, IL-4, IL-10) und Chemokine (CCL-19, -21) wurde durch die Vorbehandlung reduziert. Weiterhin zeigte sich unter Vorbehandlung eine geringere Infiltration spezifischer Entzündungszellen (ED1+, CD8+ T-Zellen, APZs, DCs, NK-T-Zellen) in das Allograft sowie signifikant geringere histomorphologische Schädigungen im Vergleich zur Kontrollgruppe. Zusätzliche Analysen ergaben eine Reduktion transplantatinfiltrierender Zellen (durchflusszytometrische Untersuchungen von Transplantatgewebeproben) sowie geringere Serumproteinkonzentrationen der Zytokine IL-6, TNFα, und IFNγ (ELISA) in der Gruppe vorbehandelter Hirntodspender im Vergleich zur Kontrolle (unveröffentliche Daten der Arbeitsgruppe). Im Unterschied zur frühen Phase nach Transplantation scheint eine einmalige Spendervorbehandlung die Ausbildung einer Chronischen Allograft Nephropathie nicht ausreichend unterdrücken zu können. Zwar zeigte sich unter Spendervorbehandlung nach 5 Monaten eine geringere Genexpression verschiedener Zyto- und Chemokine (IL-6, IFNγ, TNFα, IL-10, IP-10, und CCL19/-21) und immunhistologisch eine geringere Allograftinfiltration (MHCII+, ED1+, und CD8+ Zellen), jedoch keine suffiziente Reduktion immunkompetenter Zellen (NKT, NK-, CD4+ Zellen, APZs). Durch eine Vorbehandlung des hirntoten Spenders mit einem anti-IL-6R mAb kann zum einen die inflammatorische Immunantwort in der frühen Phase nach Transplantation im Empfänger deutlich reduziert, zum anderen der pathophysiologische Einfluss des Ischämie-/ Reperfusionsschadens erheblich gemindert werden. Empfänger von Lebendspenden scheinen dagegen nicht von einer Spendervorbehandlung zu profitieren. Eine einmalige Therapie des Spenders scheint nicht ausreichend für die Verbesserung der Transplantat- Langzeitfunktion zu sein., Brain death (BD) of the donor and the subsequent up-regulation of IL-6 production and secretion significantly influence graft immunogenicity and ischemia/reperfusion injury, thus, leading to a considerable impairment of initial and long-term graft function. In a rat kidney transplant model we evaluated the efficacy of a single pretreatment of brain dead donor rats with an anti-IL-6 receptor (IL-6R/CD126) monoclonal antibody (mAb) to decrease the inflammatory immune response and the associated ischemia/reperfusion injury (IRI) after kidney transplantation (NTx). Kidneys of BD and living (LD) F-344 donor rats were transplanted into Lewis recipient. BD was induced in the donor 6 hours (h) before organ recovery and transplantation. Both BD and LD donor rats were treated with anti-IL-6R mAb 1 h after BD induction. Recipients of grafts from untreated donors served as controls. To assess the effect of donor pretreatment was evaluated by gene expression analyses of inflammatory markers (IL-6, IL-10, IL-4, TNFa, IFNg, IP-10, CCL19/-20; RT-TaqMan PCR), by histological and immunohistochemical staining (ED1+, MHCII+, CD4+ , CD8+ cells) and flow cytometry (T- cells, antigen presenting cells (APC), NK, NKT cells) and ELISA protein analyses (IL-6, TNFa, IFNg). The long-term graft function was evaluated by serial measurement of urinary protein secretion (proteinuria) and creatinine clearance. Pretreatment of BD donors by anti-IL-6 receptor mAb minimized BD associated graft injury compared to the control group. Gene expression of specific cytokines (IL-6, TNF, IFNγ, IL-4, IL-10) and chemokines (CCL-19, -21) was reduced. Furthermore, the number of graft infiltrating cells (ED1+, CD8+ cells, APCs, DCs, NK-T cells) was significantly lower, and fewer histomorphological lesions were observed in recipients of pretreated organs, compared to the control group. Flow cytometry analysis of PBMCs revealed reduced numbers of APCs. Moreover, lower protein concentrations of IL-6, TNF, IFNγ; ELISA) were detected recipients of pretreated donor organs. While a single donor pretreatment with anti IL-6R mAbis able to reduce the early inflammatory immune response, chronic allograft nephropathy cannot be sufficiently prevented. In summary donor treatment with anti-IL-6R mAb markedly reduced the inflammatory immune response in the early phase after transplantation of grafts from BD donors, while recipients of LD grafts do not seem to benefit. Furthermore, it can be assumed that the pathophysiological effects of IRI could be significantly reduced by pretreatment. A single treatment of the donor does not seem to be sufficient for the improvement of long-term graft function.

The present first-in-human clinical trial evaluated the safety and feasibility of a newly developed and cryopreserved Cardiology Stem Cell Centre adipose-derived stromal cell (CSCC_ASC) product from healthy donors for intramyocardial injection in ten patients with ischemic heart disease and ischemic heart failure (IHF). Batches of CSCC_ASC were isolated from three healthy donors by liposuction from abdominal adipose tissue. Adipose mesenchymal stromal cells were culture expanded in bioreactors without the use of animal constituents, cryopreserved, and stored in vials in nitrogen dry-storage containers until use. Direct injection of CSCC_ASC into the myocardium did not cause any complications or serious adverse events related to either treatment or cell administration in a 6-month follow-up period. Four out of ten heart failure patients developed donor-specific de novo human leukocyte antigen (HLA) class I antibodies, and two out of ten patients had donor-specific HLA antibodies already at baseline. There were no clinical symptoms or changes in inflammatory parameters in the follow-up period that indicated an ongoing immune response. There was a tendency toward improvement in cardiac function after CSCC_ASC treatment at 6-month follow-up: left ventricular end systolic volume decreased and left ventricular ejection fraction increased. In addition, exercise capacity increased. These changes were independent of the presence or absence of HLA antibodies. It is concluded that the newly developed cryopreserved product CSCC_ASC from healthy donors was a safe and feasible treatment. We observed a tendency toward efficacy in patients with IHF. These findings have to be confirmed in larger placebo controlled clinical trials.