AIM: The therapeutic potential of adipose-derived stem cell conditioned medium (ASC-CM) was studied in the rabbit model of critical limb ischemia (CLI). METHODS: Rabbits received treatment with ASC-CM or placebo. Gastrocnemius muscle tissue was collected 35 days after ischemia induction. Ischemic changes were evaluated in hematoxylin-eosin stained tissues for early (necrotic lesions/granulation tissue) and late (fibrous scars) phases of tissue repair. The expression of proangiogenic miR-126 was also evaluated using in situ hybridization. The levels of cytokines, insulin, and C-peptide were measured in blood. RESULTS: Early repair phases were observed more often in placebo-treated samples (45.5%) than in ASC-CM-treated ones (22.2%). However, the difference was not statistically significant. We demonstrated a statistically significant positive correlation between the early healing phases in tissue samples and C-peptide levels in peripheral blood. The expression of proangiogenic miR-126 was also shown in a number of structures in all phases of ischemic tissue healing. CONCLUSION: Based on our results, we believe that treatment with ASC-CM has the potential to accelerate the healing process in ischemic tissues in the rabbit model of CLI. The whole healing process was accompanied by miR-126 tissue expression. C-peptide could be used to monitor the course of the tissue healing process.

• MeSH
• C-peptid krev   MeSH
• cytokiny krev   MeSH
• diabetická noha MeSH
• dospělí MeSH
• experimentální diabetes mellitus krev   MeSH
• fibróza MeSH
• fyziologická neovaskularizace účinky léků   MeSH
• granulační tkáň patologie   MeSH
• hojení ran účinky léků   fyziologie   MeSH
• hybridizace in situ MeSH
• inzulin krev   MeSH
• ischemie krev   MeSH
• jizva patologie   MeSH
• kosterní svaly krevní zásobení   účinky léků   patologie   MeSH
• králíci MeSH
• kultivační média speciální farmakologie   MeSH
• lidé MeSH
• mezenchymální kmenové buňky * MeSH
• mikro RNA metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• nekróza MeSH
• zadní končetina MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• králíci MeSH
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Srdeční selhání představuje jedno z hlavních chronických onemocnění s vysokou morbiditou a mortalitou. Nejčastější příčinu rozvoje srdečního selhání se sníženou systolickou funkcí představuje ischemická choroba srdeční. Ačkoliv revaskularizace ischemického myokardu vede ke zlepšení kontraktility viabilního myokardu a systolické funkce, žádná z těchto metod nemůže obnovit viabilitu již nekrotické tkáně. V našem přehledu se zabýváme možností využití autologních kmenových buněk kostní dřeně v léčbě pacientů se srdečním selháním na podkladě ischemického poškození myokardu.

Heart failure is a major chronic illness, resulting in high morbidity and mortality. The most frequent cause of heart failure withreduced ejection fraction is coronary artery disease. Although revascularisation of ischaemic myocardium leads to improvementsin contractility of viable myocardium and systolic function, it can not restore the viability of an already necrotic tissue. In our review,we focus on the use of autologous bone marrow stem cells in the treatment of patients with heart failure due to ischaemicmyocardial injury.

• MeSH
• autologní krevní transfuze * metody   trendy   MeSH
• buněčná a tkáňová terapie metody   trendy   MeSH
• hematopoetické kmenové buňky * cytologie   imunologie   účinky léků   MeSH
• ischemická choroba srdeční diagnóza   etiologie   komplikace   MeSH
• kardiomyocyty cytologie   transplantace   účinky léků   MeSH
• lidé MeSH
• metaanalýza jako téma MeSH
• reperfuzní poškození myokardu diagnóza   komplikace   terapie   MeSH
• řízená tkáňová regenerace metody   trendy   MeSH
• srdeční selhání * diagnóza   etiologie   terapie   MeSH
• statistika jako téma MeSH
• systolické srdeční selhání diagnóza   etiologie   terapie   MeSH
• způsoby aplikace léků MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

BACKGROUND Paracrine factors secreted by adipose-derived stem cells can be captured, fractionated, and concentrated to produce therapeutic factor concentrate (TFC). The present study examined whether TFC effects could be enhanced by combining TFC with a biological matrix to provide sustained release of factors in the target region. MATERIAL AND METHODS Unilateral hind limb ischemia was induced in rabbits. Ischemic limbs were injected with either placebo control, TFC, micronized small intestinal submucosa tissue (SIS), or TFC absorbed to SIS. Blood flow in both limbs was assessed with laser Doppler perfusion imaging. Tissues harvested at Day 48 were assessed immunohistochemically for vessel density; in situ hybridization and quantitative real-time PCR were employed to determine miR-126 expression. RESULTS LDP ratios were significantly elevated, compared to placebo control, on day 28 in all treatment groups (p=0.0816, p=0.0543, p=0.0639, for groups 2-4, respectively) and on day 36 in the TFC group (p=0.0866). This effect correlated with capillary density in the SIS and TFC+SIS groups (p=0.0093 and p=0.0054, respectively, compared to placebo). A correlation was observed between miR-126 levels and LDP levels at 48 days in SIS and TFC+SIS groups. CONCLUSIONS A single bolus administration of TFC and SIS had early, transient effects on reperfusion and promotion of ischemia repair. The effects were not additive. We also discovered that TFC modulated miR-126 levels that were expressed in cell types other than endothelial cells. These data suggested that TFC, alone or in combination with SIS, may be a potent therapy for patients with CLI that are at risk of amputation.

• MeSH
• extracelulární matrix metabolismus   MeSH
• ischemie genetika   patologie   terapie   MeSH
• kmenové buňky cytologie   MeSH
• končetiny krevní zásobení   patologie   MeSH
• králíci MeSH
• kůže patologie   MeSH
• laser doppler flowmetrie MeSH
• lidé středního věku MeSH
• lidé MeSH
• mikro RNA genetika   metabolismus   MeSH
• mikropartikule metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• perfuze MeSH
• regulace genové exprese MeSH
• reperfuzní poškození patologie   terapie   MeSH
• střevní sliznice fyziologie   MeSH
• tenké střevo fyziologie   MeSH
• transplantace kmenových buněk * MeSH
• tuková tkáň cytologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Transplantation of adipose-derived stem cells (ADSCs) is an emerging therapeutic option for addressing intractable diseases such as critical limb ischemia (CLI). Evidence suggests that therapeutic effects of ADSCs are primarily mediated through paracrine mechanisms rather than transdifferentiation. These secreted factors can be captured in conditioned medium (CM) and concentrated to prepare a therapeutic factor concentrate (TFC) composed of a cocktail of beneficial growth factors and cytokines that individually and in combination demonstrate disease-modifying effects. The ability of a TFC to promote reperfusion in a rabbit model of CLI was evaluated. A total of 27 adult female rabbits underwent surgery to induce ischemia in the left hindlimb. An additional five rabbits served as sham controls. One week after surgery, the ischemic limbs received intramuscular injections of either (1) placebo (control medium), (2) a low dose of TFC, or (3) a high dose of TFC. Limb perfusion was serially assessed with a Doppler probe. Blood samples were analyzed for growth factors and cytokines. Tissue was harvested postmortem on day 35 and assessed for capillary density by immunohistochemistry. At 1 month after treatment, tissue perfusion in ischemic limbs treated with a high dose of TFC was almost double (p < 0.05) that of the placebo group [58.8 ± 23 relative perfusion units (RPU) vs. 30.7 ± 13.6 RPU; mean ± SD]. This effect was correlated with greater capillary density in the affected tissues and with transiently higher serum levels of the angiogenic and prosurvival factors vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF). The conclusions from this study are that a single bolus administration of TFC demonstrated robust effects for promoting tissue reperfusion in a rabbit model of CLI and that a possible mechanism of revascularization was promotion of angiogenesis by TFC. Results of this study demonstrate that TFC represents a potent therapeutic cocktail for patients with CLI, many of whom are at risk for amputation of the affected limb.

• MeSH
• cytokiny terapeutické užití   MeSH
• fyziologická neovaskularizace účinky léků   MeSH
• hepatocytární růstový faktor metabolismus   MeSH
• injekce intramuskulární MeSH
• ischemie farmakoterapie   MeSH
• králíci MeSH
• kultivační média speciální farmakologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mezenchymální kmenové buňky metabolismus   MeSH
• mezibuněčné signální peptidy a proteiny terapeutické užití   MeSH
• průtoková cytometrie MeSH
• tuková tkáň metabolismus   MeSH
• vaskulární endoteliální růstový faktor A metabolismus   MeSH
• zadní končetina patologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKROUND: Deep skin burn injuries, especially those on the face, hands, feet, genitalia and perineum represent significant therapeutic challenges. Autologous dermo-epidermal skin grafts (DESG) have become standard of care for treating deep burns. Additionally, human autologous thrombin activated autologous platelet concentrate (APC) has gained acceptance in the setting of wounds. While each of these interventions has been independently shown to accelerate healing, the combination of the two has never been evaluated. We hypothesized that the addition of platelets (source of growth factors and inhibitors necessary for tissue repair) to the DESG (source of progenitor cells and of tissue proteases necessary for spatial and temporal control of growth regulators released from platelets) would create the optimal environment for the reciprocal interaction of cells within the healing tissues. METHODS: We used clinical examination (digital photography), standardised scales for evaluating pain and scarring, in combination with blood perfusion (laser Doppler imaging), as well as molecular and laboratory analyses. RESULTS: We show for the first time that the combination of APC and DESG leads to earlier relief of pain, and decreased use of analgesics, antipruritics and orthotic devices. Most importantly, this treatment is associated with earlier discharges from hospital and significant cost savings. CONCLUSIONS: Our findings indicate that DESG engraftment is facilitated by the local addition of platelets and by systemic thrombocytosis. This local interaction leads to the physiological revascularization at 1-3 months. We observed significant elevation of circulating platelets in early stages of engraftment (1-7 days), which normalized over the subsequent 7 and 90 days.

• MeSH
• bolest farmakoterapie   etiologie   MeSH
• hojení ran MeSH
• jizva prevence a kontrola   chirurgie   MeSH
• laser doppler flowmetrie MeSH
• lidé středního věku MeSH
• lidé MeSH
• měření bolesti MeSH
• plazma bohatá na destičky * MeSH
• popálení chirurgie   terapie   ultrasonografie   MeSH
• pruritus farmakoterapie   etiologie   MeSH
• reoperace MeSH
• transplantace kůže škodlivé účinky   metody   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH

• MeSH
• kongresy jako téma MeSH
• mínění MeSH
• regenerativní lékařství * MeSH
• Geografické názvy
• Česká republika MeSH

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• abstrakt z konference MeSH

Cell analysis is an important task in both analytical and medicinal chemistry. A pilot study of capillary electrophoresis analysis of adipose stem cells is presented. The optimum conditions for cell analysis were found in morpholinoethanesulfonic acid (MES)/NaOH buffer (pH 6.0) with an addition of NaCl. Capillary electrophoresis proved to be an effective tool in analysis of adipose stem cells. It could be also used for solving many other tasks in medicinal chemistry.

• MeSH
• diabetická noha MeSH
• elektroforéza kapilární MeSH
• finanční podpora výzkumu jako téma MeSH
• klinické laboratorní techniky MeSH
• kmenové buňky MeSH
• koncentrace vodíkových iontů MeSH
• lidé MeSH
• tuková tkáň MeSH
• Check Tag
• lidé MeSH

A contribution to the description of electrokinetic effects on the pH boundary formed by sodium borate pH 9.5 and sodium phosphate pH 2.5 electrolytes for on-line preconcentration of weak acids is presented in this article. Simulations of electrokinetic injections together with experimental studies using contactless conductivity detection verified that the preconcentration is induced mainly by dissociation changes of analytes on the pH boundary and transient ITP state. Moreover, a study of the addition of organic solvent to the injection electrolyte was performed with impressive results. Subnanomolar LODs of hydroxybenzoic acids were achieved with 80% of methanol in the injection electrolyte which represents more than 70 000-fold preconcentration in comparison with classical CZE method.

• MeSH
• elektrická vodivost MeSH
• elektroforéza kapilární metody   MeSH
• elektrolyty izolace a purifikace   MeSH
• kapilární jevy MeSH
• koncentrace vodíkových iontů MeSH
• kyselina benzoová chemie   izolace a purifikace   MeSH
• kyseliny borité chemie   MeSH
• kyseliny fosforečné chemie   MeSH
• methanol MeSH
• on-line systémy MeSH
• organické látky chemie   MeSH
• osmolární koncentrace MeSH
• oxid křemičitý MeSH
• rozpouštědla chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH