Completely decellularized matrices are promising in advanced tissue engineering, because they lose most of their immunogenicity and preserve similar mechanical properties, composition and 3D structure as the native extracellular matrix. Therefore, we intend to use decellularized pericardium and blood vessels for creating vascular patches and small-diameter vascular grafts, respectively. The decellularization will be carried out in a fully automated system under defined conditions, giving reproducible results and producing non-toxic matrices suitable for further recellularization with endothelial cells (EC), which will be supported by coating the matrices with heparin and hyaluronic acid. This “simulated glycocalyx” is also supposed to have antithrombogenic effects. The recellularization will be first tested in vitro in static and particularly dynamic cell culture systems, where the EC or their precursors will be captured from a circulating culture medium or heparinized blood. The most promising samples will be then selected for preclinical tests in vivo in a rabbit model.

Kompletně decelularizované matrice jsou vysoce perspektivní v moderním tkáňovém inženýrství, a to pro jejich výrazně sníženou imunogenní aktivitu a obdobné mechanické vlastnosti, složení a 3D strukturu jako u přirozené extracelulární matrix. V tomto projektu proto hodláme využít decelularizovaný perikard a krevní cévy pro konstrukci cévních záplat a cévních náhrad o malém průměru. Kompletní decelularizace bude dosaženo v plně automatizovaném systému za definovaných podmínek, což umožní reprodukovatelnost výsledků a vytvoření netoxických matricí vhodných pro recelularizaci endotelovými buňkami (EB), která bude dále podpořena pokrytím decelularizovaných matricí heparinem a kyselinou hyaluronovou. Očekáváme, že tato „simulovaná glykokalyx“ bude mít rovněž antitrombogenní účinky. Recelularizace bude nejprve sledována in vitro ve statických a především v dynamických kultivačních systémech, kde budou EB a jejich prekursory vychytávány z cirkulujícího kultivačního média či heparinizované králičí krve. Nejslibnější vzorky budou vybrány pro preklinické testy in vivo na modelu králíka.

• Klíčová slova
• kyselina hyaluronová, hyaluronic acid, perikard, krevní cévy, decelularizace, recelularizace, heparin, endotelové buňky, cévní záplaty, cévní náhrady, pericardium, blood vessels, decellularization, recellularization, heparin, endothelial cells, vascular patches, vascular grafts, králik in-vivo, rabbit in-vivo,

• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Perinatal derivatives are drawing growing interest among the scientific community as an unrestricted source of multipotent stromal cells, stem cells, cellular soluble mediators, and biological matrices. They are useful for the treatment of diseases that currently have limited or no effective therapeutic options by means of developing regenerative approaches. In this paper, to generate a complete view of the state of the art, a comprehensive 10-years compilation of clinical-trial data with the common denominator of PnD usage has been discussed, including commercialized products. A set of criteria was delineated to challenge the 10-years compilation of clinical trials data. We focused our attention on several aspects including, but not limited to, treated disorders, minimal or substantial manipulation, route of administration, dosage, and frequency of application. Interestingly, a clear correlation of PnD products was observed within conditions, way of administration or dosage, suggesting there is a consolidated clinical practice approach for the use of PnD in medicine. No regulatory aspects could be read from the database since this information is not mandatory for registration. The database will be publicly available for consultation. In summary, the main aims of this position paper are to show possibilities for clinical application of PnD and propose an approach for clinical trial preparation and registration in a uniform and standardized way. For this purpose, a questionnaire was created compiling different sections that are relevant when starting a new clinical trial using PnD. More importantly, we want to bring the attention of the medical community to the perinatal products as a consolidated and efficient alternative for their use as a new standard of care in the clinical practice.

• Publikační typ
• časopisecké články MeSH

PURPOSE OF THE STUDY Population aging is connected with an increased incidence of chronic diseases. A common related problem is chronic skin ulcers, which, while not life-threatening, can significantly decrease the quality of the patient's life. The present study aims to evaluate new materials and methods to improve and accelerate the treatment of leg ulcers. MATERIAL AND METHODS Twenty-five patients with chronic ulcers treated using autologous growth factors applied on a nanofiber carrier were included in the cohort. The control group consisted of 15 patients treated using standard moist wound therapy. The surface area of the ulcer was measured on the 0th, 14th, 28th, 56th, 84th, 112th, 140th, 140th, and 168th day of treatment. Ulcer depth was measured on the 0th, 5th, 28th, 84th, and 168th day of treatment. Results were statistically processed and evaluated. RESULTS During the study, the defect area decreased in both the control and experimental group. Statistically significantly better results were observed in the experimental group relative to the progress of ulcer depth. The experimental group also had more healed ulcers. DISCUSSION Moistness is necessary for chronic wounds to heal; it is needed to ensure optimal cell growth, angiogenesis, and fibrinolysis. Wounds can be treated using non-active dressings with high absorption qualities; however, these do not guarantee optimal conditions for healing, or wounds can be treated with an interactive dressing that interacts with the wound surface. The third option for treatment is the use of bioactive materials that adhere to the wound and participate directly in the individual stages of healing. CONCLUSIONS The study found that autologous growth factors had statistically significant effects on the treatment of chronic ulcers. The authors believe that this method can accelerate the healing of primary post-injury or secondary postoperative wounds of lower leg soft tissues. Key words: trophic ulcer, autologous growth factors, microangiopathy, polyneuropathy, diabetes mellitus.

• MeSH
• lidé MeSH
• nanovlákna * terapeutické užití   MeSH
• pilotní projekty MeSH
• plazma bohatá na destičky * MeSH
• vřed MeSH
• vředy dolních končetin * terapie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

• Publikační typ
• abstrakt z konference MeSH

BACKGROUND: Cardiovascular surgery is confronted by a lack of suitable materials for patch repair. Acellular animal tissues serve as an abundant source of promising biomaterials. The aim of our study was to explore the bio-integration of decellularized or recellularized pericardial matrices in vivo. METHODS: Porcine (allograft) and ovine (heterograft, xenograft) pericardia were decellularized using 1% sodium dodecyl sulfate ((1) Allo-decel and (2) Xeno-decel). We used two cell types for pressure-stimulated recellularization in a bioreactor: autologous adipose tissue-derived stromal cells (ASCs) isolated from subcutaneous fat of pigs ((3) Allo-ASC and (4) Xeno-ASC) and allogeneic Wharton's jelly mesenchymal stem cells (WJCs) ((5) Allo-WJC and (6) Xeno-WJC). These six experimental patches were implanted in porcine carotid arteries for one month. For comparison, we also implanted six types of control patches, namely, arterial or venous autografts, expanded polytetrafluoroethylene (ePTFE Propaten® Gore®), polyethylene terephthalate (PET Vascutek®), chemically stabilized bovine pericardium (XenoSure®), and detoxified porcine pericardium (BioIntegral® NoReact®). The grafts were evaluated through the use of flowmetry, angiography, and histological examination. RESULTS: All grafts were well-integrated and patent with no signs of thrombosis, stenosis, or aneurysm. A histological analysis revealed that the arterial autograft resembled a native artery. All other control and experimental patches developed neo-adventitial inflammation (NAI) and neo-intimal hyperplasia (NIH), and the endothelial lining was present. NAI and NIH were most prominent on XenoSure® and Xeno-decel and least prominent on NoReact®. In xenografts, the degree of NIH developed in the following order: Xeno-decel > Xeno-ASC > Xeno-WJC. NAI and patch resorption increased in Allo-ASC and Xeno-ASC and decreased in Allo-WJC and Xeno-WJC. CONCLUSIONS: In our setting, pre-implant seeding with ASC or WJC had a modest impact on vascular patch remodeling. However, ASC increased the neo-adventitial inflammatory reaction and patch resorption, suggesting accelerated remodeling. WJC mitigated this response, as well as neo-intimal hyperplasia on xenografts, suggesting immunomodulatory properties.

• MeSH
• allogeneické buňky MeSH
• arteriae carotides MeSH
• cévní protézy MeSH
• hyperplazie MeSH
• lidé MeSH
• ovce MeSH
• perikard MeSH
• prasata MeSH
• remodelace cév * MeSH
• skot MeSH
• tkáňové inženýrství MeSH
• transplantace hematopoetických kmenových buněk * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

An inability of the human body to heal acute wounds under certain conditions results in the formation of chronic ulcers. Chronic wounds not only cause significant pain and discomfort for patients but also serve as an entry for microorganisms into the human body, which can result in serious life-threatening problems and become a significant burden for the patients and society. The current work present results of a multicentre prospective observational study demonstrating the use of a lyophilized amniotic membrane (AM) in the treatment of chronic wounds (various etiologies). Lyophilized AM produced under the commercial brand Amnioderm® was used as an allograft material for therapy of chronic wounds, in addition to chronic ulcer standard-of-care (SoC) protocols. The duration of wounds considered for the application of AM ranged between 2 months and 11 years. In total, 16 patients were enrolled to the study, of which eight were completely healed, six demonstrated a significantly reduced ulcer size, and two did not respond to the AM therapy. The current study unambiguously demonstrates an effective alternative to the standard of chronic wound care and confirms a significant effect of the AM application for chronic wound management as a new SoC.

• Publikační typ
• kazuistiky MeSH

Background: Toxic epidermal necrolysis (TEN) is a rare life-threatening disease that mainly affects the skin and mucous membranes, resulting from a toxic delayed-type hypersensitivity (DTH) reaction (type IV reaction) to the presence of foreign antigens such as drugs. The clinical symptoms are caused by pathophysiological processes leading to massive apoptosis of keratinocytes in the dermo-epidermal junction. This results in the formation of a bulla and subsequent separation of the entire epidermis with the exposure of the dermis. The current approach in the local therapy of TEN prefers the use of biological dressings, which helps provide several critical requirements for defect healing; in particular, it helps in the acceleration of the spontaneous wound closure (re-epithelialization) of the skin defect and the reduction of the risk of development of various complications and infections, such as the risk of pathological scar maturation. This paper is a case report of the use of a lyophilized amniotic membrane (AM) for accelerating wound healing in a patient with TEN. Case Presentation: We report a case of an 8-year-old girl transferred to our center with a histologically confirmed diagnosis of TEN. Despite the application of immunosuppressive therapy consisting of corticosteroids and intravenous immunoglobulins, we have observed disease progression and exfoliation of up to 60% of the total body surface area (TBSA). In the facial area, which is cosmetically privileged, we decided to use the lyophilized amniotic membrane (Amnioderm®) to cover up approximately 2% of the TBSA. Within 2 days after the application, we observed accelerated reepithelialisation, with rapid wound closure. We have not observed any side effects nor infections during the subsequent phases of wound healing. Skin defects in non-facial areas of the body were treated with synthetic dressings. When compared to the areas covered with the lyophilized AM, the healing process was prolonged. Conclusions: To our knowledge, this is the first case study using a lyophilized amniotic membrane in the treatment of a patient with TEN. The AM application in the cosmetically-privileged area (face), proved to be very efficient in the treatment of TEN patients. The use of this allogeneic material demonstrated excellent biocompatibility and caused a unique acceleration of epithelialization and wound healing, yielding also excellent long-term results. The current study opens broad possibilities for clinical application of the used material, the improvement of current therapy of patients with TEN and better outcomes and recovery of patients.

• Publikační typ
• kazuistiky MeSH

A promising therapeutic strategy for amyotrophic lateral sclerosis (ALS) treatment is stem cell therapy. Neural progenitors derived from induced pluripotent cells (NP-iPS) might rescue or replace dying motoneurons (MNs). However, the mechanisms responsible for the beneficial effect are not fully understood. The aim here was to investigate the mechanism by studying the effect of intraspinally injected NP-iPS into asymptomatic and early symptomatic superoxide dismutase (SOD)1G93A transgenic rats. Prior to transplantation, NP-iPS were characterized in vitro for their ability to differentiate into a neuronal phenotype. Motor functions were tested in all animals, and the tissue was analyzed by immunohistochemistry, qPCR, and Western blot. NP-iPS transplantation significantly preserved MNs, slowed disease progression, and extended the survival of all treated animals. The dysregulation of spinal chondroitin sulfate proteoglycans was observed in SOD1G93A rats at the terminal stage. NP-iPS application led to normalized host genes expression (versican, has-1, tenascin-R, ngf, igf-1, bdnf, bax, bcl-2, and casp-3) and the protection of perineuronal nets around the preserved MNs. In the host spinal cord, transplanted cells remained as progenitors, many in contact with MNs, but they did not differentiate. The findings suggest that NP-iPS demonstrate neuroprotective properties by regulating local gene expression and regulate plasticity by modulating the central nervous system (CNS) extracellular matrix such as perineuronal nets (PNNs).

• MeSH
• amyotrofická laterální skleróza terapie   MeSH
• indukované pluripotentní kmenové buňky cytologie   MeSH
• krysa rodu rattus MeSH
• kultivované buňky MeSH
• lidé MeSH
• nervové kmenové buňky cytologie   metabolismus   transplantace   MeSH
• neuroplasticita * MeSH
• neurotrofní faktory genetika   metabolismus   MeSH
• periferní nervy fyziologie   MeSH
• potkani Sprague-Dawley MeSH
• proteiny regulující apoptózu genetika   metabolismus   MeSH
• regenerace nervu MeSH
• tenascin genetika   metabolismus   MeSH
• transplantace kmenových buněk metody   MeSH
• versikany genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Neurodegenerative disorders present a broad group of neurological diseases and remain one of the greatest challenges and burdens to mankind. Maladies like amyotrophic lateral sclerosis, Alzheimer's disease, stroke or spinal cord injury commonly features astroglia involvement (astrogliosis) with signs of inflammation. Regenerative, paracrine and immunomodulatory properties of human mesenchymal stromal cells (hMSCs) could target the above components, thus opening new therapeutic possibilities for regenerative medicine. A special interest should be given to hMSCs derived from the umbilical cord (UC) tissue, due to their origin, properties and lack of ethical paradigms. The aim of this study was to establish standard operating and scale-up good manufacturing practice (GMP) protocols of UC-hMSCs isolation, characterization, expansion and comparison of cells' properties when harvested on T-flasks versus using a large-scale bioreactor system. Human UC-hMSCs, isolated by tissue explant culture technique from Wharton's jelly, were harvested after reaching 75% confluence and cultured using tissue culture flasks. Obtained UC-hMSCs prior/after the cryopreservation and after harvesting in a bioreactor, were fully characterized for "mesenchymness" immunomodulatory, tumorigenicity and genetic stability, senescence and cell-doubling properties, as well as gene expression features. Our study demonstrates an efficient and simple technique for large scale UC-hMSCs expansion. Harvesting of UC-hMSCs' using classic and large scale methods did not alter UC-hMSCs' senescence, genetic stability or in vitro tumorigenicity features. We observed comparable growth and immunomodulatory capacities of fresh, frozen and expanded UC-hMSCs. We found no difference in the ability to differentiate toward adipogenic, osteogenic and chondrogenic lineages between classic and large scale UC-hMSCs expansion methods. Both, methods enabled derivation of genetically stabile cells with typical mesenchymal features. Interestingly, we found significantly increased mRNA expression levels of neural growth factor (NGF) and downregulated insulin growth factor (IGF) in UC-hMSCs cultured in bioreactor, while IL4, IL6, IL8, TGFb and VEGF expression levels remained at the similar levels. A culturing of UC-hMSCs using a large-scale automated closed bioreactor expansion system under the GMP conditions does not alter basic "mesenchymal" features and quality of the cells. Our study has been designed to pave a road toward translation of basic research data known about human UC-MSCs for the future clinical testing in patients with neurological and immunocompromised disorders. An industrial manufacturing of UC-hMSCs next will undergo regulatory approval following advanced therapy medicinal products (ATMP) criteria prior to clinical application and approval to be used in patients.

• MeSH
• bioreaktory * MeSH
• kultivované buňky MeSH
• lidé MeSH
• mezenchymální kmenové buňky fyziologie   MeSH
• nemoci nervového systému patologie   terapie   MeSH
• proliferace buněk fyziologie   MeSH
• pupečník cytologie   fyziologie   transplantace   MeSH
• transplantace mezenchymálních kmenových buněk * trendy   MeSH
• Whartonův rosol cytologie   fyziologie   transplantace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Cíl: Aterosklerotické postižení karotických tepen patří k nejčastějším příčinám ischemické CMP. Jelikož je studium progrese a vývoje nestability aterosklerotických plátů in vivo velmi limitováno, lze k objasnění těchto procesů využít data ze studií in vitro. Z těchto důvodů autoři sestrojili hemodynamický model s možností vložení aterosklerotického plátu vyjmutého během karotické endarterektomie pro studium hemodynamiky v oblasti karotické bifurkace. Cílem studie bylo zjistit, zda lze v hemodynamickém modelu nasimulovat průtokové parametry v oblasti stenózy karotické bifurkace srovnatelné se stavem in vivo před provedením karotické endarterektomie. Metody: Do studie bylo zařazeno 13 pacientů se stenózou karotidy ≥ 50 % indikovaných ke karotické endaterektomii. Během endarterektomie byly vyjmuty v celku aterosklerotické pláty z karotické tepny a vloženy do hemodynamického modelu. Výsledky: Průměrný rozdíl v naměřené průtokové rychlosti v oblasti stenózy in vivo před karotickou endarterektomií a in vitro v hemodynamickém modelu po vložení vyjmutého aterosklerotického plátu byl 18,9 cm/s u maximální systolické rychlosti, což odpovídá odchylce 7,0 %, a 8,2 cm/s u konečné diastolické rychlosti odpovídající odchylce 11,1 %. Závěr: Studie potvrdila funkčnost hemodynamického modelu a jeho možné využití při studiu hemodynamických změn v oblasti karotické stenózy.

Aim: Atherosclerotic carotid artery disease is one of the most common causes of ischemic stroke. As the study of the progression and development of instability of atherosclerotic plaques in vivo is limited, data from in vitro studies can be used to clarify these processes. For these reasons, the authors constructed a hemodynamic model with the possibility of inserting the atherosclerotic plaque removed during carotid endarterectomy for the study of hemodynamics in the carotid bifurcation. The aim of the study was to determine whether it is possible to simulate flow parameters in the area of carotid bifurcation stenosis in the hemodynamic model comparable to the in vivo state before performing carotid endarterectomy. Intact atherosclerotic plaque was removed from the carotid artery during endarterectomy and inserted into a hemodynamic model. Methods: The study included 13 patients with carotid stenosis ≥ 50% indicated for carotid endarterectomy. The atherosclerotic plaques were removed from the carotid artery during carotid endarterectomy and inserted into the hemodynamic model. Results: The mean differences in the measured maximum and end-diastolic velocities in the area of stenosis in vivo before carotid endarterectomy and in vitro in the hemodynamic model after insertion of the removed atherosclerotic plaque were 18.9 cm/s (7.0%) and 8.2 cm/s (11.1%), respectively. Conclusion: The study confirmed the functionality of the hemodynamic model and its possible use for studying the hemodynamic changes in carotid stenosis area.

• MeSH
• arteriae carotides * patofyziologie   MeSH
• aterosklerotický plát patofyziologie   MeSH
• ateroskleróza MeSH
• hemodynamika * MeSH
• karotická endarterektomie MeSH
• klinická studie jako téma MeSH
• lidé MeSH
• stenóza arteria carotis patofyziologie   MeSH
• teoretické modely MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH