Platelet-rich plasma, especially when combined with a TGF-ß inhibitor promotes proliferation, viability and myogenic differentiation of myoblasts in vitro

Regeneration of skeletal muscle after injury is limited by scar formation, slow healing time and a high recurrence rate. A therapy based on platelet-rich plasma (PRP) has become a promising lead for tendon and ligament injuries in recent years, however concerns have been raised that PRP-derived TGF-β could contribute to fibrotic remodelling in skeletal muscle after injury. Due to the lack of scientific grounds for a PRP -based muscle regeneration therapy, we have designed a study using human myogenic progenitors and evaluated the potential of PRP alone and in combination with decorin (a TGF-β inhibitor), to alter myoblast proliferation, metabolic activity, cytokine profile and expression of myogenic regulatory factors (MRFs). Advanced imaging multicolor single-cell analysis enabled us to create a valuable picture on the ratio of quiescent, activated and terminally committed myoblasts in treated versus control cell populations. Finally high-resolution confocal microscopy validated the potential of PRP and decorin to stimulate the formation of polynucleated myotubules. PRP was shown to down-regulate fibrotic cytokines, increase cell viability and proliferation, enhance the expression of MRFs, and contribute to a significant myogenic shift during differentiation. When combined with decorin further synergistc effects were identified. These results suggest that PRP could not only prevent fibrosis but could also stimulate muscle commitment, especially when combined with a TGF-β inhibitor

Effects of platelet-rich plasma derived growth factors and TGF-beta antagonists on skeletel muscle cells proliferation and differentiation

Uvod Regeneracija skeletne mišice po poškodbi je omejena s tvorbo brazgotinskega tkiva, počasnim celjenjem in relativno visoko verjetnostjo ponovitve poškodbe. Terapija, ki temelji na pripravkih avtolognih trombocitnih preparatov (ang. platelet-rich plasmaPRP) je v zadnjem času postala izjemno popularna, predvsem v primeru poškodb sklepnih vezi in mišičnih tetiv. Za zdravljenje mišičnih poškodb se PRP še ne uporablja, poglavitni pomislek je, da med ostalim vsebuje tudi rastni faktor TGF-β, ki je pomemben dejavnik brazgotinjenja v skeletno-mišičnem tkivu. V raziskavi smo želeli prirediti učinek PRP-ja za uporabo na skeletnem mišičju. Učinek rastnega faktorja TGF-β smo zavirali z njegovim zaviralcem s ciljem izločitve potencialno neželenega učinka v procesu mišične regeneracije. Materiali in metode Zaradi pomanjkanja znanstvenih temeljev o terapiji z avtologno trombocitno plazmo smo zasnovali predklinično raziskavo, v kateri smo uporabili humano, CD56-pozitivno mioblastno celično linijo. Po dodajanju PRP-ja, dekorina (zaviralca TGF-β) in njune kombinacije v gojilni medij smo preučevali stopnjo proliferacije mioblastov, njihove metabolne aktivnosti, profil izražanja fibrotičnih citokinov in ekspresijo miogenih regulatornih faktorjev. Z uporabo slikovne pretočne citometrije smo analizirali razmerje med hibernirajočimi, aktiviranimi in diferencirajočimi mioblasti ter rezultate primerjali med posameznimi skupinami. Dodatno smo opravili še vizualno analizo izražanja dezmina z uporabo visoko ločljivega konfokalnega mikroskopa ter ugotavljali stopnjo formacije več jedrnih miotubulov. Rezultati Ugotovili smo, da pride v mioblastnih celičnih kulturah pod vplivom PRP-ja do značilnega znižanja izražanja fibrotičnih citokinov, povečanja stopnje proliferacije in viabilnosti celic. Obenem pride prav tako do povečane ekspresije miogenih regulatornih faktorjev, kar pripomore k pomembnemu pomiku v miogeni diferenciaciji mioblastov. V kombinaciji PRP-ja z dekorinom smo zaznali pomembne dodatne sinergistične učinke. Razprava Izsledki raziskave dokazujejo, da PRP ne samo potencialno znižuje brazgotinjenje, temveč tudi pripomore k učinkovitejši mišični regeneraciji, še posebej v kombinaciji z zaviralcem TGF-β.Background: Muscle reinjury is often in sports medicine and as such an important target of reinjury prevention. Growth factors from platelet-rich plasma (PRP) with proven effects in tendinous and ligamentous healing have not yet been studied in skeletal muscles mainly due to the concern that exogenous TGF-β application could lead to even greater fibrosis development in an injured muscle. Therefore, only some TGF-β antagonists like decorin have shown their positive role in muscle repair so far. In our study, we investigated the effects of PRP in combination with TGF-β antagonist decorin in skeletal muscle regeneration. Objective: This study proves so far unknown positive effects of platelet-rich plasma in muscle healing, especially when combined with a TGF-β antagonist. Methodology: A novel human myoblast cell culture, defined as CD56 (NCAM)+ developed in our laboratory, was used for evaluation of potential bioactivity of PRP and decorin. The influence on the cells mitochondrial activity, expression of TGF-β was studied in parallel with cell proliferation. Further we have studied the ability of the therapeutic agents to influence the differential cascade of dormant myoblasts towards fully differentiated myotubes by monitoring step wise activation of single nuclear factors like MyoD and Myogenin via multicolor flow cytometry. Results: Our results clearly showed that PrP and decorin treated myoblasts have a significant increase in the mitochondrial activity and in the cell proliferation rate as compared to non-treated control cells. At the same time lower expression of TGF-β and MSTN was evident in PrP treated myoblasts, although PRP itself contains some amount of TGF-β. Conclusion: PrP can be a highly potential therapeutic agent for skeletal muscle regeneration and repair, especially if in combination with a TGF-β antagonist. In such way not only better healing but also lower reinjury rate could potentially be achieved

Zygote body : a new interactive 3-dimensional didactical tool for teaching anatomy

Introduction: As medical education has progressed exponentially over the past decades, there are unanimous concerns regarding the decline in students\u27 knowledge of anatomy. Teaching anatomies is becoming increasingly challenging as students are becoming more technology-dependent and have to gain an understanding of complex anatomical structures in a relatively short time. Zygote Body is a novel interface with potential to facilitate learning of human anatomy. This study investigates its relevancy and appropriateness for use in anatomy education for medical students. Methods: After integration of the web-based application, questionnaires were used to gain feedback from first year medical students (n=73). Results: Data analysis revealed significant improvements in student\u27s comprehension as well as their excitement for such innovations. Discussion: The software succeeds at engagingstudents and suggests that their comprehension of complex 3D structures was improved. However, because of its limitations, this web-based tool cannot be used as a definite tool for learning anatomy. Although multimedia may compliment the learning process, nothing can substitute living anatomy in a safe environment before handling patients

CONCEPT OF MODERN CLINICAL INFORMATION SYSTEM

Mnogo procesnih slabosti pri kliničnem in administracijskem delu zdravnikov v bolnišnicah je predvsem posledica pomakljive standardizacije in optimizacije diagnostičnih, terapevtskih in drugih postopkov v bolnišnicah. Veliko pomanjkljivosti ima skupni imenovalec v ogromnih količinah papirnatih in nesistematiziranih podatkih, česar jedro v vsakodnevni praksi predstavljajo temperaturno-terapevtski listi (TTL) bolnikov. Ti so namreč leglo nesporazumov, napak in velikih stroškovnih posledic. Tradicionalni papirnati klinični sistemi ne zadovoljujejo več potreb informacijske družbe, zato jih je potrebno nadgraditi oz. zamnjeti z modernimi kliničnimi informacijskimi sistemi. Uvedba digitalnega temperaturno-terapevtskega lista (dTTL) v splošnem prinaša: transparentnost medicinske oskrbe in s tem večjo varnost za bolnika, višjo učinkovitost dela v bolnišnici, lažje ter preglednejše vodenje podatkov v procesih zdravljenja in oskrbe ter ne nazadnje tudi znižanje skupnih stroškov. Osnovna ideja kocenpta je v zamenjavi obsotoječega temperaturnega lista v bolnišnicah z novimi, digitalnimi - ob podpori računalniških sistemov. Zdravnik mora informacijsko tehnologijo uporabljati ob fizični prisotnosti bolnika, saj je to edini način res učinkovitega zdravljenja. Programsko je potrebno integrirati že obstoječe podatkovne baze, kot so mednarodna klasifikacija bolezni, register zdravil in druge ter jih med seboj tudi povezati.Many processing difficulties in clinical and administrative work of physicians in hospitals are mainly a result of insufficient standardization and optimization of diagnostic, therapeutic and other procedures in hospitals. A lot of the weaknesses can be derived to a common denominator: a vast amount of paper and non-systemised data that in everyday routine mainly consists of patients’s charts. These are at the core of most misunderstandings, mistakes and resulting high costs. Because the traditional paper-based clinical systems fail to fulfil the expectations of today’s information society, it has to be upgraded or rather exchanged with modern clinical information systems. The introduction of the digital patient’s charts means in general: a more transparent medical care and consequently greater security for the patient, higher efficiency of work in hospitals, easier and more convenient data management in processes of treating a patient and last but not least the reduction of overall expanses. The main idea behind the concept is the exchange of the existing patient’s chart in hospitals with the new, digitalised one – with the help of computer systems. The physician has to use the information technology while the patient is present. This is the only way of making medical care truly efficient. In regard of software, the existing databases like the International Classification of Diseases and the register of medicinal products have to be integrated and also connected

Artificial intelligence in musculoskeletal oncological radiology

Due to the rarity of primary bone tumors, precise radiologic diagnosis often requires an experienced musculoskeletal radiologist. In order to make the diagnosis more precise and to prevent the overlooking of potentially dangerous conditions, artificial intelligence has been continuously incorporated into medical practice in recent decades. This paper reviews some of the most promising systems developed, including those for diagnosis of primary and secondary bone tumors, breast, lung and colon neoplasms