Effects of mesenchymal stromal cells versus serum on tendon healing in a controlled experimental trial in an equine model

Abstract Background Mesenchymal stromal cells (MSC) have shown promising results in the treatment of tendinopathy in equine medicine, making this therapeutic approach seem favorable for translation to human medicine. Having demonstrated that MSC engraft within the tendon lesions after local injection in an equine model, we hypothesized that they would improve tendon healing superior to serum injection alone. Methods Quadrilateral tendon lesions were induced in six horses by mechanical tissue disruption combined with collagenase application 3 weeks before treatment. Adipose-derived MSC suspended in serum or serum alone were then injected intralesionally. Clinical examinations, ultrasound and magnetic resonance imaging were performed over 24 weeks. Tendon biopsies for histological assessment were taken from the hindlimbs 3 weeks after treatment. Horses were sacrificed after 24 weeks and forelimb tendons were subjected to macroscopic and histological examination as well as analysis of musculoskeletal marker expression. Results Tendons injected with MSC showed a transient increase in inflammation and lesion size, as indicated by clinical and imaging parameters between week 3 and 6 (p < 0.05). Thereafter, symptoms decreased in both groups and, except that in MSC-treated tendons, mean lesion signal intensity as seen in T2w magnetic resonance imaging and cellularity as seen in the histology (p < 0.05) were lower, no major differences could be found at week 24. Conclusions These data suggest that MSC have influenced the inflammatory reaction in a way not described in tendinopathy studies before. However, at the endpoint of the current study, 24 weeks after treatment, no distinct improvement was observed in MSC-treated tendons compared to the serum-injected controls. Future studies are necessary to elucidate whether and under which conditions MSC are beneficial for tendon healing before translation into human medicine

Zdvojnásobení výsledků polymerázové řetězové reakce v reálném čase.

We added an intercallating dye to probe-based real-time PCR. One set of oligonucleotides had FAM probe while the second one was without. We were able to extract two DNA amplification curves using a single fluorescence channelPřidali jsme interkalující barvivo, aby sonda na bázi real-time PCR. Jedna sada oligonukleotidů mělo FAM sondu, zatímco druhý byl bez. Byli jsme schopni získat dva amplifikace DNA křivky pomocí jednoho fluorescenčním kanál

Superheated droplets for protein thermal stability analyses of GFP, BSA and Taq-polymerase

Here we describe a novel method for the study of protein thermal stability using superheated aqueous samples within virtual reaction chambers.</p

PCR multiplexovani pomoci jedineho fluorescenčního kanálu pro detekovani viru ptačí chřipky pomocí kvantitativní PCR s interkalačním bavivem

Since its invention in 1985 the polymerase chain reaction (PCR) has become a well-established method for amplification and detection of segments of double-stranded DNA. Incorporation of fluorogenic probe or DNA intercalating dyes (such as SYBR Green) into the PCR mixture allowed real-time reaction monitoring and extraction of quantitative information (qPCR). Probes with different excitation spectra enable multiplex qPCR of several DNA segments using multi-channel optical detection systems. Here we show multiplex qPCR using an economical EvaGreen-based system with single optical channel detection. Previously reported non quantitative multiplex realtime PCR techniques based on intercalating dyes were conducted once the PCR is completed by performing melting curve analysis (MCA). The technique presented in this paper is both qualitative and quantitative as it provides information about the presence of multiple DNA strands as well as the number of starting copies in the tested sample. Besides important internal control, multiplex qPCR also allows detecting concentrations of more than one DNA strand within the same sample. Detection of the avian influenza virus H7N9 by PCR is a well established method. Multiplex qPCR greatly enhances its specificity as it is capable of distinguishing both haemagglutinin (HA) and neuraminidase (NA) genes as well as their ratio.Od svého vynálezu v 1985 polymerázová řetězová reakce (PCR) se stal dobře zavedená metoda pro amplifikaci a detekci segmentů dvouřetězcové DNA. Začlenění barviva fluorogenní sondy nebo DNA interkalační (jako SYBR Green) do PCR směsi povoleno Monitorování v reálném čase a extrakce kvantitativních informací (qPCR) reakce. Sondy s různé excitační spektra umožňují multiplexní qPCR experimentu z několika segmentů DNA s využitím multi-kanálové optické systémy detekce. Zde vám ukážeme, multiplex qPCR experimentu pomocí ekonomické EvaGreen bázi Systém s jedním detekcí optickou kanálu. Dříve hlášeny non kvantitativní PCR techniky multiplex v reálném čase založené na interkalační barviva byly provedeny po PCR je dokončen provádění analýzy křivky tání (MCA). Technika uvedené v tomto dokumentu je jak kvalitativní a kvantitativní, neboť poskytuje informace o přítomnosti několika vláken DNA, jakož i počet spuštění kopií v testovaném vzorku. Kromě významnou vnitřní kontroly, multiplexní qPCR také umožňuje detekci koncentrace více než jednoho vlákna DNA ve stejném vzorku. Detekce viru ptačí chřipky H7N9 pomocí PCR je dobře zavedená metoda. Multiplex qPCR výrazně zvyšuje své specifičnosti jako je schopen rozlišit jak hemaglutinin (HA) a neuraminidázou (NA) genů, stejně jako jejich poměr

Polymerase chain reaction in microfluidic devices

Developments of microfluidic-based time/space domain PCR, digital PCR, and isothermal nucleic acid amplification.</p

Droplet-based synthesis of homogeneous magnetic iron oxide nanoparticles

Nanoparticles have gained large interest in a number of different fields due to their unique properties. In medical applications, for example, magnetic nanoparticles can be used for targeting, imaging, magnetically induced thermotherapy, or for any combination of the three. However, it is still a challenge to obtain narrowly dispersed, reproducible particles through a typical lab-scale synthesis when researching these materials. Here, we present a droplet capillary reactor that can be used for the synthesis of magnetic iron oxide nanoparticles. Compared to conventional batch synthesis, the particles synthesized in our droplet reactor have a narrower size distribution and a higher reproducibility. Furthermore, we demonstrate how the particle size can be changed from 5.2 ± 0.9 nm to 11.8 ± 1.7 nm by changing the reaction temperature and droplet residence time in the droplet capillary reactor

Automated droplet reactor for the synthesis of iron oxide/gold core-shell nanoparticles

AbstractCore-shell nanoparticles are promising candidates for theranostic drugs, as they combine different intrinsic properties with a small size and large surface area. However, their controlled synthesis, or the screening and optimization of synthesis conditions are often difficult and labor intensive. Through the precise control over mass and heat transfer, and automatization possibilities, microfluidic devices could be a solution to this problem in a lab scale synthesis. Here, we demonstrate a microfluidic, capillary, droplet reactor for the multi-step synthesis of iron oxide/gold core-shell nanoparticles. Through the integration of a transmission measurement at the outlet of the reactor, synthesis results can be monitored in a real-time manner. This allowed for the implementation of an optimization algorithm. Starting from three separate initial guesses, the algorithm converged to the same synthesis conditions in less than 30 minutes for each initial guess. These conditions resulted in diameter for the iron oxide core of 5.8 ± 1.4 nm, a thickness for the gold shell of 3.5 ± 0.6 nm, and a total diameter of the core-shell particles of 13.1 ± 2.5 nm. Finally, applications of the iron oxide/gold core-shell nanoparticles were demonstrated for Surface Enhanced Raman Spectroscopy (SERS), photothermal therapy, and magnetic resonance imaging (MRI).</jats:p