Bone Marrow Mesenchymal Stem Cells Expanded Inside the Nichoid Micro-Scaffold: a Focus on Anti-Inflammatory Response

Purpose Mesenchymal stem cells (MSCs) represent a promising source for stem cell therapies in numerous diseases, including pediatric respiratory system diseases. Characterized by low immunogenicity, high anti-inflammatory, and immunoregulatory features, MSCs demonstrated an excellent therapeutic profile in numerous in vitro and preclinical models. MSCs reside in a specialized physiologic microenvironment, characterized by a unique combination of biophysical, biochemical, and cellular properties. The exploitation of the 3D micro-scaffold Nichoid, which simulates the native niche, enhanced the anti-inflammatory potential of stem cells through mechanical stimulation only, overcoming the limitation of biochemical and xenogenic growth factors application.Materials and Methods In this work, we expanded pediatric bone marrow MSCs (BM-MSCs) inside the Nichoid and performed a complete cellular characterization with different approaches including viability assays, immunofluorescence analyses, RNA sequencing, and gene expression analysis.Results We demonstrated that BM-MSCs inside the scaffold remain in a stem cell quiescent state mimicking the condition of the in vivo environment. Moreover, the gene expression profile of these cells shows a significant up-regulation of genes involved in immune response when compared with the flat control.Conclusion The significant changes in the expression profile of anti-inflammatory genes could potentiate the therapeutic effect of BM-MSCs, encouraging the possible clinical translation for the treatment of pediatric congenital and acquired pulmonary disorders, including post-COVID lung manifestations

L'IMPLEMENTAZIONE DI LINEE GUIDA DI ANTIBIOTICOPROFILASSI. L'ESPERIENZA DELL' AO ISTITUTO ORTOPEDICO G.PINI

OBIETTIVI: Le infezioni del sito chirurgico (ISC) rappresentano una complicanza impegnativa sia dal punto di vista della qualit\ue0 delle cure sia dell\u2019impatto economico. La profilassi antibiotica rappresenta una pratica preventiva di comprovata efficacia testimoniata da linee guida (LG) che, tuttavia, vengono disattese nella pratica quotidiana. Tale condizione \ue8 stata verificata anche presso l\u2019AO Gaetano Pini a seguito di uno studio di sorveglianza delle ISC in interventi di artroprotesi, promosso dal gruppo GISIO-SItI. Alla luce di quanto riscontrato si \ue8 deciso di: - revisionare le LG di antibioticoprofilassi, - condividerle in azienda, - monitorarne l'applicazione. METODI: \uc8 stato istituito un gruppo multidisciplinare costituito da infettivologi, ortopedici, infermiere addetto al controllo delle infezioni, medici della direzione sanitaria, che ha revisionato le LG aziendali. Il gruppo di lavoro ha ritenuto opportuno organizzare degli incontri formativi per la diffusione delle LG revisionate e di predisporre una checklist ad hoc per il monitoraggio dell'applicazione delle stesse alla luce della considerazione che gli interventi informativi e formativi rappresentano un ottimo strumento per l'implementazione e condivisione delle LG e sono in grado di portare a miglioramenti significativi nella pratica clinica. RISULTATI: Le LG revisionate sono state presentate ai medici ortopedici e ai coordinatori infermieristici nel corso di due eventi formativi. In seguito sono stati organizzati incontri nelle singole UUOO. Durante gli incontri sono stati presentati i risultati dello studio di sorveglianza delle ISC in interventi di artroprotesi e sono state esposte le motivazioni per cui gli schemi di profilassi adottati non sono stati giudicati aderenti alle LG aziendali. \uc8 stato sottolineato che la criticit\ue0 pi\uf9 evidente e sulla quale in letteratura c'\ue8 consenso unanime \ue8 relativa alla durata della profilassi oltre le 24 ore, confermata da una seconda indagine condotta in ciascuna unit\ue0 operativa. Inoltre sono state esposte le modalit\ue0 del processo di revisione delle LG aziendali e ne \ue8 stata consegnata copia anche in formato tascabile, rimarcando come tale documento rappresenti una linea di indirizzo e pertanto sia importante motivare in cartella clinica la scelta di una profilassi diversa da quella indicata. CONCLUSIONI: I percorsi di consolidamento dei comportamenti vanno monitorati e valutati periodicamente. Nei prossimi mesi verr\ue0 verificata l'applicazione dalle LG (scelta corretta della molecola, timing di somministrazione, durata della profilassi) utilizzando la checklist predisposta a tale scopo

Efficacia analgesica di ropivacaina e bupivacaina, dopo chirurgia ortopedica nel cane, additivate all'anestesia generale mediante blocchi loco-regionali degli arti con localizzatore neurale : 159 casi

Aim of this study was to evaluate the analgesic effect of ropivacaine(RPV) and bupivacaine (BPV) used for regional limb blockade, in anaesthetised dogs undergoing orthopaedic procedures localised distally to the scapula or hip. Loco-regional limb blockade was obtained in each case by using nerve locator. A score pain, calculated with a multiparametric pain scale by a random blind operator, was given to every subject. Analgesic effects achieved by using RPV or BPV showed to be equivalent, without statistically significant differences. In this case, since the higher cardiotoxic effect induced by BPV in dogs, RPV seems to be recommended for post-operative analgesia, after orthopaedic surgery by using single nerve blockade

Phototoxicity induced in living HeLa cells by focused femtosecond laser pulses: a data-driven approach

Nonlinear optical microscopy is a powerful label-free imaging technology, providing biochemical and structural information in living cells and tissues. A possible drawback is photodamage induced by high-power ultrashort laser pulses. Here we present an experimental study on thousands of HeLa cells, to characterize the damage induced by focused femtosecond near-infrared laser pulses as a function of laser power, scanning speed and exposure time, in both wide-field and point-scanning illumination configurations. Our data-driven approach offers an interpretation of the underlying damage mechanisms and provides a predictive model that estimates its probability and extension and a safety limit for the working conditions in nonlinear optical microscopy. In particular, we demonstrate that cells can withstand high temperatures for a short amount of time, while they die if exposed for longer times to mild temperatures. It is thus better to illuminate the samples with high irradiances: thanks to the nonlinear imaging mechanism, much stronger signals will be generated, enabling fast imaging and thus avoiding sample photodamage

Imaging intracellular viscosity by a new molecular rotor suitable for phasor analysis of fluorescence lifetime

The arsenal of fluorescent probes tailored to functional imaging of cells is rapidly growing and benefits from recent developments in imaging strategies. Here, we present a new molecular rotor, which displays strong absorption in the green region of the spectrum, very little solvatochromism, and strong emission sensitivity to local viscosity. The emission increase is paralleled by an increase in emission lifetime. Owing to its concentration-independent nature, fluorescence lifetime is particularly suitable to image environmental properties, such as viscosity, at the intracellular level. Accordingly, we demonstrate that intracellular viscosity measurements can be efficiently carried out by lifetime imaging with our probe and phasor analysis, an efficient method for measuring lifetime-related properties (e.g., bionalyte concentration or local physicochemical features) in living cells. Notably, we show that it is possible to monitor the partition of our probe into different intracellular regions/organelles and to follow mitochondrial de-energization upon oxidative stress