TENOGENIC DIFFERENTIATION PROTOCOL IN XENOGENIC-FREE MEDIA ENHANCES TENDON-RELATED MARKER EXPRESSION IN ADIPOSE-DERIVED STEM CELLS

Tendon injuries are common and current therapies often are unsuccessful. Cell-based therapy using mesenchymal stem cells (MSCs) seems to be the most promising approach to heal tendon. Moreover, providing safe and regulated cell therapy products to patients requires adherence to good manufacturing practices (GMP). Adipose-derived stem cells (n=4) were cultured in 6-well plates coated with type-I collagen in a chemically defined serum-free medium (SF) or a xenogenic-free human pooled platelet lysate medium (hPL). At passage 4, ASCs were induced to tendon lineage for 14 days using 100ng/ml CTGF, 10ng/ml TGFβ3, 50ng/ml BMP12 and 50µg/ml ascorbic acid in the SF (SF-TENO) or in the hPL (hPL-TENO) medium. Cells cultured without any supplements are used as control. Morphological appearance, cell viability and FACS were performed in undifferentiated cells to evaluate the xenogenic-free culture conditions; the gene and protein expression were performed by RT-PCR and immunofluorescence to evaluate to expression of stem cell- and tendon-related markers upon cell differentiation. SF-CTRL and hPL-CTRL showed similar viability and MSC's surface proteins and expressed the stemness markers NANOG, OCT4 and Ki67. Moreover, both SF-TENO and hPL-TENO expressed significant higher levels of SCX, COL1A1, COL3A1, COMP, MMP3 and MMP13 genes already at 3d (p<0.05) respect to CTRLs. Scleraxis and collagen were also detected in both SF-TENO and hPL-TENO at protein level in higher amount than CTRLs. In conclusion, ASCs exposed to CTGF, BMP12, TGFb3 and AA in both serum and xenogenic-free media possess similar tenogenic differentiation ability moving forward the GMP-compliant approaches for the clinical use of ASCs

3D bioprinting for orthopaedic applications: Current advances, challenges and regulatory considerations

In the era of personalised medicine, novel therapeutic approaches raise increasing hopes to address currently unmet medical needs by developing patient-customised treatments. Three-dimensional (3D) bioprinting is rapidly evolving and has the potential to obtain personalised tissue constructs and overcome some limitations of standard tissue engineering approaches. Bioprinting could support a wide range of biomedical applications, such as drug testing, tissue repair or organ transplantation. There is a growing interest for 3D bioprinting in the orthopaedic field, with remarkable scientific and technical advances. However, the full exploitation of 3D bioprinting in medical applications still requires efforts to anticipate the upcoming challenges in translating bioprinted products from bench to bedside. In this review we summarised current trends, advances and challenges in the application of 3D bioprinting for bone and cartilage tissue engineering. Moreover, we provided a detailed analysis of the applicable regulations through the 3D bioprinting process and an overview of available standards covering bioprinting and additive manufacturing

Hand Held 3D Scanning for Cultural Heritage: Experimenting Low Cost Structure Sensor Scan.

In the last years 3D scanning has become an important resource in many fields, in particular it has played a key role in study and preservation of Cultural Heritage. Moreover today, thanks to the miniaturization of electronic components, it has been possible produce a new category of 3D scanners, also known as handheld scanners. Handheld scanners combine a relatively low cost with the advantage of the portability. The aim of this chapter is two-fold: first, a survey about the most recent 3D handheld scanners is presented. As second, a study about the possibility to employ the handheld scanners in the field of Cultural Heritage is conducted. In this investigation, a doorway of the Benedictine Monastery of Catania, has been used as study case for a comparison between stationary Time of Flight scanner, photogrammetry-based 3D reconstruction and handheld scanning. The study is completed by an evaluation of the meshes quality obtained with the three different kinds of technology and a 3D modeling reproduction of the case-study doorway

Electromagnetic form factors in the J/\psi mass region: The case in favor of additional resonances

Using the results of our recent analysis of e^+e^- annihilation, we plot the curves for the diagonal and transition form factors of light hadrons in the time-like region up to the production threshold of an open charm quantum number. The comparison with existing data on the decays of J/\psi into such hadrons shows that some new resonance structures may be present in the mass range between 2 GeVand the J/\psi mass. Searching them may help in a better understanding of the mass spectrum in both the simple and a more sophisticated quark models, and in revealing the details of the three-gluon mechanism of the OZI rule breaking in K\bar K channel.Comment: Formulas are added, typo is corrected, the text is rearranged. Replaced to match the version accepted in Phys Rev

Integrated three-dimensional models for noninvasive monitoring and valorization of the Morgantina silver treasure (Sicily)

The Morgantina silver treasure belonging to the Archaeological Museum of Aidone (Sicily) was involved in a three-dimensional (3-D) survey and diagnostics campaign for monitoring the collection over time in anticipation of their temporary transfer to the Metropolitan Museum of Art in New York for a period of 4 years. Using a multidisciplinary approach, a scientific and methodological protocol based on noninvasive techniques to achieve a complete and integrated knowledge of the precious items and their conservation state, as well as to increase their valorization, has been developed. All acquired data, i.e., 3-D models, ultraviolet fluorescence, x-ray images, and chemical information, will be made available, in an integrated way, within a web-oriented platform, which will present an in-progress tool to deepen existing archaeological knowledge and production technologies and to obtain referenced information of the conservation state before and after moving of the collection from its exposure site

Rho primes in analyzing e+e- annihilation, MARK III, LASS and ARGUS data

The results of an analysis are presented of some recent data on the reactions $e^+e^-\to\pi^+\pi^-\pi^+\pi^-$, $e^+e^-\to\pi^+\pi^-\pi^0\pi^0$ with the subtracted $\omega\pi^0$ events, $e^+e^-\to\omega\pi^0$, $e^+e^-\to\eta \pi^+\pi^-$, $e^+e^-\to\pi^+\pi^-$, $K^-p\to\pi^+\pi^-\Lambda$, the decays $J/\psi\to\pi^+\pi^-\pi^0$, $tau^-\to\nu_tau\pi^+\pi^-\pi^-\pi^0$ $tau^-\to\nu_\tau\omega\pi^-$, upon taking into account both the strong energy dependence of the partial widths on energy and the previously neglected mixing of the $\rho$ type resonances. The above effects are shown to exert an essential influence on the specific values of masses and coupling constants of heavy resonances and hence are necessary to be accounted for in establishing their true nature.Comment: 20 pages, ReVTeX, 9 Postscript figures As compared to hep-ph/9607398, new material concerning the analysis of the ARGUS data on the tau decays into four pion hadronic states is adde

Hypoxia promotes the inflammatory response and stemness features in visceral fat stem cells from obese subjects

Low-grade chronic inflammation is a salient feature of obesity and many associated disorders. This condition frequently occurs in central obesity and is connected to alterations of the visceral adipose tissue (AT) microenvironment. Understanding how obesity is related to inflammation may allow the development of therapeutics aimed at improving metabolic parameters in obese patients. To achieve this aim, we compared the features of 2 subpopulations of adipose-derived stem cells (ASC) isolated from both subcutaneous and visceral AT of obese patients with the features of 2 subpopulations of ASC from the same isolation sites of non-obese individuals. In particular, the behavior of ASC of obese vs non-obese subjects during hypoxia, which occurs in obese AT and is an inducer of the inflammatory response, was evaluated. Obesity deeply influenced ASC from visceral AT (obV-ASC); these cells appeared to exhibit clearly distinguishable morphology and ultrastructure as well as reduced proliferation, clonogenicity and expression of stemness, differentiation and inflammation-related genes. These cells also exhibited a deregulated response to hypoxia, which induced strong tissue-specific NF-kB activation and an NF-kB-mediated increase in inflammatory and fibrogenic responses. Moreover, obV-ASC, which showed a less stem-like phenotype, recovered stemness features after hypoxia. Our findings demonstrated the peculiar behavior of obV-ASC, their influence on the obese visceral AT microenvironment and the therapeutic potential of NF-kB inhibitors. These novel findings suggest that the deregulated hyper-responsiveness to hypoxic stimulus of ASC from visceral AT of obese subjects may contribute via paracrine mechanisms to low-grade chronic inflammation, which has been implicated in obesity-related morbidity

Isolation, characterization and osteogenic differentiation of adipose-derived stem cells : from small to large size animal models

One of the most important issues in orthopaedic surgery is the loss of bone resulting from trauma, infections, tumours or congenital deficiency. In view of the hypothetical future application of mesenchymal stem cells isolated from human adipose tissue in regenerative medicine, we have analysed and characterized adipose-derived stem cells (ASCs) isolated from adipose tissue of rat, rabbit and pig. We have compared their in vitro osteogenic differentiation abilities for exploitation in the repair of critical osteochondral defects in autologous pre-clinical models. The number of pluripotent cells per millilitre of adipose tissue is variable and the yield of rabbit ASCs is lower than that in rat and pig. However, all ASCs populations show both a stable doubling time during culture and a marked clonogenic ability. After exposure to osteogenic stimuli, ASCs from rat, rabbit and pig exhibit a significant increase in the expression of osteogenic markers such as alkaline phosphatase, extracellular calcium deposition, osteocalcin and osteonectin. However, differences have been observed depending on the animal species and/or differentiation period. Rabbit and porcine ASCs have been differentiated on granules of clinical grade hydroxyapatite (HA) towards osteoblast-like cells. These cells grow and adhere to the scaffold, with no inhibitory effect of HA during osteo-differentiation. Such in vitro studies are necessary in order to select suitable pre-clinical models to validate the use of autologous ASCs, alone or in association with proper biomaterials, for the repair of critical bone defects

Donor-matched mesenchymal stem cells from knee infrapatellar and subcutaneous adipose tissue of osteoarthritic donors display differential chondrogenic and osteogenic commitment.

Cell-based therapies have recently been proposed for the treatment of degenerative articular pathologies, such as early osteoarthritis, with an emphasis on autologous mesenchymal stem cells (MSCs), as an alternative to terminally differentiated cells. In this study, we performed a donor-matched comparison between infrapatellar fat pad MSCs (IFP-MSCs) and knee subcutaneous adipose tissue stem cells (ASCs), as appealing candidates for cell-based therapies that are easily accessible during surgery. IFP-MSCs and ASCs were obtained from 25 osteoarthritic patients undergoing total knee replacement and compared for their immunophenotype and differentiative potential. Undifferentiated IFP-MSCs and ASCs displayed the same immunophenotype, typical of MSCs (CD13+/CD29+/CD44+/CD73+/CD90+/CD105+/CD166+/CD31-/CD45-). IFP-MSCs and ASCs showed similar adipogenic potential, though undifferentiated ASCs had higher LEP expression compared to IFP-MSCs (p < 0.01). Higher levels of calcified matrix (p < 0.05) and alkaline phosphatase (p < 0.05) in ASCs highlighted their superior osteogenic commitment compared to IFP-MSCs. Conversely, IFP-MSCs pellets showed greater amounts of glycosaminoglycans (p < 0.01) and superior expression of ACAN (p < 0.001), SOX9, COMP (p < 0.001) and COL2A1 (p < 0.05) compared to ASCs pellets, revealing a superior chondrogenic potential. This was also supported by lower COL10A1 (p < 0.05) and COL1A1 (p < 0.01) expression and lower alkaline phosphatase release (p < 0.05) by IFP-MSCs compared to ASCs. The observed dissimilarities between IFP-MSCs and ASCs show that, despite expressing similar surface markers, MSCs deriving from different fat depots in the same surgical site possess specific features. Furthermore, the in vitro peculiar commitment of IFP-MSCs and ASCs from osteoarthritic donors towards the chondrogenic or osteogenic lineage may suggest a preferential use for cartilage and bone cell-based treatments, respectively

Prospect for Charge Current Neutrino Interactions Measurements at the CERN-PS

Tensions in several phenomenological models grew with experimental results on neutrino/antineutrino oscillations at Short-Baseline (SBL) and with the recent, carefully recomputed, antineutrino fluxes from nuclear reactors. At a refurbished SBL CERN-PS facility an experiment aimed to address the open issues has been proposed [1], based on the technology of imaging in ultra-pure cryogenic Liquid Argon (LAr). Motivated by this scenario a detailed study of the physics case was performed. We tackled specific physics models and we optimized the neutrino beam through a full simulation. Experimental aspects not fully covered by the LAr detection, i.e. the measurements of the lepton charge on event-by-event basis and their energy over a wide range, were also investigated. Indeed the muon leptons from Charged Current (CC) (anti-)neutrino interactions play an important role in disentangling different phenomenological scenarios provided their charge state is determined. Also, the study of muon appearance/disappearance can benefit of the large statistics of CC muon events from the primary neutrino beam. Results of our study are reported in detail in this proposal. We aim to design, construct and install two Spectrometers at "NEAR" and "FAR" sites of the SBL CERN-PS, compatible with the already proposed LAr detectors. Profiting of the large mass of the two Spectrometers their stand-alone performances have also been exploited.Comment: 70 pages, 38 figures. Proposal submitted to SPS-C, CER