Recent Advances on Surface-Modified GBM Targeted Nanoparticles: Targeting Strategies and Surface Characterization

Glioblastoma multiforme (GBM) is the most common malignant brain tumor, associated with low long-term survival. Nanoparticles (NPs) developed against GBM are a promising strategy to improve current therapies, by enhancing the brain delivery of active molecules and reducing off-target effects. In particular, NPs hold high potential for the targeted delivery of chemotherapeutics both across the blood-brain barrier (BBB) and specifically to GBM cell receptors, pathways, or the tumor microenvironment (TME). In this review, the most recent strategies to deliver drugs to GBM are explored. The main focus is on how surface functionalizations are essential for BBB crossing and for tumor specific targeting. We give a critical analysis of the various ligand-based approaches that have been used to target specific cancer cell receptors and the TME, or to interfere with the signaling pathways of GBM. Despite the increasing application of NPs in the clinical setting, new methods for ligand and surface characterization are needed to optimize the synthesis, as well as to predict their in vivo behavior. An expert opinion is given on the future of this research and what is still missing to create and characterize a functional NP system for improved GBM targeting

Raman Spectroscopy Characterization of Multi-Functionalized Liposomes as Drug-Delivery Systems for Neurological Disorders

The characterization of nanoparticle-based drug-delivery systems represents a crucial step in achieving a comprehensive overview of their physical, chemical, and biological features and evaluating their efficacy and safety in biological systems. We propose Raman Spectroscopy (RS) for the characterization of liposomes (LPs) to be tested for the control of neuroinflammation and microglial dysfunctions in Glioblastoma multiforme and Alzheimer’s disease. Drug-loaded LPs were functionalized to cross the blood–brain barrier and to guarantee localized and controlled drug release. The Raman spectra of each LP component were used to evaluate their contribution in the LP Raman fingerprint. Raman data analysis made it possible to statistically discriminate LPs with different functionalization patterns, showing that each molecular component has an influence in the Raman spectrum of the final LP formulation. Moreover, CLS analysis on Raman data revealed a good level of synthetic reproducibility of the formulations and confirmed their stability within one month from their synthesis, demonstrating the ability of the technique to evaluate the efficacy of LP synthesis using small amount of sample. RS represents a valuable tool for a fast, sensitive and label free biochemical characterization of LPs that could be used for quality control of nanoparticle-based therapeutics

Mindfulness based therapy on group of rehabilitative ABI (Acquired Brain Injury) patients: a pilot study

Introduction Mindfulness means maintaining a moment-by-moment awareness of our thoughts, feelings, bodily sensations, and surrounding environment. In the last twenty years the diffusion of Mindfulness-based intervention for rehabilitation purpose in in- and out-patient has been rapidly increasing and the research interest growing up attempting to give scientific rigor in this area. The use of this method is considering a variety of medical and psychiatric conditions (e.g chronic pain, depression etc) as reported in Didonna (2012), futhermore the group intervention has been recently proposed to obtain maximum therapeutic effect (Grossman et al 2004). So far there have been few studies describing Mindfulness-based approach for neurological and orthopedic patients (Azulay et al 2013), especially those under Rehabilitation Units assistance . These kinds of patients are frequently showing anxiety, depression and attention dysfunction as well as problem of adaptation to new situations such as hospitalization or outcome acceptance (Azulay et al 2013) As recently reported by Bédard and colleagues (2012), the symptoms above may benefit from Mindfulness-based therapy. Aim The aim of the present pilot study is to verify if the Mindfulness-based group Therapy could be usefully introduced for rehabilitative purpose with acquired brain injuries patients (e.g stroke, hemorrhagic or traumatic brain injury patients) attending the Rehabilitation Units of the Hospital of Parma. Methods Design: Observational Prospective Study Subjects: 10 individuals (5 males and 5 females) with mild to moderate brain injuries (6 patients) and stroke (4 patients), at least 1 year post-injury were recruited. The sample median age was 39,5 (SD 11,8) years while 8 to 17 years in education. Each member of the reported sample had at least one of the symptoms related to the Cognitive (attention deficits and executive dysfunctions), Physical (headache and fatigue), Psychological (anxiety, depression and post-traumatic syndrome) or Emotional-behavioural domain. Inclusion Criteria: 1) Age: 18-60; 2)education ≥ 8 years; 3) individual on-going rehabilitation program, followed by the Rehabilitation Unit of the Parma Hospital, for the presence of neuro-motor and/or neuro-psychological disabilities; 4) First ABI episode ; 5) at least 3 months from acute ABI episode; 6) Stabilization of clinical pictures; 7) Absence of significative deficits in language production and comprehension; 8) Absence of significative deficits in mnemonic functions; 9) Adequate motivation and compliance Program of intervention: group session, 90 minutes lasts, for 8 weeks. each one relied on breathing exercises, according to Trascendental Meditation Technique (Benson,1976) and was preceded by an adapted postural exercises session to improve breathing observation and environmental and body awareness. Main Outcome Measures: Mindfulness Attention Awareness Scale (MAAS), Hospital Anxiety and Depression Scale (HADS); Modified-Neurobehavioral Rating Scale, SF 12 Standard Italian Version 1.0. Results Patients have generally improved their intentional attention, a better attitude toward living in the present moment and focusing attention on required task. The pre and post Mindfulness-based therapy influence, reported into MAAS scale data, graphically represented in figure 1, showed statistically significant results (X p<0.05) in items 3 (“I find it difficult to stay focused on what is happening at present”), 10 (“I carry out work and tasks automatically without realizing what I’m doing”.) and 13 (I’m concerned about past and future). Summing up, the results indicate the effectiveness of Mindfulness-based group Therapy for rehabilitative ABI patients. This procedure may provide an alternative approach to integrate classical rehabilitation therapy in reducing post-traumatic symptoms. No relevant statistical results are been reported about the other administered scales. Conclusions The Mindfulness-based group Therapy provide a useful instrument for ABI patients, to add to the individual rehabilitation program. The particular responsiveness recorded into “attention” and “awareness” domain could suggest an increasing patient’s compliance also during the classical rehabilitation activities (e.g. physiotherapy, speech therapy etc.)

Multiplexing Biosensor for the Detection of Extracellular Vesicles as Biomarkers of Tissue Damage and Recovery after Ischemic Stroke

The inflammatory, reparative and regenerative mechanisms activated in ischemic stroke patients immediately after the event cooperate in the response to injury, in the restoration of functions and in brain remodeling even weeks after the event and can be sustained by the rehabilitation treatment. Nonetheless, patients’ response to treatments is difficult to predict because of the lack of specific measurable markers of recovery, which could be complementary to clinical scales in the evaluation of patients. Considering that Extracellular Vesicles (EVs) are carriers of multiple molecules involved in the response to stroke injury, in the present study, we have identified a panel of EV-associated molecules that (i) confirm the crucial involvement of EVs in the processes that follow ischemic stroke, (ii) could possibly profile ischemic stroke patients at the beginning of the rehabilitation program, (iii) could be used in predicting patients’ response to treatment. By means of a multiplexing Surface Plasmon Resonance imaging biosensor, subacute ischemic stroke patients were proven to have increased expression of vascular endothelial growth factor receptor 2 (VEGFR2) and translocator protein (TSPO) on the surface of small EVs in blood. Besides, microglia EVs and endothelial EVs were shown to be significantly involved in the intercellular communications that occur more than 10 days after ischemic stroke, thus being potential tools for the profiling of patients in the subacute phase after ischemic stroke and in the prediction of their recovery

Freeze Drying of Polymer Nanoparticles and Liposomes Exploiting Different Saccharide-Based Approaches

Biodegradable nanocarriers represent promising tools for controlled drug delivery. However, one major drawback related to their use is the long-term stability, which is largely influenced by the presence of water in the formulations, so to solve this problem, freeze-drying with cryoprotectants has been proposed. In the present study, the influence of the freeze-drying procedure on the storage stability of poly(lactide-co-glycolide) (PLGA) nanoparticles and liposomes was evaluated. In particular, conventional cryoprotectants were added to PLGA nanoparticle and liposome formulations in various conditions. Additionally, hyaluronic acid (HA), known for its ability to target the CD44 receptor, was assessed as a cryoprotective excipient: it was added to the nanocarriers as either a free molecule or conjugated to a phospholipid to increase the interaction with the polymer or lipid matrix while exposing HA on the nanocarrier surface. The formulations were resuspended and characterized for size, polydispersity index, zeta potential and morphology. It was demonstrated that only the highest percentages of cryoprotectants allowed the resuspension of stable nanocarriers. Moreover, unlike free HA, HA-phospholipid conjugates were able to maintain the particle mean size after the reconstitution of lyophilized nanoparticles and liposomes. This study paves the way for the use of HA-phospholipids to achieve, at the same time, nanocarrier cryoprotection and active targeting

Learning new sport actions: Pilot study to investigate the imitative and the verbal instructive teaching methods in motor education.

The aim of the project was to investigate the effects of two strategies of teaching new sport actions on performance of eight-year-old children: observational-imitative method (OIM) and descriptive-directive method (DDM). The OIM group was provided with a pre-practice instruction in the form of expert modeling observation by an expert athlete. The DDM group received only verbal explanations of few selected static images. Thirty-six children (18 males and 18 females, mean age = 8,8) participated in the experiment. Subjects were randomly assigned to the OIM or DDM groups. Participants were instructed to perform four sport motor sequences never performed before (shoulder stand, soccer action, vortex howler throw, step action). Actions were videotaped and 2D kinematic analysis performed. A 10-point Likert questionnaire was administered to blind sport experts to assess the correctness and accuracy of each action. Results suggest that the OIM is the most effective instruction method when participants have no experience with the sport action to be performed. On the contrary, if the athlete needs to learn specific aspects of an exercise (such as grasping a tool) the best method is the DDM. In fact, detailed information on how to grab the vortex helped children in throwing it. We also found gender differences which might reflect cultural influences in specific sports (e.g. soccer). Finally, repetition of the exercise also improved the DDM group's performance. This has potential applications in sport teaching, suggesting that in the absence of a model performing the action to be imitated, the DDM can be as effective as the OIM if the observer repeats the sport action many times

Microglial-Targeted nSMase2 Inhibitor Fails to Reduce Tau Propagation in PS19 Mice

The progression of Alzheimer’s disease (AD) correlates with the propagation of hyperphosphorylated tau (pTau) from the entorhinal cortex to the hippocampus and neocortex. Neutral sphingomyelinase2 (nSMase2) is critical in the biosynthesis of extracellular vesicles (EVs), which play a role in pTau propagation. We recently conjugated DPTIP, a potent nSMase2 inhibitor, to hydroxyl-PAMAM-dendrimer nanoparticles that can improve brain delivery. We showed that dendrimer-conjugated DPTIP (D–DPTIP) robustly inhibited the spread of pTau in an AAV-pTau propagation model. To further evaluate its efficacy, we tested D-DPTIP in the PS19 transgenic mouse model. Unexpectantly, D-DPTIP showed no beneficial effect. To understand this discrepancy, we assessed D-DPTIP’s brain localization. Using immunofluorescence and fluorescence-activated cell-sorting, D-DPTIP was found to be primarily internalized by microglia, where it selectively inhibited microglial nSMase2 activity with no effect on other cell types. Furthermore, D-DPTIP inhibited microglia-derived EV release into plasma without affecting other brain-derived EVs. We hypothesize that microglial targeting allowed D-DPTIP to inhibit tau propagation in the AAV-hTau model, where microglial EVs play a central role in propagation. However, in PS19 mice, where tau propagation is independent of microglial EVs, it had a limited effect. Our findings confirm microglial targeting with hydroxyl-PAMAM dendrimers and highlight the importance of understanding cell-specific mechanisms when designing targeted AD therapies