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

Terapia di Gruppo per Persone con Afasia cronica: uno Studio Pilota

Introduzione: Nel 2012 A.L.I.Ce4 ha proposto alla Medicina Rabilitativa di elaborare un Programma rivolto alle persone con afasia cronica da offrire con partecipazione gratuita ai propri associati portatori di Afasia. L’equipe interprofessionale (neuropsicologo,ricercatore,logopedista) operante c/o Medicina Riabilitativa, dopo la revisione della Letteratura che indica la Terapia di Gruppo come appropriata soprattutto nella fase cronica dell’Afasia (Cicerone et al, 2011; Salter et al, 2011; Linee Guida FLI, 2009; Marini et al., 2007; Meinzer et al., 2005; Wertz et al, 2005; Gesellshaft für Aphasiebehandlung, 2002; Pulvermuller et al., 2001), ha individuato un Programma basato sull’approccio pragmatico (Carlomagno, 2002; Davis e Wilcox, 1985). Al fine di munirsi di uno strumento di valutazione delle competenze pragmatiche le autrici hanno adattato vari questionari, producendo la SVCC (Scala di Valutazione delle Competenze Comunicative nella Conversazione in Gruppo). La Valutazione del Gradimento, sia della persona con Afasia che del familiare, è stata invece verificata somministrando un Questionario elaborato ad hoc. Obiettivo: Lo scopo dello studio pilota è verificare l’applicabilità e il gradimento del Programma proposto. Metodi: • Disegno: Studio osservazionale prospettico. • Setting: Medicina Riabilitativa di Parma – Sede Associazione A.L.I.Ce • Campione: 10 soggetti (5 maschi e 5 femmine) • Criteri di inclusione: - afasici che hanno concluso o stanno per concludere il progetto personalizzato di rieducazione logopedica presso le Strutture Riabilitative del territorio di Parma - età media 55,5 anni ( DS ± 8,3) - anni di scolarità media 11,9 (DS ± 7,3) - ad almeno un anno dall’unico evento focale (ictus o emorragia cerebrale) - adeguata motivazione e compliance • Criteri di esclusione: - deterioramento cognitivo Programma: il Programma è stato condotto da una psicologa e da una logopedista ed è stato ripetuto due volte per un totale complessivo di 40 ore (due ore a settimana per 10 sedute a ciclo). Fasi di Attuazione : 1. presentazione ai familiari e alle persone con afasia del programma 2. momento informativo su che cosa è l’afasia e su come i familiari possono aiutare praticamente le persone afasiche 3. somministrazione della SVCC (Scala di Valutazione delle Competenze Comunicative) (T0) 4. attuazione degli incontri dove il gruppo sperimenta situazioni di vita quotidiana (fare la spesa, andare dal medico, chiedere informazioni ecc) anche attraverso il role-playing 5. verifica del gradimento e Follow-up SVCC (T1) Principali misure di outcome: SVCC (adattamento interno 5) somministrata al reclutamento (T0) e al termine (T1); Questionario di Gradimento per la Persona con Afasia e per il Familiare (adattamento interno5). Risultati: 1. elevato gradimento sia per la persona affetta da afasia che per il familiare 2. aumento delle competenze pragmatiche statisticamente significativo per quanto attiene l’incremento e l’adeguatezza dell’iniziativa comunicativa spontanea e il rispetto dei ruoli parlante/ascoltatore (p=0,037) Conclusioni:: - il Programma si è dimostrato applicabile da un punto di vista organizzativo con dei costi sostenibili per A.L.I.Ce. che sponsorizzerà la ripetizione dell’iniziativa - l’applicabilità futura consentirà inoltre di ampliare il Campione al fine di approfondire l’analisi dei risultati terapeutic

Advances in the Field of Micro- and Nanotechnologies Applied to Extracellular Vesicle Research: Take-Home Message from ISEV2021

Extracellular Vesicles (EVs) are naturally secreted nanoparticles with a plethora of functions in the human body and remarkable potential as diagnostic and therapeutic tools. Starting from their discovery, EV nanoscale dimensions have hampered and slowed new discoveries in the field, sometimes generating confusion and controversies among experts. Microtechnological and especially nanotechnological advances have sped up biomedical research dealing with EVs, but efforts are needed to further clarify doubts and knowledge gaps. In the present review, we summarize some of the most interesting data presented in the Annual Meeting of the International Society for Extracellular Vesicles (ISEV), ISEV2021, to stimulate discussion and to share knowledge with experts from all fields of research. Indeed, EV research requires a multidisciplinary knowledge exchange and effort. EVs have demonstrated their importance and significant biological role; still, further technological achievements are crucial to avoid artifacts and misleading conclusions in order to enable outstanding discoveries

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.)

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

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

Characterization of the COPD Salivary Fingerprint through Surface Enhanced Raman Spectroscopy: A Pilot Study

Chronic Obstructive Pulmonary Disease (COPD) is a debilitating pathology characterized by reduced lung function, breathlessness and rapid and unrelenting decrease in quality of life. The severity rate and the therapy selection are strictly dependent on various parameters verifiable after years of clinical observations, missing a direct biomarker associated with COPD. In this work, we report the methodological application of Surface Enhanced Raman Spectroscopy combined with Multivariate statistics for the analysis of saliva samples collected from 15 patients affected by COPD and 15 related healthy subjects in a pilot study. The comparative Raman analysis allowed to determine a specific signature of the pathological saliva, highlighting differences in determined biological species, already studied and characterized in COPD onset, compared to the Raman signature of healthy samples. The unsupervised principal component analysis and hierarchical clustering revealed a sharp data dispersion between the two experimental groups. Using the linear discriminant analysis, we created a classification model able to discriminate the collected signals with accuracies, specificities, and sensitivities of more than 98%. The results of this preliminary study are promising for further applications of Raman spectroscopy in the COPD clinical field