The Effect of Electrospun Gelatin Fibers Alignment on Schwann Cell and Axon Behavior and Organization in the Perspective of Artificial Nerve Design

Electrospun fibrous substrates mimicking extracellular matrices can be prepared by electrospinning, yielding aligned fibrous matrices as internal fillers to manufacture artificial nerves. Gelatin aligned nano-fibers were prepared by electrospinning after tuning the collector rotation speed. The effect of alignment on cell adhesion and proliferation was tested in vitro using primary cultures, the Schwann cell line, RT4-D6P2T, and the sensory neuron-like cell line, 50B11. Cell adhesion and proliferation were assessed by quantifying at several time-points. Aligned nano-fibers reduced adhesion and proliferation rate compared with random fibers. Schwann cell morphology and organization were investigated by immunostaining of the cytoskeleton. Cells were elongated with their longitudinal body parallel to the aligned fibers. B5011 neuron-like cells were aligned and had parallel axon growth when cultured on the aligned gelatin fibers. The data show that the alignment of electrospun gelatin fibers can modulate Schwann cells and axon organization in vitro, suggesting that this substrate shows promise as an internal filler for the design of artificial nerves for peripheral nerve reconstruction

Long-Term Outcome After Adoptive Immunotherapy With Natural Killer Cells: Alloreactive NK Cell Dose Still Matters

Recently, many reports were published supporting the clinical use of adoptivelytransferred natural killer (NK) cells as a therapeutic tool against cancer, including acutemyeloid leukemia (AML). Our group demonstrated promising clinical response usingadoptive immunotherapy with donor-derived alloreactive KIR-ligand-mismatched NK cellsin AML patients. Moreover, the antileukemic effect was correlated with the dose of infusedalloreactive NK cells (“functional NK cell dose”). Herein, we update the results of ourprevious study on a cohort of adult AML patients (median age at enrollment 64) infirstmorphological complete remission (CR), not eligible for allogeneic stem celltransplantation. After an extended median follow-up of 55.5 months, 8/16 evaluablepatients (50%) are still off-therapy and alive disease-free. Overall survival (OS) and disease-free survival (DFS) are related with the dose of infused alloreactive NK cells (≥2×105/kg

The Effect of Electrospun Gelatin Fibers Alignment on Schwann Cell and Axon Behavior and Organization in the Perspective of Artificial Nerve Design

Electrospun fibrous substrates mimicking extracellular matrices can be prepared by electrospinning, yielding aligned fibrous matrices as internal fillers to manufacture artificial nerves. Gelatin aligned nano-fibers were prepared by electrospinning after tuning the collector rotation speed. The effect of alignment on cell adhesion and proliferation was tested in vitro using primary cultures, the Schwann cell line, RT4-D6P2T, and the sensory neuron-like cell line, 50B11. Cell adhesion and proliferation were assessed by quantifying at several time-points. Aligned nano-fibers reduced adhesion and proliferation rate compared with random fibers. Schwann cell morphology and organization were investigated by immunostaining of the cytoskeleton. Cells were elongated with their longitudinal body parallel to the aligned fibers. B5011 neuron-like cells were aligned and had parallel axon growth when cultured on the aligned gelatin fibers. The data show that the alignment of electrospun gelatin fibers can modulate Schwann cells and axon organization in vitro, suggesting that this substrate shows promise as an internal filler for the design of artificial nerves for peripheral nerve reconstruction

Biomimetic polyurethane scaffolds for a stem cell based therapy in myocardial regeneration

Scaffolds for myocardial Tissue Engineering (TE) should display biomimetic properties respect to cardiac extracellular matrix (ECM), including elastomeric properties [1]. Cardiac regeneration depends on cardiac progenitor cells (CPCs) as well as the milieu in contact with them. Laminin-1 (LN1), typical of developing heart and over-expressed in pathological heart, promotes CPC proliferation and viability [1]. In this work, a thermoplastic polyurethane (PU) was synthesized from poly(ε-caprolactone) diol (Mn = 2000 Da), 1,4-budandiisocyanate and L-lysine ethyl ester dihydrochloride [2]. Bi-layered scaffolds with 0°/90° lay-down pattern were prepared by additive-manufacturing technique [2]. Functionalisation with LN1 or gelatin (G) was performed in two steps: 1) acrylic acid grafting/polymerization by Argon plasma treatment; 2) carbodiimide-mediated coupling of proteins. Scaffolds with mean fibre diameter of 1525 m and mean spacing of 5055 m were prepared. FITR-ATR analysis of protein-coated scaffolds showed higher intensity of the absorption bands at 3370 cm-1 (-OH and –NH stretching) and 1650 cm-1 (amide I). Contact angle decreased from 90° for PU to 60-65° after G- or LN1-grafting. XPS analysis confirmed acrylic acid grafting/polymerization and protein conjugation. Scaffolds were degraded in vitro by lipase (0.3 mg/ml) in 3 weeks. CPC proliferation on PU-LN1 scaffolds was higher than on PU and PU-G scaffolds, increasing from 8,18% on day 7 to 11,8% on day 14. LN1-functionalization stimulated CPC differentiation into cardiomyocytes and endothelial cells