Comparison of the efficiency of ultrafiltration, precipitation, and ultracentrifugation methods for exosome isolation.

Extracellular vesicles (EVs) are enclosed by a lipid-bilayer membrane and secreted by all types of cells. They are classified into three groups: apoptotic bodies, microvesicles, and exosomes. Exosomes play a number of important roles in the intercellular communication and crosstalk between tissues in the body. In this study, we use three common methods based on different principles for exosome isolation, namely ultrafiltration, precipitation, and ultracentrifugation. We use field emission scanning electron microscopy (FESEM) and dynamic light scattering (DLS) analyses for characterization of exosomes. The functionality and effect of isolated exosomes on the viability of hypoxic cells was investigated by alamarBlue and Flow-cytometry. The results of the FESEM study show that the ultrafiltration method isolates vesicles with higher variability of shapes and sizes when compared to the precipitation and ultracentrifugation methods. DLS results show that mean size of exosomes isolated by ultrafiltration, precipitation, and ultracentrifugation methods are 122, 89, and 60 nm respectively. AlamarBlue analysis show that isolated exosomes increase the viability of damaged cells by 11%, 15%, and 22%, respectively. Flow-cytometry analysis of damaged cells also show that these vesicles increase the content of live cells by 9%, 15%, and 20%, respectively. This study shows that exosomes isolated by the ultracentrifugation method are characterized by smaller size and narrow size distribution. Furthermore, more homogenous particles isolated by this method show increased efficiency of the protection of hypoxic cells in comparison with the exosomes isolated by the two other methods.

Evaluating the antioxidant potential of resveratrol-gold nanoparticles in preventing oxidative stress in endothelium on a chip.

Vascular endothelial cells play a vital role in the health and maintenance of vascular homeostasis, but hyperglycemia disrupts their function by increasing cellular oxidative stress. Resveratrol, a plant polyphenol, possesses antioxidant properties that can mitigate oxidative stress. Addressing the challenges of its limited solubility and stability, gold nanoparticles (GNps) were utilized as carriers. A microfluidic chip (MFC) with dynamic flow conditions was designed to simulate body vessels and to investigate the antioxidant properties of resveratrol gold nanoparticles (RGNps), citrate gold nanoparticles (CGNps), and free Resveratrol on human umbilical vein endothelial cells (HUVEC). The 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay was employed to measure the extracellular antioxidant potential, and cell viability was determined using the Alamar Blue test. For assessing intracellular oxidative stress, the 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) assay was conducted, and results from both the cell culture plate and MFC were compared. Free Resveratrol demonstrated peak DPPH scavenging activity but had a cell viability of about 24-35%. RGNPs, both 3.0 ± 0.5 nm and 20.2 ± 4.7 nm, consistently showed high cell viability (more than about 90%) across tested concentrations. Notably, RGNPs (20 nm) exhibited antioxidative properties through DPPH scavenging activity (%) in the range of approximately 38-86% which was greater than that of CGNps at about 21-32%. In the MFC,the DCFH-DA analysis indicated that RGNPs (20 nm) reduced cellular oxidative stress by 57-82%, surpassing both CGNps and free Resveratrol. Morphologically, cells in the MFC presented superior structure compared to those in traditional cell culture plates, and the induction of hyperglycemia successfully led to the formation of multinucleated variant endothelial cells (MVECs). The MFC provides a distinct advantage in observing cell morphology and inducing endothelial cell dysfunction. RGNps have demonstrated significant potential in alleviating oxidative stress and preventing endothelial cell disorders.

Evaluation of drug-eluting nanoparticle coating on magnesium alloy for development of next generation bioabsorbable cardiovascular stents.

Since the introduction of bioabsorbable magnesium alloys into cardiovascular stent technology, many researches have been conducted to improve these metallic scaffolds. Various coatings and different coating techniques, super plastic deformation techniques and synthesizing different Mg-based alloy are examples of such efforts. In this study, a magnesium based alloy (WE43) was coated with dexamethasone loaded polymeric nanoparticles via electrospraying method. Drug release behavior, drug inhibitory effects, surface properties and cell responses to the surface were evaluated. Drug release profile was investigated and compared to drug-loaded nanoparticle on stainless steel as a control. The inhibitory effects of the drug-loaded nanoparticle coatings on smooth muscle cells was evaluated via MTT assay. Endothelial cells response to the surface was investigated by SEM. The results showed that contact angle and roughness of the surface were 131° and 600-800 nm, respectively. Drug release studies showed a burst release less than 30% after 24 h which followed by nearly zero order release kinetic. MTT assay showed that SMCs viability decreased to 60% and 25% after 24 and 72 h, respectively. SEM images indicated proper adhesion and proliferation of endothelial cells on the surface. The findings suggest that nanoparticle-coated surfaces could effectively inhibit SMC proliferation meanwhile provide desirable surface features for adhesion and proliferation of endothelial cell on magnesium alloy based stents.

Effectiveness of exosome mediated miR-126 and miR-146a delivery on cardiac tissue regeneration.

Despite advances in the treatment of acute myocardial infarction, due to the non-proliferative nature of adult cardiomyocytes, the injured myocardium is mainly replaced by fibrotic tissue, which ultimately causes heart failure. To prevent heart failure, particularly after myocardial infarction, exosome-based therapy has emerged as one of the most promising strategies to regenerate cardiac function. Exosomes can carry microRNAs in support of neovascularization, anti-inflammatory, and endogenous cardiac regeneration. This study demonstrated that animal rat models' combination treatment with microRNA-126 and microRNA-146a mimics in exosomes is desirable for cardiac regeneration after myocardial infarction. The exosomes isolated from stem cells and loaded with microRNAs were characterized their impacts in cell migration, tube formation, and vascular endothelial growth factor degree. In the following, the usefulness of loaded microRNAs in exosomes and their encapsulation within alginate derivative hydrogel was analyzed in myocardial infarction for an animal model. Exosomes isolated and loaded with microRNAs showed the synergetic impact on cell migration, tube formation, and promoted vascular endothelial growth factor folding. Moreover, microRNAs loaded exosomes and encapsulated them in alginate hydrogel could help in reducing infarct size and improving angiogenesis in myocardial infarction. The angiogenesis markers including CD31 and connexion 43 upregulated for myocardial infarction models treated with alginate-based hydrogels loaded with exosomes and microRNAs-exosomes. Histological analysis indicated that myocardial infarction model rats treated with alginate hydrogel loaded with microRNAs-exosomes possessed lower and higher degrees of fibrosis and collagen fiber, respectively. These findings have important therapeutic implications for a myocardial infarction model through angiogenesis and vascular integrity regulation.

Green synthesis of oxidized starch with a novel catalyst based on Fe3O4 nanoparticles and H2O2 reagent to form thermoplastic as a stable gel coating on the cardiovascular stents.

Oxidation of starch is one of the most commonly used approaches to improve its properties in the thermoplastic (TP) reactions. Iron oxide nanoparticle (IONP) (8.2 ± 1.5 nm) was used as a novel catalyst for this reaction. The functional groups of the carbonyl (COH) and the carboxyl (COOH) were obtained about of 7-12.2 % and 0.03-0.3 %. TP reaction and then electrospray technique of oxidized starch were used for the thin-film coating. The swelling ratio of the gelled thermoplastic structure with IONP (198 ± 9 % at 180 min) was lower than the sample without NP (193 ± 8 % at 90 min). The results from fourier transform infrared spectroscopy (FTIR), hydrogen nuclear magnetic resonance (HNMR), X-ray diffraction (XRD), and transmission electron microscopy (TEM) reveal desirable chemical and crystalline changes. Scanning electron microscopy (SEM) analysis was used to determine the thickness of the thin film (1.4 ± 0.2 µm) and the size of the electrosprayed droplets (172 ± 45 nm). Cytotoxicity studies of HUVEC and L929 cell lines against the extracts have shown appropriate biocompatibility. The blood compatibility analysis demonstrated proper results for (nanocomposite) NC. The results show that NC coated on metal surfaces can be used in medical approaches with drug delivery capability.

Improvement of sciatic nerve regeneration by multichannel nanofibrous membrane-embedded electro-conductive conduits functionalized with laminin.

Multichannel structures in the design of nerve conduits offer potential advantages for regeneration of damaged nerves. However, lack of biochemical cues and electrical stimulation could hamper satisfactory nerve regeneration. The aim of this study was to simultaneously evaluate the effects of topographical, biological, and electrical cues on sciatic nerve regeneration. Accordingly, a series of multichannel nerve conduit was made using longitudinally-aligned laminin-coated poly (lactic-co-glycolic acid) (PLGA)/carbon nanotubes (CNT) nanofibers (NF, mean diameter: 455 ± 362 nm) in the lumen and randomly-oriented polycaprolactone (PCL) NF (mean diameter: 340 ± 200 nm) on the outer surface. In vitro studies revealed that the materials were nontoxic and able to promote cell attachment and proliferation on nanofibers and on fibrin gel. To determine the influence of laminin as biological and CNT as electrical cues on nerve regeneration, either of hollow PCL conduits, PLGA NF-embedded, PLGA/CNT NF-embedded or laminin-coated PLGA/CNT NF-embedded PCL conduits were implanted in rats. A new surgery method was utilized and results were compared with an autograft. The results of motor and sensory tests in addition to histopathological examination of the regenerated nerves demonstrated the formation of nerve fibers in laminin-coated PLGA/CNT NF-embedded PCL conduits. Results suggested that these conduits have the potential to improve sciatic nerve regeneration. Graphical abstract.

Adaptive antitumor immune response stimulated by bio-nanoparticle based vaccine and checkpoint blockade.

BACKGROUND: Interactions between tumor and microenvironment determine individual response to immunotherapy. Triple negative breast cancer (TNBC) and hepatocellular carcinoma (HCC) have exhibited suboptimal responses to immune checkpoint inhibitors (ICIs). Aspartate ß-hydroxylase (ASPH), an oncofetal protein and tumor associated antigen (TAA), is a potential target for immunotherapy. METHODS: Subcutaneous HCC and orthotopic TNBC murine models were established in immunocompetent BALB/c mice with injection of BNL-T3 and 4 T1 cells, respectively. Immunohistochemistry, immunofluorescence, H&E, flow cytometry, ELISA and in vitro cytotoxicity assays were performed. RESULTS: The ASPH-MYC signaling cascade upregulates PD-L1 expression on breast and liver tumor cells. A bio-nanoparticle based λ phage vaccine targeting ASPH was administrated to mice harboring syngeneic HCC or TNBC tumors, either alone or in combination with PD-1 blockade. In control, autocrine chemokine ligand 13 (CXCL13)-C-X-C chemokine receptor type 5 (CXCR5) axis promoted tumor development and progression in HCC and TNBC. Interactions between PD-L1+ cancer cells and PD-1+ T cells resulted in T cell exhaustion and apoptosis, causing immune evasion of cancer cells. In contrast, combination therapy (Vaccine+PD-1 inhibitor) significantly suppressed primary hepatic or mammary tumor growth (with distant pulmonary metastases in TNBC). Adaptive immune responses were attributed to expansion of activated CD4+ T helper type 1 (Th1)/CD8+ cytotoxic T cells (CTLs) that displayed enhanced effector functions, and maturation of plasma cells that secreted high titers of ASPH-specific antibody. Combination therapy significantly reduced tumor infiltration of immunosuppressive CD4+/CD25+/FOXP3+ Tregs. When the PD-1/PD-L1 signal was inhibited, CXCL13 produced by ASPH+ cancer cells recruited CXCR5+/CD8+ T lymphocytes to tertiary lymphoid structures (TLSs), comprising effector and memory CTLs, T follicular helper cells, B cell germinal center, and follicular dendritic cells. TLSs facilitate activation and maturation of DCs and actively recruit immune subsets to tumor microenvironment. These CTLs secreted CXCL13 to recruit more CXCR5+ immune cells and to lyse CXCR5+ cancer cells. Upon combination treatment, formation of TLSs predicts sensitivity to ICI blockade. Combination therapy substantially prolonged overall survival of mice with HCC or TNBC. CONCLUSIONS: Synergistic antitumor efficacy attributable to a λ phage vaccine specifically targeting ASPH, an ideal TAA, combined with ICIs, inhibits tumor growth and progression of TNBC and HCC.

Hybrid PCL/chitosan-PEO nanofibrous scaffolds incorporated with A. euchroma extract for skin tissue engineering application.

Skin tissue engineering is an advanced method to repair and regenerate skin injuries. Recent research is focused on the development of scaffolds that are safe, bioactive, and cytocompatible. In this work, a new hybrid nanofibrous scaffold composed of polycaprolactone/chitosan-polyethylene oxide (PCL/Cs-PEO) incorporated with Arnebia euchroma (A. euchroma) extract were synthesized by the two-nozzle electrospinning method. Then the synthesized scaffold was characterized for morphology, sustainability, chemical structure and properties. Moreover, to verify their potential in the burn wound healing process, biodegradation rate, contact angle, swelling properties, water vapor permeability, mechanical properties, antibacterial activity and drug release profile were measured. Furthermore, cytotoxicity and biocompatibility tests were performed on human dermal fibroblasts cell line via XTT and LDH assay. It is shown that the scaffold improved and increased proliferation during in-vitro studies. Thus, results confirm the efficacy and potential of the hybrid nanofibrous scaffold for skin tissue engineering.

Bio-nanoparticle based therapeutic vaccine induces immunogenic response against triple negative breast cancer.

Triple negative breast cancer (TNBC) is more aggressive and has a poorer prognosis than other sub-types of breast tumors. This study elucidates how aspartate beta-hydroxylase (ASPH) network promotes drug resistance, and immunotherapy targeting ASPH may improve the efficacy of Doxorubicin (DOX) therapy. An orthotopic model of breast cancer generated by 4T1 cells in immunocompetent mice was used to explore efficacy of immunotherapy in combination with DOX chemotherapy. We evaluated mRNA and protein expression in cultured tumor cells and tissue, as well as assessed cell proliferation, apoptosis, soluble factors/cytokine production, immune cell population diversity and function. We observed that ASPH expression enables TNBC cells to exhibit primary resistance to DOX induced single-/double-strand breaks (SSB/DSB) and enhanced proliferation and survival. Specific bio-nanoparticle based therapeutic vaccine (BNP-TV) promoted ASPH uptake by and maturation of DCs. This BNP-TV combined with DOX induces immunogenic cell death (ICD) in orthotopic xenograft tumors and significantly suppressed primary mammary tumor growth and distant multi-organ metastases. Immunogenic cell death induced by BNP-TV targeting ASPH combined with DOX provides opportunities to treat a highly resistant and metastatic form of breast cancer.

Assessment of structural, biological and drug release properties of electro-sprayed poly lactic acid-dexamethasone coating for biomedical applications.

The efficacy of an implant is highly depends on its coating characteristics mainly determined by polymer properties and coating technique. Electro-spraying is an inexpensive and versatile coating technique with various advantages for biomedical application. In this study, the efficacy of electro-sprayed (ES) poly lactic acid (PLA)-dexamethasone (DEX) coatings for medical implants was evaluated and compared with spin-coated samples as control. Structural properties of coatings were investigated using X-ray diffraction (XRD) and differential scanning calorimetry (DSC). Confocal and scanning electron microscopy (SEM), contact angle measurement and nanoindentation tests were used to study surface properties. Coating degradation rate and drug release profile were studied for 40 days. Cell viability experiments were also performed on human endothelial (HUVEC) and smooth muscle cells (HUASMC) using MTT assay and SEM. XRD and DSC analysis showed electro-spraying significantly reduce PLA and DEX crystallinity. Surface studies showed ES coatings has significantly higher hydrophobicity and roughness with microbead-nanofiber morphology vs. micro-nanoporous structure of spin-coated samples. Initial burst release of DEX was 22% and 10% after 6 h and total release was 71% and 46% after 40 days for ES and spin-coated samples, respectively. HUVEC viability of ES samples was higher than spin-coated ones after 1 and 4 days. However, dexamethasone release profile reduced HUASMC proliferation in ES PLA-DEX samples in comparison to spin-coated after 1 and 3 days. In conclusion, in vitro results showed potential of ES PLA-DEX as a biocompatible and efficient anti-inflammatory coating with suitable drug release profile for future applications such as coronary drug eluting stents.

Macrophage reprogramming into a pro-healing phenotype by siRNA delivered with LBL assembled nanocomplexes for wound healing applications.

Excessive inflammatory responses in wounds are characterized by the presence of high levels of pro-inflammatory M1 macrophages rather than pro-healing M2 macrophages, which leads to delayed wound healing. Macrophage reprogramming from the M1 to M2 phenotype through knockdown of interferon regulatory factor 5 (irf5) has emerged as a possible therapeutic strategy. While downregulation of irf5 could be achieved by siRNA, it very much depends on successful intracellular delivery by suitable siRNA carriers. Here, we report on highly stable selenium-based layer-by-layer (LBL) nanocomplexes (NCs) for siRNA delivery with polyethyleneimine (PEI-LBL-NCs) as the final polymer layer. PEI-LBL-NCs showed good protection of siRNA with only 40% siRNA release in a buffer of pH = 8.5 after 72 h or in simulated wound fluid after 4 h. PEI-LBL-NCs also proved to be able to transfect RAW 264.7 cells with irf5-siRNA, resulting in successful reprogramming to the M2 phenotype as evidenced by a 3.4 and 2.6 times decrease in NOS-2 and TNF-α mRNA expression levels, respectively. Moreover, irf5-siRNA transfected cells exhibited a 2.5 times increase of the healing mediator Arg-1 and a 64% increase in expression of the M2 cell surface marker CD206+. Incubation of fibroblast cells with conditioned medium isolated from irf5-siRNA transfected RAW 264.7 cells resulted in accelerated wound healing in an in vitro scratch assay. These results show that irf5-siRNA loaded PEI-LBL-NCs are a promising therapeutic approach to tune macrophage polarization for improved wound healing.

Effect of Paclitaxel/etoposide co-loaded polymeric nanoparticles on tumor size and survival rate in a rat model of glioblastoma.

The aim of this work is to co-load paclitaxel (PTX) and etoposide (ETP) in methoxy poly(ethylene glycol)-poly(lactic-co-glycolic acid) nanoparticles (mPEG-PLGA NPs) to overcome pharmacokinetics and physiological limitations and enhance therapeutic efficacy for treating intracranial glioblastoma. Both drugs were loaded into mPEG-PLGA NPs by a nano-precipitation method. The resultant NPs demonstrated an enhanced cytotoxic effect indicated by lower IC50 values and augmented cell apoptosis to U87 and C6 glioma cell lines compared to both free drugs. Additionally, blood compatibility assays showed that the PTX/ETP co-loaded mPEG-PLGA NPs did not induce blood hemolysis, blood clotting, or platelet aggregation. In vivo anti-glioma efficacy evaluation in rats bearingintracranialC6glioma revealed a superior anti-glioma activity for the treatment with PTX/ETP co-loaded mPEG-PLGA NPs compared to other formulations, particularly a significantly longer median survival, 76 days compared to 36 days for free PTX and 37 days for free ETP treatment, respectively, and higher tumor regression, proved by magnetic resonance imaging (MRI).

Dosimetric effect of nanoparticles in the breast cancer treatment using INTRABEAM®system with spherical applicators in the presence of tissue heterogeneities: A Monte Carlo study.

Using the 50 kV INTRABEAM®IORT system after breast-conserving surgery: tumor recurrence and organs at risk (OARs), such as the lung and heart, long-term complications remain the challenging problems for breast cancer patients. So, the objective of this study was to address these two problems with the help of high atomic number nanoparticles (NPs). A Monte Carlo (MC) Simulation type EGSnrc C++ class library (egspp) with its Easy particle propagation (Epp) user code was used. The simulation was validated against the measured depth dose data found in our previous study (Tegaw,et al2020 Dosimetric characteristics of the INTRABEAM®system with spherical applicators in the presence of air gaps and tissue heterogeneities,Radiat. Environ. Biophys. (10.1007/s00411-020-00835-0)) using the gamma index and passed 2%/2 mm acceptance criteria in the gamma analysis. Gold (Au) NPs were selected after comparing Dose Enhancement Ratios (DERs) of bismuth (Bi), Au, and platinum (Pt) NPs which were calculated from the simulated results. As a result, 0.02, 0.2, 2, 10, and 20 mg-Au/g-breast tissue were used throughout this study. These particles were not distributed in discrete but in a uniform concentration. For 20 mg-Au/g-breast tissue, the DERs were 3.6, 0.420, and 0.323 for breast tissue, lung, heart, respectively, using the 1.5 cm-diameter applicator (AP) and 3.61, 0.428, and 0.335 forbreast tissue, lung, and heart using the 5 cm-diameter applicator, respectively. DER increased with the decrease in the depth of tissues and increase in the effective atomic number (Zeff) and concentration of Au NPs, however, there was no significant change as AP sizes increased. Therefore, Au NPs showed dual advantages such as dose enhancement within the tumor bed and reduction in the OARs (heart and lung).

Preparation and characterization of polyurethane/chitosan/CNT nanofibrous scaffold for cardiac tissue engineering.

Myocardial infarction of cardiomyocytes is a leading cause of heart failure (HF) worldwide. Since heart has very limited regeneration capacity, cardiac tissue engineering (TE) to produce a bioactive scaffold is considered. In this study, a series of polyurethane solutions (5-7%wt) in aqueous acetic acid were prepared using electrospinning. A variety of Polyurethane (PU)/Chitosan (Cs)/carbon nanotubes (CNT) composite nanofibrous scaffolds with random and aligned orientation were fabricated to structurally mimic the extracellular matrix (ECM). Electrospun nanofibers were then characterized using field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), water contact angle, degradation studies, tensile tests, electrical resistance measurement and cell viability assay. The biocompatibility of electrospun random and aligned nanofibrous scaffolds with H9C2 Cells was confirmed. The results revealed that fabricated PU/Cs/CNT composite nanofibrous scaffolds were electro-conductive and aligned nanofibers could be considered as promising scaffolds with nano-scale features for regeneration of infarcted myocardium.

Bismuth oxide nanoparticles as agents of radiation dose enhancement in intraoperative radiotherapy.

PURPOSE: Intraoperative radiotherapy (IORT) technique is an advanced radio therapeutic method used for delivery of a single high-dose radiation during surgery while removing healthy tissues from the radiation field. Nowadays, growing attention is being paid to IORT for its low-energy (kilovoltage) delivery as it requires less radiation protection, but suffers several disadvantages, including high-dose delivery and prolonged treatment time. The application of nanoparticles with high atomic number and high attenuation coefficients in kilovoltage energy may help overcome the mentioned shortcomings. This study was designed to investigate and quantify the mean dose enhancement factor (DEF) in the presence of nanoparticles using IORT method. METHODS: Bismuth oxide nanoparticles (Bi2 O3 NPs), both in sheet and spherical formats, were synthesized using a novel hydrothermal method and characterized with x-ray diffraction (XRD), transmission electron microscopy (TEM), and Brunauer-Emmett-Teller (BET) analysis. Genipin-gelatin gel dosimeter (GENIPIN) was produced in three batches of pure with sheet and with spherical nanoparticles in concentration of 46.596 µg/ml, and irradiated with 50 kV x-rays. RESULTS: Samples were scanned by a spectrophotometer, which indicated a DEF of 3.28  ±  0.37 and 2.50  ±  0.23 for sheet and spherical NPs, respectively. According to the results of this study, GENIPIN is a suitable dosimeter for the evaluation of three-dimensional dose distribution in the presence Bi2 O3 NPs. CONCLUSION: As a result, IORT along with Bi2 O3 NPs has the potential to reduce treatment time and/or normal tissue dose; moreover, it could provide localized dose enhancement.

The effect of surface modification of poly-lactide-co-glycolide/carbon nanotube nanofibrous scaffolds by laminin protein on nerve tissue engineering.

The presence of biological cues to promote the attachment, proliferation, and differentiation of neuronal cells is important in the process of nerve regeneration. In this study, laminin as a neurite promoting protein, has been used to modify poly-lactide-co-glycolide/carbon nanotube (PLGA/CNT) electrospun nanofibrous scaffolds by means of either mussel-inspired poly(dopamine) (PD) coating or via direct physical adsorption as a simple route for the functionalization of biomaterials. The laminin-modified scaffolds were characterized by a combination of field emission scanning electron microscopy (SEM), X-ray photoelectron spectroscopy, and contact angle measurements. Subsequently, various properties of scaffolds such as degradation time, amount of attached laminin and the rate of CNT release were investigated. The synergistic effect of topographical and biological cues for PC12 cell attachment, proliferation, and differentiation were then studied by SEM and confocal microscopy. The results of degradation study showed that laminin-modified scaffolds were biodegradable with good structural integrity that persisted about 4 weeks. The amount of laminin attached to the PLGA/CNT and PLGA/CNT-PD scaffolds was 3.12 ± 0.6 and 3.04 ± 071 µg per mg of the scaffold, respectively. Although laminin-modified scaffolds could improve cell proliferation identically, neurite extensions on the PLGA/CNT scaffold modified via PD coating (PLGA/CNT-PD-lam scaffold) were significantly longer than those observed on PLGA/CNT scaffold modified via physical adsorption (PLGA/CNT-lam scaffold) and unmodified scaffolds. Together, these results indicated that surface modification via PD coating could be a promising strategy to fabricate biomimetic scaffolds capable of sustaining longer neuronal growth for nerve tissue engineering.

Cell attachment effects of collagen nanoparticles on crosslinked electrospun nanofibers.

Since collagen is naturally a main extracellular matrix protein, it has been applied widely in skin's tissue engineering scaffolds to mimics the characteristics of extracellular matrix for proper transplantation of living cells. However, there are challenges that come with application of this natural polymer such as high solubility in aqueous environments which requires further consideration such as chemically cross-linking in order to stabilization. But these treatments also affect its functionality and finally cellular behaviors on scaffold. In this research we evaluated the suitability of collagen nanofibers versus collagen nanoparticles for cell adhesion and viability on glutaraldehyde cross-linked scaffolds. Appling a dual-pump electrospining machine a blend PCL-Gelatin from one side and collagen nanofibers or collagen nanoparticles from the other side were collected on the collector. The fabricated scaffolds were characterized by scanning electron microscopy, contact angle, and mechanical analysis. The cell viability, adhesion and morphology were studied respectively using MTT assay, hoechst staining and scanning electron microscopy. The results indicated significantly improvement of cell viability, adhesion and better spreading on scaffolds with collagen nanoparticles than collagen nanofibers. It seems changes in surface morphology, viscoelastic moduli and swelling ability following cross-linking with glutaraldehyde in scaffold with collagen nanoparticles are still favorable for cellular proliferation. Based on these results, in the case of glutaraldehyde cross-linking, application of collagen nanoparticles rather than collagen nanofibers in tissue regeneration scaffolds will better mimic the extracellular matrix characteristics; and preserve the viability and adhesion of seeded cells.

Aspartate ß-Hydroxylase (ASPH) Expression in Acute Myeloid Leukemia: A Potential Novel Therapeutic Target.

BACKGROUND: Aspartate ß-hydroxylase (ASPH) is an embryonic transmembrane protein aberrantly upregulated in cancer cells, associated with malignant transformation and, in some reports, with poor clinical prognosis. OBJECTIVE: To report the expression patterns of ASPH in acute myeloid leukemia (AML). METHODS: Cell surface expression of ASPH was measured via 8-color multiparameter flow cytometry in 41 AML patient samples (31 bone marrow, 10 blood) using fluorescein isothiocyanate (FITC)-conjugated anti-ASPH antibody, SNS-622. A mean fluorescent intensity (MFI) of 10 was used as a cutoff for ASPH surface expression positivity. Data regarding patient and disease characteristics were collected. RESULTS: ASPH surface expression was found on AML blasts in 16 samples (39%). Higher ASPH expression was seen in myeloblasts of African American patients (p=0.02), but no correlation was found between ASPH expression and other patient or disease characteristics. No association was found between ASPH status and CR rate (p=0.53), EFS (p=0.87), or OS (p=0.17). CONCLUSIONS: ASPH is expressed on blasts in approximately 40% of AML cases, and may serve as a new therapeutically targetable leukemia-associated antigen.

Predicting the effect of phototherapy method on breast cancer cells by mathematical modeling: UV-IR non-ionization radiation with gold nanoparticles.

Phototherapy is a minimally invasive oncological treatment strategy in which photon energy is delivered to the tumor tissue. Gold nanoparticles (GNPs) can enhance photothermal or photodynamic phenomena when excited by a wavelength beam in the range of UV-IR. GNPs are used in phototherapy for cancer cell treatment by controlling the physical and chemical conditions. Given the growing application of GNPs for the treatment of breast cancer, predicting the behavior of cancer cells during exposure to GNPs is of prime importance. However, the prediction might be far from reality due to the inherent complexities associated with the conditions of the treatment methods and the mechanisms involved in cell toxicity. This study provides general information by collecting data on the cytotoxicity of GNPs along with this process. Data mining was performed using a mathematical modeling method called SA-LOOCV-GRBF. In this study, eight parameters including particle size, zeta potential, concentration of GNPs in the cell culture medium, incubation time, light exposure time, maximum wavelength absorbance (MAW) of GNPs, irradiation beam wavelength (IW) and light source power density (PD) were measured. In this modeling, these parameters were considered as model inputs, and the cell viability of breast cancer cells after treatment was treated as the model output. As a result, the physical and chemical properties of GNPs as well as their application conditions wield influence on cytotoxicity. The results help select the desired condition for these nanoparticles in the phototherapy of breast cancer cells.