Wet-electrospun PCL/PLLA Blend Scaffolds: Effects of Versatile Coagulation Baths on Physicochemical and Biological Properties of the Scaffolds

Introduction: High surface/volume ratio and 3-dimensionality of nanofibers increases cell-scaffold interactions and promote migration and proliferation of cells. Wet electrospinning is a variant of electrospinning technology that is utilized to produce nanofibrous scaffolds. Altering the parameters governing the wet electrospinning process such as applied voltage, polymer concentration, composition and depth of the coagulation bath, and tip to bath distance can affect the morphology of the produced scaffolds. In this study, the influence of various coagulation baths on the physicochemical properties of the wet-electrospun nanofibers was investigated. Materials and Methods: Poly (ε-caprolactone)/Poly (L-lactic) acid 15% (w/v) blends under an applied voltage of 15 kV, and a tip-to-bath distance of 10 cm. were used to prepare fibrous scaffolds via wet-electrospinning technique into aqueous solution of sodium hydroxide (NaOH) (pH~13), distilled water, ethanol, water/ethanol (3:7) (v/v) and water/ethanol/methanol (6:2:2) (v/v). The final products were characterized by scanning electron microscopy (SEM), liquid displacement technique, contact angle measurement, compressive and tensile tests. As well as, cell adhesion and cell viability through human adipose-derived stem cells (hADSCs) cell culture. Results: Wet-electrospun fibers, except in the almost fully beaded structure of water/ethanol (3:7) (v/v) specimen exhibited random, dispersive and non-woven morphology under SEM observation. The coagulation bath composition significantly influenced on porosity, wettability, mechanical properties and biocompatibility of the scaffolds. The porosity measurement via liquid displacement method showed that except for the specimen in which the blend was spun into NaOH, other scaffolds could not meet the accepted ideal porosity percentage of above 80%. According to the contact angle measurement data, it was expected that all scaffolds experience low cellular attachment and proliferation. Conversely, in vitro hADSCs culture demonstrated that the scaffolds presented a non-toxic environment and enhanced cell proliferation and attachment. Conclusion: The data indicated that the scaffold spun into NaOH was the best candidate among other specimens to culture hADSCs

The Effect of Different Stabilizers on Stability of Horseradish Peroxidase- Bovine Serum Albumin-Aflatoxin B1, a Conjugated Tracer for Detection of Aflatoxin B1 in Immunoassay-Based Methods

Abstract Aflatoxins are a group of fungal toxic metabolites, which are contaminated certain food commodities. ELISA is one of the sensitive methods for detection of aflatoxins. Preparation and stabilizing of a proper conjugated tracer for detection of aflatoxins is probably the main step for designing an ELISA method. In current study, different stabilizers were applied to stabilize a newly prepared conjugated molecule, Horseradish peroxidase-Bovine serum albumin-Aflatoxin B1 (HRP-BSA-AFB1). Stabilizing effects of six different stabilizers were compared during 10 months verification at room temperature and 6 weeks verification at 37°C. Based on the results, it was concluded that trehalose-containing stabilizers especially those contain casein in their compositions show the best stabilizing effects on HRP-BSA-AFB1 conjugated tracer

Polyurethane/gelatin nanofiber neural guidance conduit in combination with resveratrol and Schwann cells for sciatic nerve regeneration in the rat model

Peripheral nerve injury is a serious challenge which influences 2.8 percent of trauma patients. Tissue engineering of peripheral nerves mainly focuses on axonal regeneration via various nerve guides. The aim of this study is to evaluate a novel polyurethane (PU)/gelatin nanofibers (GNFs) conduit’s potential combination with resveratrol (RVT) for sciatic nerve regeneration in the rat. Platelet-rich plasma (PRP) was used as a carrier for RVT. Different tests like contact angle, tensile strength etc. was used to evaluate properties of PU/GNFs conduits. In addition, the electron microscopy, MTT assay, and DAPI staining revealed its compatibility with Schwann cells. 24 male Wistar rats were allocated into four groups (n=6) (1) PU/GNF/PRP/Schwann cell, 2) PU/GNF/Schwann cell/PRP/RVT, 3) Positive control, and 4) Negative control in order to assess sciatic functional index (SFI), hot plate latency, electromyographical (EMG), the percentage of wet weight-loss of gastrocnemius muscle and histopathological studies using hematoxylin-eosin staining. The results represented sciatic functional index (SFI), hot plate latency, electromyographical improved significantly in group 1 and 2 compared to the negative control group. Histopathological findings showed remarkable improvements in myelin sheath regeneration and fibers condition in group 1 and 2 compared to the negative control group. Group 2 showed more resemblance to the normal sciatic nerve, with well-arranged fibers and an intact myelin sheath. This study successfully applied PU/GNFs/PRP/RVT conduits as a potential biocompatible nerve guide with proper mechanical properties, biocompatibility, and biodegradability that enhanced injured sciatic nerve’s recovery rate

The cardiac niche role in cardiomyocyte differentiation of rat bone marrow-derived stromal cells

Due to the restricted potential of the heart to regenerate its damaged region, stem cell therapy is a promising treatment modality for myocardial infarction. It has been shown that incubation of bone marrow-derived stromal cells (BMSCs) with initial steps of cardiac differentiation in vitro, can have a significant effect on their therapeutic opotential to treat myocardial infarction. Based on these well-established principals we were encouraged to study the direct co-culture of rat BMSCs with neonatal mouse almost pure cardiomyocytes (APCs) and cardiac niche cells (CNCs) in static 2D and microfluidic cell culture systems. Our results showed that the difference regarding the beating rate in isolated APCs and CNCs in both 2D and the microfluidic system was not statistically significant for 30 days. No beat rate could be observed in induced BMSCs in all groups during experiment time. Except for BMSCs cultured alone in both experimental culture conditions, data obtained from Real-time PCR analysis showed that differentiated BMSCs in all co-cultured groups expressed GATA4, Nkx2.5, CX43, cTnI, cTnT, and β-MHC during 4 weeks. BMSCs demonstrated a higher expression of these cardiac factors in microfluidic chips than those co-cultured in 24 well plates. Moreover, immunocytochemistry (ICC), also revealed the GATA4 expression in differentiated BMSCs in all co-cultured groups. It was found that, when combined with shear stress, co-culture with cardiomyocyte can differentiate BMSCs significantly toward cardiomyocyte rather than co-culture alone