Nanocarrier-based gene delivery for immune cell engineering.

Clinical advances in genetically modified immune cell therapies, such as chimeric antigen receptor T cell therapies, have raised hope for cancer treatment. The majority of these biotechnologies are based on viral methods for ex vivo genetic modification of the immune cells, while the non-viral methods are still in the developmental phase. Nanocarriers have been emerging as materials of choice for gene delivery to immune cells. This is due to their versatile physicochemical properties such as large surface area and size that can be optimized to overcome several practical barriers to successful gene delivery. The in vivo nanocarrier-based gene delivery can revolutionize cell-based cancer immunotherapies by replacing the current expensive autologous cell manufacturing with an off-the-shelf biomaterial-based platform. The aim of this research is to review current advances and strategies to overcome the challenges in nanoparticle-based gene delivery and their impact on the efficiency, safety, and specificity of the process. The main focus is on polymeric and lipid-based nanocarriers, and their recent preclinical applications for cancer immunotherapy.

Selenium nanoparticles incorporated in nanofibers media eliminate H1N1 activity: a novel approach for virucidal antiviral and antibacterial respiratory mask.

The consecutive viral infectious outbreaks impose severe complications on public health besides the economic burden which led to great interest in antiviral personal protective equipment (PPE). Nanofiber-based respiratory mask has been introduced as a significant barrier to eliminate the airborne transmission from aerosols toward reduction the viral infection spreading. Herein, selenium nanoparticles incorporated in polyamide 6 nanofibers coated on spunbond nonwoven were synthesized via electrospinning technique (PA6@SeNPs), with an average diameter of 180 ± 2 nm. The nanofiber-coated media were tested for 0.3 µm particulate filtration efficiency based on Standard NIOSH (42 CFR 84). PA6@SeNPs had a pressure drop of 45 ± 2 Pa and particulate filtration efficiency of more than 97.33 which is comparable to the N95 respiratory mask. The bacterial killing efficiency of these nanofibers was 91.25% and 16.67% against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli), respectively. Furthermore, the virucidal antiviral test for H1N1 infected Madin-Darby Canine Kidney cells (MDCK) exhibited TCID50 of 108.13, 105.88, and 105.5 for 2, 10, and 120 min of exposure times in comparison with 108.5, 107.5, and 106.5 in PA6 nanofibers as control sample. MTT assay indicated excellent biocompatibility of electrospun PA6@SeNP nanofibers on L292 cells. These results propose the PA6@SeNP nanofibers have a high potential to be used as an efficient layer in respiratory masks for protection against respiratory pathogens.

Co-electrospun PCL-collagen nanofibers containing saffron-synthesized gold nanoparticles as an antioxidant wound dressing.

Oxidant environment and inflammation are the leading cause of chronic wounds such as diabetic ulcers. A dressing containing antioxidants would ensure accelerated wound healing. In this study, electrospun gold nanoparticle (GNP)-embedded nanofibers were developed. GNPs (about 7 nm) were synthesized using saffron extract as a reducing and capping agent (GNP-EXT). For comparison, nanoparticles of the same size were also synthesized using citrate (GNP-CIT). Nanoparticle colloids showed a zeta potential of -27 mV. FTIR confirmed the presence of the extract molecules on the nanoparticles. DPPH assay demonstrated the significant radical scavenging properties of the GNP-EXT. The effect of nanoparticles on the viability of NIH3T3 mouse fibroblast cells was evaluated with an MTT assay that showed no significant toxicity of nanoparticles even in the highest concentration of 250 ppm. Then poly (ɛ-caprolactone) (PCL)- Collagen nanofibers containing GNPs were electrospun. By using SEM, TEM, ATR-FTIR, and contact angle measurement, the nanofibers were characterized. Proper cell adhesion and spreading was observed on nanofibers by SEM and Alamar blue assay illustrated appropriate cyto-compatibility on the obtained nanofibers after 5 days of cell seeding. Wound healing assay also confirmed the cell supporting properties and biocompatibility. The results suggest that saffron-synthesized GNP-loaded nanofibers would be considered as potential wound dressings.

Development of a High Sensitive Multiplex Lateral Flow Immunoassay (LFIA) System for Rapid Detection of Methicillin-Resistant Staphylococcus Aureus (MRSA).

Background: Methicillin-resistant Staphylococcus aureus (MRSA) has become a worldwide concern as an epidemic bacterium and a cause of nosocomial and community-acquired infections. One of the major problems in the prevention and treatment of infections caused by MRSA strains is their multi-drug resistant trait, which causes the spread of infections and increases the mortality rate. Therefore, a rapid and accurate method is needed to identify MRSA strains, initiate appropriate antibiotic therapy, and control its infection. The aim of this study was to develop a twin lateral flow immunoassay system to detect methicillin-resistant Staphylococcus aureus (MRSA). Methods: First, BSA blocked AuNPs-anti-peptidoglycan antibody and AuNPs-anti-BSA antibody were used to detect Staphylococcus aureus (S. aureus). Then, AuNPs-anti-PBP2a antibody was used to specifically detect MRSA. Sensitivity, specificity and limit of detection of this twin immunoassay system were assessed using MRSA, methicillin susceptible S. aureus and clinical samples. Results were compared to those of cefoxitin disc diffusion (FOX30) and Polymerase Chain Reaction (PCR) as gold standards. Results: The Limit of Detection (LOD) of this twin system were 103 and 104 CFU/ml for the first and second strips, respectively. Sensitivity and specificity of this innovative assay in detecting MRSA were 92.30 and 97.36%, compared to FOX30 and PCR, respectively. Conclusion: High rates of sensitivity and specificity of this initiative system show its high potentials for rapid and accurate detection of MRSA.

Cell-based wound dressing: Bilayered PCL/gelatin nanofibers-alginate/collagen hydrogel scaffold loaded with mesenchymal stem cells.

Wound dressing is applied to promote the healing process, wound protection, and additionally regeneration of injured skin. In this study, a bilayer scaffold composed of a hydrogel and nanofibers was fabricated to improve the regeneration of injured skin. To this end, polycaprolactone/gelatin (PCL/Gel) nanofibers were electrospun directly on the prepared collagen/alginate (Col/Alg) hydrogel. The bilayer scaffold was characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR), mechanical properties, and swelling/degradation time. Cytotoxicity assays were evaluated using MTT assay. Then, the nanofiber and bilayer scaffolds were seeded with Adipose-derived stem cells (ADSCs). ADSCs were isolated from rat adipose tissue and analyzed using flow cytometry, in advance. Full-thickness wounds on the backs of rats were dressed with ADSCs-seeded bilayer scaffolds and nanofibers. Histopathological evaluations were performed after 14 and 21 days using H&E (hematoxylin and eosin) staining. The results indicated that re-epithelialization, angiogenesis, and collagen remodeling were enhanced in ADSCs-seeded bilayer scaffolds and nanofibers in comparison with the control group. In conclusion, the best re-epithelialization, collagen organization, neovascularization, and low presence of inflammation in the wound area were observed in the ADSCs-seeded bilayer scaffolds.

Long-term follow-up of patients with hairy cell leukemia in the south of Iran.

BACKGROUND: Hairy cell leukemia (HCL) is an indolent chronic lymphoproliferative disorder and first-line treatment with either intravenous or subcutaneous cladribine generally leads to long-lasting remissions. METHOD: All 131 patients with hairy-cell leukemia (HCL) were analyzed, with a median follow-up of 91 months. Data is from 2007 to 2020. We evaluated the response rate to cladribine as the first line and the response rate to cladribine with or without rituximab in relapsed patients. Further, we assessed relapse-free survival, complications, and secondary malignancy. RESULTS: After a median follow-up of 91 months, the recurrence rate was 24%. The 5-year and 10-year RFS rates were 85% and 66%, respectively. Adding rituximab to 2-CDA leads to a better response rate than just cladribine (90% vs. 27.3%, p-value = 0.002) in the relapsed patients. CONCLUSION: HCL patients have long-term survival when cladribine is the first line of treatment. Furthermore, adding rituximab to cladribine leads to a higher response rate.

Nanofibers in Respiratory Masks: An Alternative to Prevent Pathogen Transmission.

Recent global outbreak of COVID-19 has raised serious awareness about our abilities to protect ourselves from hazardous pathogens and volatile organic compounds. Evidence suggests that personal protection equipment such as respiratory masks can radically decrease rates of transmission and infections due to contagious pathogens. To increase filtration efficiency without compromising breathability, application of nanofibers in production of respiratory masks have been proposed. The emergence of nanofibers in the industry has since introduced a next generation of respiratory masks that promises improved filtration efficiency and breathability via nanometric pores and thin fiber thickness. In addition, the surface of nanofibers can be functionalized and enhanced to capture specific particles. In addition to conventional techniques such as melt-blown, respiratory masks by nanofibers have provided an opportunity to prevent pathogen transmission. As the surge in global demand for respiratory masks increases, herein, we reviewed recent advancement of nanofibers as an alternative technique to be used in respiratory mask production.

Gene therapy to enhance angiogenesis in chronic wounds.

Skin injuries and chronic non-healing wounds are one of the major global burdens on the healthcare systems worldwide due to their difficult-to-treat nature, associated co-morbidities, and high health care costs. Angiogenesis has a pivotal role in the wound-healing process, which becomes impaired in many chronic non-healing wounds, leading to several healing disorders and complications. Therefore, induction or promotion of angiogenesis can be considered a promising approach for healing of chronic wounds. Gene therapy is one of the most promising upcoming strategies for the treatment of chronic wounds. It can be classified into three main approaches: gene augmentation, gene silencing, and gene editing. Despite the increasing number of encouraging results obtained using nucleic acids (NAs) as active pharmaceutical ingredients of gene therapy, efficient delivery of NAs to their site of action (cytoplasm or nucleus) remains a key challenge. Selection of the right therapeutic cargo and delivery methods is crucial for a favorable prognosis of the healing process. This article presents an overview of gene therapy and non-viral delivery methods for angiogenesis induction in chronic wounds.

Macrophages as a therapeutic target to promote diabetic wound healing.

It is well established that macrophages are key regulators of wound healing, displaying impressive plasticity and an evolving phenotype, from an aggressive pro-inflammatory or "M1" phenotype to a pro-healing or "M2" phenotype, depending on the wound healing stage, to ensure proper healing. Because dysregulated macrophage responses have been linked to impaired healing of diabetic wounds, macrophages are being considered as a therapeutic target for improved wound healing. In this review, we first discuss the role of macrophages in a normal skin wound healing process and discuss the aberrations that occur in macrophages under diabetic conditions. Next we provide an overview of recent macrophage-based therapeutic approaches, including delivery of ex-vivo-activated macrophages and delivery of pharmacological strategies aimed at eliminating or re-educating local skin macrophages. In particular, we focus on strategies to silence key regulator genes to repolarize wound macrophages to the M2 phenotype, and we provide a discussion of their potential future clinical translation.

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.

Decontamination Assessment of Nanofiber-based N95 Masks.

As the world battles with the outbreak of the novel coronavirus, it also prepares for future global pandemics that threaten our health, economy, and survivor. During the outbreak, it became evident that use of personal protective equipment (PPE), specially face masks, can significantly slow the otherwise uncontrolled spread of the virus. Nevertheless, the outbreak and its new variants have caused shortage of PPE in many regions of the world. In addition, waste management of the enormous economical and environmental footprint of single use PPE has proven to be a challenge. Therefore, this study advances the theme of decontaminating used masks. More specifically, the effect of various decontamination techniques on the integrity and functionality of nanofiber-based N95 masks (i.e. capable of at least filtering 95% of 0.3 µm aerosols) were examined. These techniques include 70% ethanol, bleaching, boiling, steaming, ironing as well as placement in autoclave, oven, and exposure to microwave (MW) and ultraviolet (UV) light. Herein, filtration efficiency (by Particle Filtration Efficiency equipment), general morphology, and microstructure of nanofibers (by Field Emission Scanning Electron microscopy) prior and after every decontamination technique were observed. The results suggest that decontamination of masks with 70% ethanol can lead to significant unfavorable changes in the microstructure and filtration efficiency (down to 57.33%) of the masks. In other techniques such as bleaching, boiling, steaming, ironing and placement in the oven, filtration efficiency dropped to only about 80% and in addition, some morphological changes in the nanofiber microstructure were seen. Expectedly, there was no significant reduction in filtration efficiency nor microstructural changes in the case of placement in autoclave and exposure to the UV light. It was concluded that, the latter methods are preferable to decontaminate nanofiber-based N95 masks.

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.

Increasing Angiogenesis Factors in Hypoxic Diabetic Wound Conditions by siRNA Delivery: Additive Effect of LbL-Gold Nanocarriers and Desloratadine-Induced Lysosomal Escape.

Impaired wound healing in people with diabetes has multifactorial causes, with insufficient neovascularization being one of the most important. Hypoxia-inducible factor-1 (HIF-1) plays a central role in the hypoxia-induced response by activating angiogenesis factors. As its activity is under precise regulatory control of prolyl-hydroxylase domain 2 (PHD-2), downregulation of PHD-2 by small interfering RNA (siRNA) could stabilize HIF-1α and, therefore, upregulate the expression of pro-angiogenic factors as well. Intracellular delivery of siRNA can be achieved with nanocarriers that must fulfill several requirements, including high stability, low toxicity, and high transfection efficiency. Here, we designed and compared the performance of layer-by-layer self-assembled siRNA-loaded gold nanoparticles with two different outer layers-Chitosan (AuNP@CS) and Poly L-arginine (AuNP@PLA). Although both formulations have exactly the same core, we find that a PLA outer layer improves the endosomal escape of siRNA, and therefore, transfection efficiency, after endocytic uptake in NIH-3T3 cells. Furthermore, we found that endosomal escape of AuNP@PLA could be improved further when cells were additionally treated with desloratadine, thus outperforming commercial reagents such as Lipofectamine® and jetPRIME®. AuNP@PLA in combination with desloratadine was proven to induce PHD-2 silencing in fibroblasts, allowing upregulation of pro-angiogenic pathways. This finding in an in vitro context constitutes a first step towards improving diabetic wound healing with siRNA therapy.

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.

pH-Sensitive PEGylated Liposomal Silybin: Synthesis, In Vitro and In Vivo Anti-Tumor Evaluation.

The drug delivery systems improve the efficacy of chemotherapeutics through enhanced targeting and controlled release however, biological barriers of tumor microenvironment greatly impede the penetration of nanomedicine within the tumor. We report herein the fabrication of a PEG-detachable silybin (SLB) pH-sensitive liposome decorated with TAT-peptide. For this, Acyl hydrazide-activated PEG2000 was prepared and linked with ketone-derivatized DPPE via an acid-labile hydrazone bond to form mPEG2000-HZ-DPPE. TAT peptide was conjugated with a shorter -PEG1000-DSPE spacer and post-inserted into PEGylated liposome (DPPC: mPEG2000-DSPE: Chol). To prepare nanoliposomes (around 100 nm), first, a novel method was used to prepare SLB-Soya PC (SLB-SPC) complex, then this complex was incorporated into nanoliposomes. The pH-sensitivity and shielding effect of long PEG chain on TAT peptide was investigated using DiI liposome and FACS analysis. Pre-treatment to the lowered pH enhanced cellular association of TAT-modified pH-sensitive liposome due to the cleavage of hydrazone bond and TAT exposure. Besides, TAT-modified pH-sensitive liposomes significantly reduced cell viability compared to the plain liposome. In vivo results were very promising with pH-sensitive liposome by detaching PEG moieties upon exposure to the acidic tumor microenvironment, enhancing cellular uptake, retarding tumor growth, and prolonging the survival of 4T1 breast tumor-bearing BALB/c mice. TAT modification of pH-sensitive liposome improved cancer cell association and cytotoxicity and demonstrated potential intracellular delivery upon exposure to acidic pH. However, in in vivo studies, TAT as a targeting ligand significantly decreased the therapeutic efficacy of the formulation attributed to an inefficient tumor accumulation and higher release rate in the circulation. The results of this study indicated that pH-sensitive liposome containing SLB, which was prepared with a novel method with a significant SLB loading efficiency, is very effective in the treatment of 4T1 breast tumor-bearing BALB/c mice and merits further investigation.

Electro-conductive carbon nanofibers containing ferrous sulfate for bone tissue engineering.

The application of electroactive scaffolds can be promising for bone tissue engineering applications. In the current paper, we aimed to fabricate an electro-conductive scaffold based on carbon nanofibers (CNFs) containing ferrous sulfate. FeSO4·7H2O salt with different concentrations 5, 10, and 15 wt%, were blended with polyacrylonitrile (PAN) polymer as the precursor and converted to Fe2O3/CNFs nanocomposite by electrospinning and heat treatment. The characterization was conducted using SEM, EDX, XRD, FTIR, and Raman methods. The results showed that the incorporation of Fe salt induces no adverse effect on the nanofibers' morphology. EDX analysis confirmed that the Fe ions are uniformly dispersed throughout the CNF mat. FTIR spectroscopy showed the interaction of Fe salt with PAN polymer. Raman spectroscopy showed that the incorporation of FeSO4·7H2O reduced the ID/IG ratio, indicating more ordered carbon in the synthesized nanocomposite. Electrical resistance measurement depicted that, although the incorporation of ferrous sulfate reduced the electrical conductivity, the conductive is suitable for electrical stimulation. The in vitro studies revealed that the prepared nanocomposites were cytocompatible and only negligible toxicity (less than 10%) induced by CNFs/Fe2O3 fabricated from PAN FeSO4·7H2O 15%. Although various nanofibrous composite fabricated with Fe NPs have been evaluated for tissue engineering applications, CNFs exhibited promising properties, such as excellent mechanical strength, biocompatibility, and electrical conductivity. These results showed that the fabricated nanocomposites could be applied as the bone tissue engineering scaffold.

Enhanced siRNA Delivery and Selective Apoptosis Induction in H1299 Cancer Cells by Layer-by-Layer-Assembled Se Nanocomplexes: Toward More Efficient Cancer Therapy.

Nanotechnology has made an important contribution to oncology in recent years, especially for drug delivery. While many different nano-delivery systems have been suggested for cancer therapy, selenium nanoparticles (SeNPs) are particularly promising anticancer drug carriers as their core material offers interesting synergistic effects to cancer cells. Se compounds can exert cytotoxic effects by acting as pro-oxidants that alter cellular redox homeostasis, eventually leading to apoptosis induction in many kinds of cancer cells. Herein, we report on the design and synthesis of novel layer-by-layer Se-based nanocomplexes (LBL-Se-NCs) as carriers of small interfering RNA (siRNA) for combined gene silencing and apoptosis induction in cancer cells. The LBL-Se-NCs were prepared using a straightforward electrostatic assembly of siRNA and chitosan (CS) on the solid core of the SeNP. In this study, we started by investigating the colloidal stability and protection of the complexed siRNA. The results show that CS not only functioned as an anchoring layer for siRNA, but also provided colloidal stability for at least 20 days in different media when CS was applied as a third layer. The release study revealed that siRNA remained better associated with LBL-Se-NCs, with only a release of 35% after 7 days, as compared to CS-NCs with a siRNA release of 100% after 48 h, making the LBL nanocarrier an excellent candidate as an off-the-shelf formulation. When applied to H1299 cells, it was found that they can selectively induce around 32% apoptosis, while significantly less apoptosis (5.6%) was induced in NIH/3T3 normal cells. At the same time, they were capable of efficiently inducing siRNA downregulation (35%) without loss of activity 7 days post-synthesis. We conclude that LBL-Se-NCs are promising siRNA carriers with enhanced stability and with a dual mode of action against cancer cells.

State-of-the-Art of Nanodiagnostics and Nanotherapeutics against SARS-CoV-2.

The pandemic outbreak of SARS-CoV-2, with millions of infected patients worldwide, has severely challenged all aspects of public health. In this regard, early and rapid detection of infected cases and providing effective therapeutics against the virus are in urgent demand. Along with conventional clinical protocols, nanomaterial-based diagnostics and therapeutics hold a great potential against coronavirus disease 2019 (COVID-19). Indeed, nanoparticles with their outstanding characteristics would render additional advantages to the current approaches for rapid and accurate diagnosis and also developing prophylactic vaccines or antiviral therapeutics. In this review, besides presenting an overview of the coronaviruses and SARS-CoV-2, we discuss the introduced nanomaterial-based detection assays and devices and also antiviral formulations and vaccines for coronaviruses.

An investigation of factors affecting the electrospinning of poly (vinyl alcohol)/kefiran composite nanofibers.

This study aimed to investigate parameters affecting the electrospinning of poly (vinyl alcohol) (PVA)/kefiran composite nanofibers. Accordingly, PVA/kefiran composite nanofibers were produced using the electrospinning of PVA, kefiran blend solutions under various electrospinning parameters (such as applied voltage, nozzle-to-collector distance, and polymer injection rate), and solution parameters (such as the ratio of polymers). PVA and kefiran solutions were prepared in 8% and 6% w/w, respectively. Kefiran was blended with PVA solution in different proportions: 70:30, 60:40, 50:50, 40:60, and 30:70. According to the scanning electron microscope (SEM) images, kefiran mixed with PVA in 40:60 ratios produced the best result in nanofiber production. Then, device parameters such as voltage (12, 15, 18, and 20 kV), distance (120, 150, 170, and 200 mm), and polymer injection rates (1, 1.5, 2, and 2.5 mL/h) were changed. The investigation of SEM images showed that the optimal condition for the fabrication of nanofibers was 18 kV, 200 mm, and 1 mL/h. The nanofibers produced in the optimal condition were uniform without knots or adhesion in a small diameter. It was also found that concentration can be regarded as the most effective parameter affecting the diameter of nanofibers. Moreover, the transmission electron microscopy (TEM) image proved that phase separation did not occur between the two polymers. Graphical abstract: Kefiran biopolymer extracted from fermented milk was used in fabrication of PVA/kefiran composite nanofibers using the electrospinning method.