Development of a Sensitive Colorimetric Indicator for Detecting Beef Spoilage in Smart Packaging.

This study aimed to fabricate and characterize a novel colorimetric indicator designed to detect ammonia (NH3) and monitor meat freshness. The sensing platform was constructed using electrospun nanofibers made from polylactic acid (PLA), which were then impregnated with anthocyanins as a natural pH-sensitive dye, extracted from red cabbage. This research involved investigating the relationship between the various concentrations of anthocyanins and the colorimetric platform's efficiency when exposed to ammonia vapor. Scanning electron microscope (SEM) results were used to examine the morphology and structure of the nanofiber mats before and after the dip-coating process. The study also delved into the selectivity of the indicator when exposed to various volatile organic compounds (VOCs) and their stability under extreme humidity levels. Furthermore, the platform's sensitivity was evaluated as it encountered ammonia (NH3) in concentrations ranging from 1 to 100 ppm, with varying dye concentrations. The developed indicator demonstrated an exceptional detection limit of 1 ppm of MH3 within just 30 min, making it highly sensitive to subtle changes in gas concentration. The indicator proved effective in assessing meat freshness by detecting spoilage levels in beef over time. It reliably identified spoilage after 10 h and 7 days, corresponding to bacterial growth thresholds (107 CFU/mL), both at room temperature and in refrigerated environments, respectively. With its simple visual detection mechanism, the platform offered a straightforward and user-friendly solution for consumers and industry professionals alike to monitor packaged beef freshness, enhancing food safety and quality assurance.

Tunable two-step shape and dimensional changes with temperature of a PNIPAM/CNC hydrogel.

PNIPAM (poly(N-isopropylacrylamide)), a well-studied thermo-responsive polymer, undergoes conformational transition around 32 °C. On the other hand, cellulose nanocrystals (CNCs), as a promising and biocompatible material, has rarely been introduced to the PNIPAM-based fibrous hydrogel system. CNCs' impact on the temperature responsive behaviors of hydrogels, either in single layer or bilayer hydrogel systems, is yet to be investigated. In this work, stable well dispersed PNIPAM/CNC suspensions (with various CNC proportions) are prepared and electrospun into nanofiber membranes. The corresponding hydrogels are then obtained via UV-induced crosslinking. CNCs are found to exert a significant constraint effect on hydrogel swelling when it exceeded 5 wt% but a negligible effect on contraction. The difference between hydrogels with various CNC proportions regarding their temperature responsive behaviors is utilized to fabricate bilayer hydrogels. These bilayer samples are capable of generating 3D geometries when they come into contact with water for the first time via anisotropic swelling between the two layers and changing their dimension reversibly in the following swelling and contraction. In addition, these geometries are found to be highly tunable via the finely tuned thickness ratio between the two layers. This promising feature would significantly extend the application of these materials in tissue engineering where a controllable geometry of the culture substrate is of great importance.

Negative In Vivo Results Despite Promising In Vitro Data With a Coated Compliant Electrospun Polyurethane Vascular Graft.

INTRODUCTION: There is a growing need for small-diameter (<6 mm) off-the-shelf synthetic vascular conduits for different surgical bypass procedures, with actual synthetic conduits showing unacceptable thrombosis rates. The goal of this study was to build vascular grafts with better compliance than standard synthetic conduits and with an inner layer stimulating endothelialization while remaining antithrombogenic. METHODS: Tubular vascular conduits made of a scaffold of polyurethane/polycaprolactone combined with a bioactive coating based on chondroitin sulfate (CS) were created using electrospinning and plasma polymerization. In vitro testing followed by a comparative in vivo trial in a sheep model as bilateral carotid bypasses was performed to assess the conduits' performance compared to the actual standard. RESULTS: In vitro, the novel small-diameter (5 mm) electrospun vascular grafts coated with chondroitin sulfate (CS) showed 10 times more compliance compared to commercial expanded polytetrafluoroethylene (ePTFE) conduits while maintaining adequate suturability, burst pressure profiles, and structural stability over time. The subsequent in vivo trial was terminated after electrospun vascular grafts coated with CS showed to be inferior compared to their expanded polytetrafluoroethylene counterparts. CONCLUSIONS: The inability of the experimental conduits to perform well in vivo despite promising in vitro results may be related to the low porosity of the grafts and the lack of rapid endothelialization despite the presence of the CS coating. Further research is warranted to explore ways to improve electrospun polyurethane/polycaprolactone scaffold in order to make it prone to transmural endothelialization while being resistant to strenuous conditions.

Surface Optimization and Design Adaptation toward Spheroid Formation On-Chip.

Spheroids have become an essential tool in preclinical cancer research. The uniformity of spheroids is a critical parameter in drug test results. Spheroids form by self-assembly of cells. Hence, the control of homogeneity of spheroids in terms of size, shape, and density is challenging. We developed surface-optimized polydimethylsiloxane (PDMS) biochip platforms for uniform spheroid formation on-chip. These biochips were surface modified with 10% bovine serum albumin (BSA) to effectively suppress cell adhesion on the PDMS surface. These surface-optimized platforms facilitate cell self-aggregations to produce homogenous non-scaffold-based spheroids. We produced uniform spheroids on these biochips using six different established human cell lines and a co-culture model. Here, we observe that the concentration of the BSA is important in blocking cell adhesion to the PDMS surfaces. Biochips treated with 3% BSA demonstrated cell repellent properties similar to the bare PDMS surfaces. This work highlights the importance of surface modification on spheroid production on PDMS-based microfluidic devices.

A 3D, Compartmental Tumor-Stromal Microenvironment Model of Patient-Derived Bone Metastasis.

Bone is a frequent site of tumor metastasis. The bone-tumor microenvironment is heterogeneous and complex in nature. Such complexity is compounded by relations between metastatic and bone cells influencing their sensitivity/resistance to chemotherapeutics. Standard chemotherapeutics may not show efficacy for every patient, and new therapeutics are slow to emerge, owing to the limitations of existing 2D/3D models. We previously developed a 3D interface model for personalized therapeutic screening, consisting of an electrospun poly lactic acid mesh activated with plasma species and seeded with stromal cells. Tumor cells embedded in an alginate-gelatin hydrogel are overlaid to create a physiologic 3D interface. Here, we applied our 3D model as a migration assay tool to verify the migratory behavior of different patient-derived bone metastasized cells. We assessed the impact of two different chemotherapeutics, Doxorubicin and Cisplatin, on migration of patient cells and their immortalized cell line counterparts. We observed different migratory behaviors and cellular metabolic activities blocked with both Doxorubicin and Cisplatin treatment; however, higher efficiency or lower IC50 was observed with Doxorubicin. Gene expression analysis of MDA-MB231 that migrated through our 3D hybrid model verified epithelial-mesenchymal transition through increased expression of mesenchymal markers involved in the metastasis process. Our findings indicate that we can model tumor migration in vivo, in line with different cell characteristics and it may be a suitable drug screening tool for personalized medicine approaches in metastatic cancer treatment.

Cellulose Nanopaper Cross-Linked Amino Graphene/Polyaniline Sensors to Detect CO2 Gas at Room Temperature.

A nanocomposite of cross-linked bacterial cellulose-amino graphene/polyaniline (CLBC-AmG/PANI) was synthesized by covalent interaction of amino-functionalized graphene (AmG) AmG and bacterial cellulose (BC) via one step esterification, and then the aniline monomer was grown on the surface of CLBC-AmG through in situ chemical polymerization. The morphological structure and properties of the samples were characterized by using scanning electron microscopy (SEM), and thermal gravimetric analyzer (TGA). The CLBC-AmG/PANI showed good electrical-resistance response toward carbon dioxide (CO2) at room temperature, compared to the BC/PANI nanopaper composites. The CLBC-AmG/PANI sensor possesses high sensitivity and fast response characteristics over CO2 concentrations ranging from 50 to 2000 ppm. This process presents an extremely suitable candidate for developing novel nanomaterials sensors owing to easy fabrication and efficient sensing performance.

Combining Electrospun Fiber Mats and Bioactive Coatings for Vascular Graft Prostheses.

The patency of small-diameter (<6 mm) synthetic vascular grafts (VGs) is still limited by the absence of a confluent, blood flow-resistant monolayer of endothelial cells (ECs) on the lumen and of vascular smooth muscle cell (VSMC) growth into the media layer. In this research, electrospinning has been combined with bioactive coatings based on chondroitin sulfate (CS) to create scaffolds that possess optimal morphological and bioactive properties for subsequent cell seeding. We fabricated random and aligned electrospun poly(ethylene terephthalate), ePET, mats with small pores (3.2 ± 0.5 or 3.9 ± 0.3 µm) and then investigated the effects of topography and bioactive coatings on EC adhesion, growth, and resistance to shear stress. Bioactive coatings were found to dominate the cell behavior, which enabled creation of a near-confluent EC monolayer that resisted physiological shear-flow conditions. CS is particularly interesting since it prevents platelet adhesion, a key issue to avoid blood clot formation in case of an incomplete EC monolayer or partial cell detachment. Regarding the media layer, circumferentially oriented nanofibers with larger pores (6.3 ± 0.5 µm) allowed growth, survival, and inward penetration of VSMCs, especially when the CS was further coated with tethered, oriented epithelial growth factor (EGF). In summary, the techniques developed here can lead to adequate scaffolds for the luminal and media layers of small-diameter synthetic VGs.

Mechanism of Action of Electrospun Chitosan-Based Nanofibers against Meat Spoilage and Pathogenic Bacteria.

This study investigates the antibacterial mechanism of action of electrospun chitosan-based nanofibers (CNFs), against Escherichia coli, Salmonella enterica serovar Typhimurium, Staphylococcus aureus and Listeria innocua, bacteria frequently involved in food contamination and spoilage. CNFs were prepared by electrospinning of chitosan and poly(ethylene oxide) (PEO) blends. The in vitro antibacterial activity of CNFs was evaluated and the susceptibility/resistance of the selected bacteria toward CNFs was examined. Strain susceptibility was evaluated in terms of bacterial type, cell surface hydrophobicity, and charge density, as well as pathogenicity. The efficiency of CNFs on the preservation and shelf life extension of fresh red meat was also assessed. Our results demonstrate that the antibacterial action of CNFs depends on the protonation of their amino groups, regardless of bacterial type and their mechanism of action was bactericidal rather than bacteriostatic. Results also indicate that bacterial susceptibility was not Gram-dependent but strain-dependent, with non-virulent bacteria showing higher susceptibility at a reduction rate of 99.9%. The susceptibility order was: E. coli > L. innocua > S. aureus > S. Typhimurium. Finally, an extension of one week of the shelf life of fresh meat was successfully achieved. These results are promising and of great utility for the potential use of CNFs as bioactive food packaging materials in the food industry, and more specifically in meat quality preservation.

Antibacterial Activity of Neat Chitosan Powder and Flakes.

This study investigates the antibacterial activity of neat chitosan powder and flakes against three different bacterial species, Escherichia coli, Listeria innocua and Staphylococcus aureus, which are frequent causes of food spoilage. The effect of chitosan concentration and purity, as well as the influence of temperature, ionic strength (salt) and impact of a solid physical support in the medium are examined. Results show that the antibacterial activity of neat chitosan: (i) requires partial solubilisation; (ii) can be promoted by environmental factors such as adequate temperature range, ionic strength and the presence of a solid physical support that may facilitate the attachment of bacteria; (iii) depends on bacterial species, with a sensitivity order of E. coli > L. innocua > S. aureus; and (iv) increases with chitosan concentration, up to a critical point above which this effect decreases. The latter may be due to remaining proteins in chitosan acting as nutrients for bacteria therefore limiting its antibacterial activity. These results on the direct use of chitosan powder and flakes as potential antimicrobial agents for food protection at pH values lower than the chitosan pKa (6.2-6.7) are promising.

Development of CdS Nanostructures by Thermal Decomposition of Aminocaproic Acid-Mixed Cd-Thiourea Complex Precursor: Structural, Optical and Photocatalytic Characterization.

The present work deals with two different CdS nanostructures produced via hydrothermal and solvothermal decompositions of aminocaproic acid (ACA)-mixed Cd-thiourea complex precursor at 175 °C. Both nanostructures were extensively characterized for their structural, morphological and optical properties. The powder X-ray diffraction characterization showed that the two CdS nanostructures present a wurtzite morphology. Scanning electron microscopy and energy-dispersive X-ray characterizations revealed that the hydrothermal decomposition produced well-shaped CdS flowers composed of six dendritic petals, and the solvothermal decomposition produced CdS microspheres with close stoichiometric chemical composition. The UV-vis absorption and photoluminescence spectra of CdS dendritic flowers and microsphere nanostructures showed that both nanostructures present a broad absorption between 200 and 700 nm and exhibit strong green emissions at 576 and 520 nm upon excitations at 290 nm and 260 nm, respectively. The transmission electron microscopy (TEM) and Brunauer-Emmett-Teller (BET) characterizations confirmed that CdS microspheres were mesoporous and were composed of small nanocrystals. A possible growth mechanism in the formation of the CdS nanostructures was proposed based on morphology evolution as a function of the reaction time. Furthermore, the as-synthesized CdS nanostructures were found to exhibit highly efficient photocatalytic activities for the degradation of methyl orange (MeO) and rhodamine B (RhB) dyes.

Development and application of low-density polyethylene-based multilayer film incorporating potassium permanganate and pumice for avocado preservation.

This study aimed to develop a low-density polyethylene-based multilayer active packaging film with three layers. The core layer was an active layer containing pumice and potassium permanganate, while the skin layer was the barrier layer impregnated with sodium chloride. The multilayer film showed an ethylene scavenging capacity of 1.6 µmol/(25 in2) within 8 d at 25 °C and was endowed with water absorption capacity. In addition, the oxygen and water vapor permeability of the multilayer film were improved in comparison to the neat one. Further, the multilayer film extended the shelf life of avocado from less than 10 d to 16 d at 25 °C, controlled ethylene and carbon dioxide concentrations, and caused a reduction in the loss of flesh firmness and weight. More importantly, according to migration testing, active agents in the core layer would not migrate to avocado peel, which ensured that avocados would not be contaminated.

Characterization of heat-treated chitosan cast films and their antimicrobial activity on the growth of natural flora of pasteurized milk.

This research aimed to evaluate the physicochemical and biocidal properties of chitosan films obtained through the solvent casting method using two different molecular weights, and thermally treated for an extended time (3 weeks) at 70 °C under vacuum condition (RH 0 %). The effect of storage time (for 30 and 180 days) under ambient conditions (23 °C and RH 40 %) on the properties of heat-treated cast films and their biocidal effectiveness was also assessed. FTIR-ATR, TGA and XRD of resulting films were analyzed to explore the dependency of antibacterial performance on the alteration in molecular and chemical structure. The results demonstrated that the solubility of treated films at 70 °C was proportionally reduced, resulting from the reduction of protonated amines and an increase in crystallinity. Likewise, increasing storage time led to a significant lowering in the solubilization of cast films. It was found that the solubilized fraction of chitosan cast films is the active fraction with the biocide behavior that can act against bacteria. In addition, the effectiveness of migrated chitosan was examined against the natural flora of pasteurized milk, such as Paenibacillus and Pseudomonas fluorescens. The results showed that cast films obtained from chitosan with lower molecular weight caused a reduction in the total count of viable cells without a significant effect on the properties of milk.

Core-shell structured PEO-chitosan nanofibers by coaxial electrospinning.

Core-shell structured PEO-chitosan nanofibers have been produced using a coaxial electrospinning setup. PEO and chitosan solutions, both in an aqueous acetic acid solvent, were used as the inner (core) and outer (shell) layer, respectively. Uniform-sized defect-free nanofibers of 150-190 nm diameter were produced. In addition, hollow nanofibers could be obtained subsequent to PEO washing of the membranes. The core-shell nanostructure and existence of chitosan on the shell layer were confirmed by TEM images obtained before and after washing the PEO content with water. The presence of chitosan on the surface of the composite nanofibers was further supported by XPS studies. The chitosan and PEO compositions in the nanofibrous mats were determined by TGA analysis, which were similar to their ratio in the feed solutions. The local compositional homogeneity of the membranes and the efficiency of the washing step to remove PEO were also verified by FTIR. In addition, DSC and XRD were used to characterize the crystalline structure and morphology of the co-electrospun nonwoven mats. The prepared coaxial nanofibers (hollow and solid) have several potential applications due to the presence of chitosan on their outer surfaces.

Synthesis of capped TiO2 nanocrystals of controlled shape and their use with MEH-PPV to develop nanocomposite films for photovoltaic applications.

The present study presents the synthesis details of titanium dioxide (TiO2) nanoparticles (NPs) of different morphologies using oleic acid (OA) and oleyl amine (OM) as capping agents. Different shapes of NPs, such as nanospheres, nanorods, and nanorhombics, were achieved. In order to develop nanocomposite thin films for photovoltaic cells, these TiO2 NPs were carefully dispersed in 2-methoxy-5-(2'-ethylhexyloxy)-p-phenylene vinylene (MEH-PPV) matrix. The properties of synthesized TiO2 NPs and MEH-PPV/TiO2 nanocomposites were characterized using transmission electron microscopy (TEM), thermogravimetric analysis (TGA), UV-Visible spectroscopy, and Photoluminescence technique. Obtained results showed promising properties for photovoltaic devices, especially solar radiation absorption properties and charge transfer at the interface of the conjugated MEH-PPV matrix and TiO2 dispersed NPs.

Crystallinity and Gas Permeability of Poly (Lactic Acid)/Starch Nanocrystal Nanocomposite.

The present work seeks to determine the impact of weight percentage (wt%) of grafted starch nanocrystals (g-SNCs) on the oxygen and water vapour permeability of poly (lactic acid), PLA. Changes in the oxygen and water vapour permeability of PLA due to changes in PLA's crystalline structures and lamellar thickness were quantified. To this end, 3, 5, and 7 wt% of g-SNC nanoparticles were blended with PLA using the solvent casting method in order to study impact of g-SNC nanoparticles on crystallization behaviour, long spacing period, melting behavior, and oxygen and water barrier properties of PLA nanocomposites. This was achieved by wide-angle X-ray diffraction (WAXD), small-angle X-ray diffraction (SAXD), differential scanning calorimetry (DSC), and oxygen and water vapour permeability machine. The results of the WAXD and SAXD analysis show that the addition of 5 wt% g-SNC in PLA induces α crystal structure at a lower crystallization time, while it significantly increases the α crystal thickness of PLA, in comparison to neat PLA. However, when g-SNC concentrations were altered (i.e., 3 or 7 wt%), the crystallization time was found to increase due to the thermodynamic barrier of crystallization. Finally, the oxygen and water vapour permeability of PLA/SNC-g-LA (5 wt%) nanocomposite film were found to be reduced by ∼70% and ~50%, respectively, when compared to the neat PLA film. This can lead to the development of PLA nanocomposites with high potential for applications in food packaging.

Uniform Tumor Spheroids on Surface-Optimized Microfluidic Biochips for Reproducible Drug Screening and Personalized Medicine.

Spheroids are recognized for resembling the important characteristics of natural tumors in cancer research. However, the lack of controllability of the spheroid size, form, and density in conventional spheroid culture methods reduces the reproducibility and precision of bioassay results and the assessment of drug-dose responses in spheroids. Nonetheless, the accurate prediction of cellular responses to drug compounds is crucial for developing new efficient therapeutic agents and optimizing existing therapeutic strategies for personalized medicine. We developed a surface-optimized PDMS microfluidic biochip to produce uniform and homogenous multicellular spheroids in a reproducible manner. This platform is surface optimized with 10% bovine serum albumin (BSA) to provide cell-repellent properties. Therefore, weak cell-surface interactions lead to the promotion of cell self-aggregations and the production of compact and uniform spheroids. We used a lung cancer cell line (A549), a co-culture model of lung cancer cells (A549) with (primary human osteoblasts, and patient-derived spine metastases cells (BML, bone metastasis secondary to lung). We observed that the behavior of cells cultured in three-dimensional (3D) spheroids within this biochip platform more closely reflects in vivo-like cellular responses to a chemotherapeutic drug, Doxorubicin, rather than on 24-well plates (two-dimensional (2D) model). It was also observed that the co-culture and patient-derived spheroids exhibited resistance to anti-cancer drugs more than the mono-culture spheroids. The repeatability of drug test results in this optimized platform is the hallmark of the reproducibility of uniform spheroids on a chip. This surface-optimized biochip can be a reliable platform to generate homogenous and uniform spheroids to study and monitor the tumor microenvironment and for drug screening.

A novel 3D co-culture platform for integrating tissue interfaces for tumor growth, migration and therapeutic sensitivity: "PP-3D-S".

Metastatic cancers can be highly heterogeneous, show large patient variability and are typically hard to treat due to chemoresistance. Personalized therapies are therefore needed to suppress tumor growth and enhance patient's quality of life. Identifying appropriate patient-specific therapies remains a challenge though, due mainly to non-physiological in vitro culture systems. Therefore, more complex and physiological in vitro human cancer microenvironment tools could drastically aid in development of new therapies. We developed a plasma-modified, electro-spun 3D scaffold (PP-3D-S) that can mimic the human cancer microenvironment for customized-cancer therapeutic screening. The PP-3D-S was characterized for optimal plasma-modifying treatment and scaffolds morphology including fiber diameter and pore size. PP-3D-S was then seeded with human fibroblasts to mimic a stromal tissue layer; cell adhesion on plasma-modified poly (lactic acid), PLA, electrospun mats vastly exceeded that on untreated controls. The cell-seeded scaffolds were then overlaid with alginate/gelatin-based hydrogel embedded with MDA-MB231 human breast cancer cells, representing a tumor-tissue interface. Among three different plasma treatments, we found that NH3 plasma promoted the most tumor cell migration to the scaffold surfaces after 7 days of culture. For all treated and non-treated mats, we observed a significant difference in tumor cell migration between small-sized and either medium- or large-sized scaffolds. In addition, we found that the PP-3D-S was highly comparable to the standard Matrigel® migration assays in two different sets of doxorubicin screening experiments, where 75% reduction in migration was achieved with 0.5 µM doxorubicin for both systems. Taken together, our data indicate that PP-3D-S is an effective, low-cost, and easy-to-use alternate 3D tumor migration model which may be suitable as a physiological drug screening tool for personalized medicine against metastatic cancers.

Effect of Polyethylene Film Thickness on the Antimicrobial Activity of Embedded Zinc Oxide Nanoparticles.

Microbial contamination of most foods occurs primarily at the surface during postprocessing and handling; therefore, preventing cross-contamination by incorporation of antimicrobial substances in contact with the surface of the product is an efficient strategy in reducing food contamination risks. Zinc oxide nanoparticles (ZnONPs) have been used widely to achieve antimicrobial films in various applications including the food industry. This work describes the fabrication of antimicrobial polymeric films containing ZnONPs produced by the coextrusion and dip-coating techniques. Effects of skin layer thicknesses containing ZnONPs on the antimicrobial effectiveness of the film by their capability to inactivate Gram-positive and Gram-negative bacteria were studied for both methods. The antimicrobial properties of the coextruded multilayer LLDPE/ZnONP nanocomposite films evidenced antimicrobial activity in the range 0.5-1.5 log reductions, while in the case of a sandblasted multilayer film, it showed high antimicrobial properties as around 99.99%. The optical properties of coextruded multilayer films were measured and discussed. Furthermore, to achieve a thinner LLDPE thickness, ZnONPs were coated with different concentrations of LLDPE solution by the dip-coating method. TEM confirmed that a homogeneous layer is formed on the surface of ZnONPs. The thickness of the LLDPE layer estimated by TEM was about 2 nm and film produced 3 log and 4 log reductions for E. coli and S. aureus, respectively. The results show that developed films have the potential to be used as food packaging films and can extend shelf life, maintain quality, and assure the safety of food. The antimicrobial mechanisms of ZnONPs were also investigated. It was found that direct contact of particles with products is necessary to assure high antibacterial activity of the films.

Effect of processing condition and antioxidants on visual properties of multilayer post-consumer recycled high density polyethylene films.

Mechanical recycling is a promising approach to reduce the environmental impact of plastic packaging waste. However, the presence of defects in recycled materials results in final products with relatively poor visual and/or mechanical properties. In this work, the origin of the visual defects in post-consumer recycled HDPE (PCR HDPE), as well as the effects of processing method, processing condition and the addition of antioxidants on the visual defects were studied in multilayer flexible polyethylene films. The nature of the defects in the film samples were investigated by combining optical microscopy, energy dispersive X-ray (EDX), hot stage microscopy, solvent extraction, and differential scanning calorimetry (DSC) techniques. It was found that the defects in PCR film samples can be mainly categorized as fiber defects and point gels. Hot stage microscopy results show that point gels can be subcategorized in two groups: (a) non-melting, non-deformable gels, and (b) melting, deformable defects. In addition, it was found that deformation of molten, deformable defects increased at higher temperatures specifically above 200 °C. Further characterizations showed that the observed deformable defects are highly entangled high molecular weight HDPE. The effect of processing temperature, processing with a twin-screw extruder and the addition of antioxidant on the visual defects in film samples were also discussed in detail. It was shown that increasing processing temperature and using twin-screw extruders were two approaches that could reduce considerably the number of defects. The addition of antioxidants was also shown to improve the film quality especially at lower processing temperatures.