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1.
Food Res Int ; 192: 114830, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39147519

ABSTRACT

The natural dual nanofibril system consisting of the rigid semicrystalline nanofibrils disintegrated from citrus fiber (CF) and soft semiflexible nanofibrils self-assembled from glycyrrhizic acid (GA) has been recently shown to be effective structural building blocks for fabrication of emulsion gels. In this work, the effect of the CF nanofibrils prepared by different mechanical disintegration approaches (i.e., high-pressure microfluidization and hydrodynamic cavitation) on the interfibrillar CF-GA interactions and the subsequent formation and properties of emulsion gels were investigated, with the aim of evaluating the potential of the dual nanofibril-stabilized emulsion gels as templates for synthesizing all-natural edible oleogels. The obtained results demonstrate that compared to the cavitation, the high-pressure microfluidization is more capable of generating CF nanofibrils with a higher degree of nanofibrillation and individualization, thus forming a denser CF-GA gel network with higher viscoelasticity and structural stability due to the stronger multiple intrafibrillar and interfibrillar interactions. The emulsion gels stabilized by the dual nanofibril system are demonstrated to be an efficient template to fabricate solid-like oleogels, and the structural properties of the oleogels can be well tuned by the mechanical disintegration of CF and the GA nanofibril concentration. The prepared oleogels possess high oil loading capacity, dense network microstructure, superior rheological and large deformation compression performances, and satisfactory thermal stability, which is attributed to the compact and ordered CF-GA dual nanofibrillar network via multiple hydrogen-bonding interactions in the continuous phase as well as at the droplet surface. This study highlights the unique use of all-natural dual nanofibrils to develop oil structured soft materials for sustainable applications.


Subject(s)
Citrus , Emulsions , Gels , Glycyrrhizic Acid , Nanofibers , Organic Chemicals , Emulsions/chemistry , Glycyrrhizic Acid/chemistry , Citrus/chemistry , Nanofibers/chemistry , Organic Chemicals/chemistry , Gels/chemistry , Rheology , Viscosity
2.
PLoS One ; 19(8): e0308207, 2024.
Article in English | MEDLINE | ID: mdl-39110684

ABSTRACT

Neurofibromatosis Type 1 (NF1) is a complex genetic disorder characterized by the development of benign neurofibromas, which can cause significant morbidity in affected individuals. While the molecular mechanisms underlying NF1 pathogenesis have been extensively studied, the development of effective therapeutic strategies remains a challenge. This paper presents the development and validation of a novel biomaterial testing model to enhance our understanding of NF1 pathophysiology, disease mechanisms and evaluate potential therapeutic interventions. Our long-term goal is to develop an invitro model of NF1 to evaluate drug targets. We have developed an in vitro system to test the cellular behavior of NF1 patient derived cells on electroconductive aligned nanofibrous biomaterials with electrical stimulatory cues. We hypothesized that cells cultured on electroconductive biomaterial will undergo morphological changes and variations in cell proliferation that could be further enhanced with the combination of exogenous electrical stimulation (ES). In this study, we developed electrospun Hyaluronic Acid-Carbon Nanotube (HA-CNT) nanofiber scaffolds to mimic the axon's topographical and bioelectrical cues that influence neurofibroma growth and development. The cellular behavior was qualitatively and quantitively analyzed through immunofluorescent stains, Alamar blue assays and ELISA assays. Schwann cells from NF1 patients appear to have lost their ability to respond to electrical stimulation in the development and regeneration range, which was seen through changes in morphology, proliferation and NGF release. Without stimulation, the conductive material enhances NF1 SC behavior. Wild-type SC respond to electrical stimulation with increased cell proliferation and NGF release. Using this system, we can better understand the interaction between axons and SC that lead to tumor formation, homeostasis and regeneration.


Subject(s)
Cell Proliferation , Electric Stimulation , Hyaluronic Acid , Nanotubes, Carbon , Schwann Cells , Schwann Cells/metabolism , Nanotubes, Carbon/chemistry , Humans , Hyaluronic Acid/chemistry , Nanofibers/chemistry , Neurofibromatosis 1/pathology , Neurofibromatosis 1/metabolism , Tissue Scaffolds/chemistry , Cells, Cultured , Biocompatible Materials/chemistry
3.
J Mater Sci Mater Med ; 35(1): 46, 2024 Aug 08.
Article in English | MEDLINE | ID: mdl-39115576

ABSTRACT

An antifungal agent, luliconazole, is commercially available in cream or gel form. The major limitation of these conventional formulations is less residence time at the infection site. The primary objective of this work was to develop luliconazole-loaded polyvinyl alcohol (Luz-PVA) nanofibers for mycotic skin conditions with a longer retention. Luz-PVA nanofibers were prepared by plate electrospinning and optimized for polymer concentration and process parameters. The optimized batch (Trial 5) was prepared by 10% PVA, processed at 22.4 kV applied voltage, and 14 cm plate and spinneret distance to yield thick, uniform, and peelable nanofibers film. There was no interaction observed between Luz and PVA in the FTIR study. DSC and XRD analysis showed that luliconazole was loaded into fabricated nanofibers with a reduced crystallinity. FESEM studies confirmed the smooth, defect-free mats of nanofibers. Luz-PVA nanofibers possessed a tensile strength of 21.8 N and a maximum elongation of 10.8%, representing the excellent elasticity of the scaffolds. For Luz-PVA nanofibers, the sustained and complete drug release was observed in 48 h. In antifungal activity using Candida albicans, the Luz-PVA nanofibers showed a greater zone of inhibition (30.55 ± 0.38 mm and 29.27 ± 0.31 mm) than marketed cream (28.06 ± 0.18 mm and 28.47 ± 0.24 mm) and pure drug (27.57 ± 0.17 mm and 27.50 ± 0.47 mm) at 1% concentration in Sabouraud dextrose agar and yeast malt agar, respectively. Therefore, Luz-PVA nanofibers exhibited good mechanical properties, longer retention time, and better antifungal activity than marketed products and, therefore, can be further examined preclinically as a potential treatment option for topical mycotic infection.


Subject(s)
Antifungal Agents , Candida albicans , Imidazoles , Microbial Sensitivity Tests , Nanofibers , Polyvinyl Alcohol , Antifungal Agents/pharmacology , Antifungal Agents/chemistry , Antifungal Agents/pharmacokinetics , Candida albicans/drug effects , Nanofibers/chemistry , Polyvinyl Alcohol/chemistry , Imidazoles/chemistry , Imidazoles/pharmacology , Administration, Topical , Spectroscopy, Fourier Transform Infrared , Tensile Strength , Delayed-Action Preparations/chemistry , Delayed-Action Preparations/pharmacology , X-Ray Diffraction
4.
ACS Appl Bio Mater ; 7(8): 5662-5678, 2024 Aug 19.
Article in English | MEDLINE | ID: mdl-39097904

ABSTRACT

Quercetin, recognized for its antioxidant, anti-inflammatory, and antibacterial properties, faces limited biomedical application due to its low solubility. Cotton, a preferred wound dressing material over synthetic ones, lacks inherent antibacterial and wound-healing attributes and can benefit from quercetin features. This study explores the potential of overcoming these challenges through the inclusion complexation of quercetin with cyclodextrins (CDs) and the development of a nanofibrous coating on a cotton nonwoven textile. Hydroxypropyl-beta-cyclodextrin (HP-ß-CD) and hydroxypropyl-gamma-cyclodextrin (HP-γ-CD) formed inclusion complexes of quercetin, with chitosan added to enhance antibacterial properties. Phase solubility results showed that inclusion complexation can enhance quercetin solubility up to 20 times, with HP-γ-CD forming a more stable inclusion complexation compared with HP-ß-CD. Electrospinning of the nanofibers from HP-ß-CD/Quercetin and HP-γ-CD/Quercetin aqueous solutions without the use of a polymeric matrix yielded a uniform, smooth fiber morphology. The structural and thermal analyses of the HP-ß-CD/Quercetin and HP-γ-CD/Quercetin nanofibers confirmed the presence of inclusion complexes between quercetin and each of the CDs (HP-ß-CD and HP-γ-CD). Moreover, HP-ß-CD/Quercetin and HP-γ-CD/Quercetin nanofibers showed a near-complete loading efficiency of quercetin and followed a fast-releasing profile of quercetin. Both HP-ß-CD/Quercetin and HP-γ-CD/Quercetin nanofibers showed significantly higher antioxidant activity compared to pristine quercetin. The HP-ß-CD/Quercetin and HP-γ-CD/Quercetin nanofibers also showed antibacterial activity, and with the addition of chitosan in the HP-γ-CD/Quercetin system, the Chitosan/HP-γ-CD/Quercetin nanofibers completely eliminated the investigated bacteria species. The nanofibers were nontoxic and well-tolerated by cells, and exploiting the quercetin and chitosan anti-inflammatory activities resulted in the downregulation of IL-6 and NO secretion in both immune as well as regenerative cells. Overall, CD inclusion complexation markedly enhances quercetin solubility, resulting in a biofunctional antioxidant, antibacterial, and anti-inflammatory wound dressing through a nanofibrous coating on cotton textiles.


Subject(s)
Anti-Bacterial Agents , Anti-Inflammatory Agents , Antioxidants , Bandages , Chitosan , Cyclodextrins , Materials Testing , Nanofibers , Quercetin , Quercetin/pharmacology , Quercetin/chemistry , Antioxidants/pharmacology , Antioxidants/chemistry , Nanofibers/chemistry , Chitosan/chemistry , Chitosan/pharmacology , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/chemistry , Cyclodextrins/chemistry , Cyclodextrins/pharmacology , Particle Size , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Microbial Sensitivity Tests , Cotton Fiber , Wound Healing/drug effects , Humans , Picrates/antagonists & inhibitors , Cell Survival/drug effects , Biphenyl Compounds
5.
J Microbiol Methods ; 224: 107010, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39098403

ABSTRACT

The quantification of cellular metabolic activity via MTT assay has become a widespread practice in eukaryotic cell studies and is progressively extending to bacterial cell investigations. This study pioneers the application of MTT assay to evaluate the metabolic activity of biofilm-forming cells within bacterial biofilms on nanofibrous materials. The biofilm formation of Staphylococcus aureus and Escherichia coli on nanomaterials electrospun from polycaprolactone (PCL), polylactic acid (PLA), and polyamide (PA) was examined. Various parameters of the MTT assay were systematically investigated, including (i) the dissolution time of the formed formazan, (ii) the addition of glucose, and (iii) the optimal wavelength for spectrophotometric determination. Based on interim findings, a refined protocol suitable for application to nanofibrous materials was devised. We recommend 2 h of the dissolution, the application of glucose, and spectrophotometric measurement at 595 nm to obtain reliable data. Comparative analysis with the reference CFU counting protocol revealed similar trends for both tested bacteria and all tested nanomaterials. The proposed MTT protocol emerges as a suitable method for assessing the metabolic activity of bacterial biofilms on PCL, PLA, and PA nanofibrous materials.


Subject(s)
Biofilms , Escherichia coli , Nanofibers , Polyesters , Staphylococcus aureus , Tetrazolium Salts , Biofilms/growth & development , Staphylococcus aureus/physiology , Nanofibers/chemistry , Escherichia coli/physiology , Tetrazolium Salts/metabolism , Tetrazolium Salts/chemistry , Polyesters/chemistry , Thiazoles/metabolism , Glucose/metabolism , Spectrophotometry/methods , Nylons/chemistry
6.
Sci Rep ; 14(1): 19256, 2024 08 20.
Article in English | MEDLINE | ID: mdl-39164352

ABSTRACT

Nanofibers show promise for wound healing by facilitating active agent delivery, moisture retention, and tissue regeneration. However, selecting suitable dressings for diverse wound types and managing varying exudate levels remains challenging. This study synthesized carbon quantum dots (CQDs) from citrate salt and thiourea using a hydrothermal method. The CQDs displayed antibacterial activity against Staphylococcus aureus and Escherichia coli. A nanoscaffold comprising gelatin, chitosan, and polycaprolactone (GCP) was synthesized and enhanced with silver nanoparticle-coated CQDs (Ag-CQDs) to form GCP-Q, while citrate addition yielded GCP-QC. Multiple analytical techniques, including electron microscopy, FT-IR spectroscopy, dynamic light scattering, UV-Vis, photoluminescence, X-ray diffraction, porosity, degradability, contact angle, and histopathology assessments characterized the CQDs and nanofibers. Integration of CQDs and citrate into the GCP nanofibers increased porosity, hydrophilicity, and degradability-properties favorable for wound healing. Hematoxylin and eosin staining showed accelerated wound closure with GCP-Q and GCP-QC compared to GCP alone. Overall, GCP-Q and GCP-QC nanofibers exhibit significant potential for skin tissue engineering applications.


Subject(s)
Anti-Bacterial Agents , Bandages , Carbon , Citric Acid , Escherichia coli , Nanofibers , Quantum Dots , Staphylococcus aureus , Wound Healing , Quantum Dots/chemistry , Nanofibers/chemistry , Wound Healing/drug effects , Carbon/chemistry , Citric Acid/chemistry , Staphylococcus aureus/drug effects , Escherichia coli/drug effects , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Animals , Chitosan/chemistry , Polyesters/chemistry , Gelatin/chemistry , Metal Nanoparticles/chemistry
7.
Methods Mol Biol ; 2835: 289-300, 2024.
Article in English | MEDLINE | ID: mdl-39105924

ABSTRACT

Hydrogels are a class of biomaterials that can provide a three-dimensional (3D) environment capable of mimicking the extracellular matrix of native tissues. In this chapter, we present a method to generate electrospun nanofibers for the purpose of reinforcing hydrogels. The addition of electrospun fibers can be used to improve the mechanical properties of hydrogels and broaden their range of applications. First, the polymer for making the electrospun fibers is formulated using chloroform/ethanol, polycaprolactone (PCL), polyethylene glycol (PEG), and polyethylene glycol diacrylate (PEGDA). Second, the polymer is used to generate thin electrospun nanofibers by an electrospinning technique using aluminum foil as a collector, which acts as the conductive substrate that collects the charged fibers. Third, the resulting electrospun fibers undergo a filtration process using nylon membrane filters, followed by lyophilization, ensuring complete removal of water from the sample.


Subject(s)
Hydrogels , Nanofibers , Polyethylene Glycols , Nanofibers/chemistry , Polyethylene Glycols/chemistry , Hydrogels/chemistry , Biocompatible Materials/chemistry , Humans , Cell- and Tissue-Based Therapy/methods , Polyesters/chemistry , Tissue Scaffolds/chemistry , Tissue Engineering/methods
8.
Methods Mol Biol ; 2835: 325-337, 2024.
Article in English | MEDLINE | ID: mdl-39105928

ABSTRACT

Gelatin, a protein derivative from collagen, is a versatile material with promising applications in tissue engineering. Among the various forms of gelatin scaffolds, nanofibrous gelatin microspheres (NFGMs) are attracting research efforts due to their fibrous nature and injectability. However, current methods for synthesizing nanofibrous gelatin microspheres (NFGMs) have limitations, such as wide size distributions and the use of toxic solvents. To address these challenges, the article introduces a novel approach. First, it describes the creation of a microfluidic device using readily available supplies. Subsequently, it outlines a unique process for producing monodispersed NFGMs through a combination of the microfluidic device and thermally induced phase separation (TIPS). This innovative method eliminates the need for sieving and the use of toxic solvents, making it a more ecofriendly and efficient alternative.


Subject(s)
Gelatin , Microspheres , Nanofibers , Gelatin/chemistry , Nanofibers/chemistry , Tissue Engineering/methods , Microfluidics/methods , Microfluidics/instrumentation , Tissue Scaffolds/chemistry , Microfluidic Analytical Techniques/instrumentation , Microfluidic Analytical Techniques/methods
9.
ACS Appl Mater Interfaces ; 16(32): 41927-41938, 2024 Aug 14.
Article in English | MEDLINE | ID: mdl-39090773

ABSTRACT

The complex microenvironment of diabetic wounds often hinders the healing process, ultimately leading to the formation of diabetic foot ulcers and even death. Dual monitoring and treatment of wounds can significantly reduce the incidence of such cases. Herein, a multifunctional Janus membrane (3D chitosan sponge-ZE/polycaprolactone nanofibers-ZP) was developed by incorporating the zinc metal-organic framework, europium metal-organic framework, and phenol red into nanofibers for diabetic wound monitoring and treatment. The directional water transport capacity of the resulting Janus membrane allows for unidirectional and irreversible drainage of wound exudate, and the multifunctional Janus membrane creates up to a 99% antibacterial environment, both of which can treat wounds. Moreover, the pH (5-8) and H2O2 (0.00-0.80 µM) levels of the wound can be monitored using the color-changing property of phenol red and the fluorescence characteristic of Eu-MOF on the obtained membrane, respectively. The healing stages of the wound can also be monitored by analyzing the RGB values of the targeted membrane images. This design can more accurately reflect the wound state and treat the wound to reduce bacterial infection and accelerate wound healing, which has been demonstrated in in vivo experiments. The results provide an important basis for early intervention in diabetic patients.


Subject(s)
Anti-Bacterial Agents , Metal-Organic Frameworks , Nanofibers , Wound Healing , Wound Healing/drug effects , Animals , Nanofibers/chemistry , Nanofibers/therapeutic use , Metal-Organic Frameworks/chemistry , Metal-Organic Frameworks/pharmacology , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Polyesters/chemistry , Chitosan/chemistry , Zinc/chemistry , Phenolsulfonphthalein/chemistry , Europium/chemistry , Mice , Humans , Membranes, Artificial , Hydrogen Peroxide/chemistry , Diabetes Mellitus, Experimental/drug therapy , Diabetic Foot/drug therapy , Diabetic Foot/pathology , Staphylococcus aureus/drug effects
10.
ACS Appl Mater Interfaces ; 16(32): 41869-41880, 2024 Aug 14.
Article in English | MEDLINE | ID: mdl-39101935

ABSTRACT

Diabetic wounds that do not heal for a long time challenge global healthcare. Mesenchymal stem cell (MSC) therapy has positive significance in promoting diabetic wound healing. However, traditional MSC therapy involves exogenous MSCs, which brings many limitations and unsatisfactory treatment. Moreover, the maintenance of MSC viability and function is difficult because of the high level of reactive oxygen species (ROS) in diabetic wounds. Therefore, we developed a nanofibrous dressing to recruit and protect endogenous MSCs while avoiding the inherent disadvantages of exogenous MSCs. Ceria nanoparticles capable of ROS scavenging are integrated into the nanofibrous dressings, together with Apt19S, a DNA aptamer with affinity and selectivity for MSCs. In addition, the homogenization and freeze-drying technology give the nanofibrous dressings good elasticity, which protects the wound from external pressure. Further experiments in diabetic mice show that the dressing has excellent endogenous MSC recruitment and anti-inflammatory properties, thereby synergistically promoting diabetic wound healing. This study is expected to explore an efficient method of stem cell therapy, providing a new way to construct high-performance wound dressings.


Subject(s)
Bandages , Diabetes Mellitus, Experimental , Mesenchymal Stem Cells , Nanofibers , Wound Healing , Animals , Wound Healing/drug effects , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Mice , Nanofibers/chemistry , Diabetes Mellitus, Experimental/therapy , Reactive Oxygen Species/metabolism , Male , Aptamers, Nucleotide/chemistry , Elasticity , Humans , Cerium
11.
ACS Synth Biol ; 13(8): 2567-2576, 2024 Aug 16.
Article in English | MEDLINE | ID: mdl-39092670

ABSTRACT

Collagen II (COL2) is the major component of cartilage tissue and is widely applied in pharmaceuticals, food, and cosmetics. In this study, COL fragments were extracted from human COL2 for secretory expression in Pichia pastoris. Three variants were successfully secreted by shake flask cultivation with a yield of 73.3-100.7 mg/L. The three COL2 variants were shown to self-assemble into triple-helix at 4 °C and capable of forming higher order assembly of nanofiber and hydrogel. The bioactivities of the COL2 variants were validated, showing that sample 205 exhibited the best performance for inducing fibroblast differentiation and cell migration. Meanwhile, sample 205 and 209 exhibited higher capacity for inducing in vitro blood clotting than commercial mouse COL1. To overexpress sample 205, the expression cassettes were constructed with different promoters and signal peptides, and the fermentation condition was optimized, obtaining a yield of 172 mg/L for sample 205. Fed-batch fermentation was carried out using a 5 L bioreactor, and the secretory protease Pep4 was knocked out to avoid sample degradation, finally obtaining a yield of 3.04 g/L. Here, a bioactive COL2 fragment was successfully identified and can be overexpressed in P. pastoris; the variant may become a potential biomaterial for skin care.


Subject(s)
Collagen Type II , Humans , Collagen Type II/genetics , Collagen Type II/metabolism , Mice , Animals , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Recombinant Proteins/biosynthesis , Fermentation , Pichia/genetics , Pichia/metabolism , Cell Movement/genetics , Fibroblasts/metabolism , Cell Differentiation , Bioreactors , Saccharomycetales/genetics , Saccharomycetales/metabolism , Nanofibers/chemistry
12.
ACS Appl Bio Mater ; 7(8): 5268-5278, 2024 Aug 19.
Article in English | MEDLINE | ID: mdl-39093691

ABSTRACT

Currently the prevalence of diabetic wounds brings a huge encumbrance onto patients, causing high disability and mortality rates and a major medical challenge for society. Therefore, in this study, we are targeting to fabricate aloe vera extract infused biocompatible nanofibrous patches to facilitate the process of diabetic wound healing. Additionally, clindamycin has been adsorbed onto the surface of in-house synthesized ceria nanoparticles and again used separately to design a nanofibrous web, as nanoceria can act as a good drug delivery vehicle and exhibit both antimicrobial and antidiabetic properties. Various physicochemical characteristics such as morphology, porosity, and chemical composition of the produced nanofibrous webs were investigated. Bacterial growth inhibition and antibiofilm studies of the nanofibrous materials confirm its antibacterial and antibiofilm efficacy against Gram-positive and Gram-negative bacteria. An in vitro drug release study confirmed that the nanofibrous mat show a sustained drug release pattern (90% of drug in 96 h). The nanofibrous web containing drug loaded nanoceria not only showed superior in vitro performance but also promoted greater wound contraction (95 ± 2%) in diabetes-induced mice in just 7 days. Consequently, it efficaciously lowers the serum glucose level, inflammatory cytokines, oxidative stress, and hepatotoxicity markers as endorsed by various ex vivo tests. Conclusively, this in-house-fabricated biocompatible nanofibrous patch can act as a potential medicated suppository that can be used for treating diabetic wounds in the proximate future.


Subject(s)
Aloe , Anti-Bacterial Agents , Bandages , Cerium , Diabetes Mellitus, Experimental , Nanofibers , Plant Extracts , Wound Healing , Cerium/chemistry , Cerium/pharmacology , Animals , Mice , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Wound Healing/drug effects , Nanofibers/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Aloe/chemistry , Diabetes Mellitus, Experimental/drug therapy , Polyurethanes/chemistry , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Polyethylene Glycols/chemistry , Materials Testing , Particle Size , Microbial Sensitivity Tests , Nanoparticles/chemistry , Hypoglycemic Agents/chemistry , Hypoglycemic Agents/pharmacology , Hypoglycemic Agents/administration & dosage , Male , Gram-Negative Bacteria/drug effects
13.
Stem Cell Res Ther ; 15(1): 262, 2024 Aug 15.
Article in English | MEDLINE | ID: mdl-39148112

ABSTRACT

BACKGROUND: Mesenchymal stem cells (MSCs)-based treatment strategy has shown promise in bolstering the healing process of chronic wounds in diabetic patients, who are at risk of amputation and mortality. To overcome the drawbacks of suboptimal cell retention and diminished cell viability at the injury site, a novel nanofibrous biomaterial-based scaffold was developed by using a controlled extrusion of a polymeric solution to deliver the cells (human adipose-derived MSCs (ADMSCs) and placenta-derived MSCs (PLMSCs)) locally to the animal model of diabetic ulcers. METHODS: The physicochemical and biological properties of the nano-bioscaffold were characterized in terms of microscopic images, FTIR spectroscopy, tensile testing, degradation and swelling tests, contact angle measurements, MTT assay, and cell attachment evaluation. To evaluate the therapeutic efficacy, a study using an excisional wound model was conducted on diabetic rats. RESULTS: The SEM and AFM images of scaffolds revealed a network of uniform nanofibers with narrow diameters between 100-130 nm and surface roughness less than 5 nm, respectively. ADMSCs and PLMSCs had a typical spindle-shaped or fibroblast-like morphology when attached to the scaffold. Desired characteristics in terms of swelling, hydrophilicity, biodegradation rate, and biocompatibility were achieved with the CS70 formulation. The wound healing process was accelerated according to wound closure rate assay upon treatment with MSCs loaded scaffold resulting in increased re-epithelialization, neovascularization, and less inflammatory reaction. Our findings unequivocally demonstrated that the cell-loaded nano-bioscaffold exhibited more efficacy compared with its acellular counterpart. In summation, our study underscores the potential of this innovative cellular scaffold as a viable solution for enhancing the healing of diabetic ulcers. CONCLUSION: The utilization of MSCs in a nanofibrous biomaterial framework demonstrates significant promise, providing a novel avenue for advancing wound care and diabetic ulcer management.


Subject(s)
Chitosan , Diabetes Mellitus, Experimental , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Nanofibers , Tissue Scaffolds , Wound Healing , Animals , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Nanofibers/chemistry , Rats , Humans , Diabetes Mellitus, Experimental/therapy , Tissue Scaffolds/chemistry , Chitosan/chemistry , Mesenchymal Stem Cell Transplantation/methods , Female , Male , Pregnancy , Adipose Tissue/cytology , Placenta/cytology
14.
PLoS One ; 19(8): e0308026, 2024.
Article in English | MEDLINE | ID: mdl-39088569

ABSTRACT

Electrospinning is a process in which high voltage creates nanostructured fibers with random orientation from a polymer solution. A novel electrospinning instrument was designed and constructed, capable of orienting and collimating the trajectory of the electrified fluid jet. The equipment collimates and adjusts the electrified fluid jet in the X-Y directions using deflector plates connected to a variable electric field. Simultaneously, different membrane thicknesses can be selected, i.e., in the Z direction. Additionally, by programming the sinusoidal function generator to perform an X-Y sweep, Lissajous figures (LF) were obtained. SEM images obtained through XYZ electrospinning of PVC and PVDF membranes were used to determine the control achieved over the orientation distribution of the processed nanofibers and the modification of their diameter, with and without applying the electric field to the deflector plates. The nanofibers obtained from the polymeric membranes, which originated after the straight segment of the Taylor cone, did not exhibit a random trajectory and position. Instead, the collimated electrified fluid jet deposited them in a cross pattern (X-Y) on the collector-cathode plate.


Subject(s)
Electricity , Nanofibers , Polymers , Nanofibers/chemistry , Polymers/chemistry , Polyvinyl Chloride/chemistry , Polyvinyls/chemistry , Fluorocarbon Polymers
15.
Int J Pharm ; 661: 124425, 2024 Aug 15.
Article in English | MEDLINE | ID: mdl-38971509

ABSTRACT

Dry powder inhalers (DPIs) are the first choice for inhalation drug development. However, some conventional DPI formulation processes require heating, which may damage high molecular weight drugs such as proteins and nucleic acids. In this study, we propose a novel DPI preparation process that avoids the use of heat. Dry powders were prepared by cryomilling nanofiber mats composed of polyvinyl alcohol, D(-)-mannitol (Man), and α-chymotrypsin (α-Chy) as the model drug using the electrospinning method. The addition of Man conferred high dispersibility and excellent in vitro aerosol performance to the nanofiber mat powder in a very short milling time (less than 0.5 min) as assessed using the Andersen cascade impactor. Powders were classified according to the degree of friability, and among these, nanofiber mats containing 15 % Man and milled for 0.25 min exhibited the highest aerosol performance. Nanofiber mats containing Man milled for less than 0.5 min also exhibited greater α-Chy enzymatic activity than a nebulized α-Chy solution. Furthermore, single inhalation induced no significant lung tissue damage as evidenced by lactate dehydrogenase activity assays of mouse bronchoalveolar lavage fluid. This novel DPI formulation process may facilitate the safe and efficient inhalational delivery of therapeutic proteins.


Subject(s)
Aerosols , Chymotrypsin , Mannitol , Nanofibers , Nanofibers/chemistry , Nanofibers/administration & dosage , Animals , Administration, Inhalation , Mannitol/chemistry , Chymotrypsin/chemistry , Mice , Dry Powder Inhalers , Polyvinyl Alcohol/chemistry , Powders , Drug Delivery Systems , Lung/metabolism , Bronchoalveolar Lavage Fluid/chemistry , Male
16.
ACS Appl Mater Interfaces ; 16(31): 40411-40427, 2024 Aug 07.
Article in English | MEDLINE | ID: mdl-39044386

ABSTRACT

The treatment of critical-sized bone defects caused by tumor removal, skeletal injuries, or infections continues to pose a major clinical challenge. A popular potential alternative solution to autologous bone grafts is a tissue-engineered approach that utilizes the combination of mesenchymal stromal/stem cells (MSCs) with synthetic biomaterial scaffolds. This approach aims to support new bone formation by mimicking many of the biochemical and biophysical cues present within native bone. Regrettably, osteocyte cells, crucial for bone maturation and homeostasis, are rarely produced within MSC-seeded scaffolds, thereby restricting the development of fully mature cortical bone from these synthetic implants. In this work, we have constructed a multimodal scaffold by combining electrospun poly(lactic-co-glycolic acid) (PLGA) fibrous scaffolds with poly(ethylene glycol) (PEG)-based hydrogels that mimic the functional unit of cortical bone, osteon (osteon-mimetic) scaffolds. These scaffolds were decorated with a novel bone morphogenic protein-6 (BMP6) peptide (BMP6p) after our findings revealed that the BMP6p drives higher levels of Smad signaling than the full-length protein counterpart, soluble or when bound to the PEG hydrogel backbone. We show that our osteon-mimetic scaffolds, in presenting concentric layers of BMP6p-PEG hydrogel overlaid on MSC-seeded PLGA nanofibers, promoted the rapid formation of osteocyte-like cells with a phenotypic dendritic morphology, producing early osteocyte markers, including E11/gp38 (E11). Maturation of these osteocyte-like cells was further confirmed by the observation of significant dentin matrix protein 1 (DMP1) throughout our bilayered scaffolds after 3 weeks, even when cultured in a medium without dexamethasone (DEX) or any other osteogenic supplements. These results demonstrate that these osteon-mimetic scaffolds, in presenting biochemical and topographical cues reminiscent of the forming osteon, can drive the formation of osteocyte-like cells in vitro from hBMSCs without the need for any osteogenic factor media supplementation.


Subject(s)
Biomimetic Materials , Mesenchymal Stem Cells , Nanofibers , Osteocytes , Osteogenesis , Tissue Scaffolds , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Mesenchymal Stem Cells/drug effects , Tissue Scaffolds/chemistry , Nanofibers/chemistry , Humans , Osteogenesis/drug effects , Biomimetic Materials/chemistry , Biomimetic Materials/pharmacology , Osteocytes/cytology , Osteocytes/metabolism , Osteocytes/drug effects , Polylactic Acid-Polyglycolic Acid Copolymer/chemistry , Bone Morphogenetic Protein 6/chemistry , Bone Morphogenetic Protein 6/pharmacology , Bone Morphogenetic Protein 6/metabolism , Polyethylene Glycols/chemistry , Cell Differentiation/drug effects , Tissue Engineering/methods , Hydrogels/chemistry , Hydrogels/pharmacology
17.
ACS Appl Mater Interfaces ; 16(31): 40767-40786, 2024 Aug 07.
Article in English | MEDLINE | ID: mdl-39047263

ABSTRACT

A modular and 3D compartmentalized microfluidic system with electrospun porous membranes (PMs) for epithelialized organ-on-a-chip systems is presented. Our novel approach involves direct deposition of polymer nanofibers onto a patterned poly(methyl methacrylate) (PMMA) substrate using electrospinning, resulting in an integrated PM within the microfluidic chip. The in situ deposition of the PM eliminates the need for additional assembly processes. To demonstrate the high throughput membrane integration capability of our approach, we successfully deposited nanofibers onto various chip designs with complex microfluidic planar structures and expanded dimensions. We characterized and tested the fully PMMA chip by growing an epithelial monolayer using the Caco-2 cell line to study drug permeability. A comprehensive analysis of the bulk and surface properties of the membrane's fibers made of PMMA and polystyrene (PS) was conducted to determine the polymer with the best performance for cell culture and drug transport applications. The PMMA-based membrane, with a PMMA/PVP ratio of 5:1, allowed for the fabrication of a uniform membrane structure along the aligned nanofibers. By modulating the fiber diameter and total thickness of the membrane, we could adjust the membrane's porosity for specific cell culture applications. The PMMA-PVP nanofibers exhibited a low polydispersity index value, indicating monodispersed nanofibers and a more homogeneous and uniform fiber network. Both types of membranes demonstrated excellent mechanical integrity under medium perfusion flow rates. However, the PMMA-PVP composition offered a tailored porous structure with modulable porosity based on the fiber diameter and thickness. Our developed platform enables dynamic in vitro modeling of the epithelial barrier and has applications in drug transport and in vitro microphysiological systems.


Subject(s)
Lab-On-A-Chip Devices , Nanofibers , Polymethyl Methacrylate , Humans , Caco-2 Cells , Porosity , Polymethyl Methacrylate/chemistry , Nanofibers/chemistry , Membranes, Artificial , Polystyrenes/chemistry
18.
Biomacromolecules ; 25(8): 5039-5047, 2024 Aug 12.
Article in English | MEDLINE | ID: mdl-39041249

ABSTRACT

In this study, we explore an approach to enhance the mechanical performance of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) by utilizing the self-reinforcing effect of ß-phase-induced PHBV electrospun nanofiber mats. This involves electrospinning combined with low-temperature postspun vapor solvent interfiber welding. Scanning electron microscopy imaging confirmed fiber alignment, while XRD diffraction revealed the presence of both α and ß crystalline phases under optimized electrospinning conditions. The resulting composite exhibited significant improvements in mechanical properties attributed to the formation of more perfectly structured α and ß polymorphs and enhanced interfacial adhesion of electrospun nanofibers after vapor solvent treatment. This approach offers entirely recyclable and biodegradable materials, presenting the potential for a new family of sustainable bioplastics.


Subject(s)
Nanofibers , Polyesters , Solvents , Polyesters/chemistry , Nanofibers/chemistry , Solvents/chemistry , Microscopy, Electron, Scanning/methods , Biocompatible Materials/chemistry , Polyhydroxybutyrates
19.
Int J Biol Macromol ; 275(Pt 1): 133609, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38960220

ABSTRACT

Developing plastic/fluorine/silicon-free and degradable water/oil-resistant coatings for paper-based packaging materials to replace disposable plastic products is a very effective way to solve the problem of 'white pollution' or microplastics pollution. A novel water/oil-resistant coating was developed by alkyl ketene dimer (AKD)-based Pickering emulsion and chitosan in this work. Cellulose nanofibrils (CNF) were used as a stabilizing solid for AKD emulsion, with the addition of chitosan as an oil-resistance agent. The coating provides excellent hydrophobicity, water/oil resistance as well as good barrier properties. The water contact angle was as high as 130° and the minimum Cobb60 value was 5.7 g/m2, which was attributed to the hydrophobicity of AKD. In addition, the kit rating reached maximum 12/12 at coating weight of 8.26 g/m2 and the water vapor transmittance rate (WVTR) was reduced to 153.4 g/(m2⋅day) at the coating weight of 10.50 g/m2. The tensile strength of the paper was increased from 28.1 to 43.6 MPa after coating. Overall, this coating can effectively improve the performance of paper-based materials, which may play an important role in the process of replacing disposable plastic packaging with paper-based materials.


Subject(s)
Cellulose , Chitosan , Emulsions , Oils , Paper , Water , Chitosan/chemistry , Cellulose/chemistry , Emulsions/chemistry , Water/chemistry , Oils/chemistry , Hydrophobic and Hydrophilic Interactions , Tensile Strength , Nanofibers/chemistry
20.
Int J Biol Macromol ; 275(Pt 1): 133598, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38960244

ABSTRACT

A novel kind of protective apparel for handicapped persons has been created with bio-based electrospun nanofibrous (NFs) membranes. Hydrophobic membranes with fine polylactic acid (PLA) NFs had a smooth, bead-less structure with an average diameter of 950 nm. The hydrophilic layer has a similar pattern but a smaller fiber diameter dispersion and an average diameter of 750 nm. The silica nanoparticle-modified super-hydrophobic top layer (contact angle, ~153°) repels water and keeps the user dry. Super-hydrophilic silver nanoparticles in the fabric's bottom layer react with perspiration to kill microorganisms. The fabric's porosity (avg. 1.2-1.5 µm) allows for breathability, while silica nanoparticles boost infrared radiation reflection, keeping users cool on hot days. The dual-layer textile has 4.9 MPa ultimate tensile strength and 68 % elongation compared to the membrane's super-hydrophobic and super-hydrophilic layers. Wearing protective clothes reduced hand temperature by 25 % in direct sunlight and 13 % in a sun simulator with 1 Sun. This fabric will work well for adult diapers, outdoor clothing, and disability accessories. Overall, the protective textiles may improve the quality of life for disabled and elderly people by providing usable textile items adapted to their needs.


Subject(s)
Disabled Persons , Hydrophobic and Hydrophilic Interactions , Nanofibers , Polyesters , Nanofibers/chemistry , Humans , Polyesters/chemistry , Membranes, Artificial , Protective Clothing , Silver/chemistry , Textiles , Silicon Dioxide/chemistry , Metal Nanoparticles/chemistry , Tensile Strength
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