UNASSIGNED: This study aims to explore a novel scaffold for osteotendinous junction regeneration and to preliminarily verify its osteogenic and tenogenic abilities in vitro.
UNASSIGNED: A polycaprolactone (PCL) scaffold with aligned and orthogonal fibers was created using melt electrowriting (MEW) and fused deposition modeling (FDM). The scaffold was coated with Type I collagen, and hydroxyapatite was carefully added to separate the regions intended for bone and tendon regeneration, before being rolled into a cylindrical shape. Human adipose-derived stem cells (hADSCs) were seeded to evaluate viability and differentiation. Scaffold characterization was performed with Scanning Electron Microscope (SEM). Osteogenesis was assessed by alkaline phosphatase (ALP) and Alizarin red staining, while immunostaining and transcription-quantitative polymerase chain reaction (RT-qPCR) evaluated osteogenic and tendogenic markers.
UNASSIGNED: Scaffolds were developed in four variations: aligned (A), collagen-coated aligned (A+C), orthogonal (O), and mineral-coated orthogonal (O+M). SEM analysis confirmed surface morphology and energy-dispersive X-ray spectroscopy (EDS) verified mineral coating on O+M types. Hydrophilicity and mechanical properties were optimized in modified scaffolds, with A+C showing increased tensile strength and O+M improved in compression. hADSCs demonstrated good viability and morphology across scaffolds, withO+M scaffolds showing higher cell proliferation and osteogenic potential, and A and A+C scaffolds supporting tenogenic differentiation.
UNASSIGNED: This study confirms the potential of a novel PCL scaffold with distinct regions for osteogenic and tenogenic differentiation, supporting the regeneration of osteotendinous junctions in vitro.

Quaternized chitosan (HTCC) was synthesized and characterized to increase chitosan solubility. Then HTCC was electrospun with poly (vinyl alcohol) (PVA) and prepared natural bioactive agent (Calendula officinalis) extract was loaded onto fibers for wound scaffold applications. Morphological, structural, and mechanical characterization of the produced wound scaffolds was performed and their in vitro bioactive component release behavior was investigated. As a result, it was observed that the degree of quaternization of chitosan was 0.89, and synthesized HTCC was soluble in acidic, basic, alkaline media and could be electrospun with PVA in the presence of a natural bioactive agent. The presence of HTCC increased Young\'s modulus and the tensile strength of the PVA scaffolds, while the presence of bioactive extract caused a decrease in Young\'s modulus and an increase in tensile strength. Calendula officinalis is released in a controlled and slow manner from the scaffolds within approximately 55 h. The release behavior was consistent with the Higuchi kinetic model. In this study, the effect of PVA cooperator on HTCC nanofiber production in the presence of a bioactive component was investigated for the first time. HTCC and Calendula officinalis extract were also used together for the first time in the composition of a fiber scaffold. The mechanical properties and release kinetics of these scaffolds were also investigated for the first time. According to the results, it is thought that the wound scaffolds produced have the potential to be used as a new treatment tool, especially for chronic wounds.

Biomaterial scaffolds boost tissue repair and regeneration by providing physical support, delivering biological signals and/or cells, and recruiting endogenous cells to facilitate tissue-material integration and remodeling. Foreign body response (FBR), an innate immune response that occurs immediately after biomaterial implantation, is a critical factor in determining the biological outcomes of biomaterial scaffolds. Electrospinning is of great simplicity and cost-effectiveness to produce nanofiber scaffolds with well-defined physicochemical properties and has been used in a variety of regenerative medicine applications in preclinical trials and clinical practice. A deep understanding of causal factors between material properties and FBR of host tissues is beneficial to the optimal design of electrospun scaffolds with favorable immunomodulatory properties. We herein prepared and characterized three electrospun scaffolds with distinct fiber configurations and investigated their effects on FBR in terms of immune cell-material interactions and host responses. Our results show that electrospun yarn scaffold results in greater cellular immune reactions and elevated FBR inin vivoassessments. Although the yarn scaffold showed aligned fiber bundles, it failed to induce cell elongation of macrophages due to its rough surface and porous grooves between yarns. In contrast, the aligned scaffold showed reduced FBR compared to the yarn scaffold, indicating a smooth surface is also a contributor to the immunomodulatory effects of the aligned scaffold. Our study suggests that balanced porousness and smooth surface of aligned fibers or yarns should be the key design parameters of electrospun scaffolds to modulate host responsein vivo.

Enhancing the mechanical properties and cytocompatibility of decellularized heart valves is the key to promote the application of biological heart valves. In order to further improve the mechanical properties, the electrospinning and non-woven processing methods are combined to prepare the polylactic acid (PLA)/decellularized heart valve nanofiber-reinforced sandwich structure electrospun scaffold. The effect of electrospinning time on the performance of decellularized heart valve is investigated from the aspects of morphology, mechanical properties, softness, and biocompatibility of decellularized heart valve. Results of the mechanical tests show that compared with the pure decellularized heart valve, the mechanical properties of the composite heart valve were significantly improved with the tensile strength increasing by 108% and tensile strain increased by 571% when the electrospinning time exceeded 2 h. In addition, with this electrospinning time, the composite heart valve has a certain promoting effect on the human umbilical vein endothelial cells proliferation behavior. This work provides a promising foundation for tissue heart valve reendothelialization to lay the groundwork for organoid.

Biomaterial-mediated, spatially localized gene delivery is important for the development of cell-populated scaffolds used in tissue engineering. Cells adhering to or penetrating into such a scaffold are to be transfected with a preloaded gene that induces the production of secreted proteins or cell reprogramming. In the present study, we produced silica nanoparticles-associated pDNA and electrospun scaffolds loaded with such nanoparticles, and studied the release of pDNA from scaffolds and cell-to-scaffold interactions in terms of cell viability and pDNA transfection efficacy. The pDNA-coated nanoparticles were characterized with dynamic light scattering and transmission electron microscopy. Particle sizes ranging from 56 to 78 nm were indicative of their potential for cell transfection. The scaffolds were characterized using scanning electron microscopy, X-ray photoelectron spectroscopy, stress-loading tests and interaction with HEK293T cells. It was found that the properties of materials and the pDNA released vary, depending on the scaffold\'s composition. The scaffolds loaded with pDNA-nanoparticles do not have a pronounced cytotoxic effect, and can be recommended for cell transfection. It was found that (pDNA-NPs) + PEI9-loaded scaffold demonstrates good potential for cell transfection. Thus, electrospun scaffolds suitable for the transfection of inhabiting cells are eligible for use in tissue engineering.

Cardiac tissue engineering (CTE) is a promising way for the restoration of injured cardiac tissue in the healthcare system. The development of biodegradable scaffolds with appropriate chemical, electrical, mechanical, and biological properties is an unmet need for the success of CTE. Electrospinning is a versatile technique that has shown potential applications in CTE. Herein, four different types of multifunctional scaffolds, including synthetic-based poly (glycerol sebacate)-polyurethane (PGU), PGU-Soy scaffold, and a series of trilayer scaffolds containing two outer layers of PGU-Soy and a middle (inner) layer of gelatin (G) as a natural and biodegradable macromolecule without simvastatin (S) and with simvastatin (GS), an anti-inflammatory agent, were fabricated in the sandwich-like structure using electrospinning technique. This approach offers a combination of the advantages of both synthetic and natural polymers to enhance the bioactivity and the cell-to-cell and cell-to-matrix intercommunication. An in vitro drug release analysis was performed after the incorporation of soybean oil (Soy) and G. Soy is used as a semiconducting material was introduced to improve the electrical conductivity of nanofibrous scaffolds. The physicochemical properties, contact angle, and biodegradability of the electrospun scaffolds were also assessed. Moreover, the blood compatibility of nanofibrous scaffolds was studied through activated partial thromboplastin time (APTT), prothrombin time (PT), and hemolytic assay. The results showed that all scaffolds exhibited defect-free morphologies with mean fiber diameters in the range of 361 ± 109 to 417 ± 167 nm. A delay in blood clotting was observed, demonstrating the anticoagulant nature of nanofibrous scaffolds. Furthermore, rat cardiomyoblast cell lines (H9C2) were cultured on scaffolds for 7 days, and the morphology and cell arrangement were monitored. Data indicated an appropriate cytocompatibility. Of note, in the PGU-Soy/GS nanofibrous scaffold, a high survival rate was indicated compared to other groups. Our findings exhibited that the simvastatin-loaded polymeric system had positive effects on cardiomyoblasts attachment and growth and could be utilized as a drug release carrier in the field of CTE.

Adverse remodeling post-myocardial infarction is hallmarked by the phenotypic change of cardiac fibroblasts (CFs) into myofibroblasts (MyoFs) and over-deposition of the fibrotic extracellular matrix (ECM) mainly composed by fibronectin and collagens, with the loss of tissue anisotropy and tissue stiffening. Reversing cardiac fibrosis represents a key challenge in cardiac regenerative medicine. Reliable in vitro models of human cardiac fibrotic tissue could be useful for preclinical testing of new advanced therapies, addressing the limited predictivity of traditional 2D cell cultures and animal in vivo models. In this work, we engineered a biomimetic in vitro model, reproducing the morphological, mechanical, and chemical cues of native cardiac fibrotic tissue. Polycaprolactone (PCL)-based scaffolds with randomly oriented fibers were fabricated by solution electrospinning technique, showing homogeneous nanofibers with an average size of 131 ± 39 nm. PCL scaffolds were then surface-functionalized with human type I collagen (C1) and fibronectin (F) by dihydroxyphenylalanine (DOPA)-mediated mussel-inspired approach (PCL/polyDOPA/C1F), in order to mimic fibrotic cardiac tissue-like ECM composition and support human CF culture. BCA assay confirmed the successful deposition of the biomimetic coating and its stability during 5 days of incubation in phosphate-buffered saline. Immunostaining for C1 and F demonstrated their homogeneous distribution in the coating. AFM mechanical characterization showed that PCL/polyDOPA/C1F scaffolds, in wet conditions, resembled fibrotic tissue stiffness with an average Young\'s modulus of about 50 kPa. PCL/polyDOPA/C1F membranes supported human CF (HCF) adhesion and proliferation. Immunostaining for α-SMA and quantification of α-SMA-positive cells showed HCF activation into MyoFs in the absence of a transforming growth factor β (TGF-β) profibrotic stimulus, suggesting the intrinsic ability of biomimetic PCL/polyDOPA/C1F scaffolds to sustain the development of cardiac fibrotic tissue. A proof-of-concept study making use of a commercially available antifibrotic drug confirmed the potentialities of the developed in vitro model for drug efficacy testing. In conclusion, the proposed model was able to replicate the main hallmarks of early-stage cardiac fibrosis, appearing as a promising tool for future preclinical testing of advanced regenerative therapies.

Fibrous polyurethane-based scaffolds have proven to be promising materials for the tissue engineering of implanted medical devices. Sterilization of such materials and medical devices is an absolutely essential step toward their medical application. In the presented work, we studied the effects of two sterilization methods (ethylene oxide treatment and electron beam irradiation) on the fibrous scaffolds produced from a polyurethane-gelatin blend. Scaffold structure and properties were studied by scanning electron microscopy (SEM), atomic force microscopy (AFM), infrared spectroscopy (FTIR), a stress-loading test, and a cell viability test with human fibroblasts. Treatment of fibrous polyurethane-based materials with ethylene oxide caused significant changes in their structure (formation of glued-like structures, increase in fiber diameter, and decrease in pore size) and mechanical properties (20% growth of the tensile strength, 30% decline of the maximal elongation). All sterilization procedures did not induce any cytotoxic effects or impede the biocompatibility of scaffolds. The obtained data determined electron beam irradiation to be a recommended sterilization method for electrospun medical devices made from polyurethane-gelatin blends.

Polyhydroxybutyrate (PHB) is a natural-source biopolymer of the polyhydroxyalkanoate (PHA) family. Nanofibrous scaffolds prepared from this biological macromolecule have piqued the interest of researchers in recent years due to their unique properties. Nonetheless, these nanofibers continue to have problems such as low surface roughness and high hydrophobicity. In this research, PHB nanofibers were produced by the electrospinning method. Following that, the surface of nanofibers was modified by atmospheric plasma. Scanning electron microscopy (SEM), water contact angle (WCA), atomic force microscopy (AFM), tensile test, and cell behavior analyses were performed on mats to investigate the performance of treated and untreated samples. The achieved results showed a lower water contact angle (from ≃120° to 43°), appropriate degradation rate (up to ≃20 % weight loss in four months), and outstanding biomineralization (Ca/P ratio of ≃1.86) for the modified sample compared to the neat PHB. Finally, not only the MTT test show better viability of MG63 osteoblast cells, but also Alizarin staining, ALP, and SEM results likewise showed better cell proliferation in the presence of modified mats. These findings back up the claim that plasma surface modification is a quick, environmentally friendly, and low-cost way to improve the performance of nanofibers in bone tissue engineering.

Although there are several methods for fabricating nanofibrous scaffolds for biomedical applications, electrospinning is probably the most versatile and feasible process. Electrospinning enables the preparation of reproducible, homogeneous fibers from many types of polymers. In addition, implementation of this technique gives the possibility to fabricated polymer-based composite mats embroidered with manifold materials, such as graphene. Flake graphene and its derivatives represent an extremely promising material for imparting new, biomedically relevant properties, functions, and applications. Graphene oxide (GO) and reduced graphene oxide (rGO), among many extraordinary properties, confer antimicrobial properties of the resulting material. Moreover, graphene oxide and reduced graphene oxide promote the desired cellular response. Tissue engineering and regenerative medicine enable advanced treatments to regenerate damaged tissues and organs. This review provides a reliable summary of the recent scientific literature on the fabrication of nanofibers and their further modification with GO/rGO flakes for biomedical applications.