Search Results (38)

Aligned fibrous scaffolds are essential for directing soft-tissue regeneration, yet synthetic polymers lack native biochemical cues. To bridge this gap, bioactive and anisotropic scaffolds were developed by combining melt electrowriting (MEW) with decellularized extracellular matrix (dECM) decoration to enhance cell–scaffold interactions for soft tissue engineering. Porous polycaprolactone (PCL) scaffolds with aligned microfibers and tunable pore architectures (aspect ratios 1:1, 1:2, and 1:3) were fabricated via MEW and subsequently coated with porcine skeletal muscle dECM using a dip-gelation method. Comprehensive surface characterization confirmed the presence and robust adhesion of the dECM coating on the PCL scaffolds, which concurrently enhanced surface hydrophilicity. Furthermore, mechanical testing demonstrated that the resulting composite scaffold retained the structural integrity required to meet the mechanical demands of tissue regeneration. In vitro studies using L929 fibroblasts demonstrated that dECM decoration significantly improved cell adhesion, proliferation, and alignment along the fiber direction. Notably, scaffolds with 1:1 and 1:2 aspect ratios supported the highest cell density and guided morphological elongation most effectively. These findings highlight the synergistic potential of topographical cues and biochemical signaling in scaffold design for functional tissue regeneration. Full article

Nanotopography refers to the intricate surface characteristics of materials at the sub-micron (<1000 nm) and nanometer (<100 nm) scales. These topographical surface features significantly influence the physical, chemical, and biological properties of biomaterials, affecting their interactions with cells and surrounding tissues. The development of nanostructured surfaces of polymeric nanocomposites has garnered increasing attention in the fields of tissue engineering and regenerative medicine due to their ability to modulate cellular responses and enhance tissue regeneration. Various top-down and bottom-up techniques, including nanolithography, etching, deposition, laser ablation, template-assisted synthesis, and nanografting techniques, are employed to create structured surfaces on biomaterials. Additionally, nanotopographies can be fabricated using polymeric nanocomposites, with or without the integration of organic and inorganic nanomaterials, through advanced methods such as using electrospinning, layer-by-layer (LbL) assembly, sol–gel processing, in situ polymerization, 3D printing, template-assisted methods, and spin coating. The surface topography of polymeric nanocomposite scaffolds can be tailored through the incorporation of organic nanomaterials (e.g., chitosan, dextran, alginate, collagen, polydopamine, cellulose, polypyrrole) and inorganic nanomaterials (e.g., silver, gold, titania, silica, zirconia, iron oxide). The choice of fabrication technique depends on the desired surface features, material properties, and specific biomedical applications. Nanotopographical modifications on biomaterials’ surface play a crucial role in regulating cell behavior, including adhesion, proliferation, differentiation, and migration, which are critical for tissue engineering and repair. For effective tissue regeneration, it is imperative that scaffolds closely mimic the native extracellular matrix (ECM), providing a mechanical framework and topographical cues that replicate matrix elasticity and nanoscale surface features. This ECM biomimicry is vital for responding to biochemical signaling cues, orchestrating cellular functions, metabolic processes, and subsequent tissue organization. The integration of nanotopography within scaffold matrices has emerged as a pivotal regulator in the development of next-generation biomaterials designed to regulate cellular responses for enhanced tissue repair and organization. Additionally, these scaffolds with specific surface topographies, such as grooves (linear channels that guide cell alignment), pillars (protrusions), holes/pits/dots (depressions), fibrous structures (mimicking ECM fibers), and tubular arrays (array of tubular structures), are crucial for regulating cell behavior and promoting tissue repair. This review presents recent advances in the fabrication methodologies used to engineer nanotopographical microenvironments in polymeric nanocomposite tissue scaffolds through the incorporation of nanomaterials and biomolecular functionalization. Furthermore, it discusses how these modifications influence cellular interactions and tissue regeneration. Finally, the review highlights the challenges and future perspectives in nanomaterial-mediated fabrication of nanotopographical polymeric scaffolds for tissue engineering and regenerative medicine. Full article

The periodontal ligament (PDL) is a fibrous connective tissue that anchors the tooth-root surface to the alveolar bone with specific orientations. It plays a crucial role in functional restoration, optimal position stabilities, biomechanical stress transmission, and appropriate tissue remodeling in response to masticatory loading conditions. This pilot study explored spatial microarchitectures to promote PDL orientations while limiting mineralized tissue formation. A computer-designed perio-complex scaffold was developed with two parts: (1) PDL-guiding architectures with defined surface topography and (2) a bone region with open structures. After SEM analysis of micropatterned topographies on PDL-guiding architectures, perio-complex scaffolds were transplanted into two-wall periodontal defects in the canine mandible. Despite the limited bone formation at the 4-week timepoint, bone parameters in micro-CT quantifications showed statistically significant differences between the no-scaffold and perio-complex scaffold transplantation groups. Histological analyses demonstrated that the PDL-guiding architecture regulated fiber orientations and facilitated the functional restoration of PDL bundles in immunohistochemistry with periostin and decorin (DCN). The perio-complex scaffold exhibited predictable and controlled fibrous tissue alignment with specific angulations, ensuring spatial compartmentalization for PDL tissues and bone regenerations. These findings highlighted that the perio-complex scaffold could serve as an advanced therapeutic approach to contribute periodontal tissue regeneration and functional restoration in tooth-supporting structures. Full article

Anterior cruciate ligament (ACL) injuries pose significant challenges, driving the need for innovative repair strategies. Tissue engineering (TE) has emerged as a promising field for ACL injuries. Wet spinning is a filament production technique that enables precise control over filament alignment, diameter, and porosity, making it suitable for developing new scaffolds for ACL injuries. This study develops fibrous scaffolds using wet spinning of polycaprolactone (PCL) reinforced with cellulose nanocrystals (CNC) to enhance mechanical properties. Wet spinning was employed to fabricate fibrous scaffolds, utilizing PCL as the primary polymer due to its favorable biocompatibility and degradability. An automated collector was developed and optimized, which allowed the stretching of the PCL filaments to diameters as low as 30 µm. Several filaments were explored and characterized using SEM, TGA, and mechanical tests. The optimized PCL/CNC filaments were used to develop 3D braided structures that mimic ACL structure. The combination of wet-spun PCL/CNC filaments (with an optimized stirring method) and the braiding procedure allowed for fully biocompatible scaffolds that mimic both the structure and the mechanical properties of native ACL. Cytotoxicity and biocompatibility tests showed cell viability and proliferation values above 99% and 81%, respectively. These findings underscore the potential of CNC-reinforced PCL scaffolds as promising candidates for ACL repair, laying the groundwork for future biomedical applications. Full article

Ligament tears can strongly influence an individual’s daily life and ability to engage in physical activities. It is essential to develop artificial scaffolds for ligament repairs in order to effectively restore damaged ligaments. In this experiment, the objective was to evaluate fibrous membranes as scaffolds for ligament repair. These membranes were created through electrospinning using piezoelectric polyvinylidene fluoride (PVDF) composites, which contained 1 wt.% and 3 wt.% of copper-impregnated nanohydroxyapatite (Cu-nHA). The proposed electrospun membrane would feature an aligned fiber structure achieved through high-speed roller stretching, which mimics the properties of biomimetic ligaments. Nanoparticles of Cu-nHA had been composited into PVDF to enhance the pirzoelectric β-phase of the PVDF crystallines. The study assessed the physicochemical properties, antibacterial activity, and biocompatibility of the membranes in vitro. A microstructure analysis revealed that the composite membrane exhibited a bionic structure with aligned fibers resembling human ligaments. The piezoelectric performance of the experimental group containing 3 wt.% Cu-nHA was significantly improved to 25.02 ± 0.68 V/g·m⁻² compared with that of the pure PVDF group at 18.98 ± 1.18 V/g·m⁻². Further enhancement in piezoelectric performance by 31.8% was achieved by manipulating the semicrystalline structures. Antibacterial and cytotoxicity tests showed that the composite membrane inherited the antibacterial properties of Cu-nHA nanoparticles without causing cytotoxic reactions. Tensile tests revealed that the membrane’s flexibility of strain was adequate for use as artificial scaffolds for ligaments. In particular, the mechanical properties of the two experimental groups containing Cu-nHA were significantly enhanced compared with those of the pure PVDF group. The favorable piezoelectric and flexible properties are highly beneficial for ligament tissue regeneration. This study successfully developed PVDF/Cu-nHA piezoelectric fibers for a biocompatible, unidirectional piezoelectric membrane with potential applications as ligament repair scaffolds. Full article

A primary challenge following severe musculoskeletal trauma is incomplete muscle regeneration. Current therapies often fail to heal damaged muscle due to dysregulated healing programs and insufficient revascularization early in the repair process. There is a limited understanding of the temporal changes that occur during the early stages of muscle remodeling in response to engineered therapies. Previous work demonstrated that nanotopographically patterned scaffolds provide cytoskeletal guidance and direct endothelial angiogenic and anti-inflammatory phenotypes. The aim of this study was to evaluate how endothelial cell (EC) patterning guides temporal and histomorphological muscle remodeling after muscle injury. In the current study, mice were treated with EC-laden engineered constructs that exhibited either aligned or random patterning of collagen nanofibrils, following a volumetric muscle loss injury (VML). Remodeling was evaluated at 2, 7, and 21 days post injury. Over the 21-day study, all groups (Acellular Aligned, EC Aligned, EC Random) demonstrated similar significant increases in vascular density and myogenesis. Animals treated with acellular controls demonstrated a two-fold decrease in muscle cross-sectional area between days 2 and 21 post injury, consistent with VML-induced muscle atrophy; however, animals treated with patterned EC-laden constructs exhibited preservation of muscle mass. The implantation of an EC-laden construct led to a 50% increase in the number of animals exhibiting areas of fibrous remodeling adjacent to the construct, along with greater collagen deposition (p < 0.01) compared to acellular controls 21 days post injury. These findings suggest that nanotopographically patterned EC-laden constructs may guide early muscle-protective programs that support muscle mass retention through myo-vascular independent pathways. Full article

Skeletal muscle tissue (SMT) has a highly hierarchical and anisotropic morphology, featuring aligned and parallel structures at multiple levels. Various factors, including trauma and disease conditions, can compromise the functionality of skeletal muscle. The in vitro modeling of SMT represents a useful tool for testing novel drugs and therapies. The successful replication of SMT native morphology demands scaffolds with an aligned anisotropic 3D architecture. In this work, a 3D PCL fibrous scaffold with aligned morphology was developed through the synergistic combination of Melt-Extrusion Additive Manufacturing (MEAM) and porogen leaching, utilizing PCL as the bulk material and PEG as the porogen. PCL/PEG blends with different polymer ratios (60/40, 50/50, 40/60) were produced and characterized through a DSC analysis. The MEAM process parameters and porogen leaching in bi-distilled water allowed for the development of a micrometric anisotropic fibrous structure with fiber diameters ranging from 10 to 100 µm, depending on PCL/PEG blend ratios. The fibrous scaffolds were coated with Gelatin type A to achieve a biomimetic coating for an in vitro cell culture and mechanically characterized via AFM. The 40/60 PCL/PEG scaffolds yielded the most homogeneous and smallest fibers and the greatest physiological stiffness. In vitro cell culture studies were performed by seeding C2C12 cells onto a selected scaffold, enabling their attachment, alignment, and myotube formation along the PCL fibers during a 14-day culture period. The resultant anisotropic scaffold morphology promoted SMT-like cell conformation, establishing a versatile platform for developing in vitro models of tissues with anisotropic morphology. Full article

An imperative requisite of tissue-engineered scaffolds is to promote host cell regeneration and concomitantly thwart microbial growth. Antibacterial agents are often added to prevent implant-related infections, which, however, aggravates the risk of bacterial resistance. For the first time, we report a fiber-based platform that selectively promotes the growth of mammalian cells and alleviates bacteria by varying fiber size, orientation, and material of polymeric yarns. The interactions of Gram-positive and -negative bacterial species with mammalian mesenchymal stem cells (MSC) were investigated on poly-€-caprolactone (PCL) yarns, polyethylene terephthalate (PET), poly-L-lactic acid (PLLA), and cotton. Various yarn configurations were studied by altering the fiber diameter (from nano- to microscale) and fiber orientations (aligned, twisted, and random) of PCL yarns. PCL nanofibrous yarn decreased the adhesion of S. aureus and E. coli, with a 2.7-fold and 1.5-fold reduction, respectively, compared to PCL microfibrous yarn. Among different fiber orientations, nanoaligned fibers resulted in an 8-fold and 30-fold reduction of S. aureus and E. coli adhesion compared to random fibers. Moreover, aligned orientation was superior in retarding the S. aureus adhesion by 14-fold compared to nanotwisted fibers. Our data demonstrate that polymeric yarns comprising fibers with nanoscale features and aligned orientation promote mammalian cell adhesion and spreading and concomitantly mitigate bacterial interaction. Moreover, we unveil the wicking of cells through polymeric yarns, facilitating early cell adhesion in fibrous scaffolds. Overall, this study provides insight to engineer scaffolds that couple superior interaction of mammalian cells with high-strength fibrous yarns for regenerative applications devoid of antibacterial agents or other surface modification strategies. Full article

Therapeutic solutions for injuries in the peripheral nervous system are limited and not existing in the case of the central nervous system. The electrical stimulation of cells through a cell-supporting conductive scaffold may contribute to new therapeutic solutions for nerve regeneration. In this work, biocompatible Polylactic acid (PLA) fibrous scaffolds incorporating Fe(III)Tosylate (FeTos) were produced by electrospinning a mixture of PLA/FeTos solutions towards a rotating cylinder, inducing fiber alignment. Fibers were coated with the conductive polymer Poly(3,4 ethylenedioxythiophene) (PEDOT) formed by vapor-phase polymerization of EDOT at 70 °C for 2 h. Different solvents (ETH, DMF and THF) were used as FeTos solvents to investigate the impact on the scaffold’s conductivity. Scaffold conductivity was estimated to be as high as 1.50 × 10⁻¹ S/cm when FeTos was dissolved in DMF. In vitro tests were performed to evaluate possible scaffold cytotoxicity, following ISO 10993-5, revealing no cytotoxic effects. Differentiation and growth of cells from the neural cell line SH-SY5Y seeded on the scaffolds were also assessed, with neuritic extensions observed in cells differentiated in neurons with retinoic acid. These extensions tended to follow the preferential alignment of the scaffold fibers. Full article

Electrospun nanofiber constructs represent a promising alternative for mimicking the natural extracellular matrix in vitro and have significant potential for cardiac patch applications. While the effect of fiber orientation on the morphological structure of cardiomyocytes has been investigated, fibers only provide contact guidance without accounting for substrate stiffness due to their deposition on rigid substrates (e.g., glass or polystyrene). This paper introduces an in situ fabrication method for suspended and well aligned nanofibrous scaffolds via roller electrospinning, providing an anisotropic microenvironment with reduced stiffness for cardiac tissue engineering. A fiber surface modification strategy, utilizing oxygen plasma treatment combined with sodium dodecyl sulfate solution, was proposed to maintain the hydrophilicity of polycaprolactone (PCL) fibers, promoting cellular adhesion. Human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs), cultured on aligned fibers, exhibited an elongated morphology with extension along the fiber axis. In comparison to Petri dishes and suspended random fiber scaffolds, hiPSC-CMs on suspended aligned fiber scaffolds demonstrated enhanced sarcomere organization, spontaneous synchronous contraction, and gene expression indicative of maturation. This work demonstrates the suspended and aligned nano-fibrous scaffold provides a more realistic biomimetic environment for hiPSC-CMs, which promoted further research on the inducing effect of fiber scaffolds on hiPSC-CMs microstructure and gene-level expression. Full article

Electrospinning has contributed substantially to the construction of nanofibrous scaffolds for potential tissue engineering and regenerative medicine applications. However, conventional electrospinning only has the ability to generate and collect nanofiber scaffolds with a randomly oriented fibrous pattern, which lack the necessary cell alignment guidance function. In this study, a novel electrospinning fiber-collecting device was designed and developed by setting a series of small pin-ring-structured collectors on a large plain plate. Specifically, we demonstrated that the pin-ring-structured collectors, which were constructed by inserting a metal pin into the center of a metal ring, could collect the as-electrospun nanofibers with radially oriented structures in an innovative manner. We first investigated the suitable polymeric concentration for electrospinning poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), and the optimum electrospinning concentration of PHBV was found to be 12% (w/v) PHBV dissolved in hexafluoroisopropyl alcohol (HFIP). Then, 12% (w/v) PHBV solution was electrospun into radially oriented nanofiber scaffolds using our novel electrospinning strategy, and their various performances were further compared with conventionally randomly oriented nanofiber scaffolds that were also produced from 12% (w/v) PHBV solution. The results showed that the radially oriented PHBV nanofiber scaffolds exhibited obviously enhanced mechanical properties and decreased hydrophobicity compared with the randomly oriented PHBV nanofiber scaffold controls. Importantly, the biological properties of radially oriented PHBV nanofiber scaffolds were also demonstrated to be enhanced, compared with randomly oriented PHBV nanofiber scaffolds, by effectively inducing cell alignment and significantly promoting cell proliferation. In sum, the present study indicates that our as-prepared nanofiber scaffolds with a radially oriented pattern are of great interest for advanced applications, such as wound dressings and tissue-engineered scaffolds. Full article

Tendon disorders are common medical conditions that could lead to significant disability, pain, healthcare costs, and a loss of productivity. Traditional approaches require long periods of treatment, and they largely fail due to the tissues weakening and the postoperative alterations of the normal joint mechanics. To overcome these limitations, innovative strategies for the treatment of these injuries need to be explored. The aim of the present work was the design of nano-fibrous scaffolds based on poly(butyl cyanoacrylate) (PBCA), a well-known biodegradable and biocompatible synthetic polymer, doped with copper oxide nanoparticles and caseinphosphopeptides (CPP), able to mimic the hierarchical structure of the tendon and to improve the tissue healing potential. These were developed as implants to be sutured to reconstruct the tendons and the ligaments during surgery. PBCA was synthetized, and then electrospun to produce aligned nanofibers. The obtained scaffolds were characterized for their structure and physico-chemical and mechanical properties, highlighting that CuO and CPP loading, and the aligned conformation determined an increase in the scaffold mechanical performance. Furthermore, the scaffolds loaded with CuO showed antioxidant and anti-inflammatory properties. Moreover, human tenocytes adhesion and proliferation to the scaffolds were assessed in vitro. Finally, the antibacterial activity of the scaffolds was evaluated using Escherichia coli and Staphylococcus aureus as representative of Gram-negative and Gram-positive bacteria, respectively, demonstrating that the CuO-doped scaffolds possessed a significant antimicrobial effect against E. coli. In conclusion, scaffolds based on PBCA and doped with CuO and CPP deserve particular attention as enhancers of the tendon tissue regeneration and able to avoid bacterial adhesion. Further investigation on the scaffold efficacy in vivo will assess their capability for enhancing the tendon ECM restoration in view of accelerating their translation to the clinic. Full article

Poly(vinyl-alcohol) hydrogels have already been successfully utilised as drug carrier systems and tissue engineering scaffolds. However, lacking mechanical strength and suturability hinders any prospects for clinical and surgical applications. The objective of this work was to fabricate mechanically robust PVA membranes, which could also withstand surgical manipulation and suturing. Electrospun membranes and control hydrogels were produced with 61 kDa PVA. Using a high-speed rotating cylindrical collector, we achieved fibre alignment (fibre diameter: 300 ± 50 nm). Subsequently, we created multilayered samples with different orientations to achieve multidirectional reinforcement. Finally, utilising glutaraldehyde as a cross-linker, we created insoluble fibrous-hydrogel membranes. Mechanical studies were performed, confirming a fourfold increase in the specific loading capacities (from 0.21 to 0.84 Nm²/g) in the case of the monolayer samples. The multilayered membranes exhibited increased resistance from both horizontal and vertical directions, which varies according to the specific arrangement. Finally, the cross-linked fibrous hydrogel samples not only exhibited specific loading capacities significantly higher than their counterpart bulk hydrogels but successfully withstood suturing. Although cross-linking optimisation and animal experiments are required, these membranes have great prospects as alternatives to current surgical meshes, while the methodology could also be applied in other systems as well. Full article

After decades of research, fully functional skin regeneration is still a challenge. Skin is a multilayered complex organ exhibiting a cascading healing process affected by various mechanisms. Specifically, nutrients, oxygen, and biochemical signals can lead to specific cell behavior, ultimately conducive to the formation of high-quality tissue. This biomolecular exchange can be tuned through scaffold engineering, one of the leading fields in skin substitutes and equivalents. The principal objective of this investigation was the design, fabrication, and evaluation of a new class of three-dimensional fibrous scaffolds consisting of poly(ε-caprolactone) (PCL)/calcium alginate (CA), with the goal to induce keratinocyte differentiation through the action of calcium leaching. Scaffolds fabricated by electrospinning using a PCL/sodium alginate solution were treated by immersion in a calcium chloride solution to replace alginate-linked sodium ions by calcium ions. This treatment not only provided ion replacement, but also induced fiber crosslinking. The scaffold morphology was examined by scanning electron microscopy and systematically assessed by measurements of the pore size and the diameter, alignment, and crosslinking of the fibers. The hydrophilicity of the scaffolds was quantified by contact angle measurements and was correlated to the augmentation of cell attachment in the presence of CA. The in vitro performance of the scaffolds was investigated by seeding and staining fibroblasts and keratinocytes and using differentiation markers to detect the evolution of basal, spinous, and granular keratinocytes. The results of this study illuminate the potential of the PCL/CA scaffolds for tissue engineering and suggest that calcium leaching out from the scaffolds might have contributed to the development of a desirable biological environment for the attachment, proliferation, and differentiation of the main skin cells (i.e., fibroblasts and keratinocytes). Full article

Fibrous shape memory scaffolds composed of thermoplastic polyurethane based on a mixture of polycaprolactone diols were fabricated. The effect of the fiber diameter and arrangement– random (rPU) or aligned (aPU), on crystallinity, mechanical properties, and shape memory was analyzed. The diameters of the fibers were controlled by changing the concentration of polyurethane (PU) solutions in the range of 5% to 16% and fibers alignment by utilization of different collectors. The chemical structure was confirmed by Fourier Transformed Infrared spectroscopy (FTIR), crystallinity was evaluated based on differential scanning calorimetry (DSC,) and mechanical properties were measured by the tensile test. Additionally, shape memory programming was performed using a dynamic mechanical analyzer (DMA), and shape recovery was evaluated in the air and in the water environment. DSC results showed that the electrospinning process did not change the crystallinity or melting temperature of synthesized thermoplastic polyurethanes. The melting temperature of the crystalline switching segments was around 26–27 °C, and the crystalline phase of hard segments was around 130 °C. Shape memory properties were analyzed in the contest of the fiber diameter and alignment of the fibers, while changes in the fibers’ diameters from 360 nm to 1760 nm did not result in significant changes in shape recovery coefficient (Rr) especially evaluated in the air. The longitudinal fiber alignment enhanced mechanical and shape recovery to up to 96% for aPU, with the highest fiber diameter when evaluated in water. Full article