Electrospun Polymer Scaffolds Research Articles

Interfacial blending in co-axially electrospun polymer core-shell fibers and their interaction with cells via focal adhesion point analysis

Electrospun polymer scaffolds have gained prominence in biomedical applications, including tissue engineering, drug delivery, and wound dressings, due to their customizable properties. As the interplay between cells and materials assumes fundamental significance in biomaterials research, understanding the relationship between fiber properties and cell behaviour is imperative. Nevertheless, altering fiber properties introduces complexity by intertwining mechanical and surface chemistry effects, challenging the differentiation of their individual impacts on cell behaviour. Core-shell fibers present an appealing solution, enabling the control of mechanical properties of scaffolds, flexibility in material and drug selection, efficient encapsulation, strong protection of bioactive drugs against harsh environments, and controlled, prolonged drug release. This study addresses a key challenge in core-shell fiber design related to the blending effect between core and shell polymers. Two types of fibers, PMMA and core-shell PC-PMMA, were electrospun, and thorough analyses confirmed the desired core-shell structure in PC-PMMA fibers. Surface chemistry analysis revealed PC diffusion to the PMMA shell of the core-shell fiber during electrospinning, subsequently prompting an investigation of the fiber’s surface potential. Conducting cellular studies on osteoblasts by super-resolution confocal microscopy provided insights into the direct influence of interfacial polymer blending and, consequently, altered fiber surface and mechanical properties on cell focal adhesion points, bridging the gap between material attributes and cell responses in core-shell fibers.

Cationic-porphyrin incorporated electrospun fibers for effective photo-inactivation of bacteria

Infectious diseases caused by pathogenic bacteria has always been a persistent concern to public health. The introduction of new microbial diseases or the re-emergence of previously controlled diseases worsens the situation. Antibacterial photodynamic therapy (APDT) is now gaining popularity in the scientific community due to its low susceptibility towards drug resistance, while bacteria often develop drug resistance rapidly in response to traditional antibiotic therapy. The present work explores the possibility of combining a biodegradable gelatin/polyurethane electrospun polymer scaffold (GP) with a cationic porphyrin (P1); to employ as a user-friendly photosensitizer in APDT. The membranes have been structurally characterized and the antibacterial action triggered by a visible light source has been investigated. The membrane GPP exhibited 97% and 96% inhibition against E. coli and S. aureus respectively in the presence of light.

Human Macrophage- and Osteoclast-Based Constructs Do Not Induce Ectopic Bone Formation

PurposeAn increasing body of evidence suggests that bone resorbing osteoclasts are important—but as yet underrated—cellular initiators of bone formation. Furthermore, macrophages also have shown stimulatory effects on the osteogenic differentiation of mesenchymal stromal cells (MSCs). Consequently, we here investigated whether human macrophage- and osteoclast-laden carrier materials can induce ectopic bone formation upon subcutaneous implantation in nude mice.MethodsHuman osteoclast precursors were isolated and differentiated toward macrophages. Subsequently, these macrophages were seeded onto two types of cell carrier materials (i.e., electrospun polymeric scaffolds and devitalized bovine bone granules) and differentiated for 14 days toward osteoclasts. DNA assay and fluorescent nuclei staining were performed. Osteoclast differentiation was assessed by a tartrate-resistant acid phosphatase (TRAP)-activity assay, TRAP, and immunocytochemical staining for β3 integrin. After 60 days of implantation into nude mice, specimens were retrieved, histologically processed, and stained with hematoxylin and eosin (HE) as well as for TRAP to study ectopic bone formation and osteoclast activity, respectively.ResultsOsteoclast precursors limitedly adhered to both material types. Osteoclast-laden samples showed increased intracellular gross TRAP-activity on both cell carrier types, TRAP staining on polymeric electrospun scaffolds, and positive β3 integrin staining on decellularized bovine bone granules compared to the macrophage-laden materials. We observed that only the positive control samples loaded with bone morphogenetic protein-2 (BMP-2) induced ectopic bone formation and TRAP signal.ConclusionWe conclude that neither human macrophage- nor osteoclast-laden constructs are capable to induce ectopic bone formation under the current experimental set-up.Lay summaryInterestingly, increasing amounts of evidence suggest that osteoclasts—the cells responsible for breaking down bone tissue—can trigger bone formation. Therefore, we here aimed to study whether blood-derived macrophages and osteoclasts can induce bone formation in vivo. Consequently, we generated human macrophage- and osteoclast-laden constructs using two types of scaffold materials and implanted them underneath the skin of nude mice. Although we confirmed the presence of macrophages and osteoclasts on the materials, we found no signs of bone formation.

Individual cells generate their own self-reinforcing contact guidance cues through local matrix fiber remodeling.

Directed cell migration arises from cells following a microenvironmental gradient (e.g. of a chemokine) or polarizing feature (e.g. a linear structure). However cells not only follow, but in many cases, also generate directionality cues by modifying their microenvironment. This bi-directional relationship is seen in the alignment of extracellular matrix (ECM) fibers ahead of invading cell masses. The forces generated by many migrating cells cause fiber alignment, which in turn promotes further migration in the direction of fiber alignment via contact guidance and durotaxis. While this positive-feedback relationship has been widely described for cells invading en masse, single cells are also able to align ECM fibers, as well as respond to contact guidance and durotaxis cues, and should therefore exhibit the same relationship. In this study, we directly tested this hypothesis by studying the migration persistence of individual HT-1080 fibrosarcoma cells migrating in photocrosslinked collagen matrices with limited remodeling potential. Our results demonstrate that this positive-feedback relationship is indeed a fundamental aspect of cell migration in fibrillar environments. We observed that the cells’ inability to align and condense fibers resulted in a decrease in persistence relative to cells in native collagen matrices and even relative to isotropic (glass) substrates. Further experiments involving 2D collagen and electrospun polymer scaffolds suggest that substrates composed of rigid, randomly oriented fibers reduce cells’ ability to follow another directionality cue by forcing them to meander to follow the available adhesive area (i.e. fibers). Finally, our results demonstrate that the bi-directional relationship between cell remodeling and migration is not a “dimensionality” effect, but a fundamental effect of fibrous substrate structure.

Gelatin Blends Enhance Performance of Electrospun Polymeric Scaffolds in Comparison to Coating Protocols.

The electrospinning of hybrid polymers is a versatile fabrication technique which takes advantage of the biological properties of natural polymers and the mechanical properties of synthetic polymers. However, the literature is scarce when it comes to comparisons of blends regarding coatings and the improvements offered thereby in terms of cellular performance. To address this, in the present study, nanofibrous electrospun scaffolds of polycaprolactone (PCL), their coating and their blend with gelatin were compared. The morphology of nanofibrous scaffolds was analyzed under field emission scanning electron microscopy (FE-SEM), indicating the influence of the presence of gelatin. The scaffolds were mechanically tested with tensile tests; PCL and PCL gelatin coated scaffolds showed higher elastic moduli than PCL/gelatin meshes. Viability of mouse embryonic fibroblasts (MEF) was evaluated by MTT assay, and cell proliferation on the scaffold was confirmed by fluorescence staining. The positive results of the MTT assay and cell growth indicated that the scaffolds of PCL/gelatin excelled in comparison to other scaffolds, and may serve as good candidates for tissue engineering applications.

Accelerated Endothelialization of Nanofibrous Scaffolds for Biomimetic Cardiovascular Implants.

Nanofiber nonwovens are highly promising to serve as biomimetic scaffolds for pioneering cardiac implants such as drug-eluting stent systems or heart valve prosthetics. For successful implant integration, rapid and homogeneous endothelialization is of utmost importance as it forms a hemocompatible surface. This study aims at physicochemical and biological evaluation of various electrospun polymer scaffolds, made of FDA approved medical-grade plastics. Human endothelial cells (EA.hy926) were examined for cell attachment, morphology, viability, as well as actin and PECAM 1 expression. The appraisal of the untreated poly-L-lactide (PLLA L210), poly-ε-caprolactone (PCL) and polyamide-6 (PA-6) nonwovens shows that the hydrophilicity (water contact angle > 80°) and surface free energy (<60 mN/m) is mostly insufficient for rapid cell colonization. Therefore, modification of the surface tension of nonpolar polymer scaffolds by plasma energy was initiated, leading to more than 60% increased wettability and improved colonization. Additionally, NH3-plasma surface functionalization resulted in a more physiological localization of cell–cell contact markers, promoting endothelialization on all polymeric surfaces, while fiber diameter remained unaltered. Our data indicates that hydrophobic nonwovens are often insufficient to mimic the native extracellular matrix but also that they can be easily adapted by targeted post-processing steps such as plasma treatment. The results achieved increase the understanding of cell–implant interactions of nanostructured polymer-based biomaterial surfaces in blood contact while also advocating for plasma technology to increase the surface energy of nonpolar biostable, as well as biodegradable polymer scaffolds. Thus, we highlight the potential of plasma-activated electrospun polymer scaffolds for the development of advanced cardiac implants.

Electrospun fibre diameter and its effects on vascular smooth muscle cells

Bypass grafting is a technique used in the treatment of vascular disease, which is currently the leading cause of mortality worldwide. While technology has moved forward over the years, synthetic grafts still show significantly lower rates of patency in small diameter bypass operations compared to the gold standard (autologous vessel grafts). Scaffold morphology plays an important role in vascular smooth muscle cell (VSMC) performance, with studies showing how fibre alignment and surface roughness can modulate phenotypic and genotypic changes. Herein, this study has looked at how the fibre diameter of electrospun polymer scaffolds can affect the performance of seeded VSMCs. Four different scaffolds were electrospun with increasing fibre sizes ranging from 0.75 to 6 µm. Culturing VSMCs on the smallest fibre diameter (0.75 µm) lead to a significant increase in cell viability after 12 days of culture. Furthermore, interesting trends were noted in the expression of two key phenotypic genes associated with mature smooth muscle cell contractility (myocardin and smooth muscle alpha-actin 1), whereby reducing the fibre diameter lead to relative upregulations compared to the larger fibre diameters. These results showed that the smallest (0.75 µm) fibre diameter may be best suited for the culture of VSMCs with the aim of increasing cell proliferation and aiding cell maturity.

Scaffold mediated delivery of dual miRNAs to transdifferentiate cardiac fibroblasts

The standard scaffold-mediated delivery of drugs/biomolecules has been successful due to the unique attributes of scaffolds, specifically the electrospun polymeric scaffolds that mimic ECM are well suited for advanced regenerative applications. Cardiac tissue engineering includes the interpretation of cellular and molecular mechanisms concerning heart regeneration and identifying an efficient reprogramming strategy to overcome the limitation of delivery systems and enhance the reprogramming efficiency. This study is a step towards developing a functional scaffold through a parallel interpretation of electrospun PLLA scaffolds with two distinct topologies to achieve sustained delivery of two muscle-specific microRNAs (miR-1 and miR-133a) to directly reprogram the adult human cardiac fibroblasts into cardiomyocyte-like cells. Polyethyleneimine was used to form stable PEI-miRNA complexes through electrostatic interactions. These complexes were immobilized on the electrospun smooth and porous scaffolds, where a loading efficiency of ~96% for the fibronectin modified and ~38% for unmodified surfaces was observed, regardless of their surface topology. The in-vitro release experiment exhibited a biphasic release pattern of PEI-miRNA polyplexes from the scaffolds. These dual miRNA scaffold systems proved to be an excellent formulation since their combinatorial effect involving the topographic cues of electrospun fibers, and dual miRNAs helped control the cardiac fibroblast cell fate precisely.

A simple route to functionalising electrospun polymer scaffolds with surface biomolecules

Surface functionalisation of polymeric electrospun scaffolds with therapeutic biomolecules is often explored in regenerative medicine and tissue engineering. However, the bioconjugation method must be carefully selected to prevent partial or full loss of activity of the biomolecule following chemical manipulation. Perfluorophenyl azide bearing a N‐hydroxysuccinimide (PFPA-NHS) active ester group is a versatile tool for UV-initiated covalent coupling of amine-containing molecules to hydrocarbon-based polymers, such as polydioxanone or polycaprolactone (PCL). This study therefore explored the feasibility of PFPA-NHS functionalisation of electrospun PCL scaffolds with model biomolecules. Protein conjugation was extensively explored using fluorescence staining and attachment studies, confirming the retention of amine coupling capability following photografting of PFPA-NHS to the PCL surface. The effect of the washing method used to remove unreacted PFPA was explored in Caco-2 cell viability studies, and it was determined that sonication washing is required to avoid cell death. A model enzyme, catalase, was then successfully attached to the surface of PCL scaffolds for potential applications in oncological photodynamic therapy. Catalase retained its enzymatic activity following attachment to the fibres and the majority of the enzyme (~60%) remained bound to the fibre after incubation in an aqueous environment for six days. The anticipated prolonged presentation and sustained release of proteins as a result of PFPA-NHS conjugation could be advantageous in progressing protein-based therapies.

Influence of surface topography on PCL electrospun scaffolds for liver tissue engineering.

Severe liver disease is one of the most common causes of death globally. Currently, whole organ transplantation is the only therapeutic method for end-stage liver disease treatment, however, the need for donor organs far outweighs demand. Recently liver tissue engineering is starting to show promise for alleviating part of this problem. Electrospinning is a well-known method to fabricate a nanofibre scaffold which mimics the natural extracellular matrix that can support cell growth. This study aims to investigate liver cell responses to topographical features on electrospun fibres. Scaffolds with large surface depression (2 μm) (LSD), small surface depression (0.37 μm) (SSD), and no surface depression (NSD) were fabricated by using a solvent–nonsolvent system. A liver cell line (HepG2) was seeded onto the scaffolds for up to 14 days. The SSD group exhibited higher levels of cell viability and DNA content compared to the other groups. Additionally, the scaffolds promoted gene expression of albumin, with all cases having similar levels, while the cell growth rate was altered. Furthermore, the scaffold with depressions showed 0.8 MPa higher ultimate tensile strength compared to the other groups. These results suggest that small depressions might be preferred by HepG2 cells over smooth and large depression fibres and highlight the potential for tailoring liver cell responses.

ECM-mimicking nanofibrous scaffold enriched with dual growth factor carrying nanoparticles for diabetic wound healing.

Polymeric nanofibrous scaffolds provide fine-tuned structures with inter-connecting pores resembling the natural extracellular matrix (ECM) in tissues, and show good potential in assisting the creation of artificial functional tissue. Additional application of growth factors helps to regulate the cellular behaviors and tissue assembly in the scaffolds, which eases the healing process. In this study, we synthesized an electrospun polymer scaffold system enriched with nanoparticles containing growth factors for accelerated healing of diabetic wounds. BSA nanoparticles were synthesized by cross-linking with PEG aldehyde. To free the amino group of BSA, heparin was conjugated by EDC/NHS chemistry. The angiogenic growth factors bFGF and VEGF were bound to heparin by electrostatic interaction. These nanoparticles were adsorbed on to electrospun collagen/PLGA/chitosan nanofibers. The synthesized nanofiber system was evaluated in vitro for its cell viability and proliferation. In vivo experiments conducted in a streptozotocin-induced diabetic mice model showed accelerated wound healing. The excellent healing efficiency of this ECM-mimicking nanofiber scaffold makes it a great candidate for therapeutic application in diabetic wounds.

Correlating in vitro performance with physico-chemical characteristics of nanofibrous scaffolds for skin tissue engineering using supervised machine learning algorithms.

The engineering of polymeric scaffolds for tissue regeneration has known a phenomenal growth during the past decades as materials scientists seek to understand cell biology and cell–material behaviour. Statistical methods are being applied to physico-chemical properties of polymeric scaffolds for tissue engineering (TE) to guide through the complexity of experimental conditions. We have attempted using experimental in vitro data and physico-chemical data of electrospun polymeric scaffolds, tested for skin TE, to model scaffold performance using machine learning (ML) approach. Fibre diameter, pore diameter, water contact angle and Young's modulus were used to find a correlation with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay of L929 fibroblasts cells on the scaffolds after 7 days. Six supervised learning algorithms were trained on the data using Seaborn/Scikit-learn Python libraries. After hyperparameter tuning, random forest regression yielded the highest accuracy of 62.74%. The predictive model was also correlated with in vivo data. This is a first preliminary study on ML methods for the prediction of cell–material interactions on nanofibrous scaffolds.

SIMPoly: A Matlab-Based Image Analysis Tool to Measure Electrospun Polymer Scaffold Fiber Diameter.

Quantifying fiber diameter is important for characterizing electrospun polymer scaffolds. Many researchers use manual measurement methods, which can be time-consuming and variable. Semi-automated tools exist, but there is room for improvement. The current work used Matlab to develop an image analysis program to quickly and consistently measure fiber diameter in scanning electron micrographs. The new Matlab method, termed “SIMPoly” (Semiautomated Image Measurements of Polymers) was validated by using synthetic images with known fiber size and was found to be accurate. The Matlab method was also applied by three different researchers to scanning electron microscopy (SEM) images of electrospun poly(lactic-co-glycolic acid) (PLGA). Results were compared with the semi-automated DiameterJ method and a manual ImageJ measurement approach, and it was found that the Matlab-based SIMPoly method provided measurements in the expected range and with the least variability between researchers. In conclusion, this work provides and describes SIMPoly, a Matlab-based image analysis method that can simply and accurately measure polymer fiber diameters in SEM images with minimal variation between users.Impact statementThis article describes Semiautomated Image Measurements of Polymers (SIMPoly), a novel image analysis tool for measuring polymer fiber diameter in scanning electron micrographs of electrospun scaffolds. The Matlab-based SIMPoly method developed is a semi-automated tool that is quick, easy to use, and absent of significant user bias. This tool can be used to accurately measure electrospun polymer fiber diameters, an important characteristic for characterizing tissue-engineered scaffolds.

Development of electrospun polymer scaffolds for the localized and controlled delivery of siponimod for the management of critical bone defects.

Sphingosine 1-phosphate (S1P) receptor modulators can influence bone regeneration owing to their positive impact on osteoblast differentiation and neovascularisation. While previous studies have utilised non-specific S1P and fingolimod, this study aims to design and characterise a controlled release vehicle to deliver the specific S1P1 & 5 receptor modulator siponimod and test its effectiveness in rat critical cranial defects. Electrospun scaffolds of poly lactide-co-glycolide (PLGA) were loaded with siponimod at drug:polymer mass ratios of 0.5:100 to 2:100. Where indicated, collagen was co-spun at a collagen:polymer mass ratio of 2:100. Thereafter, scaffolds underwent in vitro physicochemical characterisation and in vivo assessment using a rat cranial defect model. Drug-loaded scaffolds showed controlled release of siponimod, -cytocompatibility with endothelial and osteoblast cells in vitro, and furthermore, showed that released siponimod stimulated osteoblast differentiation and endothelial cell migration. The in vivo cranial defect repair study showed regeneration was occurring in the defect, although there was no significant difference in the extent of mineralisation between scaffold experimental groups. To our knowledge, this is the first study investigating siponimod in bone regeneration. In vitro studies confirm a positive impact on key cells involved in bone regeneration, however, the scaffolds did not result in significant repair of critical cranial defects.

Abstract No. 433 Dipyridamole-loaded electrospun polymer scaffold based on polyethylene glycol/polycaprolactone for localized drug delivery

One of the major causes of failure of vascular interventions is neointimal hyperplasia (NIH). To prevent the development of NIH, drugs must be delivered locally, for which drug-eluting, resorbable polymer scaffolds present a promising strategy (Liechty et.al., 2010). In this study, dipyridamole (DPA)-eluting scaffolds based on polyethylene glycol (PEG) and polycaprolactone (PCL) were fabricated and tested. PCL (MW 80,000) and PEG (MW 8000) were blended in chloroform at 3:1, 1:1, and 1:3 mass ratios for a final polymer concentration of 200 mg/mL. DPA was added at 0.1 mg/mL. Electrospinning was done with Avecta’s Spraybase system at 1 mL volume, 1 mL/hr flow rate, 15.0 kV voltage, and 15 cm distance. Bilayer scaffolds were fabricated with a layer containing DPA on a layer without DPA. Scaffolds were characterized by scanning electron microscopy (SEM) and porosity and tested for DPA release, cell proliferation, and blood biocompatibility. SEM images of the electrospun scaffolds revealed that adding PEG to PCL decreased the fiber diameter and pore size of the scaffolds. Scaffolds fabricated from 3:1 PCL:PEG demonstrated the slowest cumulative rate of release, with 9.17% cumulative release after 8 days. Scaffold fabricated from 3:1 PCL:PEG also exhibited higher levels of cell attachment and proliferation. With the 3:1 bilayer, RF24 endothelial cells proliferated on non-eluting side (luminal), while MOVAS smooth muscle cells were inhibited on DPA-eluting side (adventitial). No scaffolds showed significant hemolysis and clotting upon contact with blood. Significant differences were observed when compared to positive control (multiple t-tests, P < 0.001). PCL and PEG blended scaffolds in a 3:1 PCL:PEG mass ratio demonstrate slow and sustained drug release, adjustable cell proliferation properties, and biocompatibility, making for potential graft candidates. This approach may also be used for local and controlled release of other drugs, such as chemotherapeutic agents.

Optimized polylactic acid/polyethylene glycol (PLA/PEG) electrospun fibrous scaffold for drug delivery: effect of graphene oxide on the cefixime release mechanism

In the recent decade, there has been a growing interest in using electrospun polymeric scaffolds for sustained release of drugs. In this work, fibrous scaffolds of polylactic acid/polyethylene glycol (PLA/PEG) were prepared and characterized through scanning electron microscopy, for subsequent utilization as the drug delivery system for cefixime (CF). In this regard, the response surface experimental design (RSM) was successfully used for finding optimized electrospinning condition such as total solution concentration (), PEG content (), and applied voltage (V) on the morphology of electrospun PLA/PEG fibrous mats. While increasing the applied voltage and reducing both solution concentration and %PEG resulted in thinner fibers, Ctot was the influential factor on the fiber defects and continuity. Finally, PLA/PEG/CF fine fibers were produced under optimized electrospinning conditions and the controlled drug release profile was obtained in the presence of graphene oxide (GO). GO-CF intermolecular interactions, revealed by attenuated total reflection–Fourier transform infrared spectroscopy (ATR-FTIR) analysis, authorized the drug release mechanism and resulted in near-Fickian diffusion of cefixime from the PLA/PEG/CF/GO fibrous mats.

Photodynamically Active Electrospun Fibers for Antibiotic-Free Infection Control.

Antimicrobial biomaterials are critical to aid in the regeneration of oral soft tissue and prevent or treat localized bacterial infections. With the rising trend in antibiotic resistance, there is a pressing clinical need for new antimicrobial chemistries and biomaterial design approaches enabling on-demand activation of antibiotic-free antimicrobial functionality following an infection that are environment-friendly, flexible and commercially viable. This study explores the feasibility of integrating a bioresorbable electrospun polymer scaffold with localized antimicrobial photodynamic therapy (aPDT) capability. To enable aPDT, we encapsulated a photosensitizer (PS) in polyester fibers in the PS inert state, so that the antibacterial function would be activated on-demand via a visible light source. Fibrous scaffolds were successfully electrospun from FDA-approved polyesters, either poly(ε-caprolactone (PCL) or poly[(rac-lactide)-co-glycolide] (PLGA), with encapsulated PS (either methylene blue (MB) or erythrosin B (ER)). These were prepared and characterized with regards to their loading efficiency (UV-vis spectroscopy), microarchitecture (SEM, porometry, and BET (Brunauer-Emmett-Teller) analysis), tensile properties, hydrolytic behavior (contact angle, dye release capability, degradability), and aPDT effect. The electrospun fibers achieved an ∼100 wt % loading efficiency of PS, which significantly increased their tensile modulus and reduced their average fiber diameter and pore size with respect to PS-free controls. In vitro, PS release varied between a burst release profile to limited release within 100 h, depending on the selected scaffold formulation, while PLGA scaffolds displayed significant macroscopic shrinkage and fiber merging, following incubation in phosphate buffered saline solution. Exposure of PS-encapsulated PCL fibers to visible light successfully led to at least a 1 log reduction in Escherichia coli viability after 60 min of light exposure, whereas PS-free electrospun controls did not inactive microbes. This study successfully demonstrates the significant potential of PS-encapsulated electrospun fibers as photodynamically active biomaterial for antibiotic-free infection control.

Cancer Cells Sense Fibers by Coiling on them in a Curvature-Dependent Manner.

SummaryMetastatic cancer cells sense the complex and heterogeneous fibrous extracellular matrix (ECM) by formation of protrusions, and our knowledge of how cells physically recognize these fibers remains in its infancy. Here, using suspended ECM-mimicking isodiameter fibers ranging from 135 to 1,000 nm, we show that metastatic breast cancer cells sense fiber diameters differentially by coiling (wrapping-around) on them in a curvature-dependent manner, whereas non-tumorigenic cells exhibit diminished coiling. We report that coiling occurs at the tip of growing protrusions and the coil width and coiling rate increase in a curvature-dependent manner, but time to maximum coil width occurs biphasically. Interestingly, bundles of 135-nm diameter fibers recover coiling width and rate on 1,000-nm-diameter fibers. Coiling also coincides with curvature-dependent persistent and ballistic transport of endogenous granules inside the protrusions. Altogether, our results lay the groundwork to link biophysical sensing with biological signaling to quantitate pro- and anti-invasive fibrous environments.Video

Biocompatibility and biodegradation properties of polycaprolactone/polydioxanone composite scaffolds prepared by blend or co-electrospinning

Electrospun polymer scaffolds are regarded as an ideal tissue engineering scaffold due to similar morphological properties with the native extracellular matrix. Among these, polycaprolactone is widely used to fabricate electrospun fibrous scaffolds due to its excellent biocompatibility, good mechanical properties, and ease of manufacture. However, its low biodegradation rate has a negative influence on its application in tissue engineering scaffold. To address this issue, this study prepared hybrid scaffolds composed of polycaprolactone and polydioxanone (a fast-degrading polyether-ester) via either the blend or co-electrospinning. Subsequently, the structural characteristics, mechanical strength, in vitro/vivo degradation, cellularization, and vascularization of two kinds of hybrid scaffolds were evaluated to decide which method is more suitable for producing tissue engineering scaffolds. The incorporation of polydioxanone increased the mechanical strength of both composite scaffolds. Moreover, co-electrospun scaffolds exhibited improved hydrophilicity compared to blend scaffolds. The results of in vitro and in vivo degradation studies showed that the degradation rate of both composite scaffolds was faster than that of neat polycaprolactone scaffolds due to the incorporated polydioxanone component. Especially in co-electrospun scaffolds, the fast degradation of polydioxanone fiber gave rise to larger pore size, thus leading to faster cellularization and better vascularization compared to blend scaffolds. Therefore, co-electrospinning was demonstrated to be superior to blend electrospinning for the preparation of composite scaffolds. Co-electrospun polycaprolactone–polydioxanone scaffolds may be promising candidates for tissue engineering.

A Non-woven Path: Electrospun Poly(lactic acid) Scaffolds for Kidney Tissue Engineering

Chronic kidney disease is a major global health problem affecting millions of people; kidney tissue engineering provides an opportunity to better understand this disease, and has the capacity to provide a cure. Two-dimensional cell culture and decellularised tissue have been the main focus of this research thus far, but despite promising results these methods are not without their shortcomings. Polymer fabrication techniques such as electrospinning have the potential to provide a non-woven path for kidney tissue engineering. In this experiment we isolated rat primary kidney cells which were seeded on electrospun poly(lactic acid) scaffolds. Our results showed that the scaffolds were capable of sustaining a multi-population of kidney cells, determined by the presence of: aquaporin-1 (proximal tubules), aquaporin-2 (collecting ducts), synaptopodin (glomerular epithelia) and von Willebrand factor (glomerular endothelia cells), viability of cells appeared to be unaffected by fibre diameter. The ability of electrospun polymer scaffold to act as a conveyor for kidney cells makes them an ideal candidate within kidney tissue engineering; the non-woven path provides benefits over decellularised tissue by offering a high morphological control as well as providing superior mechanical properties with degradation over a tuneable time frame.