Photothermal driven BMSCs osteogenesis and M2 macrophage polarization on polydopamine-coated Ti3C2 nanosheets/poly(vinylidene fluoride trifluoroethylene) nanocomposite coatings.

Mild thermal stimulation plays an active role in bone tissue repair and regeneration. In this work, a bioactive polydopamine/Ti3C2/poly(vinylidene fluoride trifluoroethylene) (PDA/Ti3C2/P(VDF-TrFE)) nanocomposite coating with excellent near-infrared light (NIR)-triggered photothermal effect was designed to improve the osteogenic ability of implants. By incorporating dopamine (DA)-modified Ti3C2 nanosheets into the P(VDF-TrFE) matrix and combining them with alkali initiated in situ polymerization, the resulting PDA/Ti3C2/P(VDF-TrFE) nanocomposite coating gained high adhesion strength on Ti substrate, excellent tribological and corrosion resistance properties, which was quite important for clinical application of implant coatings. Cell biology experiments showed that NIR-triggered mild thermal stimulation on the coating surface promoted cell spreading and growth of BMSCs, and also greatly upregulated the osteogenic markers, including Runt-Related Transcription Factor 2 (RUNX2), alkaline phosphatase (ALP), osteopontin (OPN), osteocalcin (OCN). Simultaneously, the synthesis of heat shock protein 47 (HSP47) was significantly promoted by the mild thermal stimulation, which strengthened the specific interaction between HSP47 and collagen Ⅰ (COL-Ⅰ), thereby activating the integrin-mediated MEK/ERK osteogenic differentiation signaling pathway. In addition, the results also showed that the mild thermal stimulation induced the polarization of macrophages towards M2 phenotype, which can attenuate the inflammatory response of injured bone tissue. Antibacterial results indicated that the coating exhibited an outstanding antibacterial ability against S. aureus and E. coli. Conceivably, the versatile implant bioactive coatings developed in this work will show great application potential for implant osseointegration.

Preparation and performance of random- and oriented-fiber membranes with core-shell structures via coaxial electrospinning.

The cells and tissue in the human body are orderly and directionally arranged, and constructing an ideal biomimetic extracellular matrix is still a major problem to be solved in tissue engineering. In the field of the bioresorbable vascular grafts, the long-term functional prognosis requires that cells first migrate and grow along the physiological arrangement direction of the vessel itself. Moreover, the graft is required to promote the formation of neointima and the development of the vessel walls while ensuring that the whole repair process does not form a thrombus. In this study, poly (l-lactide-co-ε-caprolactone) (PLCL) shell layers and polyethylene oxide (PEO) core layers with different microstructures and loaded with sodium tanshinone IIA sulfonate (STS) were prepared by coaxial electrospinning. The mechanical properties proved that the fiber membranes had good mechanical support, higher than that of the human aorta, as well as great suture retention strengths. The hydrophilicity of the oriented-fiber membranes was greatly improved compared with that of the random-fiber membranes. Furthermore, we investigated the biocompatibility and hemocompatibility of different functional fiber membranes, and the results showed that the oriented-fiber membranes containing sodium tanshinone IIA sulfonate had an excellent antiplatelet adhesion effect compared to other fiber membranes. Cytological analysis confirmed that the functional fiber membranes were non-cytotoxic and had significant cell proliferation capacities. The oriented-fiber membranes induced cell growth along the orientation direction. Degradation tests showed that the pH variation range had little change, the material mass was gradually reduced, and the fiber morphology was slowly destroyed. Thus, results indicated the degradation rate of the oriented-fiber graft likely is suitable for the process of new tissue regeneration, while the random-fiber graft with a low degradation rate may cause the material to reside in the tissue for too long, which would impede new tissue reconstitution. In summary, the oriented-functional-fiber membranes possessing core-shell structures with sodium tanshinone IIA sulfonate/polyethylene oxide loading could be used as tissue engineering materials for applications such as vascular grafts with good prospects, and their clinical application potential will be further explored in future research.

Enhanced air filtration performances by coating aramid nanofibres on a melt-blown nonwoven.

Nanofibre membranes with a small diameter and a large specific surface area are widely used in the filtration field due to their small pore size and high porosity. To date, aramid nanofibres (ANFs) have received extensive research interest because of their high stiffness and excellent temperature resistance. However, the preparation of ANFs usually takes a long time, which greatly hampers the practical application of these fibres. Herein, we report the preparation of ANFs by a modified deprotonation method at elevated temperature. Owing to the increase of temperature, the preparation cycle of ANFs was shortened to 8 hours. The resulting ANF dispersion was further coated on a polypropylene melt-blown nonwoven to form a composite nonwoven filter. With the submicron porous structure, the filtration efficiency, pressure drop and quality factor of the filter were 95.61%, 38.22 Pa and 0.082 Pa-1, respectively. Compared to the pristine nonwoven, the filtration, mechanical, and heat insulation properties of the composite filter were also significantly improved. This work may offer a simple and efficient way for enhancing the air filtration performances of current filters.

In Situ Prevascularization Strategy with Three-Dimensional Porous Conduits for Neural Tissue Engineering.

Neovascularization is crucial for peripheral nerve regeneration and long-term functional restoration. Previous studies have emphasized strategies that enhance axonal repair over vascularization. Here, we describe the development and application of an in situ prevascularization strategy that uses 3D porous nerve guidance conduits (NGCs) to achieve angiogenesis-mediated neural regeneration. The optimal porosity of the NGC is a critical feature for achieving neovascularization and nerve growth patency. Hollow silk fibroin/poly(l-lactic acid-co-ε-caprolactone) NGCs with 3D sponge-like walls were fabricated using electrospinning and freeze-drying. In vitro results showed that 3D porous NGC favored cell biocompatibility had neuroregeneration potential and, most importantly, had angiogenic activity. Results from our mechanistic studies suggest that activation of HIF-1α signaling might be associated with this process. We also tested in situ prevascularized 3D porous NGCs in vivo by transplanting them into a 10 mm rat sciatic nerve defect model with the aim of regenerating the severed nerve. The prevascularized 3D porous NGCs greatly enhanced intraneural angiogenesis, resulting in demonstrable neurogenesis. Eight weeks after transplantation, the performance of the prevascularized 3D NGCs was similar to that of traditional autografts in terms of improved anatomical structure, morphology, and neural function. In conclusion, combining a reasonably fabricated 3D-pore conduit structure with in situ prevascularization promoted functional nerve regeneration, suggesting an alternative strategy for achieving functional recovery after peripheral nerve trauma.

In situ deposition of nano Cu2O on electrospun chitosan nanofibrous scaffolds and their antimicrobial properties.

In order to obtain a synergistic antimicrobial effect of cuprous oxide nanoparticles (Cu2O NPs) and chitosan (CS) nanofibers, the nano Cu2O/CS nanofibrous scaffolds were synthesized in situ via two subsequent steps of chelation and reduction. The Cu2+ were stably chelated on CS nanofibrous scaffolds through the coordination of amino group (-NH2) and hydroxyl group (-OH) on CS with Cu2+, and then the chelated Cu2+ were reduced to nano Cu2O by Vitamin C under alkaline conditions. And by the measurements of XRD, XPS and FTIR-ATR, the results showed that Cu2O NPs were successfully deposited on the CS nanofibrous scaffolds. SEM clarified that the particle size of Cu2O gradually decreased and the shape changed from cubic to irregular with the increase of CuSO4 concentration. With the CuSO4 concentration of 0.02 and 0.04 mol·L-1, the Cu2O/CS nanofibrous scaffolds presented outstanding hydrophilicity and antibacterial activity against Gram-negative Escherichia coli (E. coli) and Gram-positive Staphylococcus aureus (S. aureus) comparing to the CS nanofibrous scaffolds, meanwhile, they possessed good biocompatibility. This kind of nanofibrous scaffolds deposited with nano Cu2O would have broad application prospects in the field of antibacterial biomaterials.

Magnesium oxide-incorporated electrospun membranes inhibit bacterial infections and promote the healing process of infected wounds.

Bacterial infections cause severe secondary damage to wounds and hinder wound healing processes. We prepared magnesium oxide (MgO) nanoparticle-incorporated nanofibrous membranes by electrospinning and investigated their potential for wound dressing and fighting bacterial infection. MgO-Incorporated membranes possessed good elasticity and flexibility similar to native skin tissue and were hydrophilic, ensuring comfortable contact with wound beds. The cytocompatibility of membranes was dependent on the amounts of incorporated MgO nanoparticles: lower amounts promoted while higher amounts suppressed the proliferation of fibroblasts, endothelial cells, and macrophages. The antibacterial capacity of membranes was proportional to the amounts of incorporated MgO nanoparticles and they inhibited more than 98% E. coli, 90% S. aureus, and 94% S. epidermidis. MgO nanoparticle-incorporated membranes effectively suppressed bacterial infection and significantly promoted the healing processes of infected full-thickness wounds in a rat model. Subcutaneous implantation demonstrated that the incorporation of MgO nanoparticles into electrospun membranes elevated their bioactivity as evidenced by considerable cell infiltration into their dense nanofiber configuration and enhanced the remodeling of implanted membranes. This study highlights the potential of MgO-incorporated electrospun membranes in preventing bacterial infections of wounds.

A transparent hydrophilic anti-biofouling coating for intraocular lens materials prepared by "bridging" of the intermediate adhesive layer.

The attachment of bio-foulants, including unwanted cells, proteins, and bacteria, to a medical device such as an intraocular lens can lead to implantation failure. Hydrophilic polymers are often used as surface modifiers in the fabrication of anti-biofouling coatings, but a hydrophilic coating can easily become swollen and peel off the substrate. In this study, we chose polymethyl methacrylate (PMMA) as the representative material of intraocular lenses because PMMA has better biocompatibility, a higher refractive index, better optical clarity, lighter weight, more stable performance, and lower cost than other intraocular lens materials. We fabricated polyvinyl alcohol (PVA) coatings with or without a "bridge", that is, an intermediate adhesive layer (AL), to increase the adhesion bonding effect between the anti-biofouling coating and the substrate. The results indicated that the prepared coatings were transparent and noncytotoxic. Moreover, the anti-adhesion properties of the cells and the resistance properties to nonspecific protein adsorption of PMMA modified by both AL and PVA coatings were better and more durable compared with the sample only modified with a physically dipped PVA coating. The coating prepared by AL "bridging" provides a new strategy for the preparation of a transparent hydrophilic anti-biofouling coating suitable for PMMA intraocular lens materials.

Performance of PEGylated chitosan and poly (L-lactic acid-co-ε-caprolactone) bilayer vascular grafts in a canine femoral artery model.

The fabrication of a functional small-diameter vascular graft with good biocompatibility, in particular hemocompatibility, has become an urgent clinical necessity. We fabricated a native bilayer, small-diameter vascular graft using PEGylated chitosan (PEG-CS) and poly (L-lactic acid-co-ε-caprolactone; PLCL). To stabilize the inner layer, a PEG-CS blend with PLCL at ratio of 1:6 was casted on a round metal bar by a drip feed, and the outer layer, a PLCL blend with water-soluble PEG that acted as a sacrificial part to enhance pore size, was fabricated by electrospinning. The results showed excellent hemocompatibility and strong mechanical properties. In vitro, the degradation of the graft was evaluated by measuring the graft structure, mass loss rate, and changes in molecular weight. The results indicated that the graft had adequate support for the regeneration of blood vessels before collapse. An in vivo study was performed in a canine femoral artery model for up to 24 weeks, which demonstrated that the PEGylated bilayer grafts possessed excellent structural integrity, high compatibility with blood, good endothelial cell (EC) and smooth muscle cell (SMC) growth, and high expression levels of angiogenesis-related proteins, features that are highly similar to autologous blood vessels. Moreover, the results showed almost negligible calcification within 24 weeks. These findings confirm that the bilayer graft mimics native cells, thereby effectively improving vascular remodeling.

Polycaprolactone vascular graft with epigallocatechin gallate embedded sandwiched layer-by-layer functionalization for enhanced antithrombogenicity and anti-inflammation.

Small-diameter artificial vascular grafts modified with layer-by-layer (LBL) coating show promise in reducing the failure caused by thrombosis and inflammation, but undesirable stability and bioactivity issues of the coating and payload usually limits their long-term efficacy. Herein, inspired by catechol/gallol surface chemistry, a sandwiched layer-by-layer coating constructed by polyethyleneimine (PEI) and heparin with the embedding of epigallocatechin gallate (EGCG)-dexamethasone combination was used to modify the electrospun polycaprolactone (PCL) vascular grafts. Polyphenol embedding endowed the coating with abundant intermolecular interactions between each coating components, mainly contributed by the π-π stacking, weak intermolecular cross-linking and enriched hydrogen bonding, which further enhanced the coating stability and also supported the sustained release of the payloads, like polyelectrolytes and drugs. Compared with the conventional LBL coating, the loading amounts of heparin and dexamethasone in the EGCG embedded LBL coatings doubled and the drug release could be significantly prolonged without serious initial burst. The in vitro and ex vivo assays indicated that the modified PCL vascular grafts would address impressive prolonged anti-platelet adhesion/activation and anti-fibrinogen denaturation ability. Meanwhile, the dexamethasone loading entrusted the sandwiched LBL coating with mild tissue response, in terms of inhibiting the macrophage activation. These results strongly demonstrated that the sandwiched LBL coating with EGCG embedding was an effective method to improve the patency rates of PCL small artificial vascular grafts, which could also be extended to other blood-contacting materials.

Coaxial electrospinning multicomponent functional controlled-release vascular graft: Optimization of graft properties.

Small diameter vascular grafts possessing desirable biocompatibility and suitable mechanical properties have become an urgent clinic demand. Herein, heparin loaded fibrous grafts of collagen/chitosan/poly(l-lactic acid-co-ε-caprolactone) (PLCL) were successfully fabricated via coaxial electrospinning. By controlling the concentration of heparin and the ratio of collagen/chitosan/PLCL, most grafts had the heparin encapsulation efficiency higher than 70%, and the heparin presented sustained release for more than 45 days. Particularly, such multicomponent grafts had relative low initial burst release, and after heparin releasing for 3 weeks, the grafts still showed good anti-platelet adhesion ability. In addition, along with the excellent cell biocompatibility, the fabricated grafts possessed suitable mechanical properties including good tensile strength, suture retention strength, burst pressure and compliance which could well match the native blood vessels. Thus, the optimized graft properties could be properly addressed for vascular tissue application via coaxial electrospinning.

Electrospun silk fibroin/poly (L-lactide-ε-caplacton) graft with platelet-rich growth factor for inducing smooth muscle cell growth and infiltration.

The construction of a smooth muscle layer for blood vessel through electrospinning method plays a key role in vascular tissue engineering. However, smooth muscle cells (SMCs) penetration into the electrospun graft to form a smooth muscle layer is limited due to the dense packing of fibers and lack of inducing factors. In this paper, silk fibroin/poly (L-lactide-ε-caplacton) (SF/PLLA-CL) vascular graft loaded with platelet-rich growth factor (PRGF) was fabricated by electrospinning. The in vitro results showed that SMCs cultured in the graft grew fast, and the incorporation of PRGF could induce deeper SMCs infiltrating compared to the SF/PLLA-CL graft alone. Mechanical properties measurement showed that PRGF-incorporated graft had proper tensile stress, suture retention strength, burst pressure and compliance which could match the demand of native blood vessel. The success in the fabrication of PRGF-incorporated SF/PLLA-CL graft to induce fast SMCs growth and their strong penetration into graft has important application for tissue-engineered blood vessels.

A multi-layered vascular scaffold with symmetrical structure by bi-directional gradient electrospinning.

Multi-layered scaffolds are advantageous in vascular tissue engineering, in consideration of better combination of biomechanics, biocompatibility and biodegradability than the scaffolds with single structure. In this study, a bi-directional gradient electrospinning method was developed to fabricate poly(l-lactide-co-caprolactone) (P(LLA-CL)), collagen and chitosan based tubular scaffold with multi-layered symmetrical structure. The multi-layered composite scaffold showed improved mechanical property and biocompatibility, in comparison to the blended scaffold using the same proportion of raw materials. Endothelialization on the multi-layered scaffold was accelerated owing to the bioactive surface made of pure natural materials. hSMCs growth showed the similar results because of its better biocompatibility. Additionally, fibers morphology change, pH value balance and long term mechanical support results showed that the gradient structure effectively improved biodegradability.

Electrospun poly(l-lactide-co-caprolactone)-collagen-chitosan vascular graft in a canine femoral artery model.

Poly(l-lactide-co-caprolactone)-collagen-chitosan (P(LLA-CL)-COL-CS) composite grafts were electrospun in this study. Based on the test results for mechanical properties, biodegradability and in vitro cellular compatibility, the optimal weight ratio of P(LLA-CL) to COL/CS was set as 3 : 1. In vivo study was further performed in a canine femoral artery model. The results showed that the 3 : 1 grafts possessed excellent structural integrity, higher patency rate, better endothelial cell (EC) and smooth muscle cells (SMC) growth, as well as higher levels of gene and protein expression of angiogenesis-related cues than those of grafts based on P(LLA-CL). The findings confirmed that the addition of natural materials, such as collagen and chitosan, could effectively improve endothelialization, SMC incursion into the tunica media, and vascular remodeling for tissue engineering.

Fabrication of cell penetration enhanced poly (l-lactic acid-co-ɛ-caprolactone)/silk vascular scaffolds utilizing air-impedance electrospinning.

In the vascular prosthetic field, the prevailing thought is that for clinical, long-term success, especially bioresorbable grafts, cellular migration and penetration into the prosthetic structure is required to promote neointima formation and vascular wall development. In this study, we fabricated poly (l-lactic acid-co-ɛ-caprolactone) P(LLA-CL)/silk fibroin (SF) vascular scaffolds through electrospinning using both perforated mandrel subjected to various intraluminal air pressures (0-300kPa), and solid mandrel. The scaffolds were evaluated the cellular infiltration in vitro and mechanical properties. Vascular scaffolds were seeded with smooth muscle cells (SMCs) to evaluate cellular infiltration at 1, 7, and 14 days. The results revealed that air-impedance scaffolds allowed significantly more cell infiltration as compared to the scaffolds fabricated with solid mandrel. Meanwhile, results showed that both mandrel model and applied air pressure determined the interfiber distance and the alignment of fibers in the enhanced porosity regions of the structure which influenced cell infiltration. Uniaxial tensile testing indicated that the air-impedance scaffolds have sufficient ultimate strength, suture retention strength, and burst pressure as well as compliance approximating a native artery. In conclusion, the air-impedance scaffolds improved cellular infiltration without compromising overall biomechanical properties. These results support the scaffold's potential for vascular grafting and in situ regeneration.

Cell infiltration and vascularization in porous nanoyarn scaffolds prepared by dynamic liquid electrospinning.

A novel electrospun nanoyarn scaffold, aimed to improve cell infiltration and vascularization, as well as guide cell behaviors by its biomimetic structure, was fabricated for tissue engineering. Electrospun nanofibers were deposited and twisted into yarns in a water vortex before collecting on a rotating mandrel to form a nanoyarn scaffold. Field emission-scanning electronic microscope (FE-SEM) images revealed that the scaffold, composed of aligned nanoyarns (24 micro m) which were composed of a bundle of nanofibers, created a porous structure which may be conducive to cellular infiltration. Thus, we hypothesized that the biomimetic nanoyarn will have a positive influence on cell proliferation and morphology. Pig iliac endothelial cells (PIECs) and MC3T3-E1 pre-osteoblastic cells cultured on the nanoyarn scaffolds showed significantly higher proliferation rates than that on traditional electrospun nanofiber scaffolds. Histological analysis demonstrated that cells infiltrate throughout the nanoyarn scaffolds over a 10-day period, however, no cell infiltration was observed on the nanofiber scaffolds. Moreover, confocal microscopy images indicated that both PIECs and MC3T3-E1 pre-osteoblastic cells cultured on the nanoyarn scaffolds exhibit an extremely elongated morphology compared to the flattened morphology when cells were cultured on electrospun nanofiber scaffolds or tissue culture plates. Furthermore, complex capillary-like structures were observed when PIECs cultured on the nanoyarn scaffold for 7 days, indicating that the nanoyarns provide templates and topographical cues for the assembly of PIECs and the promotion of a capillary network in vitro. In conclusion, the positive cellular interactions on the nanoyarn scaffold demonstrate potential application for use in tissue engineering.

Electrospinning collagen/chitosan/poly(L-lactic acid-co-ε-caprolactone) to form a vascular graft: mechanical and biological characterization.

For blood vessel tissue engineering, an ideal vascular graft should possess excellent biocompatibility and mechanical properties. For this study, a elastic material of poly (L-lactic acid-co-ε-caprolactone) (P(LLA-CL)), collagen and chitosan blended scaffold at different ratios were fabricated by electrospinning. Upon fabrication, the scaffolds were evaluated to determine the tensile strength, burst pressure, and dynamic compliance. In addition, the contact angle and endothelial cell proliferation on the scaffolds were evaluated to demonstrate the structures potential to serve as a vascular prosthetic capable of in situ regeneration. The collagen/chitosan/P(LLA-CL) scaffold with the ratio of 20:5:75 reached the highest tensile strength with the value of 16.9 MPa, and it was elastic with strain at break values of ~112%, elastic modulus of 10.3 MPa. The burst pressure strength of the scaffold was greater than 3365 mmHg and compliance value was 0.7%/100 mmHg. Endothelial cells proliferation was significantly increased on the blended scaffolds versus the P(LLA-CL). Meanwhile, the endothelial cells were more adherent based on the increase in the degree of cell spreading on the surface of collagen/chitosan/P(LLA-CL) scaffolds. Such blended scaffold especially with the ratio of 20:5:75 thus has the potential for vascular graft applications.

Fabrication of chitosan/silk fibroin composite nanofibers for wound-dressing applications.

Chitosan, a naturally occurring polysaccharide with abundant resources, has been extensively exploited for various biomedical applications, typically as wound dressings owing to its unique biocompatibility, good biodegradability and excellent antibacterial properties. In this work, composite nanofibrous membranes of chitosan (CS) and silk fibroin (SF) were successfully fabricated by electrospinning. The morphology of electrospun blend nanofibers was observed by scanning electron microscopy (SEM) and the fiber diameters decreased with the increasing percentage of chitosan. Further, the mechanical test illustrated that the addition of silk fibroin enhanced the mechanical properties of CS/SF nanofibers. The antibacterial activities against Escherichia coli (Gram negative) and Staphylococcus aureus (Gram positive) were evaluated by the turbidity measurement method; and results suggest that the antibacterial effect of composite nanofibers varied on the type of bacteria. Furthermore, the biocompatibility of murine fibroblast on as-prepared nanofibrous membranes was investigated by hematoxylin and eosin (H&E) staining and MTT assays in vitro, and the membranes were found to promote the cell attachment and proliferation. These results suggest that as-prepared chitosan/silk fibroin (CS/SF) composite nanofibrous membranes could be a promising candidate for wound healing applications.

Genipin-crosslinked silk fibroin/hydroxybutyl chitosan nanofibrous scaffolds for tissue-engineering application.

To improve water-resistant ability and mechanical properties of silk fibroin (SF)/hydroxybutyl chitosan (HBC) nanofibrous scaffolds for tissue-engineering applications, genipin, glutaraldehyde (GTA), and ethanol were used to crosslink electrospun nanofibers, respectively. The mechanical properties of nanofibrous scaffolds were obviously improved after 24 h of crosslinking with genipin and were superior to those crosslinked with GTA and ethanol for 24 h. SEM indicated that crosslinked nanofibers with genipin and GTA vapor had good water-resistant ability. Characterization of the microstructure (porosity and pore structure) demonstrated crosslinked nanofibrous scaffolds with genipin and GTA vapor had lager porosities and mean diameters than those with ethanol. Characterization of FTIR-ATR and (13)C NMR clarified both genipin and GTA acted as crosslinking agents for SF and HBC. Furthermore, genipin could induce SF conformation from random coil or α-helix to ß-sheet. Although GTA could also successfully crosslink SF/HBC nanofibrous scaffolds, in long run, genipin maybe a better method due to lower cytotoxicity than GTA. Cell viability studies and wound-healing test in rats clarified that the genipin-crosslinked SF/HBC nanofibrous scaffolds had a good biocompatibility both in vitro and in vivo. These results suggested that genipin-crosslinked SF/HBC nanofibrous scaffolds might be potential candidates for wound dressing and tissue-engineering scaffolds.