Development of Electrospun Composite Fibers in Multiscale Structure and Investigating the Performance on Proliferation and Osteogenic Differentiation of ADSCs

Electrospun composite membranes in multiscale structures are developed for bone tissue engineering. Aligned polycaprolactone (PCL) fibers entrapping CA‐HAp microparticles (containing CaCO₃, hydroxyapatite, and casein in a hierarchical organization) are electrospun to find whether synergistic effects of fiber alignment and CA‐HAp microparticles on improving osteogenic differentiation can be obtained. CA‐HAp microparticles are in a spherical morphology of 1.42 ± 0.26 µm. Their presence increases fiber diameter and does not significantly affect fiber alignment. On all membranes, adipose derived stem cells (ADSCs) from humans spread very well. On a random group, cells distribute randomly and the presence of CA‐HAp microparticles facilitates cell proliferation, especially for the one at CA‐HAp/PCL 50 wt%; the one at CA‐HAp/PCL 20 wt% shows significantly much higher alkaline phosphatase (ALP) activity (112.0% higher) than the pure PCL membrane. On aligned samples, cells align along fibers and expression of ALP is enhanced. However, at the same composition (CA‐HAp/PCL 20 wt%), the random sample has much higher ALP activity than the aligned sample. The expressions of osteogenic marker genes are also evaluated. Combining the results and the applicability of membranes together, the random membrane at CA‐HAp/PCL 20 wt% is the best candidate for bone tissue engineering.     

Parameter Screening and Optimization for a Polycaprolactone-Based GTR/GBR Membrane Using Taguchi Design

Our objective was to determine and optimize the significant parameters affecting mechanical properties and mean fiber diameter (MFD) of a novel GTR/GBR membrane composed of polycaprolactone (PCL) and chicken eggshell membrane (ESM). For this, we prepared electrospun membrane specimens (n = 16) with varying concentrations of PCL, ESM, nano-hydroxyapatite (HAp), and altered electrospinning parameters as generated by DOE++ software. After the determination of MFD and mechanical properties for all specimens, Taguchi orthogonal array L8 design was used to screen significant factors affecting the MFD and mechanical properties. PCL wt%, ESM wt%, HAp wt%, applied voltage (AV), flow rate (FR), and spinneret-collector distance (SCD) were the independent variables investigated. The response variables analyzed were MFD, tensile strength (TS), and elastic modulus. ANOVA outlined ESM wt%, HAp wt%, AV, FR, SCD, and an interactive effect between PCL wt% and AV to be the significant factors affecting modulus values of an electrospun PCL/ESM membrane (p < 0.05). Furthermore, concentrations of PCL and ESM were the significant factors affecting MFD (p < 0.05) and there were no significant factors affecting the TS values. Optimization using DOE++ software predicted that the maximal TS of 3.125 MPa, modulus of 278.168 MPa, and MFD of 882.75 nm could be achieved.     

Preparation of electrospun nanofibers of carbon nanotube/polycaprolactone nanocomposite

Multiwalled carbon nanotube/polycaprolactone nanocomposites (MWNT/PCL) were prepared by in situ polymerization, whereby functionalized MWNTs (F-MWNTs) and unfunctionalized MWNTs (P-MWNTs) were used as reinforcing materials. The F-MWNTs were functionalized by Friedel-Crafts acylation, which introduced the aromatic amine (COC₆H₄-NH₂) groups on the side wall. The F-MWNTs were chemically bonded with the PCL chains in the F-MWNT/PCL, as indicated by the appearance of the amide II group in the FT-IR spectrum. The TGA thermograms showed that the F-MWNT/PCL had better thermal stability than PCL and P-MWNT/PCL. The PCL and the nanocomposite nanofibers were prepared by an electrospinning technique. The nanocomposites that contain more than 2 wt% of MWNTs were not able to be electrospun. The bead of the F-MWNT/PCL nanofiber was formed less than that of the P-MWNT/PCL. The nanocomposite nanofibers showed a relatively broader diameter than the pure PCL nanofibers. The MWNTs were embedded within the nanofibers and were well oriented along the axes of the electrospun nanofibers, as confirmed by transmission electron microscopy.     

Immobilization of silk fibroin on the surface of PCL nanofibrous scaffolds for tissue engineering applications

Poly(?‐caprolactone) (PCL) is explored in tissue engineering (TE) applications due to its biocompatibility, processability, and appropriate mechanical properties. However, its hydrophobic nature and lack of functional groups in its structure are major drawbacks of PCL‐based scaffolds limiting appropriate cell adhesion and proliferation. In this study, silk fibroin (SF) was immobilized on the surface of electrospun PCL nanofibers via covalent bonds in order to improve their hydrophilicity. To this end, the surface of PCL nanofibers was activated by ultraviolet (UV)–ozone irradiation followed by carboxylic functional groups immobilization on their surface by their immersion in acrylic acid under UV radiation and final immersion in SF solution. Furthermore, morphological, mechanical, contact angle, and Attenuated total reflection‐ Fourier transform infrared (ATR‐FTIR) were measured to assess the properties of the surface‐modified PCL nanofibers grafted with SF. ATR‐FTIR results confirmed the presence of SF on the surface of PCL nanofibers. Moreover, contact angle measurements of the PCL nanofibers grafted with SF showed the contact angle of zero indicating high hydrophilicity of modified nanofibers. In vitro cell culture studies using NIH 3T3 mouse fibroblasts confirmed enhanced cytocompatibility, cell adhesion, and proliferation of the SF‐treated PCL nanofibers. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 46684.     

Composite polymer electrolytes based on electrospun thermoplastic polyurethane membrane and polyethylene oxide for all‐solid‐state lithium batteries

To improve the electrochemical properties and enhance the mechanical strength of solid polymer electrolytes, series of composite polymer electrolytes (CPEs) were fabricated with hybrids of thermoplastic polyurethane (TPU) electrospun membrane, polyethylene oxide (PEO), SiO₂ nanoparticles and lithium bis(trifluoromethane)sulfonamide (LiTFSI). The structure and properties of the CPEs were confirmed by SEM, XRD, DSC, TGA, electrochemical impedance spectroscopy and linear sweep voltammetry. The TPU electrospun membrane as the skeleton can improve the mechanical properties of the CPEs. In addition, SiO₂ particles can suppress the crystallization of PEO. The results show that the TPU‐electrospun‐membrane‐supported PEO electrolyte with 5 wt% SiO₂ and 20 wt% LiTFSI (TPU/PEO‐5%SiO₂‐20%Li) presents an ionic conductivity of 6.1 × 10^?4 S cm^?1 at 60 °C with a high tensile strength of 25.6 MPa. The battery using TPU/PEO‐5%SiO₂‐20%Li as solid electrolyte and LiFePO₄ as cathode shows an attractive discharge capacity of 152, 150, 121, 75, 55 and 26 mA h g^?1 at C‐rates of 0.2C, 0.5C, 1C, 2C, 3C and 5C, respectively. The discharge capacity of the cell remains 110 mA h g^?1 after 100 cycles at 1C at 60 °C (with a capacity retention of 91%). All the results indicate that this CPE can be applied to all‐solid‐state rechargeable lithium batteries. © 2018 Society of Chemical Industry     

Electrospun Polycaprolactone/Silk Fibroin/Small Intestine Submucosa Composites for Biomedical Applications

Natural biomaterials were used to improve the biocompatibility of synthetic biopolymers. PCL was electrospun with natural biopolymers, silk fibroin, and small intestine submucosa. Due to increased electrical conductivity, the diameter of the composite fibers highly depended on the amount of SIS in the polymer solution. PCL/SF/SIS electrospun composites exhibited various synergistic effects, including enhanced mechanical properties and incredibly improved hydrophilicity compared to those of pure PCL and PCL/SF fibers. An initial cell attachment test demonstrated that the interactions between PC‐12 nerve cells and the PCL/SF/SIS composites were more favorable than those between PC‐12 cells and a PCL/SF composite.

     

Bioactive core–shell structure of TiC/TiO2/SrCO3 coating on carbon black particles for bone tissue formation: Fabrication,characterization, and biological functions

Although carbon-based nanomaterials, such as carbon nanotubes, graphene, and carbon dots, have attracted much attention for bone tissue regeneration and engineering due to the advantages of being lightweight, mechanical stability, and remarkable ability for bone repair, their toxicity and dispersity are the most concern and greatly limiting their clinical uses. In this article, the surface modification of carbon black particles based on core–shell structure design as a promising candidate material for bone tissue engineering applications is presented. TiC/TiO₂/SrCO₃-coated carbon black particles were prepared via molten salt synthesis and hydrothermal process at various temperatures to study the effects of temperature on crystal structure, morphologies, surface wettability, and biological functions. Phase composition, morphologies, and elemental distributions were studied by X-ray diffraction, field-emission scanning electron microscope, and energy-dispersive X-ray spectroscopy, respectively. Cell proliferation, cell viability, alkaline phosphatase (ALP) activity, and calcium deposition were also investigated. The investigation showed that the reaction temperature played an important role in the crystallinity, phase formation, nanotopography, and biological functions of the particles. The particles treated at 250°C offered favored surface properties of roughness, composition, crystallite size, and wettability for cell adhesion, proliferation, ALP activity, and calcium deposition. As a result, these bioactive core–shell particles would be a promising filler material for bone tissue engineering applications.     

Hybrid biomimetic electrospun fibrous mats derived from poly(ε‐caprolactone) and silk fibroin protein for wound dressing application

To achieve excellent biofunctionality of Bombyx mori silk fibroin (SF), we explored a novel hybridization method to combine the unique properties of SF with poly(ε‐caprolactone) (PCL) electrospun fibers. The hybrid electrospun fibers demonstrate excellent hydrophilicity and biocompatibility that are important to tissue engineering applications. The biomimetic fibrous structure was fabricated by conventional electrospinning of PCL. The individual surfaces of PCL electrospun fibers were coated with silk fibroin protein using a lyophilization technique. The SF coating layers were durable which were further developed by surface modification with fibronectin to improve their biological function. The hybrid electrospun fibers show excellent support for normal human dermal fibroblast (NHDF) cells adhesion and proliferation than neat PCL fibers, while the surface‐modified hybrid electrospun fibers show significantly enhanced proliferation of NHDF cells on their surface. This study indicates the new opportunity of fabrication technique that can construct a biomimetic fibrous structure while the original function as a biomaterial remained existing. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41653.     

Prevention of Intra-Abdominal Adhesion by Bi-Layer Electrospun Membrane

The aim of this study was to compare the anti-adhesion efficacy of a bi-layer electrospun fibrous membrane consisting of hyaluronic acid-loaded poly(ɛ-caprolactone) (PCL) fibrous membrane as the inner layer and PCL fibrous membrane as the outer layer with a single-layer PCL electrospun fibrous membrane in a rat cecum abrasion model. The rat model utilized a cecal abrasion and abdominal wall insult surgical protocol. The bi-layer and PCL membranes were applied between the cecum and the abdominal wall, respectively. Control animals did not receive any treatment. After postoperative day 14, a visual semiquantitative grading scale was used to grade the extent of adhesion. Histological analysis was performed to reveal the features of adhesion tissues. Bi-layer membrane treated animals showed significantly lower adhesion scores than control animals (p < 0.05) and a lower adhesion score compared with the PCL membrane. Histological analysis of the bi-layer membrane treated rat rarely demonstrated tissue adhesion while that of the PCL membrane treated rat and control rat showed loose and dense adhesion tissues, respectively. Bi-layer membrane can efficiently prevent adhesion formation in abdominal cavity and showed a significantly decreased adhesion tissue formation compared with the control.     

丝素-聚己内酯纳米纤维膜的制备工艺

以六氟异丙醇(HFIP)为溶剂,采用静电纺丝技术制备丝素(SF)-聚己内酯(PCL)复合纳米纤维膜。采用热场发射扫描电镜、Image-Pro Plus图像分析和力学拉伸的方法表征了纳米纤维膜的结构与力学性能。通过设计的三因素四水平正交试验对复合纳米纤维膜的多个指标进行了分析,采取归一化数据处理及平均权重分配的方式量化了复合纳米纤维膜的品质,确定了共混复合纳米纤维膜制备的最优工艺参数,并且采用最佳工艺参数制备了SF-PCL复合纳米纤维膜,分析了其力学性能。结果表明:在溶质质量分数为6%、溶质SF与PCL质量比为3∶2、纺丝流速1.2 mL/h时,SF-PCL复合纳米纤维膜具有较好的品质;双轴拉伸时的破坏机制与单轴不同,其断裂应力和应变只是单轴时的一半左右,膜的力学性能表现为各向同性。     

Fabrication and characterization of electrospun PCL/Antheraea pernyi silk fibroin nanofibrous scaffolds

To engineer tissue restoration, it is necessary to provide a bioactive, mechanically robust scaffold. Electrospun poly(ε‐caprolactone) (PCL) nanofiber is a promising biomaterial candidate with excellent mechanical properties, but PCL scaffolds are inert and lack natural cell recognition sites. To overcome this problem we investigated the incorporation of Antheraea pernyi silk fibroin (ASF) containing inherent RGD tripeptides with PCL in electrospinning process. The mixing ratios showed remarkable impact on the properties of hybrid nanofibers. Increasing PCL content significantly enhanced the mechanical properties of nanofibers. In particular, the mechanical properties were remarkably enhanced when PCL content increased from 50 wt% to 70 wt%. Moreover, the biological assays based on endothelial cells showed promoted cell viability when ASF content reached to 30 wt%. The data demonstrated that the nanofiber containing 70% of PCL and 30% of ASF achieved the most balanced performances for integrating the mechanical properties of PCL and the bioactivity of ASF. Furthermore, biomimetic alignment of 70PCL/30ASF nanofibers was achieved, which could support PC12 neuron‐like cell growth and guide neurite outgrowth, providing a potentially useful option for the engineering of oriented tissues. The results show that the PCL/ASF hybrid nanofibers can be considered as a promising candidate for tissue engineering scaffolds. POLYM. ENG. SCI., 57:206–213, 2017. © 2016 Society of Plastics Engineers     

Characterization and Mechanical Properties of Calcium Silicate/Citric Acid–Based Polymer Composite Materials

Optimal implants for bone tissue engineering require sufficient mechanical strength as well as apt bioactivity and biodegradability. Calcium silicate (CaSiO₃ ‐ CS) ceramics have been studied for tissue engineering and implantation for their good bioactivity properties. Elastomer poly (1.8‐octanediol citrate) (POC), one of the most biocompatible polymer, is used for biomedical application. The objective of this study is to fabricate a novel composite of calcium silicate with different ratios of POC to enhance the mechanical properties. The results showed that the compressive and the bending strengths of calcium silicate/POC biocomposite were improved remarkably at 40 wt% POC.     

Reinforcement of electrospun poly(ε‐caprolactone) scaffold using diopside nanopowder to promote biological and physical properties

Considerable efforts have been devoted toward the development of electrospun scaffolds based on poly(ε‐caprolactone) (PCL) for bone tissue engineering. However, most of previous scaffolds have lacked the structural and mechanical strength to engineer bone tissue constructs with suitable biological functions. Here, we developed bioactive and relatively robust hybrid scaffolds composed of diopside nanopowder embedded PCL electrospun nanofibers. Incorporation of various concentrations of diopside nanopowder from 0 to 3 wt % within the PCL scaffolds notably improved tensile strength (eight‐fold) and elastic modulus (two‐fold). Moreover, the addition of diopside nanopowder significantly improved bioactivity and degradation rate compared to pure PCL scaffold which might be due to their superior hydrophilicity. We investigated the proliferation and spreading of SAOS‐II cells on electrospun scaffolds. Notably, electrospun PCL‐diopside scaffolds induced significantly enhanced cell proliferation and spreading. Overall, we concluded that PCL‐diopside scaffold could potentially be used to develop clinically relevant constructs for bone tissue engineering. However, the extended in vivo studies are essential to evaluate the role of PCL‐diopside fibrous scaffolds on the new bone growth and regeneration. Therefore, in vivo studies will be the subject of our future work. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134 , 44433.     

Influence of lactic acid on degradation and biocompatibility of electrospun poly(ε‐caprolactone) fibers

Electrospinning of a poly(ε‐caprolactone) (PCL)/lactic acid (LA) blend was investigated to fabricate electrospun PCL fibers with improved biodegradability and biocompatibility for biomedical applications. Simple blending of PCL solution with various amounts of LA was used for electrospinning, and the physicochemical properties of the as‐fabricated mat were evaluated using various techniques. Scanning electron microscopy showed that fiber diameter decreased with increasing amount of LA. Fourier transform infrared spectroscopy and thermogravimetric analysis also revealed that LA was successfully incorporated in PCL fibers. The presence of LA can accelerate the biodegradation of PCL fibers and enhance the hydrophilicity of a membrane. The adhesion, viability and proliferation properties of osteoblast cells on the PCL/LA composite fibers were analyzed using in vitro cell compatibility tests which showed that LA can increase the cell compatibility of PCL fibers. Additionally, subsequent conversion of LA into calcium lactate by neutralization with calcium base can provide Ca²⁺ ions on the fiber surface to promote the nucleation of CaPO₄ particles. © 2013 Society of Chemical Industry     

Preparation and characterization of poly(ε‐caprolactone)/calcium carbonate nanocomposites and nanocomposite foams

Biodegradable poly(ε‐caprolactone) (PCL)/calcium carbonate (CaCO₃) nanocomposites were prepared and characterized. Effect of CaCO₃ on thermal and mechanical properties of PCL matrix was studied. Results showed that CaCO₃ acts as a crystallization nucleating agent and introduction of CaCO₃ leads to improved mechanical properties of the PCL matrix. PCL/CaCO₃ nanocomposite foams were prepared using chemical foaming method. Cellular parameters such as mean cell size, cell wall thickness, and cell density were collected. The cellular structure of nanocomposite foams changes with different CaCO₃ loading. Mean cell size achieved the minimum value at 5 wt% CaCO₃ loading, and cell wall thickness increased with CaCO₃ content. The changes in cellular structure and improvement of mechanical properties also enhanced the mechanical properties of PCL/CaCO₃ nanocomposite foams. Compressive moduli of PCL/CaCO₃ nanocomposite foams with similar density increased with increasing CaCO₃ loading. POLYM. COMPOS., 31:1653–1661, 2010. © 2009 Society of Plastics Engineers     

Fabrication, characterization and invitro bioactivity evaluation of QPSU/TiO2 composite membranes

Quaternized Polysulfone (QPSU) is a widely investigated material in the industry because of its unique properties such as resistance to corrosion and high mechanical properties. The ionic nature of the compound can be exploited for medical applications such as in haemodialysis, drug delivery and tissue engineering. In this study, composite membranes of QPSU with varying concentrations of Titanium oxide (TiO₂) were prepared and characterized using FT-IR, ¹H-NMR, XRD, TGA and SEM. The bioactivity of the membranes was studied by immersing them in simulated body fluid (SBF) for 7 days and subsequently observing under SEM for the formation of calcium-phosphate (Ca–PO₄) layer on the surface of the membranes. The formation of Ca–PO₄ on the samples was confirmed using FT-IR and EDAX. The results were compared with those obtained for QPSU membranes and the effect of TiO₂ concentration on the membrane properties was analyzed. It was observed that the percentage crystallinity of the composites increased upto a filler concentration of 5 wt% beyond which it decreased. TGA studies revealed an increase in the thermal stability of the composites with increasing filler concentrations. While optimum bioactivity was observed in the samples containing 5 wt% of TiO₂, higher filler content resulted in the formation of denser calcium—phosphate layer on the surface of the composites. The study shows that quaternized polysulphone/TiO₂ composites are promising bio composites having great potential for application in health care.     

Three​-dimensional porous poly(ε-caprolactone)/beta-tricalcium phosphate microsphere-aggregated scaffold for bone tissue engineering

In this study, porous scaffolds made of polycaprolactone (PCL)/β-tricalcium phosphate (BTCP) biocomposite were fabricated for bone tissue engineering (BTE) applications. The microsphere-aggregated scaffolds were prepared with various BTCP concentrations (10wt%, 20wt%, 50wt%) by the freeze-drying method. The porosity of obtained microsphere-aggregated scaffolds with various pore sizes was 80–85%, where this value was about 70% for the PCL/BTCP (50) sample with no microsphere formation. The results indicated that adding BTCP has enhanced mechanical strength, and the mineralization of PCL/BTCP composite scaffolds has been increased compared to the pure PCL scaffolds in simulated body fluid (SBF). The adhesion and proliferation of mouse bone marrow mesenchymal stem cells (mMSCs) seeded onto PCL/BTCP scaffolds were enhanced compared to the PCL. In addition, in terms of differentiation, the incorporation of BTCP led to increasing the mineral deposition and alkaline phosphatase activity of mMSCs. The synergistic effect of using microsphere-aggregated scaffolds along with BTCP as a reinforcing agent in PCL biocomposite showed that these porous biocomposite scaffolds have the potential application in BTE.     

Evaluation of ZnO-Vitamin B1 Nanoparticles on Bioactivity and Physiochemical Properties of the Polycaprolactone-Based Nanocomposites

In the present study, we investigated the use of thiamine chloride hydrochloride (vitamin B₁)-modified ZnO nanoparticles (ZnO-VB₁ NPs) to reinforce polycaprolactone matrix. The stable and bioactive PCL/ZnO-VB₁ nanocomposites were fabricated with the combination of ultrasonication and solution casting methods. Transmission electron microscope results indicated that the ZnO-VB₁ NPs were uniformly dispersed in the matrix. The nanocomposites showed high hydroxyapatite formation (high bioactivity) in the simulated body fluid. The nanocomposites with 2?wt% of the modified nanoparticles were found to have highest mechanical strength. The nanocomposites with more nanofiller concentrations exhibited high wettability.     

Preparation and characterization of Rana chensinensis skin extract/poly(ε‐caprolactone) electrospun membranes as antibacterial fibrous mats

In this study, membranes composed of Rana chensinensis skin extracts (RCSEs) and poly(ε‐caprolactone) (PCL) were fabricated by an electrospinning technique. The RCSEs were prepared by the extraction of R. chensinensis skin with acetic acid solution, and the electrospun membranes were prepared by the mixture of RCSEs and PCL in 1,1,1,3,3,3‐hexafluoro‐2‐propanol before electrospinning. The membranes were characterized by scanning electron microscopy and Fourier transform infrared spectroscopy and were subjected to mechanical tests (tensile and nanoindentation) and antibacterial evaluation. The results indicate that the surface roughness of the fibers clearly decreased with the increase in the amount of PCL in the membranes. The mechanical test indicated that PCL played a dominant role in the mechanical strength of the RCSE/PCL electrospun membranes. As a potential bactericidal packaging material for practical applications, the antibacterial activity results indicate that the membranes had antibacterial effects against Staphylococcus aureus and Escherichia coli. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 42030.     

Chitosan/CaCO3 solvent‐free nanofluid composite membranes for direct methanol fuel cells

Natural polyelectrolyte chitosan (CS) has been considered to be a promising proton‐exchange membrane material for direct methanol fuel cells due to its low cost and excellent methanol barrier ability. To further improve the ionic conductivity and mechanical property of CS, calcium‐carbonate solvent‐free nanofluids (CaCO₃‐SF) with unique flow behavior were prepared by an ion‐exchange method, and then used a novel nanofiller to modify CS to fabricate composite membranes. The surface‐grafted organic long chains on the surface of CaCO₃ nanoparticles could promote the homogeneous dispersion of CaCO₃ in the CS matrix, and thus improve the interfacial bonding and facilitate the load transfer from the matrix to stiff CaCO₃. When the content of CaCO₃‐SF was 6 wt%, the tensile strength and fracture elongation of the composite membrane were 28.25 MPa and 17.17%, respectively, which increased by 25% and 36% when compared with those of pure membrane. Moreover, the ? SO₃H groups in the structure of organic long chains could form new proton transport sites, and thus enhance the proton conductivity of the membranes. Consequently, when compared with pure CS membrane (0.0131 S cm^?1), incorporation of 6 wt% CaCO₃‐SF (0.0250 S cm^?1) exhibited about onefold increase of proton conductivity. POLYM. ENG. SCI., 59:2128–2135, 2019. © 2019 Society of Plastics Engineers