Preparation and characterization of hydroxyapatite/collagen nanocomposite gel

The self-organized hydroxyapatite/colagen (HAp/Col) nanocomposite fiber (79.6/20.4 weight ratio) was synthesized by a co-precipitation method using Ca(OH)2, H3PO4, and Col as starting substances. The gelation of the nanocomposite is essential in the application of the scaffold for bone tissue engineering. We successfully prepared HAp/Col nanocomposite gels by a facile novel method using a sodium phosphate buffer at pH 6.8. The water-insoluble nanocomposite was homogeneously dispersed in the buffer to form a viscous mixture, and gels were obtained after incubating of the mixture at 37 degrees C. The mechanical strength of the gels was analyzed against the incubation time. The demineralized gel with EDTA had the typical nanostructure of native type I Col fibers from the results of scanning electron microscopy (SEM) and atomic force microscopy (AFM); the dense network of type I Col nano-fibers below 100 nm in diameter, and the periodic pattern of 68.8+/-4.4 nm (mean +/- SD) along the fibers were observed. The gelation of the HAp/Col nanocomposite in the buffer is attributed to the physical cross-linking through entanglement of the reconstituted Col fibrils.     

Bioactive nanocomposite coatings of collagen/hydroxyapatite on titanium substrates

Collagen/hydroxyapatite (HA) nanocomposite thin films containing 10, 20, and 30 wt.% HA were prepared on commercially pure titanium substrates by the spin coating of their homogeneous sols. All of the nanocomposite coatings having a thickness of ∼7.5 μm exhibited a uniform and dense surface, without any obvious aggregation of the HA particles. A minimum contact angle of 36.5° was obtained at 20 wt.% HA, suggesting that these coatings would exhibit the best hydrophilicity. The in vitro cellular assays revealed that the coating treatment of the Ti substrates favored the adhesion of osteoblast-like cells and significantly enhanced the cell proliferation rate. The cells on the nanocomposite coatings expressed much higher alkaline phosphatase (ALP) levels than those on the uncoated Ti substrates. Increasing the amount of HA resulted in a gradual improvement in the ALP activity. The nanocomposite coatings on Ti substrates also exhibited much better cell proliferation behaviors and osteogenic potentials than the conventional composite coatings with equivalent compositions, demonstrating the greater potential of the former as implant materials for hard tissue engineering.     

Facile fabrication of the glutaraldehyde cross-linked collagen/chitosan porous scaffold for skin tissue engineering

Porous scaffold is one of the key factors in skin tissue engineering. In this study, a facile method was developed to prepare the glutaraldehyde (GA) cross-linked collagen/chitosan porous scaffold (S2). The properties of S2 were compared with the scaffolds prepared by the traditional method (S1). Compared to the rough surface and collapsed inner structure of S1, S2 showed a smooth surface and controlled size. After treated by GA with same concentration, S1 and S2 showed the similar swelling ratios, which are big enough to ensure the nutrient supply in the early stage of wound healing. The effects of the fabrication methods as well as the GA concentration on the cross-linking degree and in vitro degradation degree of the scaffolds were studied. It was found that the cross-linking degree of S2-0.25% was much higher than that of S1. Investigation of the tensile and compression properties of the scaffolds found that the mechanical property of S2-0.04% is closest to that of S1. High performance liquid chromatography (HPLC) was applied to determine the residual GA. The results proved that, compared to water rinse, oven drying is a feasible and effective method to remove the residual GA. Finally, the cytocompatibility of S2 was evaluated by in vitro culture of fibroblasts. The results of cell morphology and cell viability proved that S2-0.04% could retain the original good cytocompatibility of S1 to accelerate cell infiltration and proliferation effectively. All these results indicate that it is a feasible method to prepare the GA cross-linked collagen/chitosan scaffold.     

A novel porous scaffold fabrication technique for epithelial and endothelial tissue engineering

Porous scaffolds have the ability to minimize transport barriers for both two- (2D) and three-dimensional tissue engineering. However, current porous scaffolds may be non-ideal for 2D tissues such as epithelium due to inherent fabrication-based characteristics. While 2D tissues require porosity to support molecular transport, pores must be small enough to prevent cell migration into the scaffold in order to avoid non-epithelial tissue architecture and compromised function. Though electrospun meshes are the most popular porous scaffolds used today, their heterogeneous pore size and intense topography may be poorly-suited for epithelium. Porous scaffolds produced using other methods have similar unavoidable limitations, frequently involving insufficient pore resolution and control, which make them incompatible with 2D tissues. In addition, many of these techniques require an entirely new round of process development in order to change material or pore size. Herein we describe “pore casting,” a fabrication method that produces flat scaffolds with deterministic pore shape, size, and location that can be easily altered to accommodate new materials or pore dimensions. As proof-of-concept, pore-cast poly(ε-caprolactone) (PCL) scaffolds were fabricated and compared to electrospun PCL in vitro using canine kidney epithelium, human colon epithelium, and human umbilical vein endothelium. All cell types demonstrated improved morphology and function on pore-cast scaffolds, likely due to reduced topography and universally small pore size. These results suggest that pore casting is an attractive option for creating 2D tissue engineering scaffolds, especially when the application may benefit from well-controlled pore size or architecture.     

Zinc-doped hydroxyapatite and poly(propylene fumarate) nanocomposite scaffold for bone tissue engineering

Hydroxyapatite (HA) is a bioceramic material that shares similar crystal and chemical structures with inorganic components of the bone. However, HA lacks osteoinductive activity and has a brittle nature, making it challenging to apply for direct load-bearing bone applications. In this study, we used a wet chemical method to synthesize zinc-doped hydroxyapatite powders with different Zn/(Zn+Ca) molar ratios of 0, 0.025, 0.05, and 0.1. The corresponding Zn-HA was designated as HA, Zn2.5-HA, Zn5-HA, and Zn10-HA. The Zn-HA powders at 30 wt% were used to fabricate poly(propylene fumarate) (PPF)-based nanocomposite scaffolds (HA/PPF, Zn2.5-HA/PPF, Zn5-HA/PPF, and Zn10-HA/PPF). The physical properties of obtained scaffolds were examined by scanning electron microscopy, energy-dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), and atomic force microscopy (AFM). Live/dead cell viability assay showed that these scaffolds were biocompatible and supported excellent adhesion of MC3T3-E1 preosteoblast cells. Additionally, the proliferation of cells was detected at 1, 4, and 7 days on these scaffolds. Alkaline phosphatase (ALP) activity measurement and alizarin red staining showed good osteogenic differentiation and matrix mineralization for MC3T3-E1 cells growing on these scaffolds. Taken together, the results here indicate that Zn5-HA/PPF nanocomposite scaffolds are promising scaffold material for bone tissue engineering.

Graphical abstract

     

一种制备纳米羟基磷灰石/聚乙烯醇多孔支架材料的新方法

采用乳化发泡-冷冻法制备了聚乙烯醇和纳米羟基磷灰石/聚乙烯醇两种多孔材料.用SEM对材料进行了表征,结果表明所得样品孔径均匀,孔隙率高,且孔内外贯通,是一种新型的制备多孔材料的方法.文中还讨论了乳化剂op、聚乙烯醇、纳米羟基磷灰石、搅拌速度等因素对孔的大小和孔隙率的影响.     

A novel approach via combination of electrospinning and FDM for tri-leaflet heart valve scaffold fabrication

In this paper, a novel combination method of electrospinning and rapid prototyping (RP) fused deposition modeling (FDM) is proposed for the fabrication of a tissue engineering heart valve (TEHV) scaffold. The scaffold preparation consisted of two steps: tri-leaflet scaffold fabrication and heart valve ring fabrication. With the purpose of mimicking the anisotropic mechanical properties of the natural heart valve leaflet, electrospun thermoplastic polyurethane (ES-TPU) was introduced as the tri-leaflet scaffold material. ES-TPU scaffolds can be fabricated to have a well-aligned fiber network, which is important for applications involving mechanically anisotropic soft tissues. We developed ES-TPU scaffolds as heart valve leaflet materials under variable speed conditions and measured fiber alignment by fast Fourier transform (FFT). By using FFT to assign relative alignment values to an electrospun matrix, it is possible to systematically evaluate how different processing variables affect the structure and material properties of a scaffold. TPU was suspended at certain concentrations and electrospun from 1,1,1,3,3,3-hexafluoro-2-propanol onto rotating mandrels (200–3000 rpm). The scaffold morphological property and mechanical anisotropic property are discussed in the paper as a function of fiber diameter and mandrel RPM. The induction of varying degrees of anisotropy imparted distinctive material properties to the electrospun scaffolds. A dynamic optimum design of the heart valve ring graft was constructed by FDM. Fabrication of a 3D heart valve ring was constructed using pro-engineer based on optimum hemodynamic analysis and was converted to an STL file format. The model was then created from PCL which was sewed and glued with electrospun nanofibrous leaflets. This proposed method was proven as a promising fabrication process in fabricating a specially designed graft with the correct physical and mechanical properties.     

A self-organising biomimetic collagen/nano-hydroxyapatite-glycosaminoglycan scaffold for spinal fusion

The use of spinal fusion surgery as a treatment for degenerative spinal conditions and chronic back pain is increasing. However, this technique requires use of a bone grafting material to fuse the vertebrae, traditionally autologous bone, which consists of an optimal combination of osteogenic cell precursors, extracellular matrix proteins and mineral components. To date, this remains the ‘gold standard’ material but its supply is limited and is associated with a number of clinical and ethical difficulties; consequently, various combinations of cells with biological scaffold materials have been tested but have failed to achieve fusion rates even comparable to autologous bone. We successfully fabricated a novel collagen-based scaffold using self-organising atelocollagen combined with nano-hydroxyapatite and chondroitin sulphate, cross-linked by microbial transglutaminase. The scaffold was characterised using a range of imaging, chemical composition and thermal analysis techniques. It was found to exhibit appropriate stiffness and suitable pore size for the adhesion, growth and differentiation of MSCs. The low toxicity makes it suitable for clinical application, and its slow degradation profile would enable the scaffold to promote bone growth over an extended period. This material therefore shows promise for clinical use in spinal fusion and other procedures requiring the use of bone grafts.     

A novel approach via combination of electrospinning and FDM for tri-leaflet heart valve scaffold fabrication

In this paper, a novel combination method of electrospinning and rapid prototyping (RP) fused deposition modeling (FDM) is proposed for the fabrication of a tissue engineering heart valve (TEHV) scaffold. The scaffold preparation consisted of two steps: tri-leaflet scaffold fabrication and heart valve ring fabrication. With the purpose of mimicking the anisotropic mechanical properties of the natural heart valve leaflet, electrospun thermoplastic polyurethane (ES-TPU) was introduced as the tri-leaflet scaffold material. ES-TPU scaffolds can be fabricated to have a well-aligned fiber network, which is important for applications involving mechanically anisotropic soft tissues. We developed ES-TPU scaffolds as heart valve leaflet materials under variable speed conditions and measured fiber alignment by fast Fourier transform (FFT). By using FFT to assign relative alignment values to an electrospun matrix, it is possible to systematically evaluate how different processing variables affect the structure and material properties of a scaffold. TPU was suspended at certain concentrations and electrospun from 1,1,1,3,3,3-hexafluoro-2-propanol onto rotating mandrels (200―3000 rpm). The scaffold morphological property and mechanical anisotropic property are discussed in the paper as a function of fiber diameter and mandrel RPM. The induction of varying degrees of anisotropy imparted distinctive material properties to the electrospun scaffolds. A dynamic optimum design of the heart valve ring graft was constructed by FDM. Fabrication of a 3D heart valve ring was constructed using pro-engineer based on optimum hemodynamic analysis and was converted to an STL file format. The model was then created from PCL which was sewed and glued with electrospun nanofibrous leaflets. This proposed method was proven as a promising fabrication process in fabricating a specially designed graft with the correct physical and mechanical properties.     

A novel method for fabrication of biodegradable scaffolds with high compression moduli

It has been previously shown that, when used for meniscal reconstruction, porous copoly(L-lactide/-caprolactone) implants enhanced healing of meniscal lesions owing to their excellent adhesive properties. However, it appeared that the materials had an insufficient compression modulus to accomplish 100% fibrocartilage formation. In addition, to be used for meniscal prosthesis, the compression modulus of the porous materials should be larger than 150 kPa in order to protect the articular cartilage. A technique was developed to prepare stiff porous materials of a high molecular weight 50/50 copoly(L-lactide/-caprolactone) suitable for fibrocartilage regeneration in meniscal implants and meniscal prosthesis. Porous microspheres (50–250 m) were agglutinated in the presence of NaCl crystals (250–300 m). The microspheres were mixed with solid solvent in order to obtain a homogeneous distribution of solvent over the spheres. By changing the amount of solvent and crystals, the density and the compression modulus could be varied over a range of 0.07 g ml-1 to 0.5 g dl-1 and 40–1100 kPa, respectively.     

A novel fabrication method for titania resistive switching device

The titania showing reversible resistive switching are attractive for today's semiconductor technology in nonvolatile random-access memories. A novel fabrication method for titania resistive switching device with vertical structure is proposed. First, the Pt electrode was fabricated the bottom using conventional photolithography and chemical etching technique. Next, the titania thin films with the thickness about 50 nm was deposited on the bottom electrode by electron beam evaporation (EBE). Then, the trench of photoresist for electrode deposit was etched with mild chemical process to preserve the original structure of titania layer. After that, the platinum was deposited in the trench of photoresist using ion sputter. A final lift-off process to define the Pt top electrodes was performed with acetone in an ultrasonic bath to remove the resist. The resistive bistability was observed in this device. The on-threshold voltage is +1.5 V and the off-threshold voltage is -0.6 V. The resistance ratio between the two stable states of the device including Al electrode is approximately 1 x 10(3), the state is nonvolatile and the retention-time test performed over an hour in sweeping mode measurement. The results indicate the forming and rupture of conductive channel relate to the defects and distributing of oxygen vacancy. This method is low-cost, high-yielding, and easy to implement, which is applicable to the fabrication of nonvolatile memories.     

A novel method to measure the porosity of porous materials

ABSTRACT

Porosity is the single most important feature of porous materials. On the basis of the research work, a new method to measure the porosity of porous materials is proposed in this paper. This method is a system of equations composed of six formulas, so it is called model equation method. It is not only applicable to space holder technique but also applicable to metal prepared by pure powder metallurgy. Compared with mass volume method and immersion medium method, our new method is more simple and efficient. More importantly, it can be used as a standard measure. This study provides a fundamental basis for the measurement of porosity in porous materials.     

A novel fabrication method for the nanoscale tunneling field effect transistor

Tunneling Field Effect Transistors (TFETs) are considered as a candidate for low power applications. However, most of TFETs have been researched on only for long channels due to the misalignment problem that occurs during the source/drain doping process in device fabrication. Thus, a new method is proposed for the fabrication of TFETs in nanoscale regions. This proposed fabrication process does not need an additional mask to define the source/drain regions, and makes it possible to form a self-aligned source/drain doping process. In addition, through TCAD simulation, the electrical characteristics of a TFET with dopant engineering and a rounded gate edge shape for a higher on/off current ratio were investigated. As a result, the TFET showed the properties of a larger on-current, a lower average subthreshold swing (58.5 mV/dec), and a 30-fold smaller leakage current compared to the conventional TFET The TFET with dopant engineering and a rounded gate edge shape can also be fabricated simply through the proposed fabrication process.     

A novel method for massive fabrication of β-SiC nanowires

Silicon carbide nanowires (NWs), that were over 200 μm in length and 20–200 nm in diameter, were prepared by high-pressure reaction from SiBONC powder tablets. Annealing temperatures between 1,500 °C and 1,600 °C and Si/B molar ratios between 70:30 and 60:40 were suitable for the growth of the nanowires. The nanowires were fabricated by in situ chemical vapor growth process on the tablets. The SiC nanowires were identified as single crystal β-SiC. The analysis of X-ray diffraction (XRD) and transmission electron microscopy (TEM) showed the single crystalline nature of nanowires with a growth direction of <111>. Massive growth of single crystalline SiC nanowires is important to meet the requirements of the fabrication of SiC nanowire-based nanodevices.     

Preparation,fabrication and biocompatibility of novel injectable temperature-sensitive chitosan/glycerophosphate/collagen hydrogels

This paper introduces a novel type of injectable temperature-sensitive chitosan/glycerophosphate/collagen (C/GP/Co) hydrogel that possesses great biocompatibility for the culture of adipose tissue-derived stem cells. The C/GP/Co hydrogel is prepared by mixing 2.2% (v/v) chitosan with 50% (w/w) β-glycerophosphate at different proportions and afterwards adding 2 mg/ml of collagen. The gelation time of the prepared solution at 37°C was found to be of around 12 min. The inner structure of the hydrogel presented a porous spongy structure, as observed by scanning electron microscopy. Moreover, the osmolality of the medium in contact with the hydrogel was in the range of 310–330 mmol kg⁻¹. These analyses have shown that the C/GP/Co hydrogels are structurally feasible for cell culture, while their biocompatibility was further examined. Human adipose tissue-derived stem cells (ADSCs) were seeded into the developed C/GP and C/GP/Co hydrogels (The ratios of C/GP and C/GP/Co were 5:1 and 5:1:6, respectively), and the cellular growth was periodically observed under an inverted microscope. The proliferation of ADSCs was detected using cck-8 kits, while cell apoptosis was determined by a Live/Dead Viability/Cytotoxicity kit. After 7 days of culture, cells within the C/GP/Co hydrogels displayed a typical adherent cell morphology and good proliferation with very high cellular viability. It was thus demonstrated that the novel C/GP/Co hydrogel herein described possess excellent cellular compatibility, representing a new alternative as a scaffold for tissue engineering, with the added advantage of being a gel at the body’s temperature that turns liquid at room temperature.     

Injectable silk/hydroxyapatite nanocomposite hydrogels with vascularization capacity for bone regeneration

Localized and sustained osteogenic-angiogenic stimulation to bone defects is critical for effective bone repair.Here,desferrioxamine(DFO)was loaded on silk fibroin nanofibers and blended with hydroxyapatite nanorods(HA),forming injectable DFO-loaded silk fibroin-HA nanocomposite hydrogels.The composite hydrogels remained homogeneous distribution of HA with high ratio(60%)and also higher stiffness than that of pure silk fibroin nanofiber hydrogels,which provided stable osteogenic stimulation niches for tissue regeneration.Without the scarify of injectability,the hydrogels achieved slow delivery of DFO for above 60 days,resulting in suitable angiogenesis in vitro and in vivo and better osteogenesis than DFO-free systems.Compared to previous injectable silk fibroin-HA hydrogels,the introduction of vascularization capacity further stimulated the osteogenic differentiation of stem cells and accelerated new bone formation.Quicker and better bone healing were detected at defect sites after the injection of DFO-loaded nanocomposite hydrogels,indicating the effective synergistic effect of osteogenic and angiogenic cues.This work provides a simple and effective strategy of introducing angiogenic cues to bone matrices.We believe that the injectable nanocomposite hydrogels are suitable for the regeneration of bone tissues.     

Mineralization of osteoblasts with electrospun collagen/hydroxyapatite nanofibers

Regeneration of fractured or diseased bones is the challenge faced by current technologies in tissue engineering. The major solid components of human bone consist of collagen and hydroxyapatite. Collagen (Col) and hydroxyapatite (HA) have potential in mimicking natural extracellular matrix and replacing diseased skeletal bones. More attention has been focused on HA because of its crystallographic structure similar to inorganic compound found in natural bone and extensively investigated due to its excellent biocompatibility, bioactivity and osteoconductivity properties. In the present study, electrospun nanofibrous scaffolds are fabricated with collagen (80 mg/ml) and Col/HA (1:1). The diameter of the collagen nanofibers is around 265 ± 0.64 nm and Col/HA nanofibers are 293 ± 1.45 nm. The crystalline HA (29 ± 7.5 nm) loaded into the collagen nanofibers are embedded within nanofibrous matrix of the scaffolds. Osteoblasts cultured on both scaffolds and show insignificant level of proliferation but mineralization was significantly (p < 0.001) increased to 56% in Col/HA nanofibrous scaffolds compared to collagen. Energy dispersive X-ray analysis (EDX) spectroscopy results proved the presence of higher level of calcium and phosphorous in Col/HA nanocomposites than collagen nanofibrous scaffolds grown osteoblasts. The results of the present study suggested that the designed electrospun nanofibrous scaffold (Col/HA) have potential biomaterial for bone tissue engineering.     

Electrophoretic-deposited novel ternary silk fibroin/graphene oxide/hydroxyapatite nanocomposite coatings on titanium substrate for orthopedic applications

The combination of graphene oxide (GO) with robust mechanical property, silk fibroin (SF) with fascinating biological effects and hydroxyapatite (HA) with superior osteogenic activity is a competitive approach to make novel coatings for orthopedic applications. Herein, the feasibility of depositing ternary SF/GO/HA nanocomposite coatings on Ti substrate was firstly verified by exploiting electrophoretic nanotechnology, with SF being used as both a charging additive and a dispersion agent. The surface morphology, microstructure and composition, in vitro hemocompatibility and in vitro cytocompatibility of the resulting coatings were investigated by SEM, Raman, FTIR spectra and biocompatibility tests. Results demonstrated that GO, HA and SF could be co-deposited with a uniform, smooth thin-film morphology. The hemolysis rate analysis and the platelet adhesion test indicated good blood compatibility of the coatings. The human osteosarcoma MG63 cells displayed well adhesion and proliferation behaviors on the prepared coatings, with enhanced ALP activities. The present study suggested that SF/GO/HA nanocomposite coatings could be a promising candidate for the surface functionalization of biomaterials, especially as orthopedic implant coating.     

Effect of silica and hydroxyapatite mineralization on the mechanical properties and the biocompatibility of nanocomposite collagen scaffolds

A recently established materials concept of biomimetic composites based on silica, collagen, and calcium phosphates was adapted for the preparation of porous scaffolds suitable for tissue engineering applications. Mineralization was achieved by directed nucleation of silica on the templating organic phase during a sol-gel process with or without addition of hydroxyapatite. Both mineral phases (25 wt %, individually or combined in equal shares) influenced the scaffold's morphology at the nanoscale. Enhancement of apparent density and compressive strength was similar for silica or hydroxyapatite mineralization; however the stiffening effect of hydroxyapatite was much higher. All scaffold modifications provided proper conditions for adhesion, proliferation, and osteogenic differentiation of human bone marrow stromal cells. The open porosity allowed cells to migrate throughout the scaffolds while maintaining their viability, both confirmed by MTT staining and confocal laser scanning microscopy. Initial cell distributions were graduated due to collagen mineralization, but balanced out over the cultivation time of 28 days. RT-PCR analyses revealed higher gene expression of ALP but lower expression of BSP II and osteocalcin because of collagen mineralization. The results demonstrate that both silica and hydroxyapatite offer comparable possibilities to tailor mechanical properties of collagen-based scaffolds without being detrimental to in vitro biocompatibility.