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Fabrication and endothelialization of collagen-blended biodegradable polymer nanofibers: potential vascular graft for blood vessel tissue engineering
He, W., T. Yong, et al. (2005), Tissue Eng 11(9-10): 1574-88.
Abstract: Electrospun collagen-blended poly(L-lactic acid)-co-poly(epsilon-caprolactone) [P(LLA-CL), 70:30] nanofiber may have great potential application in tissue engineering because it mimicks the extracellular matrix (ECM) both morphologically and chemically. Blended nanofibers with various weight ratios of polymer to collagen were fabricated by electrospinning. The appearance of the blended nanofibers was investigated by scanning electron microscopy and transmission electron microscopy. The nanofibers exhibited a smooth surface and a narrow diameter distribution, with 60% of the nanofibers having diameters between 100 and 200 nm. Attenuated total reflectance-Fourier transform infrared spectra and X-ray photoelectron spectroscopy verified the existence of collagen molecules on the surface of nanofibers. Human coronary artery endothelial cells (HCAECs) were seeded onto the blended nanofibers for viability, morphogenesis, attachment, and phenotypic studies. Five characteristic endothelial cell (EC) markers, including four types of cell adhesion molecule and one EC-preferential gene (von Willebrand factor), were studied by reverse transcription-polymerase chain reaction. Results showed that the collagen-blended polymer nanofibers could enhance the viability, spreading, and attachment of HCAECs and, moreover, preserve the EC phenotype. The blending electrospinning technique shows potential in refining the composition of polymer nanofibers by adding various ingredients (e.g., growth factors) according to cell types to fabricate tissue-engineering scaffold, particularly blood vessel-engineering scaffold.

Fabrication of a cell array on ultrathin hydrophilic polymer gels utilising electron beam irradiation and UV excimer laser ablation
Iwanaga, S., Y. Akiyama, et al. (2005), Biomaterials 26(26): 5395-404.
Abstract: Most of the surface patterning methods currently applied are based on lithography techniques and microfabrication onto silicon or glass substrates. Here we report a novel method to prepare patterned surfaces on polystyrene substrates by grafting ultrathin cell-repellent polymer layers utilising both electron beam (EB) polymerisation and local laser ablation techniques for microfabrication. Polyacrylamide was grafted onto tissue culture polystyrene (TCPS) dishes using EB irradiation. Water contact angles for these PAAm-grafted TCPS surfaces were less than 10 degrees (costheta = 0.99) with PAAm grafted amounts of 1.6 microg/cm(2) as determined by ATR/FT-IR. UV excimer laser (ArF: 193 nm) ablation resulted in the successful fabrication of micropatterned surfaces composed of hydrophilic PAAm and hydrophobic basal polystyrene layers. Bovine carotid artery endothelial cells adhered only to the ablated domains after pretreatment of the patterned surfaces with 15 microg/mL fibronectin at 37 degrees C. The ablated domain sizes significantly influenced the number of cells occupying each domain. Cell patterning functionality of the patterned surfaces was maintained for more than 2 months without loss of pattern fidelity, indicating that more durable cell arrays can be obtained compared to those prepared by self-assembled monolayers of alkanethiols, as described in previous reports. The surface fabrication techniques presented here can be utilised for the preparation of cell-based biosensors as well as tissue engineering constructs.

Fabrication of calcium sulfate/PLLA composite for bone repair
Gao, C., J. Gao, et al. (2005), J Biomed Mater Res A 73(2): 244-53.
Abstract: The bone-repairing composite material CS/PLLA was fabricated by mixing poly-L-lactic acid (PLLA) and calcium sulfate hemihydrate (CSH). The structure of the composite was analyzed with Infrared spectroscope, X-ray diffraction, and scanning electron microscope. The results indicated that the crystal pattern of calcium sulfate was affected by the addition of PLLA. PLLA part impacted the development of calcium sulfate dihydrate (CSD) crystal by slowing the conversion from CSH to CSD, so the composites are actually composed of CSH, CSD, and PLLA. The absorbing test in vitro showed that CS/PLLA composite absorbed more slowly than pure CS, suggesting the addition of PLLA can adjust the absorption rate of CS to meet different requirements. The pH value changes of the media had similar trends for different composites during the absorbing test of CS/PLLA samples in aqueous medium, which was connected to the absorption of calcium sulfate. The absorption of calcium sulfate in a certain time left a porous PLLA scaffold that will enable cells to further grow in. The surface of CS/PLLA pellets was inoculated with human osteoblasts, and the primary results showed that the osteoblasts could attach and spread on the surface, which will stimulate our desire for further study.

Fabrication of collagen-coated biodegradable polymer nanofiber mesh and its potential for endothelial cells growth
He, W., Z. Ma, et al. (2005), Biomaterials 26(36): 7606-15.
Abstract: Endothelialization of biomaterials is a promising way to prevent intimal hyperplasia of small-diameter vascular grafts. The aim of this study was to design a nanofiber mesh (NFM) that facilitates viability, attachment and phenotypic maintenance of human coronary artery endothelial cells (HCAECs). Collagen-coated poly(L-lactic acid)-co-poly(epsilon-caprolactone) P(LLA-CL 70:30) NFM with a porosity of 64-67% and a fiber diameter of 470+/-130 nm was fabricated using electrospinning followed by plasma treatment and collagen coating. The structure of the NFM was observed by SEM and TEM, and mechanical property was studied by tensile test. The presence of collagen on the P(LLA-CL) NFM surface was verified by X-ray photoelectron spectroscopy (XPS) and quantified by colorimetric method. Spatial distribution of the collagen in the NFM was visualized by labelling with fluorescent probe. The collagen-coated P(LLA-CL) NFM enhanced the spreading, viability and attachment of HCAECs, and moreover, preserve HCAEC's phenotype. The P(LLA-CL) NFM is a potential material for tissue engineered vascular graft.

Fabrication of colloidal crystals with tubular-like packings
Li, F., X. Badel, et al. (2005), J Am Chem Soc 127(10): 3268-9.

Fabrication of hybrid nanocapsules by calcium phosphate mineralization of shell cross-linked polymer micelles and nanocages
Perkin, K. K., J. L. Turner, et al. (2005), Nano Lett 5(7): 1457-61.
Abstract: Self-assembled shell cross-linked poly(acrylic acid-b-isoprene) (PAA78-b-PI97) micelles or cross-linked PAA nanocages in aqueous solution were used as templates for the preparation of novel polymer-inorganic nanocapsules. The hybrid nanostructures were typically 50-70 nm in diameter and consisted of spherical polymer nanoparticles or nanocages enclosed within a continuous 10-20 nm thick surface layer of amorphous calcium phosphate. Nucleation of calcium phosphate specifically in association with the polymer nanoparticles was facilitated by low supersaturation levels and by sequestration of Ca2+ ions within the carboxylate-rich PAA domains prior to addition of HPO4(2-). Modifications in ionic concentrations were used to control the calcium phosphate surface layer thickness and prepare mineralized cross-linked PAA-b-PI micelles with variable shell permeability. The permeability of beta-carotene into the hydrophobic PI core of mineralized shell cross-linked PAA-b-PI micelles was reduced by approximately 50 or 100% respectively for hybrid nanostructures enclosed within 10 or 20 nm thick calcium phosphate layers. Our results suggest that calcium phosphate-polymer cross-linked nanocapsules could have potential applications as pH-responsive biocompatible hybrid nanostructures for use in applications such as drug delivery, bioimaging, and therapeutics.

Fabrication of hydroxyapatite sponges by dextran sulphate/amino acid templating
Gonzalez-McQuire, R., D. Green, et al. (2005), Biomaterials 26(33): 6652-6.
Abstract: We report a new template-directed method for the fabrication of hydroxyapatite (HAp) sponges by using amino-acid-coated HAp nanoparticles dispersed within a viscous polysaccharide (dextran sulfate) matrix, and describe the use of these materials for the viability and proliferation of human bone marrow stromal cells. The nanoparticles were prepared in the presence of excess amounts of aspartic acid, alanine or arginine, and subsequently organised into macroporous frameworks with typical pore sizes of 100-200 microm during thermal degradation of the dextran matrix. The sponge macrostructure was influenced by changes in the heating rate and sintering time, as well as the use of different amino acids or variations in dextran functional groups. Biocompatibility testing showed retention of cell viability, production of extracellular matrix and alkaline phosphatase expression, suggesting that it should be possible to exploit this novel fabrication method for potential applications in cartilage or soft tissue engineering.

Fabrication of implantable microelectrode arrays by laser cutting of silicone rubber and platinum foil
Schuettler, M., S. Stiess, et al. (2005), J Neural Eng 2(1): S121-8.
Abstract: A new method for fabrication of microelectrode arrays comprised of traditional implant materials is presented. The main construction principle is the use of spun-on medical grade silicone rubber as insulating substrate material and platinum foil as conductor (tracks, pads and electrodes). The silicone rubber and the platinum foil are patterned by laser cutting using an Nd:YAG laser and a microcontroller-driven, stepper-motor operated x-y table. The method does not require expensive clean room facilities and offers an extremely short design-to-prototype time of below 1 day. First prototypes demonstrate a minimal achievable feature size of about 30 microm.

Fabrication of novel biomaterials through molecular self-assembly
Zhang, S. (2003), Nat Biotechnol 21(10): 1171-8.
Abstract: Two complementary strategies can be used in the fabrication of molecular biomaterials. In the 'top-down' approach, biomaterials are generated by stripping down a complex entity into its component parts (for example, paring a virus particle down to its capsid to form a viral cage). This contrasts with the 'bottom-up' approach, in which materials are assembled molecule by molecule (and in some cases even atom by atom) to produce novel supramolecular architectures. The latter approach is likely to become an integral part of nanomaterials manufacture and requires a deep understanding of individual molecular building blocks and their structures, assembly properties and dynamic behaviors. Two key elements in molecular fabrication are chemical complementarity and structural compatibility, both of which confer the weak and noncovalent interactions that bind building blocks together during self-assembly. Using natural processes as a guide, substantial advances have been achieved at the interface of nanomaterials and biology, including the fabrication of nanofiber materials for three-dimensional cell culture and tissue engineering, the assembly of peptide or protein nanotubes and helical ribbons, the creation of living microlenses, the synthesis of metal nanowires on DNA templates, the fabrication of peptide, protein and lipid scaffolds, the assembly of electronic materials by bacterial phage selection, and the use of radiofrequency to regulate molecular behaviors.

Fabrication of reconfigurable protein matrices by cracking
Zhu, X., K. L. Mills, et al. (2005), Nat Mater 4(5): 403-6.
Abstract: The interface between extracellular matrices and cells is a dynamic environment that is crucial for regulating important cellular processes such as signal transduction, growth, differentiation, motility and apoptosis. In vitro cellular studies and the development of new biomaterials would benefit from matrices that allow reversible modulation of the cell adhesive signals at a scale that is commensurate with individual adhesion complexes. Here, we describe the fabrication of substrates containing arrays of cracks in which cell-adhesive proteins are selectively adsorbed. The widths of the cracks (120-3,200 nm) are similar in size to individual adhesion complexes (typically 500-3,000 nm) and can be modulated by adjusting the mechanical strain applied to the substrate. Morphology of cells can be reversibly manipulated multiple times through in situ adjustment of crack widths and hence the amount of the cell-adhesive proteins accessible to the cell. These substrates provide a new tool for assessing cellular responses associated with exposure to matrix proteins.

Fabrication of three-dimensional porous scaffolds of complicated shape for tissue engineering. I. Compression molding based on flexible-rigid combined mold
Wu, L., H. Zhang, et al. (2005), Tissue Eng 11(7-8): 1105-14.
Abstract: A novel method for the fabrication of complexly shaped three-dimensional porous scaffolds has been developed by combining modified compression molding and conventional particulate leaching. The resultant scaffolds of various shapes, including some shaped like auricles, were made of hydrophobic biodegradable and bioresorbable poly(D,L-lactic acid) (PDLLA) and poly(D,L-lactic-co-glycolic acid) (PLGA). A polymer-particulate mixture was first prepared by the conventional solvent casting method and then compressively molded in a specially designed flexible-rigid combined mold which facilitates shaping and mold release during the fabrication process. The molding was carried out at a moderate temperature, above the glass transition temperature and below the flow temperature of these amorphous polymers. A porous scaffold was then obtained after particulate leaching. The pores are highly interconnected and uniformly distributed both in the bulk and on the external surface of the scaffolds, and the porosity can exceed 90%. The mechanical properties of the resultant porous scaffolds are satisfactory as determined by measurements of compressive modulus and compressive stress at 10% strain. Good viability of cells seeded in the porous scaffolds was confirmed. This novel fabrication method is promising in tissue engineering because of its ability to produce precise and complexly (anatomically) shaped porous scaffolds.

Fabrication of thromboresistant multilayer thin film on plasma treated poly (vinyl chloride) surface
Tan, Q., J. Ji, et al. (2005), J Mater Sci Mater Med 16(7): 687-92.
Abstract: Layer-by-layer deposited anticoagulant multilayer films were prepared on ammonia plasma treated poly (vinyl chloride) (PVC). Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) and contact angle results revealed the presence of -NH2 on the ammonia plasma treated PVC surfaces and the layer-by-layer self-assembly process. The stability of multilayer film was studied with the radio labeled method. The remainder bovine serum albumin (BSA) in cross-linked 5(heparin/BSA) multilayer films dipped in phosphate buffered saline (PBS, pH 7.4) was more than 90% in 40 days. The static platelet adhesion result indicated the anticoagulant multilayer films deposited on the plasma treated PVC reduced platelet adhesion drastically and no thrombus forming. The plasma recalcification time revealed that the multilayer modified surfaces greatly prolonged the plasma recalcification time. Such an easy processing and shape-independent method may have good potential for surface modification of cardiovascular devices.

Fabrication of viable tissue-engineered constructs with 3D cell-assembly technique
Yan, Y., X. Wang, et al. (2005), Biomaterials 26(29): 5864-71.
Abstract: We have recently developed an organ manufacturing technique that enables us to form cell/biomaterial complex three-dimensional (3D) architectures in designed patterns. This technique employs a highly accurate 3D micropositioning system with a pressue-controlled syringe to deposit cell/biomaterial structures with a lateral resolution of 10 microm. The pressure-activated micro-syringe is equipped with a fine-bore exit needle using which a wide variety of 3D patterns with different arrays of channels (through-holes) were created. The channels can supply living cells with nutrients and allow removing the cell metabolites. The embedded cells remain viable and perform biological functions as long as the 3D structures are retained. The new technology has the potential for eventual high-throughput production of artificial human tissues and organs.

Fabrication, characterization, and biological assessment of multilayered DNA-coatings for biomaterial purposes
van den Beucken, J. J., M. R. Vos, et al. (2006), Biomaterials 27(5): 691-701.
Abstract: This study describes the fabrication of two types of multilayered coatings onto titanium by electrostatic self-assembly (ESA), using deoxyribosenucleic acid (DNA) as the anionic polyelectrolyte and poly-d-lysine (PDL) or poly(allylamine hydrochloride) (PAH) as the cationic polyelectrolyte. Both coatings were characterized using UV-vis spectrophotometry, atomic force microscopy (AFM), X-ray photospectroscopy (XPS), contact angle measurements, Fourier transform infrared spectroscopy (FTIR), and for the amount of DNA immobilized. The mutagenicity of the constituents of the coatings was assessed. Titanium substrates with or without multilayered DNA-coatings were used in cell culture experiments to study cell proliferation, viability, and morphology. Results of UV-vis spectrophotometry, AFM, and contact angle measurements clearly indicated the progressive build-up of the multilayered coatings. Furthermore, AFM and XPS data showed a more uniform build-up and morphology of [PDL/DNA]-coatings compared to [PAH/DNA]-coatings. DNA-immobilization into both coatings was linear, and approximated 3microg/cm(2) into each double-layer. The surface morphology of both types of multilayered DNA-coatings showed elevations in the nanoscale range. No mutagenic effects of DNA, PDL, or PAH were detected, and cell viability and morphology were not affected by the presence of either type of multilayered DNA-coating. Still, the results of the proliferation assay revealed an increased proliferation of primary rat dermal fibroblasts on both types of multilayered DNA-coatings compared to non-coated controls. The biocompatibility and functionalization of the coatings produced here, will be assessed in subsequent cell culture and animal-implantation studies.

Fabrication, implantation, elution, and retrieval of a steroid-loaded polycaprolactone subretinal implant
Beeley, N. R., J. V. Rossi, et al. (2005), J Biomed Mater Res A 73(4): 437-44.
Abstract: A subretinal drug delivery system was developed to overcome the limitations of current treatments for retinal disease. A rod-shaped implant was made by embedding the corticosteroid triamcinolone acetonide within a biodegradable polycaprolactone polymer matrix. The implant was fabricated by homogeneously mixing the polymer and drug in solvent. The mixture was then dried, melted, and extruded, and the prepared solid form was drawn into a filament. The rods were mechanically sectioned to a length of 2 mm with a diameter of up to 320 microm. The rods were successfully implanted into the subretinal space of six rabbits. No complications were observed during the 4-week follow-up period. Initial observations of the implantation and elution characteristics revealed that polycaprolactone is well tolerated by the retinal tissue and that the implant can elute steroid for a period of at least 4 weeks without eliciting inflammatory response or complications. In vitro drug elution rates of different polymer to drug ratios and geometries into a balanced salt solution/bovine serum albumin (1%) solution showed an early rapid-release phase and late first-order phase. Histology and device retrieval after implantation revealed minimal encapsulation and good preservation of cellular morphology during the follow-up period and a more fibrous polymer microstructure of the implant.

Facial prosthetic rehabilitation: preprosthetic surgical techniques and biomaterials
Lemon, J. C., S. Kiat-amnuay, et al. (2005), Curr Opin Otolaryngol Head Neck Surg 13(4): 255-62.
Abstract: PURPOSE OF REVIEW: Attention to detail ensuring a successful facial prosthetic rehabilitation must be considered a priority at the time of presurgery, surgery, and at every stage in fabricating the prosthesis. Teamwork between the surgeon and maxillofacial prosthodontist will ensure an optimal surgical preparation and definitive prosthesis. RECENT FINDINGS: Evidence of interaction between team members can most certainly be encouraging to the patient. During the prosthetic phase of treatment, focusing on tissue assessment, impression making, sculpting, mold fabrication, familiarity with materials, appreciation of color, delivery of instructions, and patient education will ensure a satisfactory outcome. With the desire, determination, and encouragement from the restorative team to make the most of this artificial replacement, a patient can have a higher quality of life and a more normalized lifestyle. SUMMARY: This review presents current concepts regarding facial prosthetic rehabilitation of patients with head and neck cancer and facial prosthetic biomaterials.

Factor analysis in the evaluation of the relationship between bacterial adherence to biomaterials and changes in free energy
Carballo, J., C. M. Ferreiros, et al. (1992), J Biomater Appl 7(2): 130-41.
Abstract: The relative surface charge and free energy of forty-one coagulase-negative staphylococci were found to be normally distributed; therefore, they can be considered a homogeneous group under strict statistical criteria. The adherence of these bacteria to eight different biomaterials (seven synthetic and one biologic) was found to be independent of charge and variations in free energy during adhesion. Adherence can be explained as a thermodynamic process (free energy decreased with adherence), except in the case of bovine pericardium in which free energy increases. With these biomaterials, a correlation was found between adherence and bacterial charge. Bacterial adherence and bacterial charge correlate with the surface parameters of the biomaterials. This correlation does not occur when the relationships between parameters are evaluated by means of factors analysis, thus indicating the importance of the statistical method selected for the evaluation of bacterial adherence.

Factor XII fragment and kallikrein generation in plasma during incubation with biomaterials
van der Kamp, K. W. and W. van Oeveren (1994), J Biomed Mater Res 28(3): 349-52.
Abstract: Blood biocompatibility of medical devices is in many ways dependent on surface characteristics and biochemical blood material interactions. In this study, the contact system, in which the activation of factor XII and plasma kallikrein is included, is highlighted. This article describes a simple chromogenic assay to determine the Hageman Factor fragment (HFf, or factor XIIf) and kallikrein activity in vitro. The assay is based on conversion of Z-Lys-Phe-Arg-pNA.2HCl to which human factor XIIf and kallikrein appeared to have a high affinity. To discriminate between the serine proteases factor XIIf and kallikrein to cleave this substrate, aprotinin was added to one of two complementary samples. In this in vitro study, standardized disks from glass, high-density polyethylene (HDPE), polytetrafluoro ethylene (PTFE), and polydimethyl siloxane (PDMS) were studied for their capacity to generate factor XIIf and kallikrein in plasma. Kaolin was used as positive control. On glass disks the highest and on HDPE the lowest generation of factor XIIf and kallikrein were found, both with a ratio of 1:1. On PDMS and on PTFE disks protease activities were intermediate, but with a factor XIIf and kallikrein activity ratio of 1:2 and 1:4, respectively. Apparently because of the hydrophobic surface character of PDMS and PTFE, these surfaces absorb or fail to produce the factor XIIf. This assay appeared to be discriminative even for materials that are considered mild activators of the contact system and can therefore be used as a standard method to qualify biomaterials.(ABSTRACT TRUNCATED AT 250 WORDS)

Factorial design, physicochemical characterisation and activity of ciprofloxacin-loaded PLGA nanoparticles for ocular use
Dillen, K., J. Vandervoort, et al. (2005), J Control Release 101(1-3): 369-71.

Factors affecting the free radical scavenging behavior of chitosan sulfate
Huang, R., E. Mendis, et al. (2005), Int J Biol Macromol 36(1-2): 120-7.
Abstract: Scavenging activity of hydroxyethyl chitosan sulfate (HCS) against 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl and carbon-centered radical species were studied using electron spin resonance (ESR) spectroscopy. In addition, its antioxidant activity to retard lipid peroxidation was also evaluated in a linoleic acid model system. HCS could scavenge DPPH (33.78%, 2.5 mg/mL) and carbon-centered radicals (67.74%, 0.25 mg/mL) effectively. However, chitosan sulfate did not exhibit any scavenging activity against hydroxyl radicals, but increased its generation. This was different from the published literature and was presumed due to the loss of chelating ability on Fe2+. This assumption could further confirm from the results obtained for Fe2+-ferrozine method that upon sulfation chitooligosaccharides lost its chelation properties. Therefore, HCS can be identified as antioxidant that effectively scavenges carbon centered radicals to retard lipid peroxidation.


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