Hybrid electrospun polyhydroxybutyrate/gelatin/laminin/polyaniline scaffold for nerve tissue engineering application: Preparation, characterization, and in vitro assay.

Despite the widespread central nervous system injuries, treatment of these disorders is still an issue of concern due to the complexities. Natural recovery in these patients is rarely observed, which calls for developing new methods that address these problems. In this study, natural polymers of polyhydroxybutyrate (PHB) and gelatin were electrospun into scaffolds and cross-linked. In order to modify the PHB-based scaffold for nerve tissue engineering, the scaffold surface was modified by exposure to the ammonium gas plasma under controlled conditions, and the laminin as a promoter for neural cells was coated on the sample surface. Then, polyaniline nanoparticles were inkjet-printed on a sample surface as parallel lines to induce the differentiation of stem cells into neural cells. Infrared spectroscopy, absorption of PBS, AFM, degradation rate, contact angle, electron microscopy and optical microscopy, thermal and mechanical behavior, and analysis of the viability of L929 cells were investigated for the scaffolds. The results showed gelatin decreased the contact angle from 106.2° to 38° and increased the residual weight after PBS incubation from 82 % to 38 %. The moduli of the scaffold increased from 8.78 MPa for pure PHB to 28.74 for the modified scaffold. In addition, performed methods increased cell viability from 69 % for PHB to 89 % for modified scaffold and also had a favorable effect on cell adhesion. Investigation of culturing P19 stem cells demonstrated that they successfully differentiated into neural cells. Results show that the scaffolds prepared in this study were promising for nerve tissue engineering.

Fabrication of electrospun nanofibrous thermoresponsive semi-interpenetrating poly(N-isopropylacrylamide)/polyvinyl alcohol networks containing ZnO nanoparticle mats: characterization and antibacterial and cytocompatibility evaluation.

Thermoresponsive nanofiber composites comprising biopolymers and ZnO nanoparticles with controlled release and antibacterial activity are fascinating scientific research areas. Herein, poly(N-isopropylacrylamide) (PNIPAm) was prepared and mixed with poly(vinyl alcohol) (PVA) in 75/25 and 50/50 weight ratios together with ZnO (0, 1, and 2 phr) to construct nanofiber composites. The morphology of the crosslinked nanofiber composites, ZnO content, and their mechanical behavior were assessed by SEM, EDX, and tensile analyses. The wettability results show an increment in nanofiber surface hydrophobicity by increasing the temperature above the LCST of PNIPAm. The in vitro ZnO release exhibits a faster release profile for the sample with 50 wt% PNIPAm (lower crosslinking density) compared to the one with 25 wt%. Besides, a strong interaction between PVA hydroxyl groups and ZnO can restrict the release content. However, by increasing the temperature from 28 to 32 °C, the relative ZnO release becomes half for both compositions. All crosslinked nanofiber composites demonstrated reliable biocompatibility against L929 fibroblast cells. Agar disc-diffusion and optical density methods showed thermo-controllable antibacterial activity against Staphylococcus aureus upon temperature variation between 28 and 32 °C. Furthermore, in vivo and histological results indicate the potentiality of the prepared multidisciplinary wound dressing for robust wound healing and skin tissue engineering.

Highly porous bio-glass scaffolds fabricated by polyurethane template method with hydrothermal treatment for tissue engineering uses.

Objectives: Bioglass scaffolds, which contain a significant percentage of porosity for tissue engineering purposes, have low strength. For increasing the strength and efficiency of such structures for use in tissue engineering, fabrication of hierarchical meso/macro-porous bioglass scaffolds, developing their mechanical strength by hydrothermal treatment and adjusting pH method, and achieving the appropriate mesopore size for loading large biomolecules, were considered in this study. Materials and Methods: Mesoporous bioglass (MBG) powders were synthesized using cetyltrimethylammonium bromide as a surfactant, with different amounts of calcium sources to obtain the appropriate size of the mesoporous scaffolds. Then MBG scaffolds were fabricated by a polyurethane foam templating method, and for increasing scaffold strength hydrothermal treatment (90 °C, for 5 days) and adjustment pH (pH=9) method was used to obtain hierarchical meso/macro-porous structures. The sample characterization was done by Simultaneous thermal analysis, Fourier transform infrared spectroscopy, Field Emission Scanning electron microscopy, small and wide-angle X-ray powder diffractions, transmission electron microscopy, and analysis of nitrogen adsorption-desorption isotherm. The mechanical strength of scaffolds was also determined. Results: The MBG scaffolds based on 80.28 (wt.) % SiO2- 17.89 (wt.) % CaO- 1.81 (wt.) % P2O5 presented interconnected large pores and pores in the range of 100-150 µm and 6-18 nm, respectively and 0.4 MPa compressive strength. Conclusion: The total pore volume and specific surface area were obtained from the Brunauer-Emmett-Teller theory, 0.709 cm3 g-1 and 213.83 m2 g-1, respectively. These findings could be considered in bone-cartilage tissue engineering.

In vitro and in vivo evaluation of nanofibre mats containing Calendula officinalis extract as a wound dressing.

OBJECTIVE: The present study aims to create Calendula officinalis-loaded nanofibre-based wound dressing materials to enhance the wound healing process. Calendula officinalis is an annual herb native to the Mediterranean region. It is antipyretic, antifungal, antioedema, antidiabetic, anti-inflammatory (wound, oral and pharyngeal mucosa), antispasmodic, treats chronic ocular surface diseases, acts as a stimulant and a diaphoretic. It is also used in the prevention of acute dermatitis, and in the treatment of gastrointestinal ulcers, wounds and burns. METHOD: Electrospinning is an effective method for creating nano- and microfibres for biomedical applications. Calendula officinalis (CA) of various concentrations 5%, 10% and 15%)-loaded polyvinyl alcohol (PVA)/sodium alginate (SAlg) nanofibre mats were successfully produced via blend electrospinning. Nanofibre mats were evaluated using: scanning electron microscopy (SEM); Fourier transform infrared spectroscopy (FTIR) analysis; gel content; water vapour transmission rate (WVTR); swelling ratio; in vitro drug release studies; viability evaluation (cell culture and MTT assay); and an in vivo study using male Wistar rats. Rats were divided into three groups (n=3). In each group, rats were inflicted with five full-thickness wounds on the back and were treated with sterile gauze (control), PVA/SAlg nanofibre dressing (CA-free control), PVA/SAlg/CA5%, PVA/SAlg/CA10%, and PVA/SAlg/CA15% nanofibre dressing. RESULTS: Results showed that the obtained fibres were smooth with no surface aggregates, indicating complete incorporation of Calendula officinalis. The release of Calendula officinalis from loaded PVA/SAlg fibre mats in the first four hours was burst released and then was constant. PVA/SAlg and PVA/SAlg/CA nanofibres were not toxic to L929 mouse fibroblasts and supported cell attachment and proliferation. The results of the in vivo study showed that the PVA/SAlg/CA10% nanofibre dressing had a higher full-thickness wound healing closure rate compared with the control group on days seven, 14 and 21 after treatment. CONCLUSION: The results of this evaluation showed that PVA/SAlg/CA nanofibrous mats could be a candidate as an effective wound dressing; however, the percentage of CA in this compound needs further investigation.

Platelet-rich plasma-hyaluronic acid/chondrotin sulfate/carboxymethyl chitosan hydrogel for cartilage regeneration.

In this study, the chondrogenic potential of hyaluronic acid/chondrotin sulfate/carboxymethyl chitosan hydrogels with adipose-derived mesenchymal stem cells (ADMSCs) was evaluated. Here, hyaluronic acid, chondrotin sulfate, and carboxymethyl chitosan were used as the substrate for cartilage tissue engineering in which the hydrogel is formed due to electrostatic and hydrogen bonds through mixing the polymers. Because of the instability of this hydrogel in the biological environment, 1-ethyl-3-(3-dimethylaminopropyl-carbodiimide hydrochloride/N-hydroxy-succinimide was used as a crosslinker to increase the hydrogel stability. The hydrogels showed reasonable stability due to the combined effect of self-crosslinking and chemical crosslinking. The cells were treated with the prepared hydrogel samples for 14 and 21 days in nondifferentiation medium for evaluation of the cellular behavior of ADMSCs. Gene expression evaluation was performed, and expression of specific genes involved in differentiation was shown in the crosslinked hydrogel with platelet-rich plasma (PRP) (H-EN-P) had increased the gene expression levels. Quantification of immunofluorescence intensity indicated the high level of expression of SOX9 in H-EN-P hydrogel. Based on the results, we confirmed that the presence of PRP and the similarity of the hydrogel constituents to the cartilage extracellular matrix could have positive effects on the differentiation of the cells, which is favorable for cartilage tissue engineering approaches.

Fabrication of carboxymethyl chitosan/poly(ε-caprolactone)/doxorubicin/nickel ferrite core-shell fibers for controlled release of doxorubicin against breast cancer.

The coaxial electrospinning for producing core-shell nanofibers due to control the release profile of drug by the shell layer has been developed. N-carboxymethyl chitosan (CMC)-polyvinyl alcohol (core)/poly(ε-caprolactone) (PCL) (shell) nanofibers were produced via coaxial electrospinning. Doxorubicin (DOX) and nickel ferrite nanoparticles were incorporated into the nanofibers for controlled release of DOX against MCF-7 breast cancer. The minimum CMC/PCL fiber diameter was found to be 300 nm by optimizing of three variables including voltage to distance ratio (1.5-2.5 kV/cm), CMC concentration (4-6 wt.%) and PCL concentration (8-12 wt.%). The synthesized core-shell fibers were characterized using FTIR, XRD, SEM, and TEM analysis. The extended release and controlled release of DOX from core-shell nanofibers were achieved under physiological pH without external magnetic field (EMF) and acidic pH with EMF during 25 and 7 days, respectively. The maximum cytotoxicity of MCF-7 breast cancer cells was about 83 % using CMC/PCL/nickel ferrite 10 % nanofibers and EMF.

Enhanced antimicrobial and full-thickness wound healing efficiency of hydrogels loaded with heparinized ZnO nanoparticles: In vitro and in vivo evaluation.

Nanotechnology-based fabricated wound dressings are known as appropriate substrates to enhance healing in both acute and chronic wounds. These types of materials have the ability to deliver therapeutic agents. In this study, a wound dressing including heparinized zinc oxide nanoparticles in combination with chitosan and poly(vinyl alcohol) was developed to investigate its antibacterial and regenerative properties in a rat model of full thickness skin wounds. By adding nanoparticles, the mechanical strength increased up to twice as compared to the sample without nanoparticles. In addition, heparin release profile follows the Hixson-Crowell release kinetic. Protein adsorption enhanced by adding nanoparticles in hydrogels and the prepared wound dressings were completely biocompatible. In terms of antibacterial activity, the minimum inhibitory concentration decreased by conjugation of heparin on the surface of zinc oxide nanoparticles compared to the non-functionalized nanoparticles, and, this shows the increased antibacterial synergistic effect by adding heparin to nanoparticles. Furthermore, it was found that the heparinized zinc oxide nanoparticles effectively accelerate wound closure, re-epithelialization and decrease collagen deposition compared to other groups after implantation. Hence, the prepared wound dressings have the capacity to significantly enhance healing of acute wounds.

Poly(hydroxybutyrate-co-hydroxyvalerate) Porous Matrices from Thermally Induced Phase Separation.

Thermally induced phase separation followed by freeze drying has been used to prepare biodegradable and biocompatible scaffolds with interconnected 3D microporous structures from poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) copolymers containing 5 and 12 wt % of 3-hydroxyvalerate (HV). Solutions of PHBV in 1,4-dioxane, underwent phase separation by cooling under two different thermal gradients (at -25 °C and -5 °C). The cloud point and crystallization temperature of the polymer solutions were determined by turbidimetry and differential scanning calorimetry, respectively. Parameters affecting the phase separation mechanism such as variation of both the cooling process and the composition of the PHBV copolymer were investigated. Afterwards, the influence of these variables on the morphology of the porous structure and the final mechanical properties (i.e., rigidity and damping) was evaluated via scanning electron microscopy and dynamic mechanical thermal analysis, respectively. While the morphology of the scaffolds was considerably affected by polymer crystallization upon a slow cooling rate, the effect of solvent crystallization was more evident at either high hydroxyvalerate content (i.e., 12 wt % of HV) or high cooling rate. The decrease in the HV content gave rise to scaffolds with greater stiffness because of their higher degree of crystallinity, being also noticeable the greater consistency of the structure attained when the cooling rate was higher. Scaffolds were fully biocompatible supports for cell adhesion and proliferation in 3D cultures and show potential application as a tool for tissue regeneration.

Preparation and modeling of electrospun polyhydroxybutyrate/polyaniline composite scaffold modified by plasma and printed by an inkjet method and its cellular study.

The reconstruction of the nerve tissue engineering scaffold is always of particular interest due to the inability to recover and repair neural tissues after being damaged by diseases or physical injuries. The primary purpose of this study was obtaining a model used to predict the diameter of the fibers of electrospun polyhydroxybutyrate (PHB) scaffolds. Accordingly, the range of operating parameters, namely the applied voltage, the distance between the nozzle to the collector, and solution concentration, was designed for the electrospinning process at three different levels, giving seventeen experiments. These data were modeled utilizing response surface methodology and artificial neural network method using Design Expert and Matlab software.The effect of process parameters on the diameter, as well as their interactions were investigated in detail, and the corresponding models were suggested. Both the RSM and ANN models showed an excellent agreement between the experimental and predicted response values. In the second phase of the study, PHB natural polymer was electrospun into scaffolds with high biocompatibility, resulting in a 224-360 nm diameter range .To further modify the scaffold in order to improve the compatibility of PHB, the fibrous surface of scaffolds was exposed to oxygenated plasma gas radiation under controlled conditions. Next, polyaniline (PANI) nanoparticles were then synthesized and printed on the surface of scaffolds as parallel lines. Then samples were exposed to the electric field. Fourier-transform infrared spectroscopy, water contact angle, optical and electron microscopy, tensile test, and cell viability analysis were performed to study properties of resulting scaffolds. The results indicated the fact that modification of the scaffolds by oxygen plasma and printing PANI nanoparticles in particular patterns had a favorable impact on cell adhesion and direction of cell growth, showing the potential of resulting scaffolds for nerve tissue engineering applications.

Effects of different quantities of antibody conjugated with magnetic nanoparticles on cell separation efficiency.

Antibody-conjugated magnetic nanoparticles (Ab-MNPs) have received considerable attention in bioseparation and clinical diagnostics assays due to their unique ability to detect and isolate a variety of biomolecules and cells. Because antibodies can be expensive, a key challenge for bioconjugation is to determine the optimal amount of antibodies with reasonable antigen-capturing activity. We designed an approach to determine the minimum amounts of antibodies for efficient coating. Different quantities of Herceptin (anti-human epidermal growth factor receptor 2: HER2) antibody were applied and immobilized on the surface of MNPs. Antibody binding was then checked by using an anti-human antibody conjugated with fluorochrome and flow cytometry. When the ratio of MNPs to antibodies increased from 0.79 to 795.45, mean fluorescence intensity (MFI) of conjugated MNPs decreased markedly from 185.56 to 20.07, indicating lower surface antibody coverage. We then investigated the relation between antibody content and isolation efficiency. Three Ab-MNP samples with different MFI were used to isolate SK-BR-3, a HER2-positive breast cancer cell line, from mixtures of whole blood or mononuclear cells. After isolation in a magnetic field, separation efficiency was evaluated by fluorescence microscopy and flow cytometry-based techniques. Our results collectively showed that the amount of anti-HER2 antibodies for conjugation with MNPs could be decreased by as much as one-fifteenth without compromising isolation efficiency, which in turn can reduce the cost of immunoassay biosensors.

Potential of novel electrospun core-shell structured polyurethane/starch (hyaluronic acid) nanofibers for skin tissue engineering: In vitro and in vivo evaluation.

The biomaterials with excellent biocompatibility and biodegradability ¬can lead to satisfactory wound healing. In this study, core-shell structured PU (polyurethane)/St (Starch) and PU/St (Hyaluronic Acid (HA)) nanofibers were fabricated with coaxial electrospinning technique. The morphology characterization of the core-shell structure of nanofibers was investigated by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) images. Contact-angle measurements were confirmed the core/shell structure of the electrospun nanofibers with shell and core feed rates of 0.675 L/min and <0.135 L/min, respectively. The average fiber diameter values were calculated for polyurethane nanofibers (836 ± 172.13 nm), PU/St nanofibers (612 ± 93.21 nm) and PU/St (HA) nanofibers (428 ± 78.32 nm). The average porosity values of scaffolds were determined for PU (1.251 ± 0.235 µm), PU/St (1.734 ± 0.284 µm) and PU/St (HA) (3.186 ± 0.401 µm). The core-shell PU/St and PU/St (HA) nanofibers were evaluated in vitro by using mouse fibroblasts (L929) cells. Cell morphology and viability results were exhibited significant enhancement in cell promoting and cell attachment. Furthermore, in vivo studies was indicated Core-shell PU/St (HA) wound dressing can be an appropriate candidate for skin tissue engineering and wound healing.

Design and optimization of process parameters of polyvinyl (alcohol)/chitosan/nano zinc oxide hydrogels as wound healing materials.

Composite hydrogels as wound dressings feature healing properties in treating wounds. In this study, polyvinyl (alcohol)/chitosan/nano zinc oxide nanocomposite hydrogels were formed using the freeze-thaw method and essential process parameters including thawing time, thawing temperature, and the number of freeze-thaw cycles was investigated to model nanocomposites employing response surface methodology. Critical properties including water vapor transmission rate, porosity, wound fluid absorption, and gel content were modeled using process parameters. Analysis of morphology, mechanical properties, toxicity, protein absorption, antibacterial activity, and in-vitro wound healing were also performed. Results exhibited that increased freeze-thaw cycles caused reduced pore size and increased porosity and wound fluid absorption. Besides, increased freeze-thaw cycles and reduced thawing temperature resulted in increased elastic modulus and tensile strength, while elongation at break point decreased. Antibacterial properties, biocompatibility, and in-vitro wound healing tests demonstrated that the designed system showed no toxicity and it was able to treat the wounds sufficiently.

Wound dressing based on electrospun PVA/chitosan/starch nanofibrous mats: Fabrication, antibacterial and cytocompatibility evaluation and in vitro healing assay.

Electrospun nanofibrous mats based on biopolymers have been widely investigated for tissue engineering in recent years, primarily due to remarkable morphological similarity to the natural extracellular matrix (ECM). In this research, electrospun PVA/Chitosan/Starch nanofibrous mats were fabricated using electrospinning method for wound dressing application. The prepared nanofibrous mats were then cross-linked to enhanced the water resistance and also optimize the biodegradation rate followed by characterization and evaluation of their properties as wound dressings. The morphological studies performed by SEM and AFM showed that uniform bead-free electrospun nanofibrous mats were formed. The structural properties of the fabricated mats were characterized by FTIR. The proper porosity and balanced water absorption and water vapor transmission rate (WVTR) of obtained dressings, demonstrate their ability in providing suitable moist environment for wound, result in the appropriate wound breathing and simultaneously efficient handling of wound exudates. Suitable mechanical properties of nanofibrous dressing in both dry and wet states confirm the capability of fabricated wound dressing to protect wound area against the external forces during the healing process. Antibacterial test revealed excellent antibacterial activity of nanofibrous mats against both gram negative and gram positive bacteria. Furthermore, the in vitro cytotoxicity evaluated by MTT assay, proved appropriate cytocompatibility and cell viability of the developed nanofibrous mats which were also verified with in vitro wound healing analysis performed by scratch assay, confirming the remarkable potential of the investigated nanofibrous mats for wound dressing application.

The effect of collector type on the physical, chemical, and biological properties of polycaprolactone/gelatin/nano-hydroxyapatite electrospun scaffold.

Electrospinning is considered a powerful method for the production of fibers in the nanoscale size. Small pore size results in poor cell infiltration, cell migration inhibition into scaffold pores and low oxygen diffusion. Electrospun polycaprolactone/gelatin/nano-hydroxyapatite (PCL/Gel/nHA) scaffolds were deposited into two types of fiber collectors (novel rotating disc and plate) to study fiber morphology, chemical, mechanical, hydrophilic, and biodegradation properties between each other. The proliferation and differentiation of MG-63 cells into the bone phenotype were determined using MTT method, alizarin red staining and alkaline phosphatase (ALP) activity. The rates for disc rotation were 50 and 100 rpm. The pore size measurement results indicated that the fibers produced by the disc rotation collector with speed rate 50 rpm have larger pores as compared to fibers produced by disc rotation at 100 rpm and flat plate collectors. A randomly structure with controlled pore size (38.65 ±0.33 µm) and lower fiber density, as compared to fibers collected by disc rotation with speed rate 100 rpm and flat plate collectors, was obtained. Fibers collected on the rotating disc with speed rate 50 rpm, were more hydrophilic due to larger pore size and therefore, faster infiltration of water into the scaffold and the rate of degradation was higher. These results demonstrate that PCL/Gel/nHA scaffolds made through a rotating disc collector at 50 rpm are more feasible to be used in bone tissue engineering applications due to appropriate pore size and increased adhesion and proliferation of cells, ALP activity and mineral deposits. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 933-950, 2019.

Incorporation of ZnO nanoparticles into heparinised polyvinyl alcohol/chitosan hydrogels for wound dressing application.

Available wound dressings have some major deficiencies including low water vapor transmission rate (WVTR), low absorption of wound fluids, and not providing a suitable and moist environment for wound healing. The main advantage of hydrogels is giving aid to the creation of a moist and cool environment for wound healing and providing high water vapor permeability along with preventing penetration of microbes into the wound surface. Therefore, hydrogels of heparinized polyvinyl alcohol (PVA)/chitosan (CS)/nano zinc oxide (nZnO) were prepared to be used as wound dressing. Samples were characterized via infrared spectrometry (FTIR), X-ray diffraction (XRD) and scanning electron microscope (SEM). In addition, other properties including swelling ratio, water vapor transmission rate, the size of pores, mechanical and thermal properties, cell viability, and antibacterial efficiency were investigated. Water vapor permeability, porosity, and swelling ratio showed a wide range of numerical values that facilitate the use of provided samples as ideal wound dressings. Besides, investigating mechanical and thermal properties exhibited the improvement of mentioned properties by adding nano zinc oxide. Furthermore, Heparin loading was conducted on optimum samples. Heparin release rate decreased and was more sustained by adding nanoparticles compared to hydrogel wound dressings without nZnO. Cell viability of bionanocomposite samples showed no toxicity after loading nanoparticles and this value was >70% for all samples. Antibacterial properties of hydrogel samples can effectively protect wounds especially with an increase nZnO content. Hence, these hydrogels were found applicable as robust wound dressings.

3-Dimensional cell-laden nano-hydroxyapatite/protein hydrogels for bone regeneration applications.

The ability to encapsulate cells in three-dimensional (3D) protein-based hydrogels is potentially of benefit for tissue engineering and regenerative medicine. However, as a result of their poor mechanical strength, protein-based hydrogels have traditionally been considered for soft tissue engineering only. Hence, in this study we tried to render these hydrogels suitable for hard tissue regeneration, simply by incorporation of bioactive nano-hydroxyapatite (HAp) into a photocrosslinkable gelatin hydrogel. Different cell types were also encapsulated in three dimensions in the resulting composites to prepare cell-laden constructs. According to the results, HAp significantly improves the stiffness of gelatin hydrogels, while it maintains their structural integrity and swelling ratio. It was also found that while the bare hydrogel (control) was completely inert in terms of bioactivity, a homogeneous 3D mineralization occurs throughout the nanocomposites after incubation in simulated body fluid. Moreover, encapsulated cells readily elongated, proliferated, and formed a 3D interconnected network with neighboring cells in the nanocomposite, showing the suitability of the nano-HAp/protein hydrogels for cellular growth in 3D. Therefore, the hydrogel nanocomposites developed in this study may be promising candidates for preparing cell-laden tissue-like structures with enhanced stiffness and increased osteoconductivity to induce bone formation in vivo.

Nano-hydroxyapatite reinforced polyhydroxybutyrate composites: a comprehensive study on the structural and in vitro biological properties.

Nanocomposites based on polyhydroxybutyrate (PHB) and hydroxyapatite (HAp) have recently been proposed for application in bone repair and regeneration, but very limited studies have investigated the effect of HAp on the rheological and thermal behavior of PHB. More important, the efficiency of a biomaterial depends greatly on its ability to interact with cells, but little is known about this interaction for this kind of nanocomposite. Hence, this paper dealt with some of the characteristics of solution-casted PHB/HAp nanocomposite films, and tried to explore the effect of HAp nanoparticles on cellular responses. The results showed that both rheological and thermal properties can be tailored by incorporating appropriate amounts of nanoparticles. In vitro studies showed a significant increase in proliferation and differentiation of MC3T3-E1 on nanocomposites compared to the neat polymer. Surface examination indicated that topography and chemistry of surface are important factors influencing cellular processes; while no cell differentiation was found on the neat polymer, nanocomposite with 15 wt.% filler content exhibited a pronounced differentiation resulting from high surface roughness and large amount of exposed HAp. These results suggest that HAp particles play a much more important role in determining the biological performance of PHB than has previously been supposed.

Synthesis methods for nanosized hydroxyapatite with diverse structures.

Hydroxyapatite (HAp) is the major mineral constituent of vertebrate bones and teeth. It has been well documented that HAp nanoparticles can significantly increase the biocompatibility and bioactivity of man-made biomaterials. Over the past decade, HAp nanoparticles have therefore increasingly been in demand, and extensive efforts have been devoted to develop many synthetic routes, involving both scientifically and economically new features. Several investigations have also been made to determine how critical properties of HAp can be effectively controlled by varying the processing parameters. With such a wide variety of methods for the preparation of HAp nanoparticles, choosing a specific procedure to synthesize a well-defined powder can be laborious; accordingly, in the present review, we have summarized all the available information on the preparation methodologies of HAp, and highlighted the inherent advantages and disadvantages involved in each method. This article is focused on nanosized HAp, although recent articles on microsized particles, especially those assembled from nanoparticles and/or nanocrystals, have also been reviewed for comparison. We have also provided several scientific figures and discussed a number of critical issues and challenges which require further research and development.

Differentiation of embryonic stem cells into neural cells on 3D poly (D, L-lactic acid) scaffolds versus 2D cultures.

In this study, a highly porous poly (D, L-lactic acid) (PDLLA) scaffold was designed and fabricated using dioxane and thermal-induced phase separation (TIPS) methods (liquid-liquid and solid-liquid). Additionally, we characterized the ability of mouse embryonic stem cells (ESCs) to differentiate into neural cells in PDLLA scaffold with uniform porosity, interconnectivity, and high porosity, and then compared them with cells seeded under conventional two-dimensional (2D) culture conditions. Histochemistry staining showed the migration of differentiated cells through the scaffold. Immunofluorescence analysis of the differentiated cells by counting positive cells revealed that the PDLLA scaffold resulted in a significantly greater number of neural markers, microtubule associated protein-2, ß-tubulin III, neurofilament protein, and glial fibrillary acidic protein (the astrocyte marker) when compared to those in 2D culture condition. Moreover, the expression of Nestin, Mash1, Pax6, and HB9 increased significantly in 3D scaffolds when compared with 2D cultures as detected by semi-quantitative RT-PCR. Scanning electron microscopy of differentiated neurons on scaffolds also demonstrated favorable results for neurite outgrowth. The results of this study demonstrated a promising effect of 3D scaffold culture for neural cell differentiation from ESCs in prospective tissue engineering applications.

Manufacturing of biodegradable polyurethane scaffolds based on polycaprolactone using a phase separation method: physical properties and in vitro assay.

BACKGROUND: Biodegradable polyurethanes have found widespread use in soft tissue engineering due to their suitable mechanical properties and biocompatibility. METHODS: In this study, polyurethane samples were synthesized from polycaprolactone, hexamethylene diisocyanate, and a copolymer of 1,4-butanediol as a chain extender. Polyurethane scaffolds were fabricated by a combination of liquid-liquid phase separation and salt leaching techniques. The effect of the NCO:OH ratio on porosity content and pore morphology was investigated. RESULTS: Scanning electron micrographs demonstrated that the scaffolds had a regular distribution of interconnected pores, with pore diameters of 50-300 µm, and porosities of 64%-83%. It was observed that, by increasing the NCO:OH ratio, the average pore size, compressive strength, and compressive modulus increased. L929 fibroblast and chondrocytes were cultured on the scaffolds, and all samples exhibited suitable cell attachment and growth, with a high level of biocompatibility. CONCLUSION: These biodegradable polyurethane scaffolds demonstrate potential for soft tissue engineering applications.