149 research outputs found
Preparation of poly(glycerol sebacate) fibers for tissue engineering applications
Poly(glycerol sebacate) (PGS) was discovered in the previous decade and is a promising bioelastomer with tuneable mechanical, biodegradable and biocompatible properties. Despite of these superiorities, PGS possesses solubility and processability disadvantages. To overcome these drawbacks of PGS, blends could be formed with a polymer which is soluble in a common solvent with PGS prepolymer, having a melting temperature above the crosslinking temperature and which can be removed from the structure after crosslinking. In this study, PGS fibers were fabricated for the first time using pressurized gyration as scaffolds. Fibers were obtained through blending the synthesized PGS prepolymer with poly(vinyl alcohol) (PVA) to overcome solubility/melting drawbacks of crosslinked PGS polymer. Obtained fiber diameters have a narrow size distribution which did not change after thermal crosslinking. After the washing procedure, âŒ25% decrease in the average fiber diameter was observed due to the PVA removal. Resulting PGS fibers were characterized in terms of chemical structure, morphology, and cell viability. Fibroblast cell adhesion and spreading on three-dimensional fiber networks were determined by microscopy. PGS fibers supported cell adhesion and proliferation. After 7 days of cell-PGS fiber interactions, cell proliferation and spreading increased without any toxicity
Biofabrication of Gelatin Tissue Scaffolds with Uniform Pore Size via Microbubble Assembly
The control of pore size and uniform porosity remains as an important challenge in gelatin scaffolds. The precise control in building blocks of tissue scaffolds without any additional porogen is possible with costly equipment and techniques, though some preârequirements for polymeric material, such as photoâpolymerizability or sintering ability, may be needed prior to construction. Herein, a method for the fabrication of gelatin scaffolds with homogenous porosity using simple Tâjunction microfluidics is described. The size of the microbubbles is precisely controlled with 5% deviation from the average. Porous gelatin scaffolds are obtained by buildingâup the monodispersed microbubbles in dilute crossâlinker solutions. The effect of crossâlinker density on pore diameter is also investigated. After crossâlinking, pore size of the resultant five scaffold groups are precisely controlled as 135 ± 11, 193 ± 11, 216 ± 9, 231 ± 5, and 250 ± 12 ”m. Porosity ratios above 65% are achieved in every sample group. According to the cell culture experiments, structures support high cell adhesion, viability, and migration through the porous network via interconnectivity. This study offers a practical and economical approach for the preparation of porous gelatin scaffolds with homogenous porosity which can be utilized in diverse tissue engineering applications
Self-assembled micro-stripe patterning of sessile polymeric nanofluid droplets
When sessile nanofluid droplets evaporate, solid nanoparticles can be organized in a wide variety of patterns on the substrate. The composition of the nanofluid, internal flow type of droplet and the rate of drying affect drop geometry, and the final pattern. Using poly(lactic-co-glycolic acid)-block-poly(ethylene glycol)(PLGA-b-PEG) as the example, we produced micro-stripe patterning from nanoparticles by drying of sessile fluid droplets. We investigated the nanoparticle properties and flow dynamics to clarify their effects on the patterning. Nanoparticles were prepared by hydrodynamic flow focusing using a T-junction microfluidic device with high production efficiency and the ability to generate an extremely narrow size distribution. PLGA-b-PEG was prepared as oil phase in acetonitrile and water/oil flow rate was changed from 1 to 3 at constant oil phase flow rate (50âŻÎŒL/min). Then, nanofluid was collected on the surface as sessile droplets within acetonitrile/water binary dispersed phase. Depending on size, charge and size-distribution, the nanoparticles deposited on the surface exhibited various patterns. Dynamic Light and X-ray Scattering measurements showed that, approximately 100âŻnm particles with relatively low PDI (0.04) were produced for the first time in surfactant free conditions in a microfluidic device and they generated self-assembled ordered patterns, which are regulated by the type of internal flow in the sessile nanofluid droplet during sequential evaporation of acetonitrile and water
Co-Axial Gyro-Spinning of PCL/PVA/HA Core-Sheath Fibrous Scaffolds for Bone Tissue Engineering
The present study aspires towards fabricating core-sheath fibrous scaffolds by state-of-the-art pressurized gyration for bone tissue engineering applications. The core-sheath fibers comprising dual-phase poly-Δ-caprolactone (PCL) core and polyvinyl alcohol (PVA) sheath are fabricated using a novel "co-axial" pressurized gyration method. Hydroxyapatite (HA) nanocrystals are embedded in the sheath of the fabricated scaffolds to improve the performance for application as a bone tissue regeneration material. The diameter of the fabricated fiber is 3.97 ± 1.31 ”m for PCL-PVA/3%HA while pure PCL-PVA with no HA loading gives 3.03 ± 0.45 ”m. Bead-free fiber morphology is ascertained for all sample groups. The chemistry, water contact angle and swelling behavior measurements of the fabricated core-sheath fibrous scaffolds indicate the suitability of the structures in cellular activities. Saos-2 bone osteosarcoma cells are employed to determine the biocompatibility of the scaffolds, wherein none of the scaffolds possess any cytotoxicity effect, while cell proliferation of 94% is obtained for PCL-PVA/5%HA fibers. The alkaline phosphatase activity results suggest the osteogenic activities on the scaffolds begin earlier than day 7. Overall, adaptations of co-axial pressurized gyration provides the flexibility to embed or encapsulate bioactive substances in core-sheath fiber assemblies and is a promising strategy for bone healing
Comparative study of collagen and gelatin coatings on titanium surfaces
The vast majority of studies in the bone tissue engineering field are focused on the surface modification of titanium scaffolds to obtain integration of the scaffold in the surrounding bone tissue. Our approach consisted in benefiting from the advantages of the cell-interaction capabilities of collagen and gelatin. The biopolymers were immobilised onto the Ti surface through different methods and the stability of the obtained coatings was determined. The obtained results reveal that covalent immobilisation of collagen and gelatin is required to obtain stable surface coatings
Effectiveness of Oil-Layered Albumin Microbubbles Produced Using Microfluidic T-Junctions in Series for In Vitro Inhibition of Tumor Cells
This work focuses on the synthesis of oil-layered microbubbles using two microfluidic T-junctions in series and evaluation of the effectiveness of these microbubbles loaded with doxorubicin and curcumin for cell invasion arrest from 3D spheroid models of triple negative breast cancer (TNBC), MDA-MB-231 cell line. Albumin microbubbles coated in the drug-laden oil layer were synthesized using a new method of connecting two microfluidic T-mixers in series. Double-layered microbubbles thus produced consist of an innermost core of nitrogen gas encapsulated in an aqueous layer of bovine serum albumin (BSA) which in turn, is coated with an outer layer of silicone oil. In order to identify the process conditions leading to the formation of double-layered microbubbles, a regime map was constructed based on capillary numbers for aqueous and oil phases. The microbubble formation regime transitions from double-layered to single layer microbubbles and then to formation of single oil droplets upon gradual change in flow rates of aqueous and oil phases. In vitro dissolution studies of double-layered microbubbles in an air-saturated environment indicated that a complete dissolution of such bubbles produces an oil droplet devoid of a gas bubble. Incorporation of doxorubicin and curcumin was found to produce a synergistic effect, which resulted in higher cell deaths in 2D monolayers of TNBC cells and inhibition of cell proliferation from 3D spheroid models of TNBC cells compared to the control
Generating Lifetime-Enhanced Microbubbles by Decorating Shells with Silicon Quantum Nano-Dots Using a 3-Series T-Junction Microfluidic Device
Long-term stability of microbubbles is crucial to their effectiveness. Using a new microfluidic device connecting three T-junction channels of 100 Όm in series, stable monodisperse SiQD-loaded bovine serum albumin (BSA) protein microbubbles down to 22.8 ± 1.4 Όm in diameter were generated. Fluorescence microscopy confirmed the integration of SiQD on the microbubble surface, which retained the same morphology as those without SiQD. The microbubble diameter and stability in air were manipulated through appropriate selection of T-junction numbers, capillary diameter, liquid flow rate, and BSA and SiQD concentrations. A predictive computational model was developed from the experimental data, and the number of T-junctions was incorporated into this model as one of the variables. It was illustrated that the diameter of the monodisperse microbubbles generated can be tailored by combining up to three T-junctions in series, while the operating parameters were kept constant. Computational modeling of microbubble diameter and stability agreed with experimental data. The lifetime of microbubbles increased with increasing T-junction number and higher concentrations of BSA and SiQD. The present research sheds light on a potential new route employing SiQD and triple T-junctions to form stable, monodisperse, multi-layered, and well-characterized protein and quantum dot-loaded protein microbubbles with enhanced stability for the first time
Assessment of electromechanically stimulated bone marrow stem cells seeded acellular cardiac patch in a rat myocardial infarct model
In this study, we evaluated cardiomyogenic differentiation of electromechanically stimulated rat bone marrow-derived stem cells (rt-BMSCs) on an acellular bovine pericardium (aBP) and we looked at the functioning of this engineered patch in a rat myocardial infarct (MI) model. aBP was prepared using a detergent-based decellularization procedure followed by rt-BMSCs seeding, and electrical, mechanical, or electromechanical stimulations (3 millisecond pulses of 5 V cm-1at 1 Hz, 5% stretching) to enhance cardiomyogenic differentiation. Furthermore, the electromechanically stimulated patch was applied to the MI region over 3 weeks. After this period, the retrieved patch and infarct region were evaluated for the presence of calcification, inflammatory reaction (CD68), patch to host tissue cell migration, and structural sarcomere protein expressions. In conjunction with any sign of calcification, a higher number of BrdU-labelled cells, and a low level of CD68 positive cells were observed in the infarct region under electromechanically stimulated conditions compared with static conditions. More importantly, MHC, SAC, Troponin T, and N-cad positive cells were observed in both infarct region, and retrieved engineered patch after 3 weeks. In a clear alignment with other results, our developed acellular patch promoted the expression of cardiomyogenic differentiation factors under electromechanical stimulation. Our engineered patch showed a successful integration with the host tissue followed by the cell migration to the infarct region
Poly(ester-urethane) scaffolds: effect of structure on properties and osteogenic activity of stem cells
The present study aimed to investigate the effect of structure (design and porosity) on the matrix stiffness and osteogenic activity of stem cells cultured on poly(ester-urethane) (PEU) scaffolds. Different three-dimensional (3D) forms of scaffold were prepared from lysine-based PEU using traditional salt-leaching and advanced bioplotting techniques. The resulting scaffolds were characterized by differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), mercury porosimetry and mechanical testing. The scaffolds had various pore sizes with different designs, and all were thermally stable up to 300Ăą Ă°C.Ă In vitrotests, carried out using rat bone marrow stem cells (BMSCs) for bone tissue engineering, demonstrated better viability and higher cell proliferation on bioplotted scaffolds compared to salt-leached ones, most probably due to their larger and interconnected pores and stiffer nature, as shown by higher compressive moduli, which were measured by compression testing. Similarly, SEM, von Kossa staining and EDX analyses indicated higher amounts of calcium deposition on bioplotted scaffolds during cell culture. It was concluded that the design with larger interconnected porosity and stiffness has an effect on the osteogenic activity of the stem cells.This work was supported by the European FP6 NoE EXPERTISSUES project (Grant No. NMP3-500283), the mini-project Fibrocell, TUBITAK (Grant No. TBAG 105 T508 Nanobiomat) and by METU-BAP
Cardiac tissue regeneration: A preliminary study on carbon-based nanotubes gelatin scaffold
The aim of this study was set-up and test of gelatin and carbon nanotubes scaffolds. Gelatin-based (5%) genipin cross-linked (0.2%) scaffolds embedding single-walled carbon nanotubes (SWCNTs, 0.3, 0.5, 0.7, 0.9, and 1.3% w/w) were prepared and mechanically/electrically characterized. For biological evaluation, H9c2 cell line was cultured for 10 days. Cytotoxicity, cell growth and differentiation, immunohistochemistry, and real-time PCR analysis were performed. Myoblast and cardiac differentiation were obtained by serum reduction to 1% (C1%) and stimulation with 50 nM all trans-retinoic acid (CRA), respectively. Immunohistochemistry showed elongated myotubes in C1%while round and multinucleated cells in CRAwith also a significantly increased expression of natriuretic peptides (NP) and ET-1 receptors in parallel with a decreased ET-1. On scaffolds, cell viability was similar for Gel-SWCNT0.3%/0.9%; NP and ET systems expression decreased in both concentrations with respect to control and CX-43, mainly due to a lacking of complete differentiation in cardiac phenotype during that time. Although further analyses on novel biomaterials are necessary, these results represent a useful starting point to develop new biomaterial-based scaffolds
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