1,720,972 research outputs found
A simple fabrication process for stepwise gradient wrinkle patterns with spatially controlled wavelength based on selective oxygen plasma treatment
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Robust Topographical Micro-Patterning of Nanofibrillar Collagen Gel by In Situ Photochemical Crosslinking-Assisted Collagen Embossing
No full textThe topographical micro-patterning of nanofibrillar collagen gels is promising for the fabrication of biofunctional constructs mimicking topographical cell microenvironments of in vivo extracellular matrices. Nevertheless, obtaining structurally robust collagen micro-patterns through this technique is still a challenging issue. Here, we report a novel in situ photochemical crosslinking-assisted collagen embossing (IPC-CE) process as an integrative fabrication technique based on collagen compression-based embossing and UV-riboflavin crosslinking. The IPC-CE process using a micro-patterned polydimethylsiloxane (PDMS) master mold enables the compaction of collagen nanofibrils into micro-cavities of the mold and the simultaneous occurrence of riboflavin-mediated photochemical reactions among the nanofibrils, resulting in a robust micro-patterned collagen construct. The micro-patterned collagen construct fabricated through the IPC-CE showed a remarkable mechanical resistivity against rehydration and manual handling, which could not be achieved through the conventional collagen compression-based embossing alone. Micro-patterns of various sizes (minimum feature size <10 mu m) and shapes could be obtained by controlling the compressive pressure (115 kPa) and the UV dose (3.00 J/cm(2)) applied during the process. NIH 3T3 cell culture on the micro-patterned collagen construct finally demonstrated its practical applicability in biological applications, showing a notable effect of anisotropic topography on cells in comparison with the conventional construct.11Ysciescopu
Electrospun Nanofibrous Bilayer Fabrication Technique Based on Electrolyte-Assisted Electrical Discharge
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Development of Contact Lens-Shaped Crosslinked Amniotic Membranes for Sutureless Fixation in the Treatment of Ocular Surface Diseases
No full textPurpose: To develop a new method of manufacturing contact lens-shaped crosslinked amniotic membranes (AMs) using glutaraldehyde (GA) and dialdehyde starch (DAS) as crosslinking agents.
Methods: Amniotic membranes were placed on a curved plastic mold and crosslinked with either 4.5% DAS or 1% GA, after which their physical properties and biological safety were evaluated.
Results: The tensile strength of the GA- and DAS-crosslinked samples was much increased compared with that of normal AMs. Neither crosslinking process affected AM transparency. Although the GA-crosslinked AM showed better enzymatic resistance, its physiological structure was severely damaged after the crosslinking process. On the other hand, compared with the GA-crosslinked AM, the DAS-crosslinked AM showed higher growth factor concentrations and better biocompatibility, similar to normal AMs. In addition, the DAS-crosslinked AM was effective in the recovery of corneal epithelial wounds and was well maintained over 3 days without decentration or degradation on the ocular surface in human subjects.
Conclusions: Contact lens-shaped AMs were successfully prepared with crosslinking agents. Crosslinking with DAS did not affect the structural properties or biological activ- ity of the AMs, and the improved mechanical properties helped the AM to maintain its curved shape. This crosslinking method allowed us to transplant AMs into patients' eyes without sutures.
Translational Relevance: Sutureless fixation of contact lens-shaped AMs would be very convenient and safe for the treatment of corneal surface disease.11Ysciescopu
Development of corneal lenticule-reconstituted collagen composite sheet for corneal epithelium regeneration
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Hydrogel-Assisted Electrospinning for Fabrication of a 3D Complex Tailored Nanofiber Macrostructure
No full textElectrospinning has shown great potential in tissue engineering and regenerative medicine due to a high surface-area-to-volume ratio and an extracellular matrix-mimicking structure of electrospun nanofibers, but the fabrication of a complex three-dimensional (3D) macroscopic configuration with electrospun nanofibers remains challenging. In the present study, we developed a novel hydrogel-assisted electrospinning process (GelES) to fabricate a 3D nanofiber macrostructure with a 3D complex but tailored configuration by utilizing a 3D hydrogel structure as a grounded collector instead of a metal collector in conventional electrospinning. The 3D hydrogel collector was discovered to effectively concentrate the electric field toward itself similar to the metal collector, thereby depositing electrospun nanofibers directly on its exterior surface. Synergistic advantages of the hydrogel (e.g., biocompatibility and thermally reversible sol-gel transition) and the 3D nanofiber macrostructure (e.g., mechanical robustness and high permeability) provided by the GeIES process were demonstrated in a highly permeable tubular tissue graft and a robust drug- or cell-encapsulation construct. GelES is expected to broaden potential applications of electrospinning to not only provide in vivo drug/cell delivery and tissue regeneration but also an in vitro drug testing platform by increasing the degree of freedom in the configuration of the 3D nanofiber macrostructure.11Nsciescopu
A collagen gel-coated, aligned nanofiber membrane for enhanced endothelial barrier function
Full text linkHerein, a collagen gel-coated and aligned nanofiber membrane named Col-ANM is developed, which remarkably improves endothelial barrier function by providing biochemical and topographical cues simultaneously. Col-ANM is fabricated by collagen gel coating process on an aligned polycaprolactone (PCL) nanofiber membrane, which is obtained by a simple electrospinning process adopting a parallel electrode collector. Human umbilical vein endothelial cells (HUVECs) cultured on Col-ANM exhibit remarkably enhanced endothelial barrier function with high expression levels of intercellular junction proteins of ZO-1 and VE-cadherin, a high TEER, and a cellular permeability compared with the artificial porous membranes in commercial cell culture well inserts. The enhanced endothelial barrier function is conjectured to be attributed to the synergistic effects of topographical and biochemical cues provided by the aligned PCL nanofibers and collagen gel in the Col-ANM, respectively. Finally, the reactive oxygen species is applied to the HUVEC monolayer formed on the Col-ANM to destroy the tight junctions between HUVECs. The destruction of the tight junctions is demonstrated by the decreased TEER value overtime. Results indicate the potential of Col-ANM in modeling endothelial barrier dysfunction-related diseases.11Ysciescopu
Decellularized corneal lenticule embedded compressed collagen: toward a suturable collagenous construct for limbal reconstruction
No full textRecently, compressed collagen has attracted much attention as a potential alternative for a limbal epithelial stem cell (LESC) carrier to treat limbal stem cell deficiency (LSCD), in that it can provide mechanically improved collagen fibrillar structures compared to conventional collagen hydrogel. However, its clinical efficacy as an LESC carrier has not yet been studied through in vivo transplantation due to limited mechanical strength that cannot withstand a force induced by surgical suturing and low resistance to enzymatic degradation. This study firstly presents a suturable LESC carrier based on compressed collagen in the form of a biocomposite. The biocomposite was achieved by integrating a decellularized corneal lenticule, which is a decellularized stromal tissue obtained from corneal refractive surgery, inside a compressed collagen to form a sandwich structure. A suture retention test verified that the biocomposite has a much higher suture retention strength (0.56 ± 0.12 N) compared to the compressed collagen (0.02 ± 0.01 N). The biocomposite also exhibited more than 3 times higher resistance to enzymatic degradation, indicating long-term stability after transplantation. In vitro cell culture results revealed that the biocomposite effectively supported the expansion and stratification of the LESCs with expressions of putative stem cell and differentiated corneal epithelial cell markers. Finally, the biocomposite verified its clinical efficacy by stably delivering the LESCs onto an eye of a rabbit model of LSCD and effectively reconstructing the ocular surface.11Nsciescopu
Thin and stretchable extracellular matrix (ECM) membrane reinforced by nanofiber scaffolds for developing in vitro barrier models
No full textAn extracellular matrix (ECM) membrane made up of ECM hydrogels has great potentials to develop a physiologically relevant organ-on-a-chip because of its biochemical and biophysical similarity to in vivo basement membranes (BMs). However, the limited mechanical stability of the ECM hydrogels makes it difficult to utilize the ECM membrane in long-term and dynamic cell/tissue cultures. This study proposes a thin but robust and transparent ECM membrane reinforced with silk fibroin (SF)/polycaprolactone (PCL) nanofibers, which is achieved by in situ self-assembly throughout a freestanding SF/PCL nanofiber scaffold. The SF/PCL nanofiber-reinforced ECM (NaRE) membrane shows biophysical characteristics reminiscent of native BMs, including small thickness (mu m), high permeability (<9 x 10(-5) cm s(-1)), and nanofibrillar architecture (similar to 10-100 nm). With the BM-like characteristics, the nanofiber reinforcement ensured that the NaRE membrane stably supported the construction of various types of in vitro barrier models, from epithelial or endothelial barrier models to complex co-culture models, even over two weeks of cell culture periods. Furthermore, the stretchability of the NaRE membrane allowed emulating the native organ-like cyclic stretching motions (10%-15%) and was demonstrated to manipulate the cell and tissue-level functions of the in vitro barrier model.
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