Subeom Park

Graphene-incorporated chitosan substrata for adhesion and differentiation of human mesenchymal stem cells†

A simple method that uses graphene to fabricate nanotopographic substrata was reported for stem cell engineering. Graphene-incorporated chitosan substrata promoted adhesion and differentiation of human mesenchymal stem cells (hMSCs). In addition, we proposed that nanotopographic cues of the substrata could enhance cell-cell and cell-material interactions for promoting functions of hMSCs.

Graphene-Regulated Cardiomyogenic Differentiation Process of Mesenchymal Stem Cells by Enhancing the Expression of Extracellular Matrix Proteins and Cell Signaling Molecules

The potential of graphene as a mesenchymal stem cell (MSC) culture substrate to promote cardiomyogenic differentiation is demonstrated. Graphene exhibits no sign of cytotoxicity for stem cell culture. MSCs are committed toward cardiomyogenic lineage by simply culturing them on graphene. This may be attributed, at least partially, to the regulation of expression levels of extracellular matrix and signaling molecules.

Graphene oxide flakes as a cellular adhesive: prevention of reactive oxygen species mediated death of implanted cells for cardiac repair.

Mesenchymal stem cell (MSC) implantation has emerged as a potential therapy for myocardial infarction (MI). However, the poor survival of MSCs implanted to treat MI has significantly limited the therapeutic efficacy of this approach. This poor survival is primarily due to reactive oxygen species (ROS) generated in the ischemic myocardium after the restoration of blood flow. ROS primarily causes the death of implanted MSCs by inhibiting the adhesion of the MSCs to extracellular matrices at the lesion site (i.e., anoikis). In this study, we proposed the use of graphene oxide (GO) flakes to protect the implanted MSCs from ROS-mediated death and thereby improve the therapeutic efficacy of the MSCs. GO can adsorb extracellular matrix (ECM) proteins. The survival of MSCs, which had adhered to ECM protein-adsorbed GO flakes and were subsequently exposed to ROS in vitro or implanted into the ischemia-damaged and reperfused myocardium, significantly exceeded that of unmodified MSCs. Furthermore, the MSC engraftment improved by the adhesion of MSCs to GO flakes prior to implantation enhanced the paracrine secretion from the MSCs following MSC implantation, which in turn promoted cardiac tissue repair and cardiac function restoration. This study demonstrates that GO can effectively improve the engraftment and therapeutic efficacy of MSCs used to repair the injury of ROS-abundant ischemia and reperfusion by protecting implanted cells from anoikis.

Graphene enhances the cardiomyogenic differentiation of human embryonic stem cells

Graphene has drawn attention as a substrate for stem cell culture and has been reported to stimulate the differentiation of multipotent adult stem cells. Here, we report that graphene enhances the cardiomyogenic differentiation of human embryonic stem cells (hESCs) at least in part, due to nanoroughness of graphene. Large-area graphene on glass coverslips was prepared via the chemical vapor deposition method. The coating of the graphene with vitronectin (VN) was required to ensure high viability of the hESCs cultured on the graphene. hESCs were cultured on either VN-coated glass (glass group) or VN-coated graphene (graphene group) for 21 days. The cells were also cultured on glass coated with Matrigel (Matrigel group), which is a substrate used in conventional, directed cardiomyogenic differentiation systems. The culture of hESCs on graphene promoted the expression of genes involved in the stepwise differentiation into mesodermal and endodermal lineage cells and subsequently cardiomyogenic differentiation compared with the culture on glass or Matrigel. In addition, the culture on graphene enhanced the gene expression of cardiac-specific extracellular matrices. Culture on graphene may provide a new platform for the development of stem cell therapies for ischemic heart diseases by enhancing the cardiomyogenic differentiation of hESCs.

Amphiphilic comblike polymers enhance the colloidal stability of Fe3O4 nanoparticles

Stable colloidal dispersions of magnetite (Fe(3)O(4)) nanoparticles (MNPs) were obtained with the inclusion of an amphiphilic comblike polyethylene glycol derivative (CL-PEG) as an amphiphilic polymeric surfactant. Both the size and morphology of the resulting CL-PEG-modified MNPs could be controlled and were characterized by transmission electron microscopy (TEM). The interaction between MNPs and CL-PEG was confirmed by the presence of characteristic infrared absorption peaks, and the colloidal stability of the nanoparticle dispersion in water was evaluated by long-term observation of the dispersion using UV-visible spectroscopy. SQUID measurements confirmed the magnetization of CL-PEG-modified MNPs. The zeta potential of the CL-PEG-modified MNPs showed a dramatic conversion from positive to negative in response to the pH of the surrounding aqueous medium due to the presence of carboxyl groups at the surface. These carboxyl groups can be used to functionalize the MNPs with biomolecules for biotechnological applications. However, regardless of surface electrostatics, the flexible, hydrophilic side chains of CL-PEG-modified MNPs prevented the approach of adjacent nanoparticles, thereby resisting aggregation and resulting in a stable aqueous colloid. The cytotoxicity of MNPs and CL-PEG-modified MNPs was evaluated by a MTT assay.

Bacterial Cellulose Nanofibrillar Patch as a Wound Healing Platform of Tympanic Membrane Perforation

Bacterial cellulose (BC)-based biomaterials on medical device platforms have gained significant interest for tissue-engineered scaffolds or engraftment materials in regenerative medicine. In particular, BC has an ultrafine and highly pure nanofibril network structure and can be used as an efficient wound-healing platform since cell migration into a wound site is strongly meditated by the structural properties of the extracellular matrix. Here, the fabrication of a nanofibrillar patch by using BC and its application as a new wound-healing platform for traumatic tympanic membrane (TM) perforation is reported. TM perforation is a very common clinical problem worldwide and presents as conductive hearing loss and chronic perforations. The BC nanofibrillar patch can be synthesized from Gluconacetobacter xylinus; it is found that the patch contained a network of nanofibrils and is transparent. The thickness of the BC nanofibrillar patch is found to be approximately 10.33 ± 0.58 μm, and the tensile strength and Youngs modulus of the BC nanofibrillar patch are 11.85 ± 2.43 and 11.90 ± 0.48 MPa, respectively, satisfying the requirements of an ideal wound-healing platform for TM regeneration. In vitro studies involving TM cells show that TM cell proliferation and migration are stimulated under the guidance of the BC nanofibrillar patch. In vivo animal studies demonstrate that the BC nanofibrillar patch promotes the rate of TM healing as well as aids in the recovery of TM function. These data demonstrate that the BC nanofibrillar patch is a useful wound-healing platform for TM perforation.

Amphiphilic comb-like polymer for harvest of conductive nano-cellulose

In this study, electrically conductive bacterial cellulose (BC) was prepared by culturing Gluconacetobacter xylinus in a carbon nanotube (CNT)-dispersed medium. The CNTs were dispersed by adopting a non-covalent approach in the presence of non-ionic amphiphilic comb-like polymer (CLP). Specifically, the hydrophobic backbone of CLP was chemophysically attached to the surface of the CNTs and the hydrophilic side chains were released freely toward the medium in an aqueous environment. CLP-modified CNTs were stable and did not show any noticeable sediment, even after centrifugation at 15,000 rpm for 30 min. Notably, the dispersion solution of CLP-modified CNTs was stable at room temperature for several months because the long-range entropic repulsion among polymer-decorated tubes acted as a barrier to aggregation. The morphology of the BC membrane was studied by field-emission scanning electron microscopy. The presence of CLP bound to the CNT surface was characterized by Fourier transform infrared spectroscopy and the conductivity of the CNT-incorporated BC membrane was measured by four-probe measurements.

Quantitative comparison of tumor vascularity of hepatocellular carcinoma after intravenous contrast agent: conventional versus harmonic power Doppler US.

AbstractBackground: The purpose of this study was to make a quantitative comparison between conventional and harmonic power Doppler (PD) ultrasound (US) in depicting vascularity of hepatocellular carcinoma (HCC). Methods: Ten nodular HCCs in 10 patients were prospectively examined using a 2–4-MHz convex transducer and a standardized examination protocol. Serial US images were obtained before and 20, 30, 40, 50, 60, 90, 120, 150, 180, 240, and 300 s after intravenous injection of 2 g of contrast agent using conventional and harmonic PD US. The percentage of area with Doppler signal within each HCC nodule (%PDA) was calculated in each image with a PC-based image analysis program, and the results with both US techniques were compared. Results: In the majority of cases, %PDA was greater on conventional PD US than on harmonic PD US. Mean %PDA of 10 HCCs was significantly higher on conventional PD US than on harmonic PD US except at 20 s after injection. The highest values of mean %PDA were 34.9% in conventional PD US and 19.5% in harmonic PD US at 60 s after injection. Conclusion: Area with PD signals within the HCC is smaller and the duration of effective enhancement is shorter in harmonic PD US than in conventional PD US.

Engineering structures and functions of mesenchymal stem cells by suspended large-area graphene nanopatterns

Inspired by the hierarchical nanofibrous and highly oriented structures of natural extracellular matrices, we report a rational design of chemical vapor deposition graphene-anchored scaffolds that provide both physical and chemical cues in a multilayered organization to control the adhesion and functions of cells for regenerative medicine. These hierarchical platforms are fabricated by transferring large graphene film onto nanogroove patterns. The top graphene layer exhibits planar morphology with slight roughness (~20 nm between peaks) due to the underlying topography, which results in a suspended structure between the nanoridges. We demonstrate that the adhesion and differentiation of human mesenchymal stem cells were sensitively controlled and enhanced by the both the nanotopography and graphene cues in our scaffolds. Our results indicate that the layered physical and chemical cues can affect the apparent cell behaviors, and can synergistically enhance cell functionality. Therefore, these suspended graphene platforms may be used to advance regenerative medicine.

Use of magnetic nanoparticles to manipulate the metabolic environment of bacteria for controlled biopolymer synthesis.

Magnetic nanoparticles (MNPs) were covalently immobilized on the surface of Acetobacter xylinus and the location of the bacteria was controlled to manipulate bacterial bioactivation. The bacteria were positioned in the middle of an incubation tube by applying an external magnetic field, and the cellulose produced at the different metabolizing locations was characterized by X-ray diffraction, electron microscopy, and differential scanning calorimetry. To the best of our knowledge, this is the first experiment in which MNPs were employed in the control of cell metabolism.