Kyung Jae Jeong

Synthetic ligand-coated magnetic nanoparticles for microfluidic bacterial separation from blood.

Bacterial sepsis is a serious clinical condition that can lead to multiple organ dysfunction and death despite timely treatment with antibiotics and fluid resuscitation. We have developed an approach to clearing bacteria and endotoxin from the bloodstream, using magnetic nanoparticles (MNPs) modified with bis-Zn-DPA, a synthetic ligand that binds to both Gram-positive and Gram-negative bacteria. Magnetic microfluidic devices were used to remove MNPs bound to Escherichia coli , a Gram-negative bacterium commonly implicated in bacterial sepsis, from bovine whole blood at flows as high as 60 mL/h, resulting in almost 100% clearance. Such devices could be adapted to clear bacteria from septicemic patients.

Nanocomposite Gold-Silk Nanofibers

Cell-biomaterial interactions can be controlled by modifying the surface chemistry or nanotopography of the material, to induce cell proliferation and differentiation if desired. Here we combine both approaches in forming silk nanofibers (SNFs) containing gold nanoparticles (AuNPs) and subsequently chemically modifying the fibers. Silk fibroin mixed with gold seed nanoparticles was electrospun to form SNFs doped with gold seed nanoparticles (SNF(seed)). Following gold reduction, there was a 2-fold increase in particle diameter confirmed by the appearance of a strong absorption peak at 525 nm. AuNPs were dispersed throughout the AuNP-doped silk nanofibers (SNFs(Au)). The Youngs modulus of the SNFs(Au) was almost 70% higher than that of SNFs. SNFs(Au) were modified with the arginine-glycine-aspartic acid (RGD) peptide. Human mesenchymal stem cells that were cultured on RGD-modified SNF(Au) had a more than 2-fold larger cell area compared to the cells cultured on bare SNFs; SNF(Au) also increased cell size. This approach may be used to alter the cell-material interface in tissue engineering and other applications.

Polydopamine coatings enhance biointegration of a model polymeric implant

The biointegration of implants affects their function, stability and safety. Although most research on this topic has focused on bone and other hard tissues, biointegration with soft tissues is important in numerous applications, such as in prosthetic corneas. Here, we have adapted polydopamine-based adhesive surface chemistry to enhance the biointegration with soft tissue of a model polymer—poly(methyl methacrylate) (PMMA), commonly used in prosthetic corneas. Polydopamine coating (PDA) and subsequent modification with the cell-adhesive peptide RGD (PDA-PEG-RGD) significantly enhanced cellular proliferation of corneal epithelial cells and keratocytes without causing excessive secretion of pro-inflammatory cytokines (e.g.IL-6) by either cell type. PDA adhered tightly to collagen gels, while PDA-PEG-RGD and uncoated PMMA did not. PDAs adhesion to collagen was greatly reduced by preincubation in serum. Tissue reaction to both polydopamine-coated surfaces was benign after 45 days of subcutaneous implantation. However, in contrast to the findings with collagen gels, PDA-PEG-RGD bound much more tightly to tissue than did PDA—although both bound better than unmodified PMMA. Polydopamine-based surface chemistries are potentially useful in enhancing tissue integration of implants with soft tissues.

Hydroxyapatite for Keratoprosthesis Biointegration

PURPOSE Integration of keratoprosthesis with the surrounding cornea is very important in preventing bacterial invasion, which may cause ocular injury. Here the authors investigated whether hydroxyapatite (HAp) coating can improve keratoprosthesis (KPro) biointegration, using polymethyl methacrylate (PMMA)--the principal component of the Boston KPro--as a model polymer. METHODS HAp coatings were induced on PMMA discs after treatment with concentrated NaOH and coating with poly-dopamine (PDA) or polydopamine and then with 11-mercaptoundecanoic acid (11-MUA). Coatings were characterized chemically (Fourier transform infrared spectroscopy [FTIR], energy dispersive X-ray spectroscopy [EDX]) and morphologically (SEM) and were used as substrates for keratocyte growth in vitro. Cylinders of coated PMMA were implanted in porcine corneas ex vivo for 2 weeks, and the force required to pull them out was measured. The inflammatory reaction to coated discs was assessed in the rabbit cornea in vivo. RESULTS FTIR of the coatings showed absorption bands characteristic of phosphate groups, and EDX showed that the Ca/P ratios were close to those of HAp. By SEM, each method resulted in morphologically distinct HAp films; the 11-MUA group had the most uniform coating. The hydroxyapatite coatings caused comparable enhancement of keratocyte proliferation compared with unmodified PMMA surfaces. HAp coating significantly increased the force and work required to pull PMMA cylinders out of porcine corneas ex vivo. HAp coating of implants reduced the inflammatory response around the PMMA implants in vivo. CONCLUSIONS These results are encouraging for the potential of HAp-coated surfaces for use in keratoprostheses.

Depth-resolved holographic optical coherence imaging using a high-sensitivity photorefractive polymer device

We present coherence-gated holographic imaging using a highly sensitive photorefractive (PR) polymer composite as the recording medium. Due to the high sensitivity of the composite holographic recording at intensities as low as 5 mW/cm2 allowed for a frame exposure time of only 500ms. Motivated by regenerative medical applications, we demonstrate optical depth sectioning of a polymer foam for use as a cell culture matrix. An axial resolution of 18 μm and a transverse resolution of 30 μm up to a depth of 600 μm was obtained using an off-axis recording geometry.

Raman Spectroscopic Investigation of Peptide—Glycosaminoglycan Interactions

Protein–glycosaminoglycan (GAG) interactions play a central role in tissue engineering and drug delivery. A rapid and efficacious method for screening these interactions is essential. Raman spectroscopy was used to identify chemical interactions and conformational changes occurring upon binding between a synthetic peptide (QRRFMQYSARRF) and two glycosaminoglycans (GAGs), heparin and chondroitin 6-sulfate (C6S). The results identify three main chemical groups that are involved in the binding of the synthetic peptide with heparin and C6S. Tyrosine formed hydrogen bonds with the GAGs via its hydroxyl group. The amide I band demonstrated substantial shifts in Raman wavenumbers when bound to heparin and C6S (Δω = —10.2 ± 0.7 cm−1 and Δω = —11.9 ± 0.3 cm−1, respectively), suggesting that the peptide underwent planar conformational changes after binding occurred. Upon binding to the peptide, the sulfate peak of heparin displayed a substantially greater shift in the Raman wavenumber (—7.5 ± 0.5 cm−1) than that of C6S (—2.6 ± 0.5 cm−1). The greater amide I and sulfate band shifts seen during peptide–heparin interactions are indicative of a stronger association compared to that between the peptide and C6S. This observation was confirmed by capillary electrophoresis, which demonstrated a lower dissociation constant (KD) between the peptide and heparin (KD of 19.2 ± 3.3 μM) than between the peptide and C6S (26.7 ± 2.5 μM). We conclude that the shift in the Raman wavenumbers of amide I and sulfate groups can be used for high-throughput screening of interaction affinities between libraries of peptides and GAGs.

A novel assay to probe heparin-peptide interactions using pentapeptide-stabilized gold nanoparticles.

In this article, we present a novel assay to probe the interactions between heparin and heparin-binding peptides based on CALNN pentapeptide-stabilized gold nanoparticles. This assay relies on rapid aggregation of gold nanoparticles and dramatic retardation in the presence of a large excess of heparin due to the binding of peptides to heparin. Using this method, the dissociation constant ( K d) and melting temperature ( T m) of three different peptides against heparin were determined. The results from capillary electrophoresis demonstrated that K d values measured by this method were comparatively accurate. It was found that the peptide with the lowest K d did not have the highest T m. Structural analysis by circular dichroism was performed to explain this phenomenon. A comparison with the results from affinity chromatography indicates that electrostatic interactions only are not the major determinant of the affinity between heparin and peptide, but other interactions such as hydrogen-bonding and hydrophobic interactions may play important roles in the overall interactions. This novel assay is inexpensive, label-free, and easy to implement in the laboratories, does not suffer precipitation of the heparin-peptide complex or their conformational changes caused by surface immobilization, and is expected to be a useful complement to other existing methods.

Stable and reproducible electronic conduction through DNA molecular junctions

This letter presents the observation of stable and reproducible electronic conduction through double stranded (ds) DNA molecules in a nominally dry state. Stable conduction was realized by immobilizing 15 base-pair guanine:cytosine rich dsDNA within gold nanogap junctions, stabilizing the dsDNA with a polycation, and characterizing in nitrogen. In air, the current levels decrease with successive voltage scans likely due to oxidation of the guanine bases under bias. In nitrogen, reproducible current-voltage traces are observed and the current levels at specific bias points are stable with time. The stability allows comprehensive electrical studies and could enable conductance-based DNA sensors.This letter presents the observation of stable and reproducible electronic conduction through double stranded (ds) DNA molecules in a nominally dry state. Stable conduction was realized by immobilizing 15 base-pair guanine:cytosine rich dsDNA within gold nanogap junctions, stabilizing the dsDNA with a polycation, and characterizing in nitrogen. In air, the current levels decrease with successive voltage scans likely due to oxidation of the guanine bases under bias. In nitrogen, reproducible current-voltage traces are observed and the current levels at specific bias points are stable with time. The stability allows comprehensive electrical studies and could enable conductance-based DNA sensors.

Electronic conduction in DNA attached to gold electrodes

Experimental studies of electronic conduction in DNA have produced widely varying electronic properties and the debate is open on DNA electron transfer/transport mechanisms. We have advanced work towards quantifying the electrical conductivity of DNA oligonucleotides by studying the conductivity of double stranded DNA that is immobilized in a well defined manner in break junctions. To this end an 18 base pair long (approx. 5 nm) guanine rich double stranded DNA molecule has been designed to be attached to gold electrodes. The oligonucleotides used here have been synthesized using thiol (-SH) end groups that can form strong bonds to gold surfaces, and thus should provide low-resistance coupling to the electrodes. For a number of devices, conductivity between the contact pads increases significantly following exposure to a solution containing the DNA double strands. This increase in conductivity is thought to be due to electronic conduction through DNA double strands which are bonded to the two contacts. This hypothesis is confirmed by the loss in conductivity that is measured upon the denaturation of the double strand in deionized water.

Design and fabrication of bio-hybrid materials using inkjet printing

The integration of biomolecules such as proteins, carbohydrates, or enzymes into functional materials, whether through physical or chemical coupling, remains a critical processing step in the fabrication of engineered biosensors or tissue scaffolds, where anisotropy and composition can directly impact material function and host integration. A means to achieve these features is through the selective patterning of biomolecules, which is used to recruit and direct cell growth in vitro. The authors describe the design of protein-based materials using inkjet printing and discuss how fluid physical properties of the formulated inks influence pattern formation and material performance. When interfaced with carbon nanotubes, the biohybrid films retain their chemical signature but with enhanced structural stability and electrical conductivity over time. These structures also support the adhesion and proliferation of human dermal fibroblasts. Together, these properties demonstrate the utility of printed biohybrid f...