Hyukjin Lee

Hyaluronic Acid-Paclitaxel Conjugate Micelles : Synthesis, Characterization, and Antitumor Activity

Chemical conjugates of paclitaxel and hyaluronic acid (HA) were synthesized by utilizing a novel HA solubilization method in a single organic phase. Hydrophilic HA was completely dissolved in anhydrous DMSO with addition of poly(ethylene glycol) (PEG) by forming nanocomplexes. Paclitaxel was then chemically conjugated to HA in the DMSO phase via an ester linkage without modifying extremely hydrophilic HA. A series of HA-paclitaxel conjugates with different conjugation percentages were synthesized and characterized. HA-paclitaxel conjugates self-assembled in aqueous solution to form nanosized micellar aggregates, as characterized by dynamic light scattering (DLS), atomic force microscopy (AFM), and transmission electron microscopy (TEM). An intact form of paclitaxel was regenerated from HA-paclitaxel conjugate micelles at acidic pH conditions. HA-paclitaxel conjugate micelles exhibited more pronounced cytotoxic effect for HA receptor overexpressing cancer cells than for HA receptor deficient cells, suggesting that they can be potentially utilized as tumor-specific nanoparticulate therapeutic agents.

Synthesis, characterization, and in vivo diagnostic applications of hyaluronic acid immobilized gold nanoprobes

Herein we describe a new class of multifunctional gold nanoprobes for ultra-sensitive optical detection of reactive oxygen species (ROS) and hyaluronidase (HAdase). The nanoprobes were fabricated by end-immobilizing near-infrared fluorescence (NIRF) dye labeled hyaluronic acid (HA) onto the surface of gold nanoparticles (AuNPs). The nanoprobes effectively induced nanoparticle surface energy transfer (NSET) between NIRF dyes and AuNPs. When the surface immobilized HA was cleaved by ROS and HAdase, strong fluorescence recovery signals were attained with extreme sensitivity. In live animal models of rheumatoid arthritis (RA) and metastatic tumor, local arthritic inflammation and tumor sites were clearly identified upon systemic injection of the nanoprobes. These results suggest that the gold nanoprobes can be exploited not only as in vitro molecular and cellular imaging sensors for ROS and HAdase, but also as in vivo optical imaging agents for detection of local HA degrading diseases such as RA and tumor.

Poly[lactic-co-(glycolic acid)]-Grafted Hyaluronic Acid Copolymer Micelle Nanoparticles for Target-Specific Delivery of Doxorubicin

PLGA-grafted HA copolymers were synthesized and utilized as target specific micelle carriers for DOX. For grafting hydrophobic PLGA chains onto the backbone of hydrophilic HA, HA was solubilized in an anhydrous DMSO by nano-complexing with dimethoxy-PEG. The carboxylic groups of HA were chemically grafted with PLGA, producing HA-g-PLGA copolymers. Resultant HA-g-PLGA self-assembled in aqueous solution to form multi-cored micellar aggregates and DOX was encapsulated during the self-assembly. DOX-loaded HA-g-PLGA micelle nanoparticles exhibited higher cellular uptake and greater cytotoxicity than free DOX for HCT-116 cells that over-expressed HA receptor, suggesting that they were taken up by the cells via HA receptor-mediated endocytosis.

Catechol-Grafted Poly(ethylene glycol) for PEGylation on Versatile Substrates

We report on catechol-grafted poly(ethylene) glycol (PEG-g-catechol) for the preparation of nonfouling surfaces on versatile substrates including adhesion-resistant PTFE. PEG-g-catechol was prepared by the step-growth polymerization of PEO to which dopamine, a mussel-derived adhesive molecule, was conjugated. The immersion of substrates into an aqueous solution of PEG-g-catechol resulted in robust PEGylation on versatile surfaces of noble metals, oxides, and synthetic polymers. Surface PEGylation was unambiguously confirmed by various surface analytical tools such as ellipsometry, goniometry, infrared spectroscopy, and X-ray photoelectron spectroscopy. Contrary to existing PEG derivatives that are difficult-to-modify synthetic polymer surfaces, PEG-g-catechol can be considered to be a new class of PEGs for the facile surface PEGylation of various types of surfaces.

Heparin immobilized gold nanoparticles for targeted detection and apoptotic death of metastatic cancer cells

In the present study, heparin immobilized, multifunctional gold nanoparticles (AuNPs) were developed as a new class of theragnostic nanomaterials for metastatic cancer cell imaging and apoptosis. AuNPs were surface modified with fluorescent dye labeled heparin molecules to detect a metastatic stage of cancer cells that over-express heparin-degrading enzymes. The heparin immobilized AuNPs exhibited enhanced fluorescence signals by specific cleavage of heparin molecules from the surface of AuNPs by heparinase or heparanase secreted from metastatic cancer cells. In addition, heparin immobilized AuNPs that were additionally tethered with RGD peptides on the surface demonstrated highly specific apoptotic activities for selective cancer cells over-expressing RGD receptors on the membrane, revealing that internalized heparin within cells clearly triggered an apoptotic event. These results suggest that heparin immobilized AuNPs can be usefully exploited for optical imaging agents for metastatic tumors as well as therapeutic cancer treatment.

Photo-crosslinkable, biomimetic, and thermo-sensitive pluronic grafted hyaluronic acid copolymers for injectable delivery of chondrocytes

Hyaluronic acid (HA) grafted with Pluronic F127 copolymer was used as biomimetic hydrogels for cell delivery. The graft copolymer was synthesized by conjugating amine end-capped Pluronic F127 to carboxylic groups of HA using coupling agents. The synthesized HA-g-Pluronic exhibited thermo-sensitive sol-gel transition behaviors over the temperature range of 20-40 degrees C. HA-g-Pluronic copolymers with vinyl groups were photo-crosslinked to prepare more robust hydrogels for cell cultivation. For improved cellular adhesion and proliferation, cell adhesive peptide (Arg-Gly-Asp (RGD)) was additionally conjugated to the HA backbone. The resultant thermo-sensitive, photocrosslinkable, and RGD modified HA-g-Pluronic copolymers were used to encapsulate and cultivate bovine chondrocytes in vitro. A tissue containing cartilage-like components such as GAG and type II collagen was successfully produced within the hydrogels, indicating that the synthesized HA-g-Pluronic copolymers can be potentially used as an injectable cell carrier.

Optical imaging of intracellular reactive oxygen species for the assessment of the cytotoxicity of nanoparticles

The generation of intracellular reactive oxygen species (ROS) was optically monitored using ROS-sensitive gold nanoprobes in response to an exposure of nanoparticles (NPs). Fluorescent dye-labeled hyaluronic acid was grafted onto the surface of gold nanoparticles (HF-AuNPs) for imaging intracellular ROS. The ultrasensitive detection of intracellular ROS was utilized as a powerful analytical tool to assess early cellular toxicities of monodisperse polystyrene (PS) particles with different sizes and different functional groups on the surface. The effect of PEGylation on the surface of PS NPs was also investigated by evaluating intracellular ROS generation. For various PS NPs, the extent of intracellular ROS was well correlated with cellular uptake, apoptosis inducing activity, and cytotoxic effect of NPs. In addition to the nanoparticles, commonly used polymeric gene carriers such as linear and branched polyethylenimine (PEI) were tested to analyze their extent of intracellular ROS generation related to cellular toxicity. This study demonstrated that sensitive and optical detection of intracellular ROS generation can provide a valuable toxicity index value for a wide range of NPs as an early indicator for cellular responses.

Perspectives on: Local and sustained delivery of angiogenic growth factors

This review emphasizes the role of angiogenesis in tissue engineering, introduces various angiogenic growth factors, and highlights current status of delivery systems for angiogenic growth factors using natural and synthetic biomaterials. A short overview of angiogenic growth factors is presented, followed by the introduction of emerging strategies for designing smart delivery carriers.

Surface PEGylation via Native Chemical Ligation

Native chemical ligation (NCL) is an emerging chemoselective chemistry that forms an amide bond by trans-thioesterification followed by intramolecular nucleophilic rearrangement between thioester and cysteine. The reaction is simple, occurs in a mild aqueous solution, and gives near-quantitative yields of a desired product. Since the first report in 1994, most studies involving the use of NCL have focused on the total synthesis of proteins to address fundamental questions pertaining to many aspects of protein science, such as folding, mirror images, and site-specific labeling of proteins, but applications of the NCL reaction for other areas remain largely unexplored. Herein, we present a facile strategy for surface immobilization of poly(ethylene glycol) (PEG) utilizing the NCL reaction. Surface immobilization of PEG (i.e., PEGylation) plays a key role in preventing nonspecific protein adsorption on surfaces, which is crucial in a wide variety of medical devices. Using cysteine-PEG and thioester-containing phosphonic acid conjugates, we achieved efficient surface PEGylation on titanium surfaces. Ellipsometry, goniometry, and X-ray photoelectron spectroscopy (XPS) unambiguously confirmed the presence of PEGs, which provided nonfouling effects of surfaces. This study indicates that the NCL reaction will be a useful toolkit for surface bioconjugation and functionalization.

Controlling mechanical properties of bio-inspired hydrogels by modulating nano-scale, inter-polymeric junctions

Summary Quinone tanning is a well-characterized biochemical process found in invertebrates, which produce diverse materials from extremely hard tissues to soft water-resistant adhesives. Herein, we report new types of catecholamine PEG derivatives, PEG-NH-catechols that can utilize an expanded spectrum of catecholamine chemistry. The PEGs enable simultaneous participation of amine and catechol in quinone tanning crosslinking. The intermolecular reaction between PEG-NH-catechols forms a dramatic nano-scale junction resulting in enhancement of gelation kinetics and mechanical properties of PEG hydrogels compared to results obtained by using PEGs in the absence of amine groups. Therefore, the study provides new insight into designing new crosslinking chemistry for controlling nano-scale chemical reactions that can broaden unique properties of bulk hydrogels.