Jue-Yeon Lee

Enhanced bone formation by controlled growth factor delivery from chitosan-based biomaterials.

For the purpose of obtaining high bone forming efficacy, development of chitosan was attempted as a tool useful as a scaffolding device. Porous chitosan matrices, chitosan-poly(L-lactide) (PLLA) composite matrices and chitosan coated on PLLA matrices were dealt with in this research. Porous chitosan matrix was fabricated by freeze-drying and cross-linking aqueous chitosan solution. Porous chitosan matrix combined with ceramics and constituents of extracellular matrices were prepared and examined for their bone regenerative potential. Composite porous matrix of chitosan-PLLA was prepared by mixing polylactide with chitosan and freeze-drying. All chitosan based devices demonstrated improved bone forming capacity by increasing mechanical stability and biocompatibility. Release of platelet-derived growth factor-BB (PDGF-BB) from these matrices exerted significant osteoinductive effect in addition to the high osteoconducting capacity of the porous chitosan matrices. The hydrophobic surface of PLLA matrices was modified by chitosan to enhance cell affinity and wettability. The chitosan coated PLLA matrix induced increased osteoblast attachment as compared with intact PLLA surface. Overall results in this study demonstrated the usefulness of chitosan as drug releasing scaffolds and as modification tools for currently used biomaterials to enhance tissue regeneration efficacy. These results may expand the feasibility of combinative strategy of controlled local drug delivery concept and tissue engineered bone formation in reconstructive therapy in the field of periodontics, orthopedics and plastic surgery.

Osteoblastic differentiation of human bone marrow stromal cells in self-assembled BMP-2 receptor-binding peptide-amphiphiles.

Self-assembled nanostructures consisting of BMP receptor-binding peptides, termed osteopromotive domains (OPDs), and hydrophobic alkyl chains were fabricated with the aim of developing three-dimensional scaffolding materials for osteoblastic differentiation. OPD peptide was identified from BMP-2 and had an affinity for BMP receptors thereby inducing differentiation of human bone marrow stromal cells into osteoblastic cells. The peptide-hydrophobic alkyl chain amphiphiles were designed to mimic nanofibrous extracellular structures and to add osteogenic ligands to enhance osteoblastic cell function. The OPD peptide-amphiphiles (OPDAs) that end with the alkylation of the N-terminus of the OPD peptide were synthesized by standard solid phase chemistry. The self-assembly was triggered by mixing OPDA solution with calcium ions. Observation using scanning electron microscopy (SEM) revealed the formation of nanofibrous structures with extremely high aspect ratios and high surface areas. The FT-IR and circular dichroism (CD) spectrophotometry demonstrated that self-assembled nanofibers have a beta-sheet structure. The activation of Smad, an osteoblastic differentiation marker, was obtained in the cell culture gel of self-assembled OPDA; therefore, the intracellular signal transduction for osteogenesis was performed like an OPD peptide. Cell survival was supported in the OPDA gel for 10 days, and osteoblastic differentiation of human bone marrow stromal cells (hBMSCs) was evident as demonstrated by calcein staining and ALP activity measurement. These results revealed that self-assembled OPDA maintained osteogenic activity by the surface-exposed OPD peptide. Taken together, the self-assembled OPDA nanofibrous gel can be utilized as a cell culture scaffold in bone regeneration.

Transforming Growth Factor (TGF)-β1 Releasing Tricalcium Phosphate/Chitosan Microgranules as Bone Substitutes

AbstractPurpose. Tricalcium phosphate (TCP)/chitosan composite microgranules were developed as bone substitutes and tissue engineering scaffolds with the aim of obtaining a high bone forming efficacy. The microgranules have the ability to fill various types of defect sites with closer packing. In addition, the transforming growth factor-beta 1 (TGF-β1) was added to the microgranules in order to improve bone-healing efficacy. Methods. TCP/chitosan microgranules were fabricated by dropping a TCP suspended chitosan solution into a NaOH/ethanol solution. TGF-β1 was incorporated into the TCP/chitosan microgranules by soaking the microgranules into the TGF-β1 solution. Scanning electron microscopy (SEM) observations as well as experiments examining the release of TGF-β1 from chitosan and TCP/chitosan microgranules were performed. SEM was used to examine the cell morphologies on the microgranules, and the extent of cell proliferation was evaluated using a dimethyl-thiazol tetrazolium bromide (MTT) assay. The differentiated cell function was assessed by measuring the alkaline phosphatase activity as well as performing an osteocalcin assay. Results. The size of the prepared microgranules was 350-500μm and TCP powders were observed on the surface of the microgranules. TGF-β1 was released from the TCP/chitosan microgranules at a therapeutic concentration for 4 weeks. The proliferation of osteoblasts on the TGF-β1 loaded microgranules was the highest among the microgranules. SEM indicated that the seeded osteoblastic cells were firmly attached to the microgranules and proliferated in a multilayer fashion. The ALPase activity and osteocalcin content of all the samples increased during the culture period. Conclusions. These results suggest that the TCP/chitosan microgranules are potential bone substitutes with a drug releasing capacity and a osteoblastic cells culture scaffold.

Investigations of the space environment aboard the Universitetsky-Tat’yana and Universitetsky-Tat’yana-2 microsatellites

The first results obtained through the university small satellites program developed at Moscow State University (MSU) are presented. The space environment was investigated aboard two MSU microsatellites designed for scientific and educational purposes, Universitetsky-Tat’yana and Universitetsky-Tat’yana-2. The scientific equipment is described to study charged particles in near Earth space and atmospheric radiations in ultraviolet, red, and infrared optical wavelength ranges. The dynamic properties of fluxes of charged particles in microsatellite orbits are studied and findings are presented regarding specific parameters of solar proton penetration during the geomagnetic disturbances. Experimental results are considered concerning flashes of ultraviolet (UV), red (R), and infrared (IR) radiation that are transient light phenomena in the upper atmosphere. The space educational MSU program developed on the basis of the Universitetsky-Tat’yana projects is reviewed.

Bioactive peptide-modified biomaterials for bone regeneration.

Bioactive biomaterials are desirable as tissue engineering scaffolds by virtue of their capability to mimic the natural environment of the extracellular matrix. Bioactive biomaterials have been achieved by incorporating synthetic short peptide sequences into suitable materials either by surface modification or by bulk incorporation. The goal is to enhance cell attachment and other basic functions. Bioactive peptides can be obtained from biological or chemically synthesized sources, increasing their specific cellular responses for tissue growth and development. Compared to using an entire growth factor in regenerative therapy, these peptides demonstrate potential advantages such as overcoming possible immunogenicity, being less susceptible to degradation, and producing fewer tumor-related side effects. Biomaterial scaffolds modified with peptides can provide biological ligands for cell-scaffold interactions that promote cell attachment, proliferation, and differentiation. Peptide-based biomaterial scaffolds can be fabricated to form two- and three-dimensional structures. This review discusses cell-binding, biominerailization inducing peptides, and receptor-binding peptides for bone regeneration. This review also addresses issues related to peptide immobilization as well as potential complications that may develop as a result of using these versatile bioactive peptides. The development of self-assembled peptide amphiphiles with the goal of generating new three-dimensional scaffolds for tissue engineering is also summarized.

MEASUREMENTS OF THE RELATIVE ABUNDANCES OF HIGH-ENERGY COSMIC-RAY NUCLEI IN THE TeV/NUCLEON REGION

We present measurements of the relative abundances of cosmic-ray nuclei in the energy range of 500-3980 GeV/nucleon from the second flight of the Cosmic Ray Energetics And Mass balloon-borne experiment. Particle energy was determined using a sampling tungsten/scintillating-fiber calorimeter, while particle charge was identified precisely with a dual-layer silicon charge detector installed for this flight. The resulting element ratios C/O, N/O, Ne/O, Mg/O, Si/O, and Fe/O at the top of atmosphere are 0.919 ? 0.123stat ? 0.030syst, 0.076 ? 0.019stat ? 0.013syst, 0.115 ? 0.031stat ? 0.004syst, 0.153 ? 0.039stat ? 0.005syst, 0.180 ? 0.045stat ? 0.006syst, and 0.139?? 0.043stat ? 0.005syst, respectively, which agree with measurements at lower energies. The source abundance of N/O is found to be 0.054 ? 0.013stat ? 0.009syst+0.010esc ?0.017. The cosmic-ray source abundances are compared to local Galactic (LG) abundances as a function of first ionization potential and as a function of condensation temperature. At high energies the trend that the cosmic-ray source abundances at large ionization potential or low condensation temperature are suppressed compared to their LG abundances continues. Therefore, the injection mechanism must be the same at TeV/nucleon energies as at the lower energies measured by HEAO-3, CRN, and TRACER. Furthermore, the cosmic-ray source abundances are compared to a mixture of 80% solar system abundances and 20% massive stellar outflow (MSO) as a function of atomic mass. The good agreement with TIGER measurements at lower energies confirms the existence of a substantial fraction of MSO material required in the ~TeV per nucleon region.

Effect of immobilized cell-binding peptides on chitosan membranes for osteoblastic differentiation of mesenchymal stem cells

Two cell‐binding domains from FGF‐2 (fibroblast growth factor‐2) were shown to increase cell attachment and osteoblastic differentiation. Two synthetic peptides derived from FGF‐2, namely residues 36–41 (F36; PDGRVD) and 77–83 (F77; KEDGRLL), were prepared and their N‐termini further modified for ease of surface immobilization. Chitosan membranes were used in the present study as mechanical supportive biomaterials for peptide immobilization. Peptides could be stably immobilized on to the surface of chitosan membranes. The adhesion of mesenchymal stem cells to the peptide (F36 and F77)‐immobilized chitosan membrane was increased in a dose‐dependent manner and completely inhibited by soluble RGD (Arg‐Gly‐Asp) and anti‐integrin antibody, indicating the existence of an interaction between F36/F77 and integrin. Peptide‐immobilized chitosan supported human bone‐marrow‐derived mesenchymal‐stem‐cell differentiation into osteoblastic cells, as demonstrated by alkaline phosphate expression and mineralization. Taken together, the identified peptide‐immobilized chitosan membranes were able to support cell adhesion and osteoblastic differentiation; thus these peptides might be useful as bioactive agents for osteoblastic differentiation and surface‐modification tools in bone regenerative therapy.

Synthetic peptide‐coated bone mineral for enhanced osteoblastic activation in vitro and in vivo

A 15-mer synthetic peptide, designated P1, was derived from the bone morphogenetic protein (BMP) receptor I and BMP receptor II binding domains of BMP-2 for the purpose of enhancing bone regeneration capacity of inorganic bovine bone mineral. A second peptide, denoted P2, was designed by adding seven glutamic acid residues to the N-terminal of P1 to increase the surface coating efficiency onto bone mineral. The coating efficiency of P1 increased with the concentration of peptide. P2 peptide, in contrast, had a higher coating efficiency at lower peptide concentrations. The peptides properly transduced intracellular signals properly via the Smad and ERK pathways, thereby increasing mineralization in vitro, implying that the peptides alone can induce osteoblastic differentiation. Adhesion of cells to bone mineral was greater when peptides were present than in bone mineral alone. P1- and P2-coated bone mineral increased osteoblastic differentiation, as demonstrated by ALPase activity. P1-coated bone mineral stimulated more new bone regeneration in bone defect sites after 2 weeks than the peptide-free control. These peptides, in combination with bone grafts or implants, have the potential to enhance osteoblastic differentiation and bone regeneration.

A New Type of Space Telescope for Observation of Extreme Lightning Phenomena in the Upper Atmosphere

A new type of space telescope with a 3 mm × 3 mm Micro-Electro-Mechanical System (MEMS) micromirror array has been fabricated and launched into space. This telescope has unique features: a wide field of surveillance view, and fast zoom-in and tracking capabilities. Although the micromirror array area is small, the space telescope was capable of observing the space-time development of extreme lightning in the upper atmosphere. It fulfilled its purpose by proving the principles of a space telescope. The concept and technologies used in this telescope can be extended to large MEMS space telescopes for future missions for earth and space science, including gamma ray bursts and ultra high energy cosmic rays. The performance of the space telescope during the ground test before launch as well as its performance in space are here presented to demonstrate the fast zoom-in and tracking capabilities of the telescope.

Selective osteogenesis by a synthetic mineral inducing peptide for the treatment of osteoporosis

Mineralization in mammalian cells is accomplished by concerted regulation of protein-based extracellular matrix (ECM) components, such as non-collagenous proteins and collagen fibrils. In this study, we investigated the ability of a collagen-binding motif (CBM) peptide derived from osteopontin to selectively affect osteogenic or adipogenic differentiation in vitro and in vivo. In particular, increased osteogenic differentiation and decreased adipogenic differentiation were observed in human mesenchymal stem cells (hMSCs). Osteocalcin (OCN) protein expression in MC3T3-E1 cells without osteogenic inducers was then investigated following treatment with the CBM peptide. In ovariectomized (OVX) mice, estrogen deficiency induced osteoporosis and increased fat tissue deposition. However, after the CBM peptide or estradiol was injected into the OVX mice for 2 months, the increased serum OCN concentration and alkaline phosphate (ALP) activity were decreased in the estradiol-treated group (OVX-E) and the high-concentration CBM peptide-treated group (OVX-HP). Significant bone loss was also observed in the ovariectomized mice (OVX-PBS). In particular, the bone volume per total volume (BV/TV) and bone mineral density (BMD) were significantly decreased in the OVX mice; however, both of these markers were restored in the OVX-HP group, which also had significantly well-developed bone structure and bone formation. In contrast to the bone structural change, adipose tissue was increased in the OVX-PBS. However, a significant decrease in total fat and subcutaneous fat was observed in the low-concentration CBM peptide-treated group (OVX-LP) and the estradiol-treated group (OVX-E). Taken together, these results suggest that the CBM peptide could be an effective therapeutic agent for osteoporosis due to its selective stimulation of osteogenic differentiation, rather than adipogenesis.