• Journal of Korean Neurosurgical Society
• /
• v.56 no.5
• /
• pp.383-389
• /
• 2014

Objective : Neural tissue transplantation has been a promising strategy for the treatment of Parkinson's disease (PD). However, transplantation has the disadvantages of low-cell survival and/or development of dyskinesia. Transplantation of cell aggregates has the potential to overcome these problems, because the cells can extend their axons into the host brain and establish synaptic connections with host neurons. In this present study, aggregates of human brain-derived neural stem cells (HB-NSC) were transplanted into a PD animal model and compared to previous report on transplantation of single-cell suspensions. Methods : Rats received an injection of 6-OHDA into the right medial forebrain bundle to generate the PD model and followed by injections of PBS only, or HB-NSC aggregates in PBS into the ipsilateral striatum. Behavioral tests, multitracer (2-deoxy-2-[$^{18}F$]-fluoro-D-glucose ([$^{18}F$]-FDG) and [$^{18}F$]-N-(3-fluoropropyl)-2-carbomethoxy-3-(4-iodophenyl)nortropane ([$^{18}F$]-FP-CIT) microPET scans, as well as immunohistochemical (IHC) and immunofluorescent (IF) staining were conducted to evaluate the results. Results : The stepping test showed significant improvement of contralateral forelimb control in the HB-NSC group from 6-10 weeks compared to the control group (p<0.05). [$^{18}F$]-FP-CIT microPET at 10 weeks posttransplantation demonstrated a significant increase in uptake in the HB-NSC group compared to pretransplantation (p<0.05). In IHC and IF staining, tyrosine hydroxylase and human ${\beta}2$ microglobulin (a human cell marker) positive cells were visualized at the transplant site. Conclusion : These results suggest that the HB-NSC aggregates can survive in the striatum and exert therapeutic effects in a PD model by secreting dopamine.

• Nutrition Research and Practice
• /
• v.11 no.3
• /
• pp.190-197
• /
• 2017

BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of $50-250{\mu}g/mL$. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to $250{\mu}g/mL$ and were 149% and 129% at $250{\mu}g/mL$ concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of $500{\mu}g/mL$. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.16 no.1
• /
• pp.585-592
• /
• 2015

The objective of this study is to determine if when chymopapain is loaded onto a nano-carrier, an injection of it reduces the spreading range of the drug within the discs. The materials for the experiment, which were conducted for three weeks, included fifteen intervertebral discs taken from two cadavers, which were divided according to the types of injected chymopapain solutions as follows: ordinary chymopapain group and nano-carrier system group. The nano-carrier system group was again divided into two subgroups according to the types of pluronics, the basic material for the nano-carriers: Pluronic F 127(DA-PF 127) in the nano-carrier group and Pluronic F 68(DA-PF 68) in the nano-carrier group. The results showed that the action of chymopapain using a pluronic-based nano-carrier system was localized around the center of the injection site instead of broad spreading, compared to that of the ordinary chymopapain group (p<0.01). This characteristic suggests a possible application to effective agents for minimally invasive spinal treatment through which disc lesions were removed selectively.

• Archives of Plastic Surgery
• /
• v.36 no.3
• /
• pp.247-253
• /
• 2009

Purpose: Hydroxyapatite(HA) has been widely used due to its chemical similarity to bone and good biocompatibility. HA is composed of macropores and micropores. Too much irregularities of the micropores are ineffective against the adhesion and proliferation of osteoblast. Many efforts have been tried to overcome these drawbacks. HA crystal coating on the irregular surface of HA scaffold, crystallized HA, is one of the method to improve cell adhesion. Meanwhile, the collagen has been incorporated with HA to create composite scaffold that chemically resembles the natural extracellular matrix components of bone. The authors proposed to examine the effect of collagen - coated crystallized HA on the adhesion and proliferation of osteoblast. Method: HA powder containing $10{\mu}m$ pore size was manufactured as 1 cm pellet size. For the making crystallized HA, 0.1 M EDTA solution was used to dissolve HA powder and heated $100^{\circ}C$ for 48 hours. Next, the crystallized HA pellets were coated with collagen (0.1, 0.5, and 1%). The osteoblasts were seeded into HA pellets and incubated for the various times (1, 5, and 9 days). After the indicating days, methylthiazol tetrazolium (MTT) assay was performed for cell proliferation and alkaline phosphatase (ALP) activty was measured for bone formation. Result: In SEM study, the surface of crystallized HA pellet was more regular than HA pellet. MTT assay showed that the proliferation of osteoblasts increased in a collagen dose - dependent and time - dependent manner and had a maximum effect at 1% collagen concentration. ALP activity also increased in a collagen dose - dependent manner and had a highest effect at 1% collagen concentration. Conclusion: These data showed that crystallization and collagen coating of HA was effective for osteoblast proliferation and ALP activity. Therefore, our results suggest that crystallized - HA scaffold with collagen coating is may be a good strategy for tissue engineering application for bone formation.

• Archives of Plastic Surgery
• /
• v.33 no.5
• /
• pp.612-615
• /
• 2006

Purpose: An ideal bony construct can be divided into two broad categories: (1) the design and fabrication of biodegradable, biomimetic scaffolds that provide correct signals to induce osteogenesis: (2) the identification of an ideal source of osteoprogenitor cells to seed onto the scaffold. We selected poly-glycolic acid as a synthetic scaffold among various scaffolds because of these properties. Meanwhile, culture medium is supplemented with fetal bovine serum(FBS): such serum contains essential elements such as proteins, hormones, growth factors and trace minerals. The composition of FBS can be ideal for various cell growth in vitro. We supposed that we could enhance bone growth at a fractured site if FBS was mixed with synthetic scaffold-PGA. Methods: We cultured human osteoblasts in five different prepared culture dishes made with FBS and PGA mixture. The mixtures contained different ratio of FBS, that is, 0, 1.5, 3, 7, and 10%. We cultured human osteoblasts for seven days and examined the growth and attachment of the cells at the 1st, 3rd, 5th, 7th days, respectively. Results: In the mixture of 0% FBS and PGA, the growth of the cells lasted for one day. In 1.5 and 3% FBS and PGA, the growth of the cells was examined at the 3rd day, then minimally declined at the 5th and 7th days. In 7% FBS and PGA, the growth of the cells lasted for 5 days, then declined at the 7th day. In 10% FBS and PGA, the growth of the cells lasted for 5 days, then declined at the 7th day. Staining status of the osteoblasts with alkaline phosphatase showed pale pink color in 0% FBS and PGA groups, but bright pink color in 1.5, 3, 7, 10% FBS and PGA groups, especially in 3%, 7%. Conclusion: In consequence, the growth of human osteoblast was higher in the mixture of FBS and PGA groups than in pure PGA ones. It is assumed that the mixture of FBS and PGA affects the proliferation of human osteoblasts.

• Journal of Chest Surgery
• /
• v.37 no.6
• /
• pp.474-481
• /
• 2004

Background: Tracheal transplantation is necessary in patients with extensive tracheal stenosis, congenital lesions and other oncologic conditions but bears. many critical problems compared to other organ transplantations. The purpose of this study was to develop intestine-cartilage composite grafts for potential application in tracheal reconstruction by free intestinal graft. Material and Method: Hyaline cartilage was harvested from trachea of 2 weeks old New Zealand White Rabbits. Chondrocytes were isolated and cultured for 8 weeks. Cultured chondrocytes were seeded in the PLGA scaffolds and mixed in pluronic gel Chondrocyte bearing scaffolds and gel mixture were embedded in submucosal area of stomach and colon of 3 kg weighted New Zealand White Rabbits under general anesthesia. 10 weeks after implantation, bowels were harvested for evaluation. Result: We identified implantation site by gross examination and palpation. Developed cartilage made a good frame for shape memory. Microscopic examinations included special stain s howed absorption of scaffold and cartilage formation even though it was not fully matured. Conclusion: Intestine-cartilage composite graft could be applicable in the future as tracheal substitute and should be further investigated.

• Microbiology and Biotechnology Letters
• /
• v.36 no.1
• /
• pp.12-20
• /
• 2008

Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to minute virus of mice(MVM), and there are several reports of MVM contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the MVM safety, a real-time PCR method was developed for quantitative detection of MVM in cell lines, raw materials, manufacturing processes, and final products as well as MVM clearance validation. Specific primers for amplification of MVM DNA was selected, and MVM DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $6{\times}10^{-2}TCID_{50}/mL$. The real-time PCR method was proven to be reproducible and very specific to MVM. The established real-time PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with MVM. MVM DNA could be Quantified in CHO cell as well as culture supernatant. When the real-time PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of MVM.

• Journal of the Korea Organic Resources Recycling Association
• /
• v.13 no.3
• /
• pp.51-62
• /
• 2005

The effects of the processing mixture of food wastes and various organic wastes when vermicomposted on earthworm(Eisenia foefida) growth, the cast production amounts and the chemical properties of casts were evaluated. The substrates used in this experiments were cow manure, pig manure sludge, fermented pig manure with sawdust, nightsoil sludge, and sewage sludge and were respectively mixed with food wastes at a ratios of 50:50(v/v). The control consisted of food wastes alone without other wastes. All of earthworm died in the food wastes 100%, therefore the process of food wastes alone by vermicomposting was impossible in this experiment. Worm cast produced sufficiently contained quantities of available phosphorus, exchangeable potassium, exchangeable magnesium, and cation exchange capacity. The increase of earthworm's biomass occured on the mixtures of food wastes and cow manure, fermented pig manure with sawdust. Dry weight of worm cast was the highest on the mixtures of food wastes and fermented pig manure with sawdust and the proportion of cast weight after vermicomposting was significantly the highest on the mixtures of food wastes and cow manure($p{\leq}0.05$). Also, the mixtures of food wastes and cow manure, and fermented pig manure with sawdust showed a positive values of conversion rate and conversion efficiency rate of organic matter to earthworm tissue than that of other treatments. These results suggested that cow manure and fermented pig manure with sawdust are adequate to process with food wastes by vermicomposting.

• KIPS Transactions on Software and Data Engineering
• /
• v.7 no.10
• /
• pp.377-382
• /
• 2018

This study deals with internal state and tissue density analysis methods for red ginseng grade determination. For internal measurement of red ginseng, there have been various studies on nondestructive testing methods since the 1990s, It was difficult to grasp the most important inner hole and inside whites in the grading. So in this study, we developed a closed capturing device for infra-red illumination environment, and developed an internal measurement system that can detect the presence and diameter of inner hole and inside whites. Made devices consisted of infrared lights with a high transmission rate of red ginseng in 920 nanometer wave band, a infra-red camera and a Y axis actuator with a red ginseng automatically controlled focus on the camera. The proposed algorithm performs an auto-focus system on the Y-axis actuator to automatically adjust the sharp focus of the object according to the size and thickness. Then red ginseng is rotated $360^{\circ}$ at $1^{\circ}$ intervals and 360 total images are acquired, and reconstructed as a sinogram through Radon transform and Back-projection algorithm was performed to acquire internal images of red ginseng. As a result of the algorithm, it was possible to acquire internal cross-sectional image regardless of the thickness and shape of red ginseng. In the future, if more than 10,000 different shapes and sizes of red ginseng internal cross-sectional image are acquired and the classification criterion is applied, it can be used as a reliable automated ginseng grade automatic measurement method.

• Polymer(Korea)
• /
• v.30 no.3
• /
• pp.196-201
• /
• 2006

Poly (ethylene glycol)-based diblock and triblock thermo- sensitive polyester copolymers were investigated for application on tissue engineering and injectable biomaterials in drug delivery system due to their nontoxicity, biocompatibility and biodegradability. We synthesized the diblock copolymers consisting of methoxy poly (ethylene glycol) (MPEG) (Mn=750 g/mole) and poly $(\varepsilon-caprolactone)$ (PCL) by ring opening polymerization of $\varepsilon-CL$ with MPEG as an initiator in the presence of HCl $Et_2O$. The effect of diblock copolymers on in vivo osteogenic differentiation of rat bone marrow stromal cells (BMSCS) with and without the presence of osteogenic supplements (dexamethasone) was investigated. Thin sections were cut from paraffin embedded tissues and histological sections were stained by H&E, von Kossa, and immunohistochemical staining for osteocalcin. In conclusion, dexamethasone containing thermo- sensitive hydrogel might be improved osteogenic differentiation of BMSCs. We expect the osteoinduction effect to be excellent when it uses stem cell or other osteogenic materials.