• Biomedical Science Letters
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• v.15 no.1
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• pp.61-66
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• 2009

Leflunomide is an immunomodulatory agent used for the treatment of rheumatoid arthritis (RA). Leflunomide known as a regulator of iNOS synthesis which largely decreases NO production in diverse cell type. However, the effect of leflunomide on chondrocyte is still poorly understood. In our previous studies, we have shown that direct production of Nitric oxide (NO) by treating chondrocytes with NO donor, sodium nitroprusside (SNP), causes apoptosis via p38 mitogen-activated protein kinase in association with elevation of p53 protein level, caspase-3 activation. In this study, we characterized the molecular mechanism by which A77 1726 inhibit apoptosis. We found that A77 1726 inhibit NO-induced apoptosis as determined by MTT (Thiazolyl Blue Tetrazolium Bromide) assay and DNA fragmentation. The inhibition of apoptosis by A77 1726 was accompanied by increased PI-3 kinase and AKT activities. So, inhibition of phosphatidylinositol (PI)-3kinase with LY294002 rescued apoptosis. Triciribine, the specific inhibitor of AKT, also abolished anti-apoptotic effect. Our results indicate that A77 1726, the active metabolite of leflunomide, mediates NO-induced apoptosis in chondrocytes by modulating up-regulation of PI-3 kinase and AKT.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.11 no.9
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• pp.3358-3365
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• 2010

This study was carried out to investigate the effect of Sorbus commixta (SC), Geranium thunbergii (GT) and their mixture (SC:GT=1:1, MIX) on inhibition of bone loss and chondral defect. To examine their activities, we measured the alkaline phosphatase (ALP) activity in human osteoblast-like MG-63 cells and performed tartrate-resistant acid phosphate (TRAP) staining in osteoclast differentiated from Raw264.7 cells. To investigate the influence on chondrocyte differentiation, we performed alcian-blue staining in chondrocyte differentiated from ATDC5 cells. All of SC, GT and MIX did not increase ALP activity in MG-63 cells. However, SC and mixture (SC:GT=1:1, MIX) significantly inhibited osteoclastic differentiation. And they also induced chondrocyte differentiation. These results suggest that SC and GT may have a potential for the treatment of bone loss and chondral defect by suppression of osteoclast differentiation and stimulation of chondrocyte differentiation. Therefore, clarification of their mechanisms and active components will be needed.

• Journal of the Korean Arthroscopy Society
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• v.11 no.1
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• pp.1-6
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• 2007

Purpose: The purpose of this preliminary report is to investigate the short term outcome of performing gel type fibrin matrix autologous chondrocyte implantation to patients who have damaged knee joint cartilage using secondary arthroscopy. Material and Methods: Six patients who have damaged knee joint cartilage were involved. The average size of defect was $5.13\;cm^2$. While performing primary arthroscopy, whole layer of cartilage bone was obtained either from the margin of damaged cartilage or the bilateral margin of a trochlea. The cartilaginous cells were obtained for culture for four to six weeks. While performing secondary minimal invasive arthrotomy, gel type fibrin matrix autologous chondrocyte was implanted on the chondral defect site. Results: 4 among 6 patients to be more than good in Modified Cincinnati Knee Scoring system. Lysholm function score was 59.5 preoperatively, and it improved to 76.25. ICRS grading by performing secondary arthroscopy revealed 4 out of 6 patients to be nearly normal. Conclusion: Gel type fibrin matrix autologous chondrocyte implantation is a treatment for cartilage defect, which takes less time to operate than the conventional implantation. In addition, this method minimizes the size of incision and allows arthroscopic surgery. However, long term follow up and more case study is thought to be necessary.

• Tribology and Lubricants
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• v.21 no.5
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• pp.236-240
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• 2005

In order to prepare for the suitable surfaces of implants or medical devices, quantitative evaluation of adhesion between cells and biomaterials is essential. To better understand adhesion formation between cells and biomaterials, we used the cytodetachment technique which measures the adhesive force of a single cell through changing the, culture time and detachment speed. The results showed that the adhesive force could be affected by the culture time of cells on the surface of materials and the detachment speed. Moreover, there was a large discrepancy among the adhesion strength measured by similar techniques conducted on the different cells and substrates. It can be 'concluded that the variation of the force measurement technique can seriously alter the level of the force required to detach a cell on the surface of materials.

• Archives of Plastic Surgery
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• v.47 no.5
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• pp.392-403
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• 2020

Severe cartilage defects and congenital anomalies affect millions of people and involve considerable medical expenses. Tissue engineering offers many advantages over conventional treatments, as therapy can be tailored to specific defects using abundant bioengineered resources. This article introduces the basic concepts of cartilage tissue engineering and reviews recent progress in the field, with a focus on craniofacial reconstruction and facial aesthetics. The basic concepts of tissue engineering consist of cells, scaffolds, and stimuli. Generally, the cartilage tissue engineering process includes the following steps: harvesting autologous chondrogenic cells, cell expansion, redifferentiation, in vitro incubation with a scaffold, and transfer to patients. Despite the promising prospects of cartilage tissue engineering, problems and challenges still exist due to certain limitations. The limited proliferation of chondrocytes and their tendency to dedifferentiate necessitate further developments in stem cell technology and chondrocyte molecular biology. Progress should be made in designing fully biocompatible scaffolds with a minimal immune response to regenerate tissue effectively

• Biomedical Science Letters
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• v.15 no.1
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• pp.67-72
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• 2009

Microtubule-interfering agents (MIAs), including paclitaxel, have been attributed in part to interference with microtubule assembly, impairment of mitosis, and changes in cytoskeleton. But the signaling mechanisms that link microtubule disarray to destructive or protective cellular responses are poorly understood. This study investigated the effect of paclitaxel on differentiation such as type II collagen expression and sulfated proteoglycan accumulation in rabbit articular chondrocytes. Paclitaxel caused differentiated chondrocyte phenotype as demonstrated by increment of type II collagen expression and proteoglycan synthesis Paclitaxel treatment stimulated activation of ERK-1/2 and p38 kinase. Inhibition of ERK-1/2 with PD98059 enhanced paclitaxel-induced differentiation, whereas inhibition of p38 kinase with SB203580 suppressed paclitaxel-induced differentiation. Our findings suggest that ERK-1/2 and p38 kinase oppositely regulate paclitaxel-induced differentiation in chondrocytes.

• Proceedings of the KSME Conference
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• 2005.11a
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• pp.155.1-155.1
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• 2005

• Archives of Plastic Surgery
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• v.40 no.6
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• pp.676-686
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• 2013

Background To overcome the potential drawbacks of a short half-life and dose-related adverse effects of using active transforming growth factor-beta 1 for cartilage engineering, a cell-mediated latent growth factor activation strategy was developed incorporating latent transforming growth factor-${\beta}$1 (LTGF) into an electrospun poly(L-lactide) scaffold. Methods The electrospun scaffold was surface modified with NH3 plasma and biofunctionalised with LTGF to produce both random and orientated biofunctionalised electrospun scaffolds. Scaffold surface chemical analysis and growth factor bioavailability assays were performed. In vitro biocompatibility and human nasal chondrocyte gene expression with these biofunctionalised electrospun scaffold templates were assessed. In vivo chondrogenic activity and chondrocyte gene expression were evaluated in athymic rats. Results Chemical analysis demonstrated that LTGF anchored to the scaffolds was available for enzymatic, chemical and cell activation. The biofunctionalised scaffolds were non-toxic. Gene expression suggested chondrocyte re-differentiation after 14 days in culture. By 6 weeks, the implanted biofunctionalised scaffolds had induced highly passaged chondrocytes to re-express Col2A1 and produce type II collagen. Conclusions We have demonstrated a proof of concept for cell-mediated activation of anchored growth factors using a novel biofunctionalised scaffold in cartilage engineering. This presents a platform for development of protein delivery systems and for tissue engineering.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.5
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• pp.1262-1269
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• 2008

Low intensity pulsed ultrasound(LIPUS) is known to accelerate bone regeneration, but the precise cellular signaling mechanism is still unclear. The purpose if this study was to determine the effect of LIPUS on the signaling mechanism of rat chondrocyte. In the explant culture condition, there was inhibition effect of 1 $W/cm^2$ intensity LIPUS on chondrocytes proliferation but chondrocytes proliferation was increased at 0.25 $W/cm^2$ intensity. In addition, western blot analysis of MAPKs showed that LIPUS increased ERK1/2 activity from the 10 min treatment of LIPUS. Hydrogen peroxide($H_2O_2$), resulted in a time- and dose-dependent cell proliferation, which was largely attributed to apoptosis. $H_2O_2$ treatment caused marked sustained nucleus condensation in Hoechst stain. LIPUS and $H_2O_2$ activates phosphorylation of p-ERK1/2 and PD 98059($10^{-5}M$) blocked the effect of LIPUS and $H_2O_2$. Moreover, the synergistic phosphorylation of p44/42 MAPK by $H_2O_2$, LIPUS was selectively inhibited by PD 98059, ERK1/2 inhibitor. In order to determine whether the increase in cell proliferation caused by $H_2O_2$ and LIPUS could be explained by changes in the level of the prostaglandin $E_2$. Our study demonstrated that LIPUS stimulate the cell proliferation via activated phosphorylation of ERK1/2 in condrocyte. LIPUS has anabolic effects on rat cartilage in explant cultures, indicating a potential important method for the treatment of osteoarthritic cartilarge.

• Proceedings of the KSLP Conference
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• 2003.11a
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• pp.183-183
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• 2003

Background and Objectives : Cartilage reconstruction is one of medical issue in otolaryngology. Tissue engineering is presently being utilized in part of cartilage repair. Sources of cells for tissue engineering are chondrocyte from mature cartilage and bone marrow mesenchymal stem cells that are able to differentiate into chondrocyte. Recent studies have shown that adipose tissue have mesenchymal stem cells which can differentiate into adipogenic, chondrogenic myogenic osteogenic cells and neural cell in vitro. In this study, we have examined chondrogenic potential of the canine adipose tissue-derived mesenchymal stem cell(ATSC). Materials and Methods : We harvested canine adipose tissue from inguinal area. ATSCs were enzymatically released from canine adipose tissue. Under appropriate culture conditions, ATSCs were induced to differentiate into the chondrocyte lineages using micromass culture technique. We used immunostain to type II collagen and toluidine blue stain to confirm chondrogenic differentiation of ATSCs. Results : We could isolate ATSCs from canine adipose tissue. ATSCs expressed CD29 and CD44 which are specific surface markers of mesenchymal stem cell. ATSCs differentiated into micromass that has positive response to immunostain of type II collagen and toluidine blue stain. Conclusion : In vitro, ATSCs differentiated into cells that have characteristic cartilage matrix molecules in the presence of lineage-specific induction factors. Adipose tissue may represent an alternative source to bone marrow-derived MSCs.