• The Korean Journal of Zoology
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• v.34 no.4
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• pp.460-468
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• 1991

계배 limb bud간충직 연골원성 세포로부터 연골원세포로의 분화과정에서 Ca2+의 역할을 추구해 보기 위하여 Hamburger-Hamilton stage 23/24의 계배 limb bud간충직세포들을 미세배양법으로 배양하면서, Ca2+, calcium ionophore인 A23187, calcium channel blocker인 D600을 각각 농도 및 처리기간을 변화시켜 처리하면서 연골화의 정도를 검정하여, 연골세포 분화에 미치는 Ca2+의 작용양상을 분석하였다. 그 결과 Ca2+은 3 mM의 농도로 배양초기에 처리하였을 때 가장 효과적으로 연골화를 촉진하였으며, A23187(0.05 $\mu$ M) 처리는 세포내로 Ca2+유입을 증가시켜 연골화를 촉진시킨 반면, D6OO(30 $\mu$ M이하) 처리는 세포내로 Ca2+ 유입을 차단시킴으로서 연골화를 억제하는 것으로 나타났다. 이러한 결과에 따른 세포내로의 칼슘 유입 변화는 45Ca로 확인하였다. 그러므로 Ca2+에 의한 간충직세포의 연골세포로의 분화촉진 작용은 Ca2+이 연골화의 응집시기에 세포간의 접촉을 유도할 뿐만 아니라 배양 초기에 Ca2+이 세포 내로 들어감으로써 수반되는 일련의 기작에 의한 것임을 알았다.

• Archives of Plastic Surgery
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• v.34 no.4
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• pp.413-419
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• 2007

Purpose: In this study, porous type I collagen scaffolds were cross-linked using dehydrothermal(DHT) treatment and/or 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide(EDC), in the presence and absence of chondroitin-6-sulfate(CS) and cultured autologous chondrocytes(Chondro) for cartilage regeneration. Methods: Cartilage defects were created in the proximal part of the ear of New Zealand rabbits. Four prepared types of scaffolds(n=4) were inserted. The groups included Chondro-Collagen-DHT(Group 1), Chondro- Collagen-DHT-EDC(Group 2), Chondro-CS-Collagen- DHT(Group 3), and Chondro-CS-Collagen-DHT-EDC (Group 4). Histomorphometric analysis and cartilage-specific gene expression of the reconstructed tissues were evaluated 4, 8, and 12 weeks after implantation. Results: EDC cross-linked groups 2 and 4 regenerated more cartilage than other groups. However, calcification was observed in the 4th week after implantation. CS did not increase chondrogenesis in all groups. Cartilage-specific type II collagen mRNA expression increased in the course of time in all groups.Conclusion: EDC cross-linking methods maintain the scaffold and promote extracellular matrix production of chondrocytes.

• Journal of Korean Foot and Ankle Society
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• v.27 no.2
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• pp.71-74
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• 2023

A medial opening wedge supramalleolar osteotomy (SMO) introduced by Takakura et al. is a useful realignment procedure for patients with ankle joint arthritis and varus malalignment by shifting the weight-bearing axis laterally and redistributing the loads on the ankle joint. When pain persists after arthroscopic microfracture in patients with medial osteochondral lesion of the talus (OLT), redo arthroscopy, osteochondral autograft transplantation, autologous chondrocyte implantation, or matrix-induced chondrogenesis might be indicated. On the other hand, there is insufficient scientific evidence for realignment surgery through SMO, while the effect of realignment surgery has been studied consecutively for osteochondral lesions of the knee. Therefore, this paper reports a patient with medial OLT who underwent redo arthroscopy combined with SMO for persistent pain after primary arthroscopic microfracture.

• Molecules and Cells
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• v.24 no.3
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• pp.343-350
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• 2007

Mesenchymal stem/progenitor cells (MPCs) were isolated from porcine umbilical cord blood (UCB) and their morphology, proliferation, cell cycle status, cell-surface antigen profile and expression of hematopoietic cytokines were characterized. Their capacity to differentiate in vitro into osteocytes, adipocytes and chondrocytes was also evaluated. Primary cultures of adherent porcine MPCs (pMPCs) exhibited a typical fibroblast-like morphology with significant renewal capacity and proliferative ability. Subsequent robust cell growth was indicated by the high percentage of quiescent (G0/G1) cells. The cells expressed the mesenchymal surface markers, CD29, CD49b and CD105, but not the hematopoietic markers, CD45 and CD133 and synthesized hematopoietic cytokines. Over 21 days of induction, the cells differentiated into osteocytes adipocytes and chondrocytes. The expression of lineage specific genes was gradually upregulated during osteogenesis, adipogenesis and chondrogenesis. We conclude that porcine umbilical cord blood contains a population of MPCs capable of self-renewal and of differentiating in vitro into three classical mesenchymal lineages.

• Biomolecules & Therapeutics
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• v.24 no.4
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• pp.395-401
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• 2016

Endochondral bone formation is the process by which mesenchymal cells condense into chondrocytes, which are ultimately responsible for new bone formation. The processes of chondrogenic differentiation and hypertrophy are critical for bone formation and are therefore highly regulated. The present study was designed to investigate the effect of aloe-emodin on chondrogenic differentiation in clonal mouse chondrogenic ATDC5 cells. Aloe-emodin treatment stimulated the accumulation of cartilage nodules in a dose-dependent manner. ATDC5 cells were treated with aloe-emodin and stained with alcian blue. Compared with the control cells, the ATDC5 cells showed more intense alcian blue staining. This finding suggested that aloe-emodin induced the synthesis of matrix proteoglycans and increased the activity of alkaline phosphatase. Aloe-emodin also enhanced the expressions of chondrogenic marker genes such as collagen II, collagen X, BSP and RunX2 in a time-dependent manner. Furthermore, examination of the MAPK signaling pathway showed that aloe-emodin increased the activation of extracellular signal-regulated kinase (ERK), but had no effect on p38 and c-jun N-terminal kinase (JNK). Aloe-emodin also enhanced the protein expression of BMP-2 in a time-dependent manner. Thus, these results showed that aloe-emodin exhibited chodromodulating effects via the BMP-2 or ERK signaling pathway. Aloe-emodin may have potential future applications for the treatment of growth disorders.

• The Journal of Korean Medicine
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• v.33 no.2
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• pp.1-10
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• 2012

Objectives: This study was aimed to investigate the effect of Bojungikgitang-gagambang (BJIG) on longitudinal bone growth in rats. Methods: The BJIG treated group (300 mg/kg) and the control group (vehicle) were administered orally twice daily for 4 days. To investigate the effects of BJIG we measured body weight gain. The bone growth effect was analyzed by measuring between fluorescent lines marked with tetracycline, which plays the role of fluorescent dye on the surface of the tibia. Tetracycline was intraperitoneally injected. The height of growth plates in the epiphyseal plate was measured. The expression of bone morphogenetic protein-2 (BMP-2) and insuline-like growth factor-1 (IGF-1) was investigated by immunohistochemistry. Results: BJIG caused a significant acceleration of longitudinal bone growth of $349.7{\pm}15.9{\mu}m/day$ compared to control ($319.8{\pm}21.4{\mu}m/day$). The height of overall growth plate was not significantly more compared to the control, but the size of cells in the proliferative zone and hypertrophic zone were. In the immunohistochemistry, BMP-2 and IGF-1 were expressed markedly in the proliferative or hypertrophic zone, respectively. Conclusions: BJIG stimulated the chondrocyte hypertrophy and chondrogenesis in the growth plate and directly increased the longitudinal tibia length of rats.

• Archives of Plastic Surgery
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• v.33 no.6
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• pp.723-731
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• 2006

Purpose: Collagen is the principal structural biomolecule in cartilage extracellular matrix, which makes it a logical target for cartilage engineering. In this study, porous type I collagen scaffolds were cross-linked using dehydrothermal(DHT) treatment and/or 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide(EDC), in the presence and absence of chondroitin-6-sulfate(CS) for cartilage regeneration. Methods: Cartilage defects were created in the proximal part of the ear of New Zealand rabbits. Four types of scaffolds(n=4) were inserted. The types included DHT cross-linked(Group 1), DHT and EDC cross- linked(Group 2), CS added DHT cross-linked(Group 3), and CS added DHT and EDC cross-linked(Group 4). Histomorphometric analysis and cartilage-specific gene expression of the reconstructed tissues were evaluated respectively 4, 8, and 12 weeks after implantation. Results: The largest quantity of regenerated cartilage was found in DHT cross-linked groups 1 and 3 in the 8th week and then decreased in the 12th week, while calcification increased. Calcification was observed from the 8th week and the area increased in the 12th week. Group 4 was treated with EDC cross-linking and CS, and the matrix did not degrade in the 12th week. Cartilage-specific type II collagen mRNA expression increased with time in all groups. Conclusion: CS did not increase chondrogenesis in all groups. EDC cross-linking may prevent chondrocyte infiltration from the perichondrium into the collagen scaffold.

• Proceedings of the Polymer Society of Korea Conference
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• 2006.10a
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• pp.268-268
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• 2006

Due to their multipotency, stem cells can differentiate into a variety of specialized cell types, such as chondrocytes, osteoblasts, myoblasts, and nerve cells. As an alternative to mature tissue cells, stem cells are of importance in tissue engineering and regenerative medicine. Since interactions between scaffold and cells play an important role in the tissue development in vitro, synthetic oligopeptides have been immobilized onto polymeric scaffolds to improve specific cell attachment and even to stimulate cell differentiation. In this study, chondrogenic differentiation of stem cells was evaluated using surface-modified PLLA scaffolds, i.e., either hydrophilic acrylic acid (AA)-grafted PLLA or RGD-immobilized one. Porous PLLA scaffolds were prepared using a gas foaming method, followed by plasma treatment and subsequent grafting of AA to introduce a hydrophilicity (PLLA-PAA). This was further processed to fix RGD peptide to make an RGD-immobilized scaffold (PLLA-PAA-RGD). Stem cells were seeded at $1{\times}10^{6}$ cells per scaffold and the cell-PLLA constructs were cultured for up to 4 weeks in the chondrogenic medium. Using these surface-modified scaffolds, adhesion, proliferation, and chondrogenic differentiation of stem cells were evaluated. The surface of PLLA scaffolds turned hydrophilic (water contact angle, 45 degrees) with both plasma treatment and AA grafting. The hydrophilicity of RGD-immobilized surface was not significantly altered. Cell proliferation rate on the either PLLA-PAA or PLLA-PAA-RGD surface was obviously improved, especially with the RGD-immobilized one as compared to the control PLLA one. Chondrogenic differentiation was clearly identified with Safranin O staining of GAG in the AA- or RGD-grafted PLLA substrates. This study demonstrated that modified polymer surfaces may provide better environment for chondrogenesis of stem cells.

• Molecules and Cells
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• v.24 no.1
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• pp.139-147
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• 2007

Although transforming growth factors (TGFs) are implicated in the process of endochondral ossification, which is initiated by the differentiation of mesenchymal cells into chondrocytes, it is not clear how $TGF-{\beta}3$ regulates the chondrogenic differentiation of limb bud mesenchymal cells. Here, differential display polymerase chain reaction (DD-PCR) screening and RT-PCR analysis revealed that transcripts of A Disintegrin And Metalloprotease 10 (ADAM 10) decreased during the chondro-inhibitory action of $TGF-{\beta}3$ on cultured chick leg bud mesenchymal cells. Electroporation of ADAM 10 morpholino antisense oligonucleotides inhibited the ectodomain shedding of delta-1, and cell proliferation and subsequent precartilage condensation, in a manner similar to that caused by $TGF-{\beta}3$. The suppression of mesenchymal cell proliferation induced by $TGF-{\beta}3$ and ADAM 10 morpholino antisense oligonucleotides was reversed by activation of ADAM 10 with phorbol 12-myristate 13-acetate (PMA) or knockdown of Notch-1 with siRNA. Collectively, these data indicate that, in cultured chick leg bud mesenchyme cells, $TGF-{\beta}3$ downregulates ADAM 10 and inhibits cell proliferation and subsequent precartilage condensation by inhibiting the ectodomain shedding of delta-1, and that this results in the activation of Notch signaling.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.4
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• pp.588-595
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• 2013

Mesenchymal stem cells (MSCs) are often known to have a therapeutic potential in the cell-mediated repair for fatal or incurable diseases. In this study, canine umbilical cord MSCs (cUC-MSCs) were isolated from umbilical cord matrix (n = 3) and subjected to proliferative culture for 5 consecutive passages. The cells at each passage were characterized for multipotent MSC properties such as proliferation kinetics, expression patterns of MSC surface markers and self-renewal associated markers, and chondrogenic differentiation. In results, the proliferation of the cells as determined by the cumulative population doubling level was observed at its peak on passage 3 and stopped after passage 5, whereas cell doubling time dramatically increased after passage 4. Expression of MSC surface markers (CD44, CD54, CD61, CD80, CD90 and Flk-1), molecule (HMGA2) and pluripotent markers (sox2, nanog) associated with self-renewal was negatively correlated with the number of passages. However, MSC surface marker (CD105) and pluripotent marker (Oct3/4) decreased with increasing the number of subpassage. cUC-MSCs at passage 1 to 5 underwent chondrogenesis under specific culture conditions, but percentage of chondrogenic differentiation decreased with increasing the number of subpassage. Collectively, the present study suggested that sequential subpassage could affect multipotent properties of cUC-MSCs and needs to be addressed before clinical applications.