• 대한방사선기술학회지:방사선기술과학
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• 제25권2호
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• pp.77-82
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• 2002

이미 분화된 연골세포의 성숙과정에 미치는 X-선의 작용에 대해서는 잘 알려져 있다. 그러나 연골세포나 섬유아세포, 근육세포로 분화될 수 있는 미분화 간충직 세포의 분화과정에 미치는 X-선의 영향에 대해서는 잘 알려져 있지 않아, 본 연구에서는 초기 분화연구에 좋은 대상이 되는 계배 미분화 간충직세포를 이용하여 선량(1-10Gy)에 따라 연골세포 분화 과정에 X-선이 어떤 영향을 미치는가를 조사하였다. 연구결과 선량 의존적으로 연골세포분화가 억제됨을 alcian blue로 sulfated proteoglycan을 염색한 결과를 통해 알 수 있었다. 이는 X-선이 간충직세포와 같은 성숙 이전의 연골성 세포들에게는 모두 영향을 미침을 보여주는 것이다. 또한 이미 알려진 바와 같이 X-선은 분화된 연골세포의 성숙과정에 영향을 주기도 하지만 상기 연구를 통해서 간충직세포로부터 연골세포로 분화하는 과정을 억제시키기도 함을 보여줌으로써 간충직세포로부터 성숙된 연골세포로 되는 전과정에 X-선이 영향을 미친다는 사실과, 분화가 이루어지지 않은 세포일수록 X-선 조사의 영향을 크게 받음을 알 수 있다.

• 한국식품영양과학회지
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• 제33권5호
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• pp.797-802
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• 2004

L-Ascorbic acid (AsA), commonly known as vitamin C, which is one of the antioxidant vitamins, plays a role in cellular oxidant quenching. Some of the biochemical reactions in which it takes part have been traced through organ culture technique. But in cell cultured system, views on stimulatory and inhibitory action of AsA on cell growth are conflicting. Therefore, this study aimed to clarify the inhibitory action of high concentration AsA on the cell growth in Primary chondrocyte isolated from rat ribs. Cells were exposed to ascorbate at various concentrations. Supplement of AsA induced stimulation of cell growth in primary cultured cells of chondrocytes. Most remarkable stimulation of cell growth by AsA was found in primary cultured chondrocytes. However, it showed that they were dead in the medium which contained AsA at the concentration higher than 1.0 mM. This lethal effect of AsA causing the cell death was inhibited by the addition of catalase in the medium. This supposed that hydroxyl radical (ㆍOH) induced from $H_2O$$_2$ was actively cytotoxic agent. Based on the results, when AsA was added in medium at normal concentrations, the cell growth was stimulated by inducing the formation of extracellular matrix. On the contrary, if added in medium at excess concentrations, the cell growth was inhibited because $H_2O$$_2$ were generated from AsA in medium. Therefore, addition of AsA at the normal concentrations stimulates cell growth, but excess concentrations of AsA induces cell death.

• Journal of Microbiology and Biotechnology
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• 제31권10호
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• pp.1401-1408
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• 2021

This study examined whether the oral administration of low-molecular-weight collagen peptide (LMCP) containing 3% Gly-Pro-Hyp with >15% tripeptide (Gly-X-Y) content could ameliorate osteoarthritis (OA) progression using a rabbit anterior cruciate ligament transection (ACLT) model of induced OA and chondrocytes isolated from a patient with OA. Oral LMCP administration (100 or 200 mg/kg/day) for 12 weeks ameliorated cartilage damage and reduced the loss of proteoglycan compared to the findings in the ACLT control group, resulting in dose-dependent (p < 0.05) improvements of the OARSI score in hematoxylin & eosin (H&E) and Safranin O staining. In micro-computed tomography analysis, LMCP also significantly (p < 0.05) suppressed the deterioration of the microstructure in tibial subchondral bone during OA progression. The elevation of IL-1β and IL-6 concentrations in synovial fluid following OA induction was dose-dependently (p < 0.05) reduced by LMCP treatment. Furthermore, immunohistochemistry illustrated that LMCP significantly (p < 0.05) upregulated type II collagen and downregulated matrix metalloproteinase-13 in cartilage tissue. Consistent with the in vivo results, LMCP significantly (p < 0.05) increased the mRNA expression of COL2A1 and ACAN in chondrocytes isolated from a patient with OA regardless of the conditions for IL-1β induction. These findings suggest that LMCP has potential as a therapeutic treatment for OA that stimulates cartilage regeneration.

• 대한수의학회지
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• 제58권4호
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• pp.211-217
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• 2018

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy. Physiological cell environment not only connects cells to each other, but also connects cells to the extracellular matrix that provide mechanical support, thus exposing the entire cell surface and activating signaling pathways. Hydrogel is a polymeric material that swells in water and maintains a distinct 3-dimensional (3D) network structure by cross linking. In this study, we investigated the optimized cellular function for canine adipose tissue-derived MSCs (cAD-MSCs) using hydrogel. We observed that the expression levels of Ki67 and proliferating cell nuclear antigen, which are involved in cell proliferation and stemness, were increased in transwell-hydrogel (3D-TN) compared to the transwell-normal (TN). Also, transforming growth factor-${\beta}1$ and SOX9, which are typical bone morphogenesis-inducing factors, were increased in 3D-TN compared to the TN. Collagen type II alpha 1, which is a chondrocyte-specific marker, was increased in 3D-TN compared to the TN. Osteocalcin, which is a osteocyte-specific marker, was increased in 3D-TN compared to the TN. Collectively, preconditioning cAD-MSCs via 3D culture systems can enhance inherent secretory properties that may improve the potency and efficacy of MSCs-based therapies for bone regeneration process.

• International Journal of Oral Biology
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• 제44권3호
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• pp.115-123
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• 2019

Among the environmental chemicals that may be able to disrupt the endocrine systems of animals and humans are polychlorinated biphenyls (PCBs), a chemical class of considerable concern. PCB consists of two six-carbon rings linked by a single carbon bond, and theoretically, 209 congeners can form, depending on the number of chlorines and their location on the biphenyl rings. Furthermore, 3,3',4,4',5-pentachlorobiphenyl (PCB126) exposure also increases nitric oxide production and nuclear factor kappa-light-chain-enhancer of activated B cells binding activity in chondrocytes, thus contributing as an initiator of chondrocyte apoptosis and resulting in thymic atrophy and immunosuppression. This study identified whether cardiac and immune abnormalities from PCB126 were caused by the Kv1.3 and Kv1.5 channels. PCB126 did not affect either the steady-state current or peak current of the Kv1.3 and Kv1.5 channels. However, PCB126 right-shifted the steady-state activation curves of human Kv1.3 channels. These results suggest that PCBs can affect the heart in a way that does not block voltage-dependent potassium channels including Kv1.3 and Kv1.5 directly.

• Journal of Veterinary Science
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• 제19권6호
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• pp.735-743
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• 2018

We investigated the effect of transforming growth factor beta 1 ($TGF-{\beta}1$) on equine hyaluronan synthase 2 (HAS2) gene expression and hyaluronan (HA) synthesis in culture models of articular chondrocytes. Equine chondrocytes were treated with $TGF-{\beta}1$ at different concentrations and times in monolayer cultures. In three-dimensional cultures, chondrocyte-seeded gelatin scaffolds were cultured in chondrogenic media containing 10 ng/mL of $TGF-{\beta}1$. The amounts of HA in conditioned media and in scaffolds were determined by enzyme-linked immunosorbent assays. HAS2 mRNA expression was analyzed by semi-quantitative reverse transcription polymerase chain reaction. The uronic acid content and DNA content of the scaffolds were measured by using colorimetric and Hoechst 33258 assays, respectively. Cell proliferation was evaluated by using the alamarBlue assay. Scanning electron microscopy (SEM), histology, and immunohistochemistry were used for microscopic analysis of the samples. The upregulation of HAS2 mRNA levels by $TGF-{\beta}1$ stimulation was dose and time dependent. $TGF-{\beta}1$ was shown to enhance HA and uronic acid content in the scaffolds. Cell proliferation and DNA content were significantly lower in $TGF-{\beta}1$ treatments. SEM and histological results revealed the formation of a cartilaginous-like extracellular matrix in the $TGF-{\beta}1$-treated scaffolds. Together, our results suggest that $TGF-{\beta}1$ has a stimulatory effect on equine chondrocytes, enhancing HA synthesis and promoting cartilage matrix generation.

• 대한정형외과학회지
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• 제54권6호
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• pp.478-489
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• 2019

골관절염은 관절연골 침식의 진행을 특징적으로 보이는 질환으로 관절운동 중에 통증을 증가시키고 기계적 스트레스를 견디는 능력을 감소시켜 결과적으로 관절의 가동성과 기능을 저하시킨다. 외상 또는 퇴행성으로 인한 관절연골의 손상이 일반적 관절염의 주요 원인으로 생각되며 이러한 관절연골 손상의 재생에 관한 수많은 연구와 시도들이 현재까지 진행되어 오고 있다. 현재까지 연골 손상의 경우 미세골절술과 자가연골세포 이식술이 일반적인 수술적 치료방법으로 제시되어 왔으나 비교적 양호한 임상 결과에도 불구하고 정상 유리연골의 생성이 미흡하여 시간이 경과하면서 결과가 악화되는 등 단점이 있다. 이를 보완하기 위하여 줄기세포 기반 치료법이 개발되었다. 이 종설에서는 현재 사용되는 다양한 연골 재생 방법들의 장단점 및 결과에 대해 요약하고 특히 중간엽 줄기세포(mesenchymal stem cells) 기반 연골 재생 치료법을 논하고 나아가 이상적인 미래 연골 재생 치료법에 대해서도 고민해보고자 한다.

• IMMUNE NETWORK
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• 제22권2호
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• pp.14.1-14.17
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• 2022

Osteoarthritis (OA) is a common degenerative joint disease characterized by breakdown of joint cartilage. Mitochondrial dysfunction of the chondrocyte is a risk factor for OA progression. We examined the therapeutic potential of mitochondrial transplantation for OA. Mitochondria were injected into the knee joint of monosodium iodoacetate-induced OA rats. Chondrocytes from OA rats or patients with OA were cultured to examine mitochondrial function in cellular pathophysiology. Pain, cartilage destruction, and bone loss were improved in mitochondrial transplanted-OA rats. The transcript levels of IL-1β, TNF-α, matrix metallopeptidase 13, and MCP-1 in cartilage were markedly decreased by mitochondrial transplantation. Mitochondrial function, as indicated by membrane potential and oxygen consumption rate, in chondrocytes from OA rats was improved by mitochondrial transplantation. Likewise, the mitochondrial function of chondrocytes from OA patients was improved by coculture with mitochondria. Furthermore, inflammatory cell death was significantly decreased by coculture with mitochondria. Mitochondrial transplantation ameliorated OA progression, which is caused by mitochondrial dysfunction. These results suggest the therapeutic potential of mitochondrial transplantation for OA.

• Anatomy and Cell Biology
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• 제57권3호
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• pp.400-407
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• 2024

Intrauterine alcohol exposure delays bone maturation and intensifies osteoporosis and fracture risk. As most studies emphasize the neurological aspects of intrauterine alcohol exposure, there is a lack of research on the implications pertaining to osseous tissue. Previous studies investigated these effects in fetuses, with limited studies on postnatal life. Postnatal studies are crucial since peak bone growth occurs during adolescence. This study aimed at assessing the effects of prenatal alcohol exposure on the humerus proximal and distal growth plate chondrocytes in 3-week-old rats. Sprague Dawley rats (n=9) were assigned to either the ethanol group (n=3), saline (n=3), and untreated (n=3) group and time-mated. Once pregnant, as confirmed by the presence of a copulation plug, the former 2 groups were treated with 0.015 ml/g of 25.2% ethanol and 0.9% saline. The untreated group received no treatment. The left humeri belonging to 6 pups per group were used. Serial sections were cut with a microtome at 5 ㎛ thickness. These sections were stained with haematoxylin and eosin for assessment of normal morphology or immunolabeled with anti-Ki-67 and transforming growth factor β-1 (TGFβ-1) antibody. Prenatal alcohol exposure adversely effected the growth plate sizes and the number of cells in the proliferative zone. Fewer TGFβ-1 immunopositive and proliferative chondrocytes were found using the anti-Ki-67 antibody. This may explain the growth retardation in offspring exposed to gestational alcohol, showing that gestational alcohol exposure inhibits cell proliferation, aiding the diminished stature.

• 폴리머
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• 제36권6호
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• pp.768-775
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• 2012

해조류로부터 얻어지는 알지네이트는 캡슐화된 세포의 생존율에 긍정적인 영향을 끼치며 살아있는 세포를 신속하게 포접하여 캡슐화할 수 있어 세포이식을 위한 생체재료 분야에 널리 쓰인다. 탈미네랄화된 골분(DBP)은 천연 뼈조직으로부터 유래되어 조직과의 반응정도가 낮고 항원성 또한 낮아 임상에 적용되어 사용되어 왔다. 알지네이트에 비율별 DBP을 포함시켜 연골세포를 파종한 뒤 미세캡슐을 제조한 후 MTT 분석을 통하여 세포의 부착 및 증식률을 관찰하였고 glycosaminoglycan(sGAG)와 콜라겐 함량 측정과 연골세포의 특정유전자 표현형을 확인하기 위하여 PCR을 실시하였다. 또한 연골세포가 파종된 알지네이트 미세캡슐을 누드마우스의 피하에 이식한 뒤 적출하여 면역화학적 염색을 실시하였다. 실험 결과 1%의 DBP를 함유한 알지네이트 미세캡슐에서 가장 높은 세포 증식률을 보였고 표현형 유지에도 긍정적인 영향을 미치는 것을 확인하였다. 이번 연구 결과를 토대로 알지네이트와 DBP를 이용한 미세캡슐을 제조함으로써 DBP내의 성장인자와 알지네이트의 상호작용으로 인하여 연골세포의 성장에 긍정적인 영향을 미쳐 생체공학적 지지체로 적합할 것으로 예상된다.