• 생명과학회지
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• 제19권3호
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• pp.327-333
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• 2009

건선(psoriasis)은 각화(keratinization)의 장애에 의한 질병으로 잘 알려져 있다. 이 질병에있어서, 몇몇 보고에 따르면 피부 미세혈관이 증가되고, 혈관 신생 유도 인자들인 내피 성장 인자(VEGF) 및 섬유아세포 성장 인자(bFGF)가 과발현된다고 보고 되어져 있다. 신생혈관(angiogenesis)은 건선의 진행성에 있어 중요한 역할을 할지도 모른다. Acitretin은 keratinocyte 성장과 분화에 그것의 잠재적인 활동 때문에 anti-psoriatic 약으로 널리 이용된다. 그러나 신생혈관에 대한 효과는 여전히 불명확하다. 본 면역혈청학적 연구 목표는 건선염 피부에 있는 VEGF의 발현에 acitretin의 효과를 조사하기 위한 것이었다. 우리는 10명의 건선염 피부환자와 대조군으로 3명의 정상인 피부에 acitretin의 치료 전과 후의 VEGF의 발현량을 비교하였다. 치료 전 대조군인 정상인 피부에서보다 건선염 피부환자에서 VEGF의 발현은 명백히 높은 수준이었다. 건선염 피부환자에서 VEGF의 발현량은 acitretin 치료 후 치료 전과 비교시 감소되었다. Acitretin은 건선 피부에서 VEGF와 같은 혈관 신생 유도 인자의 발현을 감소 시켜 신생혈관형성에 있어 억제의 효과가 있음을 나타낸다. 우리는 신생혈관 관점에서 acitretin이 건선에 대한 치료 수단이 될 수 있다는 것을 제안한다.

• 생명과학회지
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• 제17권5호
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• pp.687-693
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• 2007

사람의 섬유아세포인 GM10을 사용하여 glucose의 배양조건에 다른 물질대사 및 IGFBP-3 발현을 살펴본 결과, glucose 농도에 따른 glucose 소비와 triglyceride 축적 수준은 고농도 glucose 배양 조건에서 증가한 반면, 총 단백질 함량은 고농도 glucose배양 조건에서 시간의 경과에 따라 감소하였다. 또한 고농도 glucose 배양 조건에서 5일 동안 배양한 세포 배양액내의 유리아미노산 함량은 저농도보다 고농도 glucose 배양 조건에서 높게 나타났다. IGFBP-3 단백질 수준과 mRNA수준은 저농도 glucose배양 조건에서 증가하였으나, IGFBP-3 단백질 분해효소에 따른 영향은 없었다. 이상의 결과에서 glucose 배양조건에 따른 물질대사는 부분적으로 세포를 이용한 당뇨병 모델 실험에서 in vivo와 같은 결과를 얻지 못하였지만, IGFs와 같은 세포 성장인자에 대한 연구를 통해 세포수준의 당뇨병 모델화가 가능하다고 여겨진다.

• 대한후두음성언어의학회지
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• 제32권1호
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• pp.15-23
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• 2021

Background and Objectives During speech, the vocal folds oscillate at frequencies ranging from 100-200 Hz with amplitudes of a few millimeters. Mechanical stimulation is an essential factor which affects metabolism of human vocal folds. The effect of mechanical vibration on the cellular response in the human vocal fold fibroblasts cells (hVFFs) was evaluated. Materials and Method We created a culture systemic device capable of generating vibratory stimulations at human phonation frequencies. To establish optimal cell culture condition, cellular proliferation and viability assay was examined. Quantitative real time polymerase chain reaction was used to assess extracellular matrix (ECM) related and growth factors expression on response to changes in vibratory frequency and amplitude. Western blot was used to investigate ECM and inflammation-related transcription factor activation and its related cellular signaling transduction pathway. Results The cell viability was stable with vibratory stimulation within 24 h. A statistically significant increase of ECM genes (collagen type I alpha 1 and collagen type I alpha 2) and growth factor [transforming growth factor β1 (TGF-β1) and fibroblast growth factor 1 (FGF-1)] observe under the experimental conditions. Vibratory stimulation induced transcriptional activation of NF-κB by phosphorylation of p65 subunit through cellular Mitogen-activated protein kinases activation by extracellular signal regulated kinase and p38 mitogen-activated protein kinases (MAPKs) phosphorylation on hVFFs. Conclusion This study confirmed enhancing synthesis of collagen, TGF-β1 and FGF was testified by vibratory stimulation on hVFFs. This mechanism is thought to be due to the activation of NF-κB and MAPKs. Taken together, these results demonstrate that vibratory bioreactor may be a suitable alternative to hVFFs for studying vocal folds cellular response to vibratory vocalization.

• Tuberculosis and Respiratory Diseases
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• 제64권3호
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• pp.200-205
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• 2008

연구배경: 혈관신생은 종양의 성장과 유지 및 전이에 필수적이며 따라서 종양조직은 혈관신생을 위해 많은 종류의 혈관형성 촉진인자들을 생성하고 있다. VEGF와 bFGF는 혈관신생과 관련되는 물질로 본 연구에서는 폐암환자에서 조직 내 VEGF와 bFGF의 발현에 대해 알아보고자 하였다. 방법: 조직학적으로 샘암종 또는 편평상피세포암종으로 진단받고 완치의 목적으로 수술을 시행 받은 35명의 폐암환자 조직에서 VEGF 및 bFGF의 농도를 ELISA 방법으로 측정하였으며 이에 대한 임상적 양상을 후향적으로 분석하였다. 결과: VEGF 및 bFGF의 농도는 종양조직이 대조조직보다 유의하게 높았으며 T2+T3의 종양조직이 T1의 종양 조직보다 유의하게 VEGF의 농도가 높았다. 림프절 전이가 있었던 경우가 없었던 경우보다 종양조직에서 VEGF의 농도가 증가한 경향을 보였다(p=0.06). 하지만 VEGF 및 bFGF의 농도는 환자의 병기 및 생존율에 통계적으로 유의한 차이를 보이지 않았다. 결론: VEGF 및 bFGF 모두 종양조직에서 증가하였으나 VEGF만이 종양크기, 림프절 전이 등의 임상양상과 연관성을 보여주었다. 하지만 각각의 VEGF 및 bFGF의 종양조직 내 농도는 예후와 관련되지는 않았다.

• Fisheries and Aquatic Sciences
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• 제16권3호
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• pp.177-185
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• 2013

The development of species-specific fish cell lines has become a valuable tool for biological research. In recent years, marine medaka Oryzias dancena has been recognized as a good experimental model fish but there are no reports of establishment of cell lines from this fish. In this study, two cell lines from O. dancena blastula embryos were established from 41 total trials (4.9%). The two cell lines displayed typical in vitro morphology and have been cultured for >121 passages, which corresponds to 293 days. The doubling times of the cell lines were 29.84 and 28.59 h, respectively, and both possessed the potential to expand in a clonal manner, albeit with significant differences between the two cell lines. The absence of any of the four main medium supplements; i.e., fish serum, fetal bovine serum, basic fibroblast growth factor, and medaka embryo extract, significantly inhibited growth. The proportion of cells possessing normal chromosome number was 45% and 46.7% of the cell lines, respectively. Taken together, two cell lines that proliferate continuously were established from marine medaka and these cell lines may provide a basic tool for characterizing the unique features of this fish species.

• The Korean Journal of Physiology and Pharmacology
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• 제19권1호
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• pp.35-42
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• 2015

In cardiovascular disorders, understanding of endothelial cell (EC) function is essential to elucidate the disease mechanism. Although the mouse model has many advantages for in vivo and in vitro research, efficient procedures for the isolation and propagation of primary mouse EC have been problematic. We describe a high yield process for isolation and in vitro culture of primary EC from mouse arteries (aorta, braches of superior mesenteric artery, and cerebral arteries from the circle of Willis). Mouse arteries were carefully dissected without damage under a light microscope, and small pieces of the vessels were transferred on/in a Matrigel matrix enriched with endothelial growth supplement. Primary cells that proliferated in Matrigel were propagated in advanced DMEM with fetal calf serum or platelet-derived serum, EC growth supplement, and heparin. To improve the purity of the cell culture, we applied shearing stress and anti-fibroblast antibody. EC were characterized by a monolayer cobble stone appearance, positive staining with acetylated low density lipoprotein labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate, RT-PCR using primers for von-Willebrand factor, and determination of the protein level endothelial nitric oxide synthase. Our simple, efficient method would facilitate in vitro functional investigations of EC from mouse vessels.

• Plant Resources
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• 제8권3호
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• pp.209-216
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• 2005

Persicaria thunbergii has been utilized for the treatment of cancer as a folk medicine. We examined the effect of isorhamnetin, a flavonoid isolated from Persicaria thunbergii, on angiogenesis in vitro and in vivo. Basic fibroblast growth factor (bFGF) is a potent angiogenic factor found in various tumors. In this study, we found that the isorhamnetin decreased bFGF-induced human umbilical vein endothelial cells (HUVECs) proliferation and migration in a concentration-dependent manner (5, 10 and $20\;{\mu}M$) whereas, it did not inhibit bFGF-induced capillary-like formation of HUVECs. The chicken chorioallantoic membrane assay revealed that addition of isorhamnetin (10, 20 and $40\;{\mu}M$) displayed an antiangiogenic effect in vivo. These results suggest that the isorhamnetin inhibits the proliferation and migration of endothelial cells induced by bFGF, which may explain its anti-angiogenic properties.

• Asian-Australasian Journal of Animal Sciences
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• 제21권2호
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• pp.277-290
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• 2008

The process of wound repair involves an ordered sequence of events such as overlapping biochemical and cellular events that, in the best of circumstances, result in the restoration of both the structural and functional integrity of the damaged tissue. An important event during wound healing is the contraction of newly formed connective tissues by fibroblasts. The polypeptide growth factors, like transforming growth factor-${\beta}$(TGF-${\beta}$, insulin-like growth factor I (IGF- I) and epidermal growth factor (EGF), play very important mediator roles in the process of wound contraction. Deer antlers, as models of mammalian regeneration, are cranial appendages that develop after birth as extensions of a permanent protuberance (pedicle) on the frontal bone. Antlers contain various growth factors which stimulate dermal fibroblast growth. They are involved in digestion and respiration and are necessary for normal wound healing and skin health. In order to investigate and evaluate the effects of red deer antlers on skin wound site, the speed of full-thickness skin wound healing and the expression of IGF-I, TGF-${\beta}$ and EGF in skin wounds, three groups of skin full-thickness rat models with a high concentration of antler ointment, a low concentration of antler ointment and without antler ointment were compared. At post-injury days 0, 2, 4, 8, 16, 20, 32, 40 and 60, the skin wound area was measured, the expressions of IGF-I, TGF- ${\beta}$ and EGF mRNA were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and collagen formation by sirius red dye and the localization of IGF-I, TGF-${\beta}$ and EGF peptides were inspected by histological immunohistochemical techniques. Wound healing was significantly more rapid in antler treated skins. In addition, the wound treated with a high concentration antler ointment, a low concentration antler ointment, and the control closed completely at post-injury day 40, day 44 and day 60, respectively. Via RT-PCR, the expressions of IGF-I (day 8 and day 16), TGF-${\beta}$(day 8, day 16 and day 20) and EGF (day 4, day 8, day 16, and day 32) were obviously up-regulated in high concentration antler-treated skins compared to control skins. Similar results could be seen in the histological detection of collagen dye and immunohistochemical methods using the corresponding polyclone antibodies of IGF-I, TGF-${\beta}$ and EGF. These results illustrate that antlers stimulate and accelerate the repair of cutaneous wounds.

• The Journal of Advanced Prosthodontics
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• 제6권5호
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• pp.379-386
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• 2014

PURPOSE. These days, mesenchymal stem cells (MSCs) have received worldwide attention because of their potentiality in tissue engineering for implant dentistry. The purpose of this study was to evaluate various growth inducing factors in media for improvement of acquisition of bone marrow mesenchymal stem cells (BMMSCs) and colony forming unit-fibroblast (CFU-F). MATERIALS AND METHODS. The mouse BMMSCs were freshly obtained from female C3H mouse femur and tibia. The cells seeded at the density of $10^6$/dish in media supplemented with different density of fetal bovine serum (FBS), $1{\alpha}$, 25-dihydroxyvitamin (VD3) and recombinant human epidermal growth factor (rhEGF). After 14 days, CFU-F assay was conducted to analyze the cell attachment and proliferation, and moreover for VD3, the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was additionally conducted. RESULTS. The cell proliferation was increased with the increase of FBS concentration (P<.05). The cell proliferation was highest at the density of 20 ng/mL rhEGF compared with 0 ng/mL and 200 ng/mL rhEGF (P<.05). For VD3, although the colony number was increased with the increase of its concentration, the difference was not statistically significant (P>.05). CONCLUTION. FBS played the main role in cell attachment and growth, and the growth factor like rhEGF played the additional effect. However, VD3 did not have much efficacy compare with the other two factors. Improvement of the conditions could be adopted to acquire more functional MSCs to apply into bony defect around implants easily.

• 생명과학회지
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• 제28권3호
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• pp.275-283
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• 2018

인간 섬유아세포 성장인자는 조직 생성 및 상처 치료 효과로 인해 상업적으로 중요한 치료제 또는 화장품소재로서 가능성이 높다. 피부 조직에 침투성을 부여하여 치료효과를 높이기 위해서 단백질 전달 도메인인 PTD를 FGF에 융합을 시도하고 있으며 피부로의 투과능력과 그로인한 치료 효과에 대한 연구도 진행되고 있다. 그러나, PTD가 FGF 단백질의 N- 또는 C-말단에 결합 되었을 때 PTD의 위치가 대장균 발현시스템에서 재조합단백질 접힘 및 안정성, 그리고 결국 피부로의 도입능력에 상당한 영향을 미치는지에 대해서는 알려져 있지 않다. 여기에서 우리는 대조군으로 PTD가 융합되지 않은 인간 염기성 섬유아세포 성장인자(FGF2)와 PTD가 FGF2의 N-말단 또는 C-말단에 융합된 FGF2 복합체를 중합효소연쇄반응(OE-PCR)을 통해 클로닝 하였다. 그 다음 이들 재조합 FGF2의 단백질 발현 및 특성을 확인하고 마우스 등 피부를 이용하여 조직 내로의 도입 능력을 조사 하였다. 결과적으로, 불용성 PTD-FGF2 (N 말단 융합)와는 달리 대조군 FGF2와 FGF2-PTD 융합 단백질(C- 말단 융합)은 가용성 형태로 발현되어 재조합단백질 획득이 용이하였고, 마우스 피부 도입능력은 FGF2-PTD 융합단백질에서만 나타내 보였다. 우리의 결과는 C-말단에 융합된 FGF2-PTD 융합단백질이 발현, 정제, 피부 투과능력의 측면에서 다른 두 옵션들보다 노동, 비용, 시간면에서 보다 더 효율적일 수 있음을 시사한다.