• 대한소아치과학회지
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• 제44권3호
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• pp.350-357
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• 2017

본 연구는 상악 정중과잉치에서 얻은 치수유래 줄기세포(human dental pulp stem cells from supernumerary tooth, sDPSCs)를 대상으로 분화능의 변화 특성을 알아보고자 하였다. 전신 병력이 없는 6세 남자아이 상악 중절치와 측절치 사이에 매복 된 과잉치를 발치하여 sDPSCs를 얻었다. 세포들을 16계대까지 배양하였고, 1 - 8계대는 Young군으로 9 - 16계대는 Old군으로 나누었다. 각 계대 배양에 소요된 시간은 Young군에서는 $2.25{\pm}0.46$일, Old군에서는 $3.25{\pm}0.46$일을 보였으며, 이는 통계적으로 유의한 차이를 보였다(p < 0.05). 각 계대 별로 세포 형태를 관찰하고 분화를 유도한 후 Alizarin-red solution 염색을 통해 골모세포(odontoblast)로 분화되는 정도를 관찰하였다. 1, 8, 9계대에서 분화제를 처리하지 않은 세포의 형태에서는 방추형의 섬유모세포와 유사한 길쭉한 형태로 과립(nodule)이 적었지만, 16계대에서는 세포의 크기가 커지고 넓적한 형태로 변하고 과립도 많아졌다. 1계대는 분화 7일부터 과립이 관찰되며, 8계대에서는 14일 동안 분화제를 처리한 후 과립이 명확히 관찰되었다. 그러나 9계대 이후에서는 과립의 빈도가 상당히 감소되었다. ARS 염색에서는 1, 8계대는 진한 붉은색으로 염색되었으나, 9, 16계대는 염색이 옅게 되었다. 이를 통해 sDPSCs는 8계대 이전의 세포를 줄기세포의 원천으로 우선 고려하는 것이 좋다고 사료된다.

• 대한화장품학회지
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• 제30권1호
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• pp.79-83
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• 2004

파슬리추출물이 피부에 미치는 개선효과를 조사하기 위하여, 배양 인체 섬유아세포에서 total Collagen, type I procollagen을 각질형성세포주인 HaCaT 세포에서 prostaglandin E$_2$(PGE$_2$), interleukin l$\alpha$ (IL-l$\alpha$)와 tumor necrosis factor $\alpha$ (TNF$\alpha$)를 무모생쥐(Female albino hairless mice, Skh:hr-1)에서는 진피의 두께와 밀도를 측정하였다. 그 결과, 1 $\mu\textrm{g}$/mL 농도의 파슬리 추출물은 total collagen은 23%, type I procollagen은 18% 증가시켰고, 자외선 B에 의한 PGE$_2$의 생합성은 약 60% 정도 감소시켰다. 10uM RA, 100 $\mu\textrm{g}$/mL SLS와 자외선 B 30 mJ/$\textrm{cm}^2$로 조사했을 때, IL-1$\alpha$ 및 TNF $\alpha$의 생합성 역시 1 $\mu\textrm{g}$/mL 파슬리추출물 처리 시 감소되었다. 4일 동안 1% 파슬리추출물로 폐쇄첩포한 무모생쥐의 진피 두께는 대조군에 비해 약 1.5배 정도 두꺼워지고, 밀도도 훨씬 촘촘해졌다. 본 연구의 결과는 파슬리추출물이 피부에서 노화방지 효과 및 자극완화 효과가 있음을 시사하고 있다.

• 대한화장품학회지
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• 제37권1호
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• pp.75-81
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• 2011

본 연구에서는 유산균 발효에 의한 인삼열매 추출물의 여러 가지 생리활성을 평가하였다. 고성능액체크로마토그래피를 수행하여 인삼열매 추출물과 유산균 발효 인삼열매 추출물의 ginsenoside Re, Rc 및 Rb1의 함량분석 결과 유산균 발효에 의해 ginsenoside Re, Rc 및 Rb1의 함량이 모두 증가하였다. 인삼열매 추출물과 유산균 발효 인삼열매 추출물의 DPPH free radical 소거활성과 SOD 유사활성 측정결과 1.00 %의 농도에서 유산균 발효 인삼열매 추출물은 각각 86.34, 76.82 %의 DPPH free radical 소거활성과 SOD 유사활성을 나타내었고, 인삼열매 추출물은 각각 49.78, 40.80 %의 DPPH free radical 소거활성과 SOD 유사활성을 나타내어 유산균 발효에 의해 항산화 활성이 증가하는 것을 확인하였다. 또한 유산균 발효 인삼열매 추출물 0.50 %의 농도로 처리한 피부 섬유아세포에서 procollagen type I (COL1A1)은 828.13 % 증가하였으며, matrix metalloproteinase (MMP)-1은 87.88 %, tumor necrosis factor (TNF)-${\alpha}$는 99.92 %감소하는 효과를 보여주었다. 이처럼 항산화, 주름, 항염에 효능을 보이는 유산균 발효에 의한 인삼열매 추출물은 기능성 화장품 소재로 개발될 수 있는 가능성이 있다.

• 동의생리병리학회지
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• 제31권6호
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• pp.374-379
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• 2017

Hair loss affects interpersonal relationships and causes psychological stress. In this study, we investigated the antioxidant activity of Cynanchi Wilfordii Radix (CWR) and its effects on dermal papilla (DP) cells. Antioxidant efficacy was examined by ABTS assay. To confirm the effect on cell activity, MTS assay was performed and cell count was directly measured by hemocytometer. The mRNA expression of genes involved in hair formation and hair loss formation was confirmed by quantitative RT-PCR. CWR has a strong antioxidant activity. Cell viability of DP cells was increased to 118.5% by treatment of 0.5 mg/ml CWR for 24 hours, but the effect on the cell number was insignificant. These results suggest that CWR increases mitochondrial activity without promoting cell proliferation. Treatment of DP cells with 0.5 mg/ml CWR resulted in 48.5% reduction of mRNA expression of type 2 $5{\alpha}$-reductase, a major cause of male hair loss. In addition, mRNA expression of bone morphogenetic pretein (BMP), fibroblast growth factor (FGF)7, and FGF10, which are closely related to hair growth, was also decreased. Reactive oxygen species (ROS) acts as a cause of hair loss. The excellent antioxidant efficacy of CWR is thought to be able to effectively remove ROS. The dihydrotestosterone produced by type 2 $5{\alpha}$-reductase in DP cells is a potent inducer of male pattern hair loss. The inhibitory effect of type 2 $5{\alpha}$-reductase mRNA on DP cells induced by CWR may induce a positive therapeutic effect of male pattern hair loss.

• 대한치과교정학회지
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• 제26권5호
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• pp.591-599
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• 1996

본 연구는 널리 이용되고 있는 두 종류의 교정용 호선에 다양한 처리를 가한 후 호선의 세포독성을 비교, 평가하고자 시행하였다. 018x025 inch 굵기의 stainless steel 호선과 Co-Cr 호선을 실험 재료로 선택하여 stainless steel 호선을 A 호선, Co-Cr호선을 B 호선이라 칭하였으며 각각의 호선을 다시 가해진 처리에 따라 4군으로 나누었다. A-1군과 B-1군은 제작된 상태 그대로의 호선을 이용하였으며 A-2, B-2군은 기기를 이용하여 $850^{\circ}\;F$에서 4분간 열처리하였다. A-3, B-3군은 같은 방법으로 열처리한 후 표면의 불순물을 제거하기 위해 전해연마를 시행하였고 A-4, B-4군은 소량의 은납(Ag-solder)을 납착(soldering) 하였다. 사람의 치은 섬유아세포를 배양하고 agar overlay법을 이용하여 각군의 호선의 세포독성을 검사하였으며 세포독성을 반응지수(탈색지수/용해지수)로 평가하여 다음의 결론을 얻었다. 1. stainless steel 호선과 Co-Cr 호선 모두 제작된 그대로의 상태에서는 세포독성을 나타내지 않았다. 2. 두 호선에 대한 열처리나 전해연마는 호선의 세포독성에 영향을 미치지 못하였다. 3. 은납이 납착된 stainless steel호선은 은납이 납착된 Co-Cr호선에 비해 더 넓은 범위의 탈색을 나타냈으나 탈색지수와 세포독성(반응지수)에서는 차이를 보이지 않았다. 4. 은납이 납착된 호선은 두 호선 모두에서 중증도의 세포독성을 나타냈다.

• 한국약용작물학회지
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• 제22권6호
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• pp.475-482
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• 2014

This study was indicated to enhance the anti-inflammation activities by the fermentation of the fruits of Aronia melanocarpa (Michx.) Elliott. The extracts by 70% ethanol (EE) showed better biological activities than those by hot water (WE) from campared result of the effect of extraction solvents. Then, the extract from 70% ethanol extraction was further fermented by lactic acid, denoted as FEE. For antioxidant activities, the FEE had showed the highest value as 0.832 of reducing powder, in comparison with those of EE and WE. Cytotoxicity of the water extraction (WE) was measured for 12.06% in addition of $1.0mg/m{\ell}$ of FEE. For anti-inflammation activities, NO production from the macrophage, RAW 264.7 was observed as $7.24{\mu}M$ and $8.52{\mu}M$ from FEE and EE, respectively. Prostaglandin $E_2$ ($PGE_2$) production from human fibroblast cell, CCD-986sk, was also estimated for $152pg/m{\ell}$ in addition of $1.0mg/m{\ell}$ of the FEE. The lowest production of both IL-6 and TNF-${\alpha}$ were $3.5pg/m{\ell}$ and $865.5pg/m{\ell}$, respectively in addition of $1.0mg/m{\ell}$ of the FEE, whereas $74.5pg/m{\ell}$ and $982.4pg/m{\ell}$ in treated with same concenrations of the EE. It was also found that the FEE was higher amounts than other extracts through HPLC analysis of the anthocyanins. These results strongly indicate that fermentation process of the lactic acid could enhance anti-inflammation activities of extracts by increasing the amounts of the anthocyanins, especially cyanidin-galactoside. Our results suggest that the application of the fermentation process for other medicinal herbs can be improved their biological activities.

• 생명과학회지
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• 제19권12호
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• pp.1731-1736
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• 2009

최근의 연구에서 생쥐에 장기간 서구식 사료를 급여했을 때 내인성 SHP 단백질의 수준이 증가함을 보고하였다. 또한 HepG2 세포배양을 통한 실험에서, CDCA 처리가 내인성 SHP 단백질의 수준을 증가시킬 뿐만 아니라 외인성으로 발현된 flag-SHP의 분해율을 감소시켰다. 그리고 HepG2 세포를 ad-flag-SHP로 유전자 형질전환 시켰을 때, 담즙산에 의해 유도되어진 소장 FGF-19이 외인성으로 발현된 flag-SHP 단백질의 반감기를 증가시켰다. 그러나 cholic acid와 FGF-19에 의한 내인성 SHP 단백질의 발현수준과 분해율은 생쥐 또는 배양된 간암세포주에서 아직 명확히 이해되고 있지 않다. 이 연구는 cholic acid의 처리가 생쥐에서 내인성 SHP 단백질의 수준에 미치는 영향과, FGF-19이 HepG2 세포주에서 내인성 SHP 단백질의 분해율에 미치는 영향을 조사하였다. 정상적인 사료를 급여한 대조군 생쥐에서의 내인성 SHP 단백질 수준과 비교하여, 0.5%의 cholic acid를 첨가한 사료를 급여한 생쥐에서는 12시간과 24시간의 처리기간 동안에 내인성 SHP 단백질의 수준이 증가하였다. 배양된 인간 간암세포주인 HepG2에서 CDCA의 처리는 CDCA를 처리하지 않은 대조군 세포주와 비교하여 내인성 SHP 단백질의 분해율을 유의성 있게 변화시키지 않았다. 한편 외인성 ad-flag-SHP 단백질에 대한 이전의 연구와 일치하게, HepG2 세포에 cyclohexamide를 처리하였을 때 FGF-19는 내인성 SHP 단백질의 분해율을 현저히 감소시켰다. 이러한 결과는 담즙산과 FGF-19 모두 생쥐의 간과 HepG2 세포주에서 내인성 SHP 단백질의 수준을 증가시킴을 제시한다.

• 한방재활의학과학회지
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• 제21권2호
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• pp.31-48
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• 2011

Objectives : In order to investigate the anti-obesity effects of Wolbi-tang(here in after referred to WBT) on the obese gene and obese inhibitory, C57BL/6 mice were induced by high-fat diet. Methods : C57BL/6 mice were divided into 5 groups(normal, only high-fat diet, high-fat diet with Reductil, high-fat diet with WBT 400, 200 mg/kg extract) and fed for 5 weeks. And observed body weight change, total cholesterol, low density lipoprotein cholesterol(LDL-cholesterol), high density lipoprotein cholesterol (HDL-cholesterol), triglyceride, glucose, leptin change, alanine transaminase(ALT), aspartate transaminase(AST), serum creatinine, the expression of ${\beta}3$-adrenergic receptor(${\beta}3AR$), leptin, uncoupling protein(UCP2) gene in 3T3-L1 adipocyte, 3T3-L1 adipocyte proliferation, histological analysis of adipose tissue and liver tissue. Results : 1. Refer to cell cytotoxicity, viability of human fibroblast cells(hFCs) showed not significant changes. 2. The amount of ALT, AST was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. The amount of creatinine showed not significant changes. 3. Body weight was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. 4. The amount of total cholesterol and triglyceride was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. LDL-cholesterol was decreased and HDL-cholesterol was increased significantly in WBT 400 mg/kg groups. 5. The amount of glucose was decreased significantly in WBT 400 mg/kg groups. 6. The amount of serum leptin was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. 7. The revelation of ${\beta}3AR$ in 3T3-L1 adipocyte was increased significantly in WBT $100{\mu}g/ml$, $50{\mu}g/ml$ groups. The revelation of leptin was decreased significantly in WBT $100{\mu}g/ml$, $50{\mu}g/ml$ groups. The revelation of UCP2 was decreased significantly in WBT $100{\mu}g/ml$ group. 8. 3T3-L1 adipocyte proliferation was decreased significantly in WBT $100{\mu}g/ml$, $50{\mu}g/ml$ groups. The size of adipocyte was decreased relative to the control group in WBT 400 mg/kg group. 9. The adipose vacuoles in liver tissue was decreased relative to the control group. Conclusions : These results suggested that WBT has inhibitory effects of obesity. WBT might be applicated on treatment of obesity and metabolic syndrome. Further studies analysing its effects were needed.

• 한방재활의학과학회지
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• 제21권2호
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• pp.63-85
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• 2011

Objectives : This study was carried out to know the effects of Danggwisayeokgaohsuyusaenggang-tang(hereinafter referred to DST) on arthritis induced by collagen on DBA/1 OlaHsd mice. Methods : For this purpose, DST was orally administered to mouse with arthritis induced by collagen II. Cytotoxicity, high performance liquid chromatograph(HPLC) analysis, arthritis index, value of immunocyte in draining lymph node and paw joint, cytokine were measured in vivo. Results : 1. The cytotoxicity against human fibroblast cells(hFCs) was not measured in any concentration. 2. In HPLC analysis, There are high peak patterns at 8 minute(min), 12 min, 35 min, 45 min. 3. The arthritis index was decreased significantly. 4. The degree of arthritis induced damage of joint of DST group is slight compared with control group in histopathologic observation(Hematoxylin and eosin stain(H&E), Masson's trichrome(M-T) staining). 5. In total cell counts of draining lymph node(DLN) and paw joint, the cells in DLN decreased significantly on DST 200 mg/kg and the cells in paw joint decreased significantly on 200 mg/kg and 50 mg/kg. 6. In DLN, $CD4^+/CD25^+$, $CD3^+/CD69^+$, major histocompatibility complex(MHC), class-II/$CD11c^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg $CD3^+/CD8^+$ cells decreased significantly on DST 200 200 mg/kg, $CD4^+$, $CD3^+/CD44^+$ cells decreased. 7. In paw joints, $CD4^+$, $CD11b^+/Gr-1^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg. 8. In joints, levels of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, cyclo-oxygenase-2(COX-2), NOS-II were decreased on DST 200 mg/kg and DST 50 mg/kg. 9. In analysing of cytokine in CD3/CD28 activated spleen, IL-17 was decreased significantly, IL-4 was increased significantly $INF-{\gamma}$ was decreased on DST 200 mg/kg. 10. In analysing of cytokine in collagen activated spleen, IL-17 were decreased significantly, IL-4 was increased significantly. Conclusions : This results demonstrated that DST suppressed the inflammatory progression of collagen-induced arthritis(CIA) mice and supported further studies are required to survey continuously in looking for the effective substance and mechanism in the future.

• Journal of Periodontal and Implant Science
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• 제32권4호
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• pp.733-744
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• 2002

The fibroblasts are the principal cells in the periodontal ligament of peridontium. As the periodontal ligament fibroblasts (PDLF) show similar phenotype with osteoblasts, the PDLF are thought to play an important role in alveolar bone remodeling. Cell-to-cell contacted signaling is crucial for osteoclast formation. Recently it has been reported that PDLJ enhance the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The aims of this study were to $clarify\;^{1)}$ the mechanism of PDLF-induced osteoclastogenesis $and\;^{2)}$ whether we can use preosteoclast cell line instead of primary hematopoietic preosteoclast cells for studying the mechanism of PDLF-induced osteoclastogenesis. Osteoclastic differentiation of mouse macrophage cell line RAW264.7 was compared with that of mouse bone marrow-derived M-CSF dependent cell (MDBM), a well-known hematopoietic preosteoclast model, by examining, 1) osteoclast-specific gene expression such as calcitonin receptor, M-CSF receptor (c-fms), cathepsin K, receptoractivator nuclear factor kappa B (RANK) ,2) generation of TRAP(+) multinucleated cells (MNCs), and 3) generation of resorption pit on the $OAAS^{TM}$ plate. RAW264.7 cultured in the medium containing of soluble osteoclast differentiation Factor (sODF) showed similar phenotype with MDBM-derived osteoclasts, those are mRNA expression pattern of osteoclast-specific genes, TRAP(+) MNCs generation, and bone resorbing abivity. Formation of resorption pits by osteoclastic MNCs differentiated from sODF-treated RAW264.7, was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for ODF, to the sODF-containing culture me야um. The effects of PDLF on differentiation of RAW264.7 into　the TRAP(+) multinucleated osteoclast-like cells were examined using coculture system. PDLF were fxed with paraformaldehyde, followed by coculture with RAW264.7, which induced formation of TRAP(+) MNCs in the absence of additional treatment of sODF. When compared with untreated and fixed PDLF (fPDLF), IL-1 ${\beta}$-treated, or lipopolysaccha-ride-treated and then fixed PDLF showed two-folld increase in the supporting activity of osteoclastogenesis from RAW264.7 coculture system. There were no TRAP(+) MNCs formation in coculture system of RAW264.7 with PDLF of no fixation. These findigs suggested that we can replace the primary hematopoietic preosteoclasts for RAW264. 7 cell line for studying the mechanism of PDLF-induced osteoclastogenesis, and we hypothesize that PDLF control osteoclastogenesis through ODF expression which might be enhanced by inflammatory signals.