• The Korean Journal of Physiology and Pharmacology
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• 제24권6호
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• pp.453-462
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• 2020

Chronic joint pain due to loss of cartilage function, degradation of subchondral bone, and related conditions are common plights of an arthritis patient. Antioxidant compounds could solve the problems in arthritic condition. The objective of this study was to evaluate the anti-arthritic activity of D-carvone against complete Freund's adjuvant (CFA)-induced arthritis in rats. D-carvone was orally administered for 25 days at the doses of 30 and 60 mg/kg against CFA-induced arthritic rats. Changes in body weight, paw swelling, organ index, hematological parameters, oxidative stress markers, inflammatory cytokines, and histopathology were recorded. Oral treatment of D-carvone significantly improved the body weight, reduced the paw swelling, edema formation, and organ index in arthritic rats. The levels of white blood cells were reduced, red blood cells and hemoglobin levels were improved in D-carvone treated arthritic rats. Lipid peroxidation levels were lowered whereas enzymatic and non-enzymatic antioxidants were significantly elevated by D-carvone administration against arthritic rats. D-carvone significantly modulated inflammatory cytokine levels and improved the ankle joint pathology against CFA-induced arthritic inflammation. In conclusion, D-carvone proved significant anti-arthritic activity against CFA-induced arthritis in rats.

• The Korean Journal of Physiology and Pharmacology
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• 제25권2호
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• pp.139-146
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• 2021

Mitochondrial NADP+-dependent isocitrate dehydrogenase 2 (IDH2) produces NADPH, which is known to inhibit mitochondrial oxidative stress. Ureteral obstruction induces kidney inflammation and fibrosis via oxidative stress. Here, we investigated the role and underlying mechanism of IDH2 in unilateral ureteral obstruction (UUO)-induced kidney inflammation using IDH2 gene deleted mice (IDH2-/-). Eight- to 10-week-old female IDH2-/- mice and wild type (IDH2+/+) littermates were subjected to UUO and kidneys were harvested 5 days after UUO. IDH2 was not detected in the kidneys of IDH2-/- mice, while UUO decreased IDH2 in IDH2+/+ mice. UUO increased the expressions of markers of oxidative stress in both IDH2+/+ and IDH2-/- mice, and these changes were greater in IDH2-/- mice compared to IDH2+/+ mice. Bone marrow-derived macrophages of IDH2-/- mice showed a more migrating phenotype with greater ruffle formation and Rac1 distribution than that of IDH2+/+ mice. Correspondently, UUO-induced infiltration of monocytes/macrophages was greater in IDH2-/- mice compared to IDH2+/+ mice. Taken together, these data demonstrate that IDH2 plays a protective role against UUO-induced inflammation through inhibition of oxidative stress and macrophage infiltration.

• 생명과학회지
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• 제28권12호
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• pp.1448-1454
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• 2018

최근 들어 곤충을 식품 및 바이오 소재로 이용한 연구가 활발히 진행되고 있다. 그러나 곤충을 이용한 조골세포 활성 및 분화에 따른 골 형성 촉진 효과에 대한 연구는 아직 미흡한 실정이다. 뿐만 아니라 풀무치를 이용한 기능성 연구는 거의 이루어지지 않고 있다. 따라서 본 연구에서는 골 형성 촉진 효능을 가진 새로운 천연물 소재 개발을 위해 풀무치 추출물의 MG-63 조골세포의 분화 촉진 효과를 연구하였다. 조골세포에서 풀무치 추출물의 독성 및 증식 효과를 평가하기 위하여 MTS assay를 진행한 결과, $1,000{\mu}g/ml$ 농도까지 세포 독성이 나타나지 않았으며, 48시간 배양했을 때 $500-1,000{\mu}g/ml$ 농도에서 105%와 116%의 세포 증식 효능을 확인 하였다. 풀무치 추출물이 조골세포 분화에 미치는 영향을 확인하기 위하여 3일 및 5일간 풀무치 추출물을 MG-63 조골세포에 처리한 후 ALP 활성을 측정하였다. 그 결과 $100{\mu}g/ml$ 농도에서 positive control로 사용한 조골세포 분화배지(DM)군과 유사한 정도로 분화가 증가하였으며 500 및 $1,000{\mu}g/ml$ 농도에서는 2-3배까지 조골세포 분화가 촉진되었다. 이 결과는 ALP staining에서도 유사하게 나타났다. mRNA 발현량의 변화를 측정한 결과, Alpl과 Runx2 유전자 발현량이 증가하였고, 단백질 발현량을 측정했을 때에도 유사한 결과를 확인하였다. 이를 통해 ALP와 Runx2 유전자 및 단백질 발현에 의해서 ALP 활성이 증가하고 조골세포 분화가 촉진되었을 것으로 판단되며, 풀무치 추출물을 이용한 골 형성 촉진에 따른 골다공증 예방 및 치료 기능성 소재 개발에 대한 가능성을 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권11호
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• pp.1837-1847
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• 2020

Objective: To evaluate the pancreatic differentiation potential of α-1,3-galactosyltransferase knockout (GalTKO) pig-derived bone marrow-derived mesenchymal stem cells (BM-MSCs) using epigenetic modifiers with different pancreatic induction media. Methods: The BM-MSCs have been differentiated into pancreatic β-like cells by inducing the overexpression of key transcription regulatory factors or by exposure to specific soluble inducers/small molecules. In this study, we evaluated the pancreatic differentiation of GalTKO pig-derived BM-MSCs using epigenetic modifiers, 5-azacytidine (5-Aza) and valproic acid (VPA), and two types of pancreatic induction media - advanced Dulbecco's modified Eagle's medium (ADMEM)-based and N2B27-based media. GalTKO BM-MSCs were treated with pancreatic induction media and the expression of pancreas-islets-specific markers was evaluated by real-time quantitative polymerase chain reaction, Western blotting, and immunofluorescence. Morphological changes and changes in the 5'-C-phosphate-G-3' (CpG) island methylation patterns were also evaluated. Results: The expression of the pluripotent marker (POU class 5 homeobox 1 [OCT4]) was upregulated upon exposure to 5-Aza and/or VPA. GalTKO BM-MSCs showed increased expression of neurogenic differentiation 1 in the ADMEM-based (5-Aza) media, while the expression of NK6 homeobox 1 was elevated in cells induced with the N2B27-based (5-Aza) media. Moreover, the morphological transition and formation of islets-like cellular clusters were also prominent in the cells induced with the N2B27-based media with 5-Aza. The higher insulin expression revealed the augmented trans-differentiation ability of GalTKO BM-MSCs into pancreatic β-like cells in the N2B27-based media than in the ADMEM-based media. Conclusion: 5-Aza treated GalTKO BM-MSCs showed an enhanced demethylation pattern in the second CpG island of the OCT4 promoter region compared to that in the GalTKO BM-MSCs. The exposure of GalTKO pig-derived BM-MSCs to the N2B27-based microenvironment can significantly enhance their trans-differentiation ability into pancreatic β-like cells.

• 생명과학회지
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• 제29권9호
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• pp.1027-1033
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• 2019

현재 우리나라는 고령화 사회에 접어들었으며, 이와 함께 골 대사 질환이 큰 문제로 대두되고 있다. 그 중 가장 흔하게 발생하는 골다공증은 여러 요인에 의해 발생하고 있으며 이의 예방 및 치료를 위한 소재 개발 연구가 활발하게 진행되고 있다. 그러나 곤충을 소재로 한 골다공증 예방 및 치료 연구는 미흡한 실정이다. 따라서 본 연구에서는 골 형성 촉진 효능을 가진 새로운 소재 개발을 위해 갈색거저리 유충 오일의 MG-63 조골세포의 분화 촉진 효과를 연구하였다. 조골세포에서 갈색거저리 유충 오일의 독성 및 증식 효과를 평가하기 위하여 MTS assay를 진행한 결과 $80{\mu}g/ml$ 농도까지 세포 독성이 나타나지 않았으며, 48시간 배양했을 때 $40-80{\mu}g/ml$ 농도에서 120% 정도의 세포 증식 효능을 확인 하였다. 갈색거저리 유충 오일이 조골세포 분화에 미치는 영향을 확인하기 위하여 5일간 갈색거저리 유충 오일을 MG-63 조골세포에 처리한 후 ALP 활성을 측정하였다. 그 결과 $80{\mu}g/ml$ 농도에서 무처리 군에 비해 약 180%의 조골세포 분화 촉진 효능을 관찰 하였다. 이 결과는 ALP staining에서도 유사하게 나타났다. mRNA 발현량의 변화를 측정한 결과, Alpl과 Runx2 유전자 발현량이 증가하였고, 단백질 발현량을 측정했을 때에도 유사한 결과를 확인하였다. 이를 통해 ALP와 Runx2 유전자 및 단백질 발현에 의해서 ALP 활성이 증가하고 조골세포 분화가 촉진되었을 것으로 판단되며, 갈색거저리 유충 오일을 이용한 골 형성 촉진에 따른 골다공증 예방 및 치료 기능성 소재 개발에 대한 가능성을 확인하였다.

• Reproductive and Developmental Biology
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• 제34권1호
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• pp.1-6
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• 2010

Mesenchymal stem cells (MSCs) have the multipotent capacity and this potential can be applied for obtaining valuable cell types which can use for cell therapy on various regenerative diseases. However, insufficient availability of cellular source is the major problem in cell therapy field using adult stem cell sources. Recently, human embryonic stem cells (hESCs) have been highlighted to overcome a limitation of adult cellular sources because they retain unlimited proliferation capacity and pluripotency. To use of hESCs in cell therapy, above all, animal pathogen free culture system and purification of a specific target cell population to avoid teratoma formation are required. In this study, we describe the differentiation of a mesenchymal stem cell-like cells population from feeder-free cultured hESCs(hESC-MSCs) using direct induction system. hESC-MSCs revealed characteristics similar to MSCs derived from bone marrow, and undifferentiated cell markers were extremely low in hESC-MSCs in RT-PCR, immunostaining and FACS analyses. Thus, this study proffer a basis of effective generation of specialized human mesenchymal stem cell types which can use for further clinical applications, from xenofree cultured hESCs using direct induction system.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권4호
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• pp.299-305
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• 2010

Purpose: Adipose tissue is located beneath the skin, around internal organs, and in the bone marrow in humans. Its main role is to store energy in the form of fat, although it also cushions and insulates the body. Adipose tissue also has the ability to dynamically expand and shrink throughout the life of an adult. Recently, it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including, osteogenic, chondrogenic, endothelial cells, and myogenic lineages. Materials and Methods: This study focused on endothelial cell culture from the adipose tissue. Adipose tissues were harvested from buccal fat pad during bilateral sagittal split ramus osteotomy for surgical correction of mandibular prognathism. The tissues were treated with 0.075% type I collagenase. The samples were neutralized with DMEM/and centrifuged for 10 min at 2,400 rpm. The pellet was treated with 3 volume of RBC lysis buffer and filtered through a 100 ${\mu}m$ nylon cell strainer. The filtered cells were centrifuged for 10 min at 2,400 rpm. The cells were further cultured in the endothelial cell culture medium (EGM-2, Cambrex, Walkersville, Md., USA) supplemented with 10% fetal bovine serum, human EGF, human VEGF, human insulin-like growth factor-1, human FGF-$\beta$, heparin, ascorbic acid and hydrocortisone at a density of $1{\times}10^5$ cells/well in a 24-well plate. Low positivity of endothelial cell markers, such as CD31 and CD146, was observed during early passage of cells. Results: Increase of CD146 positivity was observed in passage 5 to 7 adipose tissue-derived cells. However, CD44, representative mesenchymal stem cell marker, was also strongly expressed. CD146 sorted adipose tissue-derived cells was cultured using immuno-magnetic beads. Magnetic labeling with 100 ${\mu}l$ microbeads per 108 cells was performed for 30 minutes at $4^{\circ}C$ a using CD146 direct cell isolation kit. Magnetic separation was carried out and a separator under a biological hood. Aliquous of CD146+ sorted cells were evaluated for purity by flow cytometry. Sorted cells were 96.04% positivity for CD146. And then tube formation was examined. These CD146 sorted adipose tissue-derived cells formed tube-like structures on Matrigel. Conclusion: These results suggest that adipose tissue-derived cells are endothelial cells. With the fabrication of the vascularized scaffold construct, novel approaches could be developed to enhance the engineered scaffold by the addition of adipose tissue-derived endothelial cells and periosteal-derived osteoblastic cells to promote bone growth.