• Title/Summary/Keyword: fibroblasts

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Ell3 Modulates the Wound Healing Activity of Conditioned Medium of Adipose-derived Stem Cells

  • Lee, Jae-Yong;Oh, Nuri;Park, Kyung-Soon
    • Development and Reproduction
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    • v.21 no.3
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    • pp.335-342
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    • 2017
  • While adipose-derived stem cell-conditioned medium (ADSC-CM) has been demonstrated to promote skin wound healing, the mechanism regulating this effect remains unelucidated. In this study, we aimed to investigate the role of Ell3 in the wound healing activity of ADSC-CM. In vitro analysis revealed that Ell3 suppression in ADSCs impairs the promotive activity of ADSC-CM on the proliferation and migration of mouse embryonic fibroblasts (MEF) and normal human dermal fibroblasts (NHDF). Consistently, the expression of MMP family genes, which regulate cell proliferation and migration, was significantly suppressed in MEF and NHDF treated with siEll3-transfected ADSC-CM. Proinflammatory cytokines, such as interleukin-1 and interleukin-6, were highly expressed in MEF treated with siEll3-transfected ADSC-CM. The wound healing activity of siEll3-transfected ADSC-CM was significantly lower than that of the control in vivo. Our results suggest that Ell3 may contribute to the inhibition of inflammatory response during skin wound healing.

v-Crk Induces Rac-dependent Membrane Ruffling and Cell Migration in CAS-deficient Embryonic Fibroblasts

  • Sung, Bong Hwan;Yeo, Myoung Gu;Oh, Hye Jin;Song, Woo Keun
    • Molecules and Cells
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    • v.25 no.1
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    • pp.131-137
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    • 2008
  • Crk-associated substrate (CAS) is a focal adhesion protein that is involved in integrin signaling and cell migration. CAS deficiency reduces the migration and spreading of cells, both of which are processes mediated by Rac activation. We examined the functions of v-Crk, the oncogene product of the CT10 virus p47gag-crk, which affects cell migration and spreading, membrane ruffling, and Rac activation in CAS-deficient mouse embryonic fibroblasts (CAS-/- MEFs). CAS-/- MEFs showed less spreading than did CAS+/+ MEFs, but spreading was recovered in mutant cells that expressed v-Crk (CAS-/-v-Crk MEF). We observed that the reduction in spreading was linked to the formation of membrane ruffles, which were accompanied by Rac activation. In CAS-/- MEFs, Rac activity was significantly reduced, and Rac was not localized to the membrane. In contrast, Rac was active and localized to the membrane in CAS-/-v-Crk MEFs. Lamellipodia protrusion and ruffle retraction velocities were both reduced in CAS-/- MEFs, but not in CAS-/-v-Crk MEFs. We also found that microinjection of anti-gag antibodies inhibited the migration of CAS-/-v-Crk MEFs. These findings indicate that v-Crk controls cell migration and membrane dynamics by activating Rac in CAS-deficient MEFs.

Cytotoxicity Effects of Fraction and Chloroform Extracts from Corn is fructus on Cancer Cell Lines (산수유 클로로포름 추출물과 분획물의 암세포주에 대한 세포독성)

  • Yang Hyun Ok;Choi Won Hyung;Kim Young Hyun;Baek Seung Hwa;Chun Hyun Ja
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.5
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    • pp.1343-1346
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    • 2004
  • Cornis fructus were extracted by successive extractions and then fractionated with chloroform extract to get active fractions. This study was performed to determine the cytotoxic effect of chloroform extract from Corn is fructus on NIH 3T3 fibroblasts and cancer cell lines using MTT assay. All extracts did not exhibit cytotoxicity in NIH 3T3 fibroblasts. Chloroform extract exhibited antitumor activity in A549, MDA-MB-123, B16 melanoma and SNU-C4 cells. Futher fractionation with chloroform extract was performed to obtain effective fractions. 3 fraction showed the strongest cytotoxic effect against A549, MDA-MB-123, B16 melanoma and SNU-C4 cells. These results suggest that 3 fraction of the chloroform extract from Cornis fructus possessed bioactive material of antitumorous agents.

ELECTRON MICROSCOPIC STUDY ON THE PULP OF HUMAN PRIMARY TOOTH IN THE SHEDDING STAGE (탈락기(脫落期) 유치치수(乳齒齒髓)의 미세구조(微細構造)에 관(關)한 전자현미경적(電子顯微鏡的) 연구(硏究))

  • Kim, Woo-Chul
    • Journal of the korean academy of Pediatric Dentistry
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    • v.10 no.1
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    • pp.25-33
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    • 1983
  • With electron microscope, author studied on the pulp structure of human primary tooth in shedding stage. Non-carious human primary molar teeth were selected for this study. Using standard methods, specimens were sectioned and examined by light and electron microscope, The results were as follows; 1. In coronal pulp, odontoblasts were replaced by multinucleated odontoclasts, which contained a large number of mitochondria of varying shape and vacuoles in cytoplasm. Where odontoclasts were in contact with tooth surface, the characteristic ruffled border and clear zone were observed. 2. Fibrous tissue with plentiful collagen fibers and fibroblasts was observed adjacent to the dentin in the pulp. Fibroblast contained a number of mitochondria and well-developed rough-surfaced endoplasmic reticulum. 3. Inflammatory cells were observed in the pulp and active fibroblasts could be seen between inflammatory cells. In many cases, cervical epithelium proliferated toward absorbed area. 4. Inflammatory cells consisted of a number of lymphocytes, polymorphonuclear leukocytes, plasma cells and macrophages. Macrophage containing lysosomes in digestive state or phagocyting PMN could be seen. 5. In the primary molar of delayed root resorption, odontoblast layer, zone of Weil and cell-rich zone could be seen at roof of pulp chamber and odontoblast in this area cont과ained some lipid droplets.

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The Effect of Lipid Concentration in Culture Medium on Senescence and Lipid Peroxides Production of Fibroblast from Neonate Rats (배양액내 지방함량의 변화가 신생흰쥐 피부섬유아세포의 노화와 지질과산화물 생성에 미치는 영향)

  • 장영애
    • Journal of Nutrition and Health
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    • v.29 no.1
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    • pp.97-103
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    • 1996
  • This study was performed to investigate the effects of lipid on cellular senescence, lipid peroxide production, and morphological changes. For this study we used primary skin fibroblasts from neonate rats grown in media various lipid contents. Fibroblasts were cultured until they lost their proliferation potential either in control medium (Dulbecco's modified Eagle's medium supplement with 10% fetal bovine serum) or in media supplemented with various concentrations of lipid-cholesterol rice component from bovine serum. Cumulative population doublings(CPD, as an index of cellular life span), and cellular thiobarbituric acid reactive substances (TBARS, as an index of lipid peroxide) concentrations were measured and morphological changes were observed. CPD were shortened with increasing lipid concentration in media ; 28.12 for cells grown in control medium and 13.42, 11.42, and 6.19 for those grown in 0.1%, 1% and 5% lipid rich components containing media, respectively. Cellular proliferation ratios were those grown in 5% lipid rich components containing media were delayed and they were degenerated soon. TBARS concentrations were increased with increasing concentration of lipid in media. Morphological changes were observed in cells grown in control medium by cellular senescence. Especially lipid droplets were observed in cells grown in 5% lipid rich components containing media. Therefore it seems that lipid contents in media had an effect on cellular proliferation and cellular life span, possibly via lipid peroxide production.

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Effects of He-Ne Laser Irradiation on the Activity of the Cultured Fibroblast (He-Ne 레이저 조사가 배양 섬유모세포의 활성에 미치는 영향)

  • Song In-Young;Lee Jae-Hyoung
    • The Journal of Korean Physical Therapy
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    • v.9 no.1
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    • pp.71-79
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    • 1997
  • The purpose of this study were to determined the effect of laser irradiation on the fibroblast activity. Cultures of 3T3 fibroblasts were subjected to Helium Neon laser(632.8 nm) irradiation of various energy density. On one, two and three consecutive days the fibroblast monolayers wert irradiated for period from 0 to 32 minutes with 8 mW of average output power. The fibroblast activity was determined by the quantitative assay of MTT, SRB and NR after incubation of the fibroblasts for 24 hours. Results show that exposure duration from 2 min to 32 min could increase MTT at three consecutive days, whereas control and 1 min, one and two days irradiation had were not inclosed. The SRB and NR were inclosed at two and three consecutive days from 2 min to 32 min, whereas control and 1 min, and once radiation were not increased. These result demonstrate that energy density from 0.48 to 7.64 J/cm could increase cellular protein contents and fibroblast activity at more than twice irradiation of laser, whereas low energy density (less than 0.24 J/m) and once irradiation of laser had no effect. The results suggest that the beneficial effect of the He-Ne laser with adequate dose on fibroblast activity in vitro.

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Evaluation of neutral red uptake assay using Balb/c 3T3 cells as a screening test to predict skin phototoxicity potential

  • Lee, Jong-Kwon;Lee, Eun-Hee;Kim, Ju-Hwan;Hong, Jin-Tae;Kim, Hyung-Soo;Park, Ki-Sook;Ahn, Kwang-Soo;Cho, Dae-Hyun;Lee, Sun-Hee
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.05a
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    • pp.109-109
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    • 2001
  • In order to evaluate the neutral red uptake assay as an alternative method for phototoxicity test, we compared the potential of phototoxicity in vitro in cultured human fibroblasts and 3T3 fibroblast cells derived from Balb/c mice. Both fibroblasts were exposed to various known phototoxic chemicals (promethazine, neutral red, chlorpromazine, chlortetracycline, amiodarone, bithionol, 8-methoxypsoralen) and non-phototoxic chemical (ammonium laureth sulfate) and irradiated with 5 J/cm$^2$ of UVA.(omitted)

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Characterization of X-linked RNA Transcripts in Matured Bovine Spermatozoa

  • Jeon, Byeong-Gyun;Kumar, B. Mohana;Rho, Gyu-Jin
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.209-214
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    • 2011
  • Although the function and utility of RNA transcripts derived from matured spermatozoa remains unclear, they might play important roles in the establishment of a paternal genome and subsequently embryo development. Herein, we investigated the expression of X-chromosome linked RNA transcripts in matured bovine spermatozoa. The total RNA was extracted from the matured spermatozoa, and then converted to cDNA. Autosomal genes (ACT-${\beta}$ and H-2A) and X-chromosome linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) were analyzed for the characterization of X-chromosome linked RNA transcripts and compared to female fibroblasts by RT-PCR. The transcripts of autosomal genes (ACT-${\beta}$ and H2A) and X-chromosome linked genes (ANT3, HPRT, MeCP2, RPS4X and ZFX) were not detected in spermatozoa. However, XIAP (X-linked inhibitor of apoptosis protein) and XIST (X-chromosome inactive-specific transcript, a kind of paternal imprinted gene) transcripts were detected in spermatozoa, and relative levels of XIAP and XIST transcripts were similar and 0.5-fold lower when compared to female fibroblasts, respectively. Based on the findings, it is summarized that the presence of RNA transcripts of XIAP and XIST in the isolated spermatozoa may imply their role in inhibition of apoptosis and induction of X-chromosome inactivation in embryo development.

Protective Effect of Gryllus Bimaculatus Methanol Extract on UVB-induced Photoaging in Human Skin Fibroblasts (UVB로 손상된 피부 섬유아세포에서 쌍별귀뚜라미 메탄올 추출물의 보호효과)

  • Jeong, Taekyoung;Yu, Myeongnam;Heo, Huijin;Yang, Jinwoo;Jeong, Heonsang;Lee, Junsoo
    • Journal of the Korean Society of Food Culture
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    • v.35 no.5
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    • pp.478-482
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    • 2020
  • We evaluated the protective effects of cricket methanol extract (CME) on ultra-violet B (UVB)-induced photoaging in human skin fibroblasts. The fibroblast cells were treated with 10, 50, and 100 ㎍/mL of CME for 24 h, and then exposed to UVB (30 mJ/㎠). CME showed a dose-dependent cytoprotective effect without any observable cytotoxicity. CME reduced UVB-induced production of reactive oxygen species (ROS) by 34.4, 34.9, 40.6% at concentrations of 10, 50, 100 ㎍/mL respectively. CME inhibited the release of matrix metalloproteinase (MMP) 1 and 3. Furthermore, CME also reduced UVB-induced collagen degradation in the fibroblast cells. Taken together, our data suggests that CME has a significant protective effect on UVB-induced photoaging of the skin. This benefit occurs through multiple mechanisms. The results also suggest a potential role for CME as an ingredient in anti-photoaging cosmetic products in the future.

MODULATION OF INSULIN-STIMULATED DNA SYNTHESIS BY CHOLERA TOXIN IN BOVINE MAMMARY FIBROBLASTS

  • Yuh, I.S.;Park, C.K.;Han, J.Y.;Sheffield, L.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.4
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    • pp.483-489
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    • 1993
  • Bovine fibroblasts were cultured in Dulbecco's Modified Eagle's Medium and then treated with control, insulin (I, $1{\mu}g/ml$), cholera toxin (CT, 0.1-100 ng/ml) or CT (0.1-100 ng/ml) + I ($1{\mu}g/ml$). Cholera toxin, an activator of adenylate cyclase, significantly decreased insulin induced DNA synthesis (p<0.05). The modulation of DNA synthesis apparently involves events occurring in early stage of cell growth, at least between the first 4 and 8 hour of CT treatment. Insulin induced collagen as well as noncollagen synthesis in cell layer, however, these syntheses were reduced by addition of cholera toxin (p<0.05) but were not completely reduced. It is not clear whether the reduction of insulin-induced cell layer collagen or noncollagen proteins by CT is involved in the inhibitory effect on insulin-induced DNA synthesis. However, we could rule out the hypothesis that insulin-induced DNA synthesis is reduced by CT-induced cellular differentiation.