Cell Viability of Normal Human Skin Fibroblast and Fibroblasts Derived from Granulation Tissue: Effects of Nutraceuticals

Borawska, M.H.,Czechowska, S.K.,Markiewicz, R.,Hayirli, A.,Olszewska, E.,Kazim Sahin, D.V.M The Korean Society of Food Science and Nutrition 2009 Journal of medicinal food Vol.12 No.2

The effects of lycopene, genistein, and epigallocatechin-3-gallate (EGCG) on cell viability were tested in vitro using a normal human skin fibroblast (NHSF) cell line (CRL-1474) and granulation tissue fibroblasts (GTFs) obtained from a patient with middle ear cholesteatoma. Cell cultures were added with lycopene (1, 5, and $10\;{\mu}M$), genistein (1, 5, 10, 25, and $50\;{\mu}M$), and EGCG (1, 5, 10, 25, and $50\;{\mu}M$) and their respective control cultures were established by adding 5 mL/L tetrahydrofuran (THF), 5 mL/L dimethyl sulfoxide (DMSO), and 5 mL/L DMSO. A colorimetric assay was employed for determining cell viability using thiazolyl blue tetrazolium bromide. Cell viability was expressed as a percentage of the control. Data were analyzed using two-way analysis of variance separately for each compound. Lycopene addition decreased viability of NHSFs and GTFs compared with THF addition (64.1%, 60.5%, and 100%, respectively, P < .0001). Genistein addition also increased viability of both NHSFs and GTFs compared with DMSO addition (P < .02). Increasing EGCG concentration tended to cause a linear increase in viability of NHSFs but did not alter viability of GTFs (P < .10). Our data suggest that genistein and EGCG but not lycopene could help maintaining or improving skin health through enhancing viability of skin fibroblasts.

Cell Viability of Normal Human Skin Fibroblast and Fibroblasts Derived from Granulation Tissue: Effects of Nutraceuticals

M.H. Borawska,S.K. Czechowska,R. Markiewicz,A. Hayirli,E. Olszewska,K. Sahin 한국식품영양과학회 2009 Journal of medicinal food Vol.12 No.2

The effects of lycopene, genistein, and epigallocatechin-3-gallate (EGCG) on cell viability were tested in vitro using a normal human skin fibroblast (NHSF) cell line (CRL-1474) and granulation tissue fibroblasts (GTFs) obtained from a patient with middle ear cholesteatoma. Cell cultures were added with lycopene (1, 5, and 10 μM), genistein (1, 5, 10, 25, and 50 μM), and EGCG (1, 5, 10, 25, and 50 μM) and their respective control cultures were established by adding 5 mL/L tetrahydrofuran (THF), 5 mL/L dimethyl sulfoxide (DMSO), and 5 mL/L DMSO. A colorimetric assay was employed for determining cell viability using thiazolyl blue tetrazolium bromide. Cell viability was expressed as a percentage of the control. Data were analyzed using two-way analysis of variance separately for each compound. Lycopene addition decreased viability of NHSFs and GTFs compared with THF addition (64.1%, 60.5%, and 100%, respectively, P < .0001). Genistein addition also increased viability of both NHSFs and GTFs compared with DMSO addition (P < .02). Increasing EGCG concentration tended to cause a linear increase in viability of NHSFs but did not alter viability of GTFs (P < .10). Our data suggest that genistein and EGCG but not lycopene could help maintaining or improving skin health through enhancing viability of skin fibroblasts.

Effect of Ferulic Acid on Cell Viability and Cell Adhesion Activity in Normal Human Gingival Fibroblasts

Joo-Hyun Lee,Byung-Jo Jin,Il-Hong Son,Du-Seok Han 대한의생명과학회 2004 Biomedical Science Letters Vol.10 No.3

This study was designed to investigate the effect of ferulic acid on cell viability and cell adhesion activity in normal human gingival fibroblasts. The cell viability and cell adhesion activity of ferulic acid was measured by MTT assay or XTT assay, respectively, after normal human gingival fibroblasts were treated with or without ferulic acid for 48 hours. The cell viability of ferulic acid on normal human gingival fibroblasts did not show any decreasement by MTT assay and also, cell adhesion activity did not decreased by XTT assay, respectively, compared with control after cells were treated with various concentrations of ferulic acid for 48 hours. MTT?? and XTT?? were 2,130.0 μM and 1,773.7 μM ferulic acid, respectively. These results suggest that ferulic acid is non-toxic to normal human gingival fibroblasts by showing no significant differences in the cell viability and the adhesion activity compared with control by colorimetric assay.

Effect of Ferulic Acid on Cell Viability and Cell Adhesion Activity in Normal Human Gingival Fibroblasts

Lee Joo-Hyun,Jin Byung-Jo,Son Il-Hong,Han Du-Seok 대한의생명과학회 2004 Biomedical Science Letters Vol.10 No.3

This study was designed to investigate the effect of ferulic acid on cell viability and cell adhesion activity in normal human gingival fibroblasts. The cell viability and cell adhesion activity of ferulic acid was measured by MTT assay or XTT assay, respectively, after normal human gingival fibroblasts were treated with or without ferulic acid for 48 hours. The cell viability of ferolic acid on normal human gingival fibroblasts did not show any decreasement by MTT assay and also, cell adhesion activity did not decreased by XTT assay, respectively, compared with control after cells were treated with various concentrations of ferolic acid for 48 hours. MTT/sub 50/ and XTT/sub 50/ were 2,130.0 μM and 1,773.7 μM ferolic acid, respectively. These results suggest that ferolic acid is non-toxic to normal human gingival fibroblasts by showing no significant differences in the cell viability and the adhesion activity compared with control by colorimetric assay.

녹차추출물 (-)-Epigallocatechin-3-gallate가 사람치주인대섬유모세포 활성 유지에 미치는 영향

정임희,윤정호,정의원,김창성,김남희,정원균,최성호 한국생체재료학회 2010 생체재료학회지 Vol.14 No.3

The purpose of this study was to evaluate the effect of (-)-Epigallocatechin-3-gallate (EGCG) on the viability of human periodontal ligament fibroblast (hPDLF). Firstly, various concentration of EGCG were applied to hPDLF for 8 days, and then 10 μM EGCG was put for the first 6 days to the experimental group and was stopped for the next 8days. For the positive control group, the 10 μM EGCG was supplied during the whole period, while any EGCG was not supplied for the negative control. Cell viability was analyzed with MTT assay. In the first experiment, the group which treated 10 μM EGCG for 6days showed highest cell viability. In the second treatment, the negative control showed continuous growing up in cell proliferation until 12th day and then a little decreased. After stopping EGCG supply, the experimental group’s proliferation was slightly cut down and then showed big increases. Lastly, the positive control°Øs increase was lasted during the whole period, but there was no a large variance and it showed lowest cell viability on 14th day. From these results, we concluded that EGCG do not affect such a lot on the increase of hPDLF proliferation,but it is possible for EGCG to make hPDLF hibernate with proper concentration of EGCG.

Effect of ferulic Acid on Cell Viability and Cell Adhesion Activity in Normal Human Gingival Fibrobalsts

Lee, Joo-Hyun,Jin, Byung-Jo,Son, Il-Hong,Han, Du-Seok THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 2004 Journal of biomedical laboratory sciences Vol.10 No.3

This study was designed to investigate the effect of ferulic acid on cell viability and cell adhesion activity in normal human gingival fibroblasts. The cell viability and cell adhesion activity of ferulic acid was measured by MTT assay or XTT assay, respectively, after normal human gingival fibroblasts were treated with or without ferulic acid for 48 hours. The cell viability of ferulic acid on normal human gingival fibroblasts did not show any decreasement by MTT assay and also, cell adhesion activity did not decreased by XTT assay, respectively, compared with control after cells were treated with various concentrations of ferulic acid for 48 hours. MTT_(50) and XTT_(50) were 2,130.0 μM and 1,773.7 μM ferulic acid, respectively. These results suggest that ferulic acid is non-toxic to normal human gingival fibroblasts by showing no significant differences in the cell viability and the adhesion activity compared with control by colorimetric assay.

MICROPATTERNED GROOVES AND ACID-ETCHING ON TITANIUM SUBSTRATA ALTER VIABILITY AND GENE EXPRESSION OF ADHERED HUMAN GINGIVAL FIBROBLASTS: A PILOT STUDY

Lee, Suk-Won,Kim, Su-Yeon,Lee, Keun-Woo The Korean Academy of Prosthodonitics 2007 대한치과보철학회지 Vol.45 No.3

Statement of problem. Prior to determining an optimal width of micropatterned grooves provided on titanium substrata, we have done a pilot study using surface topographies in combined microm and submicrom levels. Purpose. The purpose of this study was twofold 1) to assess the proliferation and 2) to analyze the expression of genes encoding the intracellular signaling proteins involved in cell-substratum adhesions and adhesion-dependent G1 phase cell cycle progression of human gingival fibroblasts plated on smooth and microgrooved/acid-etched titanium substrata. Material and methods. Three groups of titanium discs as NE0 (smooth Ti substrata), E15 (Ti substrata with microgrooves of $15{\mu}m$ of spacing and $3.5{\mu}m$ in depth and with further acidetching), and E30 (Ti substrata with microgrooves of $30{\mu}m$ spacing and $3.5{\mu}m$ in depth and with further acid-etching) served as the human gingival fibroblasts' substrata. Viability and proliferation of fibroblasts were determined using an XTT assay. Gene expressions of fibronectin, ${\alpha}5$ integrin, CDK4, and $p27^{kip}$ were analyzed in RT-PCR. Cell-substratum interactions were analyzed in SEM. Results. From the XTT assay at 24 h incubation, the mean optical density (OD) value of E15 was significantly greater than the values of E30 and NE0. At 48 and 96 h however, the mean OD values of E30 were significantly greater than the values of E15 and NE0. No differences in the expression of PCR transcripts at 96 h incubations were noted between groups, whereas at 48 h, an unexpected increase in the expression of all the transcripts were noted in E15 compared with other two groups. Fibroblasts were observed to orient and adhere inside the microgrooves. Conclusion. Micropatterned grooves and acid-etching on Ti substrata alter viability and gene expression of adhered human gingival fibroblasts.

Increased poly(dimethylsiloxane) stiffness improves viability and morphology of mouse fibroblast cells

박중열,유성주,이은중,이대호,김지영,이상훈 한국바이오칩학회 2010 BioChip Journal Vol.4 No.3

We monitored the viability and morphology of mouse fibroblast cells cultured on PDMS sub strates with different degrees of polymer stiffness. The stiffness was controlled by varying the ratio between base and crosslinker agent during mixing. Although the standard PDMS mixing ratio is 10 : 1 (base to crosslinker; Young’s modulus, E=580 kPa), we found that a PDMS substrate with a high stiffness (mixing ratio of 5 : 1, E=1,000 kPa) was more favora ble as a substrate for fibroblast cell growth. It is important to note that an extracellular matrix coating was not applied to the PDMS so that the effect of stiffness on cell growth could be studied in isolation. A stiffness reduction of 40% (from a mixing ratio of 5 : 1 to 10 : 1)produced a significant reduction in survival rate (viability was reduced by 15%), and viability worsened (was reduced by 45%) for a substrate stiffness of 280 kPa (a mixing ratio of 20 : 1). The rate of spreading for the cells was measured to show that stiffer materials promoted more prolific fibroblast growth. These results provide PDMS stiffness guidelines for cell culture substrates.

Cytotoxicity of Impregnated Dental Gingival Retraction Cord Extracts in Immortalized Human Oral Fibroblasts and Keratinocytes in vitro

이명진,양송이 한국재료학회 2023 한국재료학회지 Vol.33 No.4

This study evaluated cell viability and cytokine release in immortalized human oral fibroblasts (hTERT-hNOFs) and keratinocytes (IHOK) exposed to a dental-impregnated gingival retraction cord. To prepare the extracts, dental gingival retraction cords impregnated with aluminum chloride hexahydrate were immersed in a cell culture medium for 24 h at 37 °C. hTERT-hNOFs and IHOK were cultured for 24 h. The cell culture medium was removed and extracts of the dental gingival retraction cords were added. After incubation with the extract solution, cell viability was evaluated using an MTT assay. The levels of the cytokines IL-1α and IL-8 were measured in the supernatants of each cell type. The cell viability after exposure to the extract solution for 10 min exceeded 70 % in both cell types. The ET50 values for hTERT-hNOF and IHOK were 35.75 and 28.98 min, respectively. For IHOK, the IL-1α level was (5.35 ± 5.22) pg/mL at 10 min, (3.58 ± 5.38) pg/mL at 20 min, and (2.85 ± 4.28) pg/mL at 60 min of exposure (p > 0.05). The IL-8 level in IHOK was (67.16 ± 18.70) pg/mL at 10 min, (78.36 ± 7.50) pg/mL at 20 min, and (111.9 ± 26.10) pg/mL at 60 min of exposure (p > 0.05). Cytokine release was not observed from hTERThNOFs. Based on these results, cell viability and cytokine release were confirmed in cells exposed to the impregnated gingival retraction cord. In addition, the application of the extracts to hTERT-hNOF and IHOK during the actual contact time and determination of ET50 may be beneficial for evaluating the biocompatibility of dental-impregnated gingival retraction cords.

Valproic Acid-induced PPAR-alpha and FGF21 Expression Involves Survival Response in Hepatocytes

아자모프 바커부딘,강여원,이찬희,심완석,이광민,송박용 한국생명과학회 2024 생명과학회지 Vol.34 No.4

Hepatocyte damage caused by medications or herbal products is one of the important problem when these compounds are chronically administrated. Thus, improving hepatocyte survival during treatment offers a wide range of opportunities. Valproic acid (VPA), a branched short-chain fatty acid derived from naturally occurring valeric acid, is commonly used to treat epilepsy and seizures. Although VPA exerts numerous effects in cancer, HIV therapy, and neurodegenerative disease, its effects on the liver and its mechanism of action have not been fully elucidated. Here, we demonstrated that VPA caused moderate liver cell toxicity and apoptosis. Interestingly, VPA treatment increased transcription levels of PPAR alpha (PPAR-α) and fibroblast growth factor 21 (FGF21) in murine (Hepa1c1c7) hepatoma cells in a time and concentration dependent manner. VPA-induced FGF21 expression was significantly weaker under PPAR-α silencing condition than in cells transfected with non-targeting control siRNA. Subsequent experiments showed that cell viability was significantly lowered when the FGF21 signaling pathway was blocked by FGF receptor antagonist. Finally, we further determined that AMPK phosphorylation was not responsible for VPA-induced FGF21 expression and PPAR- increments. These results indicate that increases of FGF21 expression alleviate VPA-induced hepatic toxicity, thereby making FGF21 a potential biomarker for predicting liver damage during VPA treatments.