• Journal of the korean academy of Pediatric Dentistry
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• v.45 no.3
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• pp.271-279
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• 2018

Previous studies to elucidate the etiology of cyclosporine(Cs)-induced gingival overgrowth in children have not completely excluded all factors that may cause differences among individuals. This study examined the effect of cyclosporine on the metabolism of type 1 collagen(CoL-I) in experimental models that controlled the effects of biological variations on individuals. Five 5-week-old male Sprague-Dawley rats were administered Cs by gastric feeding for 6 weeks. Gingival specimens were harvested from the mandibular posterior area before beginning Cs administration and at 2, 4, and 6 weeks thereafter. Gingival fibroblasts were cultured from all the 20 biopsies collected from the gingiva. Half of the fibroblasts collected prior to the Cs administration were designated as Control. The other half of the fibroblasts were treated with Cs in vitro and called in vitro test group(Tt). The fibroblasts collected 2, 4, and 6 weeks after the Cs administration were called in vivo test groups : T2, T4, T6, respectively. Immunofluorescence microscopy was used to detect CoL-I in all the fibroblasts. CoL-I was analyzed at both the gene and protein expression levels by real-time polymerase chain reaction and western blotting. Changes in CoL-I before and after Cs treatment were evaluated from the gingiva of each rat. There was no significant difference in gene expression of CoL-I in the control and test groups. CoL-I protein expression levels of fibroblasts increased in in vitro Cs treatment for each individual, and also increased in in vivo Cs treatment. In this study, the experimental method that control biological variations that can occur due to differences among individuals was useful. Subsequent studies on other factors besides CoL-I and in-depth studies in humans are needed.

• The Korean Journal of Pain
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• v.34 no.1
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• pp.19-26
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• 2021

Background: Prolotherapy is a proliferation therapy as an alternative medicine. A combination of dextrose solution and lidocaine is usually used in prolotherapy. The concentrations of dextrose and lidocaine used in the clinical field are very high (dextrose 10%-25%, lidocaine 0.075%-1%). Several studies show about 1% dextrose and more than 0.2% lidocaine induced cell death in various cell types. We investigated the effects of low concentrations of dextrose and lidocaine in fibroblasts and suggest the optimal range of concentrations of dextrose and lidocaine in prolotherapy. Methods: Various concentrations of dextrose and lidocaine were treated in NIH-3T3. Viability was examined with trypan blue exclusion assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Migration assay was performed for measuring the motile activity. Extracellular signal-regulated kinase (Erk) activation and protein expression of collagen I and α-smooth muscle actin (α-SMA) were determined with western blot analysis. Results: The cell viability was decreased in concentrations of more than 5% dextrose and 0.1% lidocaine. However, in the concentrations 1% dextrose (D1) and 0.01% lidocaine (L0.01), fibroblasts proliferated mildly. The ability of migration in fibroblast was increased in the D1, L0.01, and D1 + L0.01 groups sequentially. D1 and L0.01 increased Erk activation and the expression of collagen I and α-SMA and D1 + L0.01 further increased. The inhibition of Erk activation suppressed fibroblast proliferation and the synthesis of collagen I. Conclusions: D1, L0.01, and the combination of D1 and L0.01 induced fibroblast proliferation and increased collagen I synthesis via Erk activation.

• Journal of Oral Medicine and Pain
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• v.20 no.1
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• pp.53-65
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• 1995

The growth and synthetic activities of fibroblasts are regulated by cytokines and growth factors derived from activated inflammatory cells. Stimulatory effect of low level laser (LLL) radiation on wound healing seems to be in part due to direct stimulatory action on cell proliferation and synthetic activities of fibroblasts. Also indirect stimulatory effect on the fibroblast function through inflammatory or immune cells is another possible mechanism of biostimulatory action of LLL. This study was performed to determine the growth rate of human gingival fibroblasts obtained biopsy and culture, fibroblast cell line, and immune cell line by using $[^3H]-$ thymidine incorporation test. And gene expression pattern was also analyzed by using the DNA probe such as Hsp70, IL-1$\beta$, MIP-1$\alpha$ and actin cDNA. Proliferation rate of gingival fibroblast was increased by LLL irradiation, but no more effect was added by LPS or IL-1$\beta$ pretreatment Enhanced Hsp70 gene expression was found from gingival fibroblasts and fibroblast cell line COS by LLL irradiation., which was not more increased by LPS or IL-1$\beta$ pretreatment. LLL-irradiated promyelcytic cell line HL-60 and macrophage cell line RAW264.7 showed significant stimulatory effect of proliferation rate when compared with respective control. However there were no changes in growth rate of other immune cell tested in this study, such as B cell line WR19n.l and 230, helper T cell line Jurkat and Hut78, cytolytic T cell line CTLL-r8. By LLL-irradiation Hsp70 gene expression was increased in RAW246.7 and HL-60, not in CTLL-R8. And IL-1$\beta$ and MIP-1$\alpha$ gene expression were induced only from LLL-irradiated RAW264.7. These results led us to presume that LLL radiation may affect to the immune cells, especially to macrophage, through which it might promote wound healing process.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.6
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• pp.951-957
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• 2004

Hypoglycemic effect of Prunus mume (PM) extract containing in Sangjinyangheul-tang and Hwangkeumtang, one of the diabetic herbal medicines, was determined by investigating insulin-like action, insulin sensitizing action and a-glucoamylase suppressing action. Insulin-like activity of 3T3-L1 fibroblast was not shown with the treatment of PM methanol extracts. However, treatment with 20% or 40% PM methanol extracts and differentiation inducers significantly decreased the differentiation of 3T3-L1 fibroblasts to adipocytes. A significant insulin sensitizing activity was observed in 3T3-L1 adipocytes, giving PM extracts (60%, 80% and 100%) with 1 ng/mL insulin to reach glucose uptake level increased by 50 ng/mL of insulin alone. In addition, 20% and 40% methanol extracts of PM suppressed the a-glucoamylase activity by 30% in vitro. However, there was no significant differences in the peak of serum glucose levels and area under the curve in Sprague Dawley male rats treated with PM ethanol extract or cellulose and 2 g maltose or dextrin/kg body weight. These data suggested that PM extracts contain effective insulin sensitizing compounds, lipid synthesis suppressing compounds and possibly a-glucoamylase suppressing compounds. Therefore, PM extracts are beneficial for anti-diabetic treatment in obese diabetic patients.

• Applied Biological Chemistry
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• v.47 no.2
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• pp.258-264
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• 2004

Hypoglycemic effect of Schizandrae Fructus (SF) extract containing in Okchun-san was determined on 3T3-L1 fibroblasts and adipocytes by investigating insulin-like activity, insulin sensitizing activity and ${\alpha}-glucoamylase$ suppressing activity. SF were extracted by using 70% ethanol followed by XAD-4 column chromatography with a mixture solvent of methanol and water, and the fractional extractions were utilized for assaying hypoglycemic effect. No inhibition of ${\alpha}-glucoamylase$ activity of SF was observed. Insulin-like activity 3T3-L1 adipocytes was not shown by SF. A significant insulin sensitizing activity of SF extractions was observed in 3T3-L1 adipocytes, giving SF extractions with 1 ng/ml insulin to reach glucose uptake level increased by 50 ng/ml of insulin alone. When cells were treated with SF (Fr. 4 or 5) plus 1 ng/ml insulin, glucose uptake was increased more than seven times as compared to 1 ng/ml of insulin alone, suggesting that SF extracts increased GLUT4 content by enhancing insulin signaling. These data suggest that SF extracts (especially Fr. 4 and 5) contains an effective insulin sensitizing compounds for hypoglycemic activity in 3T3-L1 adipocytes.

• The Journal of Korean Medicine
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• v.23 no.1
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• pp.83-91
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• 2002

Objectives : In this study, water extracts from Coix lachryma-jobi Linne var. mayuen Stapf. were investigated for their effects on insulin-like action and glucose uptake in 3T3-Ll cells. Methods : We examined the effects of insulin-like action on the differentiation of 3T3-Ll fibroblasts. The Coicis Semen was treated with hot water and the extract was freeze-dried. The hot water extract was chromatographed on nonionic polymer resin (Amberlite XAD-4, Sigma) with distilled water (Fr. 1), 20% (Fr. 2), 40% (Fr. 3), 60% (Fr. 4), 80% (Fr. 5), and 100% EtOH (Fr. 6), successively. Results : Total extract of Coicis Semen was fractionated into 0 to 100% MeOH with Amberlite XDA-4 column. Treatment of cells with $10{\;}\mu\textrm{g}/ml$ of total extracts of Coicis Semen significantly increased the differentiation (p<0.05). At $1{\;}\mu\textrm{g}/ml$ level of insulin, the differentiation was accelerated (p<0.01). The effect of extracts plus insulin on the differentiation was greater than that of insulin alone. In 3T3-Ll adipocytes, glucose uptake was higher by 2.7 times with 5 uM of total extract in low dosage of insulin (3 ng/ml) than without total extract. 5 and 50 uM of water and 40% MeOH fractions increased glucose uptake by 3.5 times in 3T3-Ll adipocytes (p<0.00l). Conclusions : Coicis Semen contains compounds which playa role of insulin-like action and insulin sensitizer.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.440-447
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• 2003

Up until today, the key to contouring has been resumed in these two alternatives, either limiting the adipocyte storing capacity by modulating lipogenesis, or by stimulating lipolysis to eliminate adipocyte lipid content. Another interesting way could be the regulation of adipocyte differentiation. In this work, we have evaluated the effect of a brown algal extract of Sphacelaria scoparia (SSE) on the differentiation of pre-adipocytes into adipocytes. A pre-adipocyte line (3T3-L 1) was used. The differentiation was evaluated by the measure of produced lipids thanks to red oil coloration and spectrophotometry, and also by the expression of adipocyte differentiation markers: enzymes such as fatty acid synthase (FAS) and stearoyl CoA desaturase (SCD), or membrane proteins such as glucose transporters (GLUT -4) and fatty acid transporters (FAT) expressed on the surface of human adipocytes. These genes are under control of two transcription factors: CAAT-enhancer binding protein (c/EBP alpha) and sterol response element binding protein (SREBP1). All these markers were analysed at different stages of differentiation by RT -PCR. Sphacelaria extract (SSE) inhibits pre-adipocytes differentiating into adipocytes following a dose-dependant relation, using a kinetics similar to retinoic acid. It decreases the expression of mRNA specific to FAS, FAT, GLUT -4, SCD1, c/EBP alpha and SREBP1. Moreover, SSE regulated on collagen 1 and collagen 4 expression. A stimulation of collagen 1 was also measured in human skin fibroblasts. Thus, SSE performs as a genuine differentiation inhibitor and not only as a lipogenesis inhibitor, and could be used in slimming products.

• Food Science and Preservation
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• v.16 no.1
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• pp.115-121
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• 2009

We prepared five different kinds of herb mixtures prescribed for hypoglycemic effect. And the physicochemical properties of their water extracts were assessed to identify functional materials. Yields were in the range $19.52{\sim}29.79%$. Total phenolics and flavonoid contents were $349.24{\sim}1,752.21\;mg%$ and $163.06{\sim}1,118.47\;mg%$, respectively, and herb mixtures No. 2, 3 and 5 showed particularly high levels greater than 1,000 mg%. Electron-donating ability was best in herb mixtures showing high levels of total phenolics and flavonoids. Nitrite-scavenging abilities were more than 70% in herb mixtures No. 2 and 5, and decreased as pH increased. Herb mixture extracts strongly inhibited differentiation of 3T3-L1 fibroblasts, with potencies ranked in the herb mixture order 5, 1, 4, 3, and 2. The five different kinds of herb mixtures prescribed for their hypoglycemic effects may be useful as functional food materials.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.1
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• pp.42-47
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• 2006

L-Ascorbic acid (AsA) is an essential nutrient for prevention of scurvy in humans, primates and guinea pigs that lack $L-gulono-\gamma-lactone$ oxidase which is required for the final step of AsA biosynthesis. AsA participates in various hydroxylation reactions involved in the biosynthesis of collagen. The purpose of this study is to clarify the role of AsA on collagen synthesis in 3T6 fibroblasts and primary cultured cells of chondrocytes. Cells were cultured in medium supplemented with catalase and AsA at various concentration. Supplement of AsA induced collagen synthesis in 3T6 fibroblasts and primary cultured cells of chondrocytes. The most remarkable induction of collagen synthesis by AsA was found in primary cultured chondrocytes. The content of collagen representing the amounts of extracellular matrix significantly increased in the cells of which growth was stimulated by AsA, while it decreased with increasing passage numbers of subculture in cells. It showed that the content of collagen decreased in the medium which contained AsA at the concentration higher than 5.0 mM. However, the contents of collagen to DNA were not different among various AsA concentrations. Supplementing with AsA resulted in enhancement of collagen formation and extracellular matrix. Therefore, there might be a Positive correlation between the activity of catalase and the AsA concentration. Moreover, it can be assumed that AsA stimulates the collagen synthesis by optimizing the cell-culture environment.

• Nutrition Research and Practice
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• v.8 no.6
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• pp.613-617
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• 2014

BACKGROUND/OBJECTIVES: Adipogenesis is part of the cell differentiation process in which undifferentiated fibroblasts (pre-adipocytes) become mature adipocytes with the accumulation of lipid droplets and subsequent cell morphological changes. Several transcription factors and food components have been suggested to be involved in adipogenesis. The aim of this study was to determine whether mulberry leaf ethanol extract (MLEE) affects adipogenesis in 3T3-L1 adipocytes. MATERIALS/METHODS: The 3T3-L1 adipocytes were treated with different doses of MLEE for 8 days starting 2 days post-confluence. Cell viability, fat accumulation, and adipogenesis-related factors including CCAAT-enhancer-binding protein alpha ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$), $PPAR{\gamma}$ coactivator 1 alpha (PGC-$1{\alpha}$), fatty acid synthase (FAS), and adiponectin were analyzed. RESULTS: Results showed that MLEE treatments at 10, 25, 50, and $100{\mu}g/ml$ had no effect on cell morphology and viability. Without evident toxicity, all MLEE treated cells had lower fat accumulation compared with control as shown by lower absorbances of Oil Red O stain. MLEE at 50 and $100{\mu}g/ml$ significantly reduced protein levels of $PPAR{\gamma}$, PGC-$1{\alpha}$, FAS, and adiponectin in differentiated adipocytes. Furthermore, protein level of $C/EBP{\alpha}$ was significantly decreased by the treatment of $100{\mu}g/ml$ MLEE. CONCLUSION: These results demonstrate that MLEE treatment has an anti-adipogenic effect in differentiated adipocytes without toxicity, suggesting its potential as an anti-obesity therapeutic.