• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.21 no.4
• /
• pp.332-344
• /
• 1999

Cytokines are hormone-like proteins which mediate and regulate inflammatory and immune responses. Interleukin-6 (IL-6) is involved in the final differentiation of B cells into antibody-producing cells. Interleukin-8 (IL-8) is a neutrophil chemotactic factor that plays an important role in the recruitment of neutrophil to inflammatory loci. Inflammatory mediators by cells in the gingiva have been implicated in the initiation and progression of periodontitis and oral infection. The purpose of this study was conducted to investigate the effect of lipopolysaccharide (LPS), staphylococcus enterotoxin B (SEB) on production of IL-6 and IL-8 by human gingival and facial dermal fibroblasts. Primary cultured human gingival and facial dermal fibroblasts were incubated with LPS (0.01, 0.1, $1.0{\mu}g/ml$), SEB (0.01, 0.1, $1.0{\mu}g/ml$) or LPS $(0.1{\mu}g/ml)$ plus SEB $(0.1{\mu}g/ml)$. Culture supernatants were collected at 24, 48, and 72 hrs and assessed for IL-6 and IL-8 production by enzyme-linked immunosorbent assay. IL-6 production in gingival fibroblasts stimulated with LPS was higher than that with SEB. IL-6 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-6 production in facial dermal fibroblasts was increased only by stimulation with a high concentration of LPS $(1.0{\mu}g/ml)$. Its production in facial dermal fibroblasts by exposure with SEB was decreased in comparison with control, nontreated cells. Therefore, gingival fibroblasts showed higher sensitivity than facial dermal fibroblasts in response to low concentration of LPS. Also, IL-6 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-8 production in gingival fibroblasts was enhanced greatly only by stimulation of high concentration of LPS $(1.0{\mu}g/ml)$. That by exposure with SEB was increased only in 24 hrs cultivation. IL-8 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-8 production in facial dermal fibroblasts was decreased by LPS and increased only in 48 hrs cultivation by SEB. IL-8 production by double exposure with LPS plus SEB was enhanced only in 48 hrs cultivation in comparison with single exposure of LPS or SEB. therefore, IL-6 and IL-8 production were released at various quantities according to bacterial toxin applied and site of fibroblast harvested. These results suggest that gingival fibroblasts may be concerned with IL-6 and IL-8 related inflammatory response more than facial dermal fibroblasts.

• Tuberculosis and Respiratory Diseases
• /
• v.77 no.2
• /
• pp.73-80
• /
• 2014

Background: Low levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells. Methods: HFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-$1{\beta}$ (IL-$1{\beta}$) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D ($1,25(OH)_2D$) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction. Results: IL-$1{\beta}$ significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and $1,25(OH)_2D$ (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and $1,25(OH)_2D$. Conclusion: Vitamin D, 25(OH)D, and $1,25(OH)_2D$ play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-$1{\beta}$ stimulated MMP-9 production and conversion to its active form but also inhibiting IL-$1{\beta}$ inhibition on TIMP-1 and TIMP-2 production.

• Journal of Ginseng Research
• /
• v.44 no.2
• /
• pp.341-349
• /
• 2020

Background: The replicative senescence of human dermal fibroblasts (HDFs) is accompanied by growth arrest. In our previous study, the treatment of senescent HDFs with Rg3(S) lowered the intrinsic reactive oxygen species (ROS) levels and reversed cellular senescence by inducing peroxiredoxin-3, an antioxidant enzyme. However, the signaling pathways involved in Rg3(S)-induced senescence reversal in HDFs and the relatedness of the stereoisomer Rg3(R) in corresponding signaling pathways are not known yet. Methods: We performed senescence-associated β-galactosidase and cell cycle assays in Rg3(S)-treated senescent HDFs. The levels of ROS, adenosine triphosphate (ATP), and cyclic adenosine monophosphate (cAMP) as well as the mitochondrial DNA copy number, nicotinamide adenine dinucleotide (NAD)⁺/1,4-dihydronicotinamide adenine dinucleotide (NADH) ratio, and NAD-dependent sirtuins expression were measured and compared among young, old, and Rg3(S)-pretreated old HDFs. Major signaling pathways of phosphatidylinositol 3-kinase/Akt, 5' adenosine monophosphate-activated protein kinase (AMPK), and sirtuin 1/3, including cell cycle regulatory proteins, were examined by immunoblot analysis. Results: Ginsenoside Rg3(S) reversed the replicative senescence of HDFs by restoring the ATP level and NAD⁺/NADH ratio in downregulated senescent HDFs. Rg3(S) recovered directly the cellular levels of ROS and the NAD⁺/NADH ratio in young HDFs inactivated by rotenone. Rg3(S) mainly downregulated phosphatidylinositol 3-kinase/Akt through the inhibition of mTOR by cell cycle regulators like p53/p21 in senescent HDFs, whereas Rg3(R) did not alter the corresponding signaling pathways. Rg3(S)-activated sirtuin 3/PGC1α to stimulate mitochondrial biogenesis. Conclusion: Cellular molecular analysis suggests that Rg3(S) specifically reverses the replicative senescence of HDFs by modulating Akt-mTOR-sirtuin signaling to promote the biogenesis of mitochondria.

• Korean Journal of Food Science and Technology
• /
• v.41 no.4
• /
• pp.441-445
• /
• 2009

Recent research has revealed that hydrolyzed collagen peptides have beneficial effects in various diseases such as osteoarthritis and human rheumatoid arthritis and also play a protective role in skin by improving the activity of antioxidants. In this study, we investigated the effects of a novel mixture (AP-CPM01) containing collagen peptides and elastin peptides on photoaged hairless mice skin both in vivo and in vitro. To evaluate the effects of AP-CPM01 on UVBinduced skin wrinkle formation in vivo, the hairless mice were exposed to UVB irradiation and orally administered the AP-CPM01 at 333 mg/kg per day for 10 weeks. The effects on skin appearance and epidermal thickness were measured using bioengineering and histochemical methods. In addition, the influence of AP-CPM01 on collagen metabolism in human skin fibroblasts was also investigated. The skin of mice in the AP-CPM01 treated group had better appearance and less wrinkling than that of mice in the control group. In the human fibroblast cells, the amount of de novo procollagen synthesis was increased after AP-CPM01 treatment, reflecting that AP-CPM01 can induce de novo procollagen synthesis and reduce UVB-induced skin wrinkle formation. These results suggest that AP-CPM01 is a potent candidate for antiphotoaging functions.

• The Korean Journal of Physiology and Pharmacology
• /
• v.10 no.5
• /
• pp.289-295
• /
• 2006

Sodium fluoride (NaF) has been shown to be cytotoxic and elicit inflammatory response in human. However, the cellular mechanisms underlying NaF-induced cytotoxicity in periodontal tissues have not yet been elucidated. This study is aimed to investigate the mechanisms of NaF-induced apoptosis in human gingival fibroblast (HGF). NaF decreased the cell viability of HGF in a dose- and time-dependent manner. NaF gave rise to apoptotic morphological changes including cell shrinkage, chromatin condensation, and DNA fragmentation. However, NaF did not affect the production of ROS. In addition, NaF augumented cytochrome c release from mitochondria into the cytosol, and enhanced caspase -9 and -3 activities., cleavage (85 kDa fragments) of poly (ADP-ribose) polymerase (PARP) and upregulation of voltage-dependent anion channel (VDAC) 1. These results demonstrated that NaF-induced apoptosis in HGF may be mediated with mitochondria. Furthermore, NaF elevated caspase-8 activity and upregulated Fas-ligand (Fas-L), suggesting involvement of death receptor mediated pathway in NaF-induced apoptosis. Expression of Bcl-2, an anti-apoptotic Bcl-2 family, was downregulated, whereas expression of Bax, a pro-apoptotic Bcl-2 family, was not affected in NaF-treated HGF. These results suggest that NaF induces apoptosis in HGF through both mitochondria- and death receptor-mediated pathway mediated by Bcl-2 family.

• Journal of Life Science
• /
• v.17 no.5 s.85
• /
• pp.687-693
• /
• 2007

Insulin-like growth factor-I(IGF-I) has significant insulin-like anabolic effects which include the stimulation of glucose and amino acid uptake, as well as protein and glycogen synthesis. IGFs exist in serum and other biological fluids as complexes bound to a family of structurally related insulin-like growth factor binding proteins(IGFBPs). Six human IGFBPs can modulate the effects of IGFs on target tissues by several mechanisms, including altering the serum's half-life and the transcapillary transport of IGFs, as well as changing the availability of IGFs to specific cell surface receptors. Human fibroblasts secrete IGFBPs that can modify IGF-I action. Previous to our study using either Northern blotting, and Western blotting have shown that fibroblasts express mRNA IGFBP-3, -4, and -5, and synthesize these proteins. In addition, fibroblast cell lysates revealed that the IGFBP-3 was most abundant. For these reasons, we undertook to gain further insight into the effects of high and low glucose incubation condition on metabolism and IGFBP-3 expression. In results of metabolites and IGFBP-3 expression in GM10 cells cultivated with various glucose concentration, the consumption of glucose and accumulation of triglyceride were increased in condition of high glucose, and total protein level was decreased. in the course of time. After 5 days incubation, levels of free amino acid in medium containing glucose of high concentration glucose were higher than in conditions of low glucose. Although the levels of IGFBP-3 protein and mRNA levels were increased in low glucose, and IGFBP-3 was not affected by any pretense. Taken together, we suggest that the study of growth factors, like IGFs, might be a possible model of diabetes militus in cell, although the results in cell models were not in accord with in vivo.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.31 no.1 s.49
• /
• pp.103-109
• /
• 2005

UV irradiation leads to distinct changes in skin connective tissue, which is degradation of collagen. Many of these alterations in the extracellular matrix are mediated by matrix metalloproteinases. In this study, to develop a new anti-aging agent, we screened the antioxidant activity of solvent fractions from ethanolic extract of Melothria Heterophylla. Among the four solvent fractions tested, the EtOAc fraction exhibited the highest antioxidant activity. It was investigated the inhibitory effect of the EtOAc fraction on the expression and activity of MMP-1 in UVA-irradiated human dermal fibroblasts. The EtOAc fraction inhibited the activity of MMP-1 in a dose dependent manner with the $IC_{50}$ values of $9{\mu}g/mL$. Also, UVA-induced MMP-1 expression was reduced about $90\%$ by $100{\mu}g/mL$ of the EtOAc fraction but MMP-1 mRNA expression was not inhibited. Therefore, we conclude that the EtOAc fraction significantly inhibits MMP-1 expression at the protein level. From these results, we suggest that the EtOAc fraction from M. heterophylla could be used as a new anti-aging agent for the photo-damaged skin.

• Journal of Periodontal and Implant Science
• /
• v.25 no.3
• /
• pp.741-755
• /
• 1995

Currently titanium is the material of choice for implants because of its biological acceptance. This high degree of biocompatibility is thought to result, in part, from the protective and stable oxide layer that presumably aids in the bonding of the extracellular matrix at the implant-tissue interface. Endosseous dental implants are interfaced with bone, connective tissue, and epithelium when implanted into the jaw bone. The soft tissue interface including connective tissue and epithelium is one of the most critical factors in the determination of implant maintenance and prognosis. For maintenance of failing or failed implants, it is essential to treat the implant fixture surface to remove bacterial endotoxins and make a surface tolerated by surrounding soft and hard tissues. In this study, the effect of mechanical treatment on titanium plasma sprayed implant on adhesiveness and proliferation of human gingival fibroblasts and changed surface characteristics were studied. titanium plasma sprayed discs manufactured by Friedrichsfeld company were treated with loaw speed stone bur, a rubber point and a jetpolisher. Its surface components were analyzed with Energy dispersive X-ray spectroscopy to evaluate whether the surface characteristics were altered or not. To observe the spreading pattern of the human gingival fibroblasts which attached to the all specimens author used the scanning electron microscope. The results were as follows : Pure titanium and plasma sprayed titanium, stone polished titanium showed titanium peak and small amout of aluminum, so there was no alteration on surface characteristics. Under the scanning electron microscopic examination in the initial attachment of human gingival fibroblast, there was a slight enhancement in pure titanium, stone polished titanium than plasma sprayed titanium. After 6 hours, the pure titanium and stone polished titanium showed human gingival fibroblasts were elongated and connected with numerous processes. Human gingival fibroblasts were more intimately attached on the pure titanium discs than on the other discs. The human gingival fibroblasts attached on the plasma sprayed titanium by thin and elongated processes. After 24 hours, the human gingival fibroblasts connected with each other via numerous processes and compeletly covered the pure titanium and stone polshed titanium discs. Human gingival fibroblasts had multiple point contacts with more long and thin lamellopodia and showed a little bare surface on plasma sprayed titanium discs.

• Asian-Australasian Journal of Animal Sciences
• /
• v.19 no.11
• /
• pp.1574-1579
• /
• 2006

This study was performed to determine the developmental potentials of nuclear transfer (NT) embryos using life-span extended cells transfected with a foreign gene as donor cells. A life-span extended bovine embryonic fibroblast cell line was transfected with an expression vector in which the human type II collagen (BOMAR) and ear fibroblasts were used as a donor cell. Cytogenetic analysis was performed to analyze the chromosomal abnormality of donor cells. The fusion rate of 1.8 kV/cm for $15{\mu}sec$ given twice was significantly higher than that of other groups (p<0.05) and the embryos lysed were significantly higher after 1.8 kV/cm for $20{\mu}sec$ given once compared to other groups (p<0.01). The blastocyst development in the ear cell group was statistically significant compared to both BOMAR groups (p<0.01). Both BOMAR groups cultured more than 40 passages (>40 passages) had a lower number of chromosomes; however, fresh granulosa cell (GC) and BOMAR groups cultured less than 20 passages had normal chromosome numbers. Both >40 passages BOMAR groups had numerous obscure debris in metaphase spreads. The transfected foreign gene was expressed in all BOMAR groups, but not in the GC group. Based on these results, the lower developmental potential of NT embryos using life-span extended donor cells transfected with a foreign gene might be a cause of chromosomal abnormality in donor cells.

• Journal of Animal Reproduction and Biotechnology
• /
• v.34 no.3
• /
• pp.232-239
• /
• 2019

In general, cloned pigs have been produced using the somatic cell nuclear transfer (SCNT) technique with various types of somatic cells; however, the SCNT technique has disadvantages not only in its low efficiency but also in the development of abnormal clones. This study aimed to compare early embryonic development and quality of SCNT embryos with those of induced pluripotent stem cells (iPSCs) NT embryos (iPSC-NTs). Ear fibroblast cells were used as donor cells and iPSCs were generated from these cells by lentiviral transduction with human six factors (Oct4, Sox2, c-Myc, Nanog, Klf4 and Lin28). Blastocyst formation rate in iPSC-NT (23/258, 8.9%) was significantly lower than that in SCNT (46/175, 26.3%; p < 0.05). Total cell number in blastocysts was similar between two groups, but blastocysts in iPSC-NT had a lower number of apoptotic cells than in SCNT (2.0 ± 0.6 vs. 9.8 ± 2.9, p < 0.05). Quantitative PCR data showed that apoptosis-related genes (bax, caspase-3, and caspase-9) were highly expressed in SCNT than iPSC-NT (p < 0.05). Although an early development rate was low in iPSC-NT, the quality of cloned embryos from porcine iPSC was higher than that of embryos from somatic cells. Therefore, porcine iPSCs could be used as a preferable cell source to create a clone or transgenic animals by using the NT technique.