Soy isoflavones have been reported to possess many physiological activities such as antioxidant activity and inhibition of cancer cell proliferation. This study investigated the photoprotective effects of soybean extract in human fibroblast cell line and hairless mice model. Human fibroblast was treated with soybean extract before and after ultraviolet B (UVB; 290-302 nm) irradiation. In the soybean extract treated group, the cells showed better resistance to ultraviolet (UV) than control group. The amount of type I collagen recovered from the soybean treated group was higher than the vehicle group exposed to UV-induced damage. Moreover, increased expression of metalloproteinases-1 as a result of UV irradiation was suppressed by the soybean extract. Female mice were orally administered soybean extract and irradiated with UVB light for 8 weeks. The effects of the soybean extract on the skin appearance, collagen deposition and epidermal thickness in the UV-damaged mouse skin were analyzed using histopathological methods. In soybean extract treated group, the skin had a better morphology than that of the control group. Furthermore, the amount of type I collagen was increased and overexpression of MMP-1 was reduced in the soybean extract group compared to vehicle group. Additionally, up-regulation of pro-inflammatory cytokines induced by UV irradiation was suppressed by dietary soybean extract treatment. It appears that soybean extract had a photoprotective effect, including anti-aging and anti-inflammatory effect, from UV-induced damage in not only human fibroblast, but also hairless mice. We confirmed that these effects were possibly due to promotion of collagen synthesis and inhibition of MMP-1 expression.

Ultraviolet B (UVB) exposure is a risk factor for skin damage resulting in oxidative stress, inflammation, and cell death. The purpose of this study was to investigate the physicochemical properties of Platycodon grandiflorum (PG) to improve its biological activities using a three-step steaming process. We investigated the protective effects of PG and steamed PG extracts on human dermal fibroblasts (HDFs) against UVB radiation-induced oxidative stress and inflammation as well as the underlying mechanisms. The antioxidant potential of the PG extracts was evaluated by measuring the 2,2-diphenyl-1- picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) scavenging activity. ABTS and DPPH were shown by the 0, 30, and 70% ethanol extracts of 2S-PG and 3S-PG (IC50, 28~45 and 27~30 μg/mL, respectively). Treatment of UVB-irradiated cells with steamed PG (25~400 μg/mL) did not affect their viability. The streamed PG extract suppressed UVB-induced generation of reactive oxygen species (ROS). In addition, streamed PG extract reduced cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) protein expression in UVB-irradiated HDF, regulating nuclear factor (NF)-κB expression. These findings suggest that steamed PG extract may be potentially effective against inflammation associated with UVB-induced oxidation stress.

본 연구는 예부터 약용으로 사용되었거나, 현재 식품공 전에 식품원료로 사용이 가능한 것으로 등록된 국내 산림 지역에 자생하는 식물을 식품산업에 활용하고자 선행연구에서 우수한 항산화 활성을 보인 갈참나무 잎을 본 연구 에서 사용하였다. 70% 에탄올을 이용하여 추출한 갈참나무 잎을 이용하여, hydrogen peroxide로 산화적 스트레스를 유도한 피부 섬유아세포에서의 세포 보호효과, 세포내 항산화 효과 및 항노화 효과를 측정하였다. 세포 독성을 평가한 결과 25, 50 및 100 μg/mL의 갈참나무 잎 추출물을 처리하였을 때 모두 독성을 나타내지 않았으며, hydrogen peroxide로 산화적 스트레스를 유도한 상태에서는 세포를 보호하여 농도 유의적으로 세포생존율이 증가하였다. 특히, 100 μg/mL의 농도에서는 양성대조군으로 사용한 50 μM ascorbic acid 수준까지 세포 생존율이 증가하였다. 세포내 항산화 효과를 확인 하기위해 사용한 H2-DCFDA assay에서는 형광현미경과 형광흡광도 측정에서 모두 농도 유의적으로 세포내 ROS 저감 활성을 확인하였고 갈참나무 잎 추출물을 100 μg/mL 농도로 처리했을 때는 50 μM ascorbic acid와 비슷한 세포내 항산화 효과를 나타내었다. 또한 SA- β-galactosidase assay를 이용한 갈참나무 잎 추출물의 피부 섬유아세포에 대한 항노화활성은 ROS 생성 억제 효과 와 유사한 경향으로 갈참나무 잎 추출물의 농도 유의적으로 세포 노화 억제효과를 확인하였다. 이상의 결과를 종 합하여 볼 때 갈참나무 잎 추출물이 hydrogen peroxide로 인한 산화적 스트레스 상태에서 세포 보호효과, 항산화 효 과 및 항노화 효과가 관찰되어 기능성 식품원료로서의 활용도가 매우 넓을 것으로 판단된다.

Adhesive capsulitis of the shoulder is a common cause of pain that occurs during shoulder movement, thereby restricting shoulder rotation in clinical practice. Although most patients respond to pain relief treatment (NSAID or corticosteroids) by improving their range of motion, it remains poorly understood without any definitive treatment algorithm. In addition to immune cells, synoviocytes, chondrocytes and osteoblasts in the joint are known to produce pro-inflammatory mediators such as reactive oxygen species (ROS), inflammatory cytokines and lipid mediators, presumably contributing to the pathogenesis of osteoarthritis (OA) and adhesive capsulitis. Although inflammation and also fibrosis are proposed to be the basic pathological changes of a frozen shoulder, there is a lack of information regarding the downstream targets of the pro-inflammatory ROS signaling pathway in the synoviocytes and also how these ROS targets are modulated at the transcription level by a corticosteroid - dexamethasone. In this study, we used human fibroblast like synoviocytes (HFLS) to characterize the signaling targets of ROS by employing a human DNA microarray tool and studied the role of dexamethasone in this process. Our data suggest that several genes such as FOS, FOSB and NFkBIZ, which are known to be involved in pro- or anti- inflammation response, are modulated at the transcription level by ROS and dexamethasone.

현재 식품공전에 식품원료로 사용이 가능한 것으로 등 록된 국내 산림지역 자생 식물을 식품산업에 활용하고자 국내 자생식물 45종을 선별하였고, 본 연구팀의 선행연구 에서 45종의 항산화 활성을 평가한 결과, 다른 종들과 비 교하여 우수한 항산화 활성을 보인 밤나무 잎을 본 연구 에서 사용하였다. 본 연구에서는 hydrogen peroxide로 산화적 스트레스를 유도한 상태에서 밤나무 잎 추출물의 세포 보호효과, 세포내 항산화 효과 및 항노화 효과를 관찰 하였다. 세포 보호효과를 측정한 결과 밤나무 잎 추출물 의 농도 유의적으로 세포를 보호하는 효과를 확인하였고, 특히 100 μg/mL에서는 양성대조군으로 사용한 항산화 물질인 ascorbic acid 수준까지 세포를 보호하는 효과를 나타내었다. H2-DCFDA 염색을 통한 세포내 항산화 효과를 확인한 결과, 형광현미경과 형광흡광도 측정에서 모두 밤나무 잎 추출물이 농도 유의적으로 세포내 ROS를 저감을 확인하였고, 세포 보호효과와 마찬가지로 농도 100 μg/mL 에서는 ascorbic acid와 비슷한 수준을 보였다. SA-β- galactosidase 염색을 통하여 관찰한 세포노화 억제 효과는 ROS 생성 억제 효과와 유사한 경향으로 밤나무 잎 추출 물의 농도 유의적으로 세포노화 억제를 관찰되었다. 이상 의 결과를 종합하여 볼 때 밤나무 잎 추출물은 산화적 스트레스로 인한 세포노화를 억제하는 효능을 보였으며, 이는 천연물에서 유래한 항산화 및 항노화 활성 첨가제로 활용도가 매우 넓을 것으로 판단된다.

Several human leukocyte subsets including natural killer (NK) cells, cytotoxic T lymphocytes (CTL), and polymorphonuclear neutrophils (PMN) participate in cellular immune responses directed against vascularized pig-to-human xenografts. As these leukocytes express the death receptor Fas either constitutively (PMN) or upon activation (NK, CTL), we explored in vitro whether the transgenic expression of membrane-bound human Fas ligand (mFasL) on porcine fetal fibroblasts is a valuable strategy to protect porcine xenografts. cDNA of mFasL carrying the deletion at the cleavage site with metalloproteinase and lacking the death domain in its cytoplasmic tail was subcloned into pCAGGS expression vector driven by the chicken β-actin promoter containing blastidin- resistance cassette. The mFasL expression vector was transfected into mini-pig fetal fibroblasts by lipofection method. Blastidin-resistant cells were screened by PCR and FISH. The expression of mFasL was confirmed by Western blot and FACS with the mouse anti-human FasL antibody. Interaction of two transgenic clonal cell lines with human leukocytes was analyzed using functional assay for cytotoxicity. mFasL expressed on porcine fetal fibroblasts protected porcine fetal fibroblasts against killing mediated by human NK cells. The rate of NK cell mediated cytotoxicity was significantly reduced in transgenic clonal cells (54±10.80%) compared to normal minipig fetal fibroblasts. This result indicated that grafts of transgenic pigs expressing mFasL could control the cellular immune response to xenografts, and create a window of opportunity to facilitate xenograft survival.

Human gingival fibroblasts (hGFs) were reported to play an important role in inflammatory reactions to lipopolysaccharide (LPS) from P.gingivalis in the periodontal connective tissue. Although the biostimulatory effects of hyperbaric oxygen therapy, such as anti-inflammatory activity, have been reported, the pathological mechanism is not completely understood. This study examined the changes in the inflammatory cytokine profiles, which are produced after exposure to hyperbaric oxygen in P.gingivalis LPS-treated human gingival fibroblasts, and subsequently to examine the mitogen activated protein kinase (MAPK) pathway involved in cytokine production. Gingival fibroblasts with or without P.gingivalis LPS were exposed to hyperbaric oxygen, and the cytokine profiles in the supernatant were observed using a human inflammation antibody array. The expression of cyclooxyginase-2 (COX-2) protein, phosphorylation of extracellular signal-regulated kinase (ERK1/2), p38, and c-Jun-N-terminal kinase (JNK) MAPK by western blot analysis, and the amount of prostaglandin E2 (PGE2) in the supernatant by an enzyme-linked immunoassay were determined. COX-2 protein expression and PGE2productionwereincreasedsignificantlyintheP. gingivalis LPS-treated group, and were decreased by treating P. gingivalis LPS with hyperbaric oxygen. Treatment of P. gingivalis LPS in the gingival fibroblasts led an increase in the amount of pro-inflammatory-related cytokines interleukin-6 (IL-6) and IL-8 released, whereas hyperbaric oxygen inhibits the irrelease. Ananalysis of the MAPK signal transduction showed that hyperbaric oxygen induced a significant decrease in the level of P38 phosphorylation regardless of the presence or absence of LPS. In addition, hyperbaric oxygen promoted JNK phosphorylation, significantly in the presence of LPS. Hyperbaric oxygen can inhibit pro-inflammatory cytokines and mediate the MAPK signal pathway, and appears to be useful as an anti-inflammatory tool.

To investigate the differential expression of genes by 635nm LEDs irradiation in arachidonic acid-treated human gingival fibroblasts, cDNA microarray was carried out. Human gingival fibroblasts were primary cultured and arachidonic acid was treated to induce inflammation. 635nm of wave length was used for LEDs irradiation. The experimental group was categorized into four group ; control, only LEDs irradiation group, only arachidonic acid-treated group and arachidonic acid-treated with LEDs irradiation group. The expression of 8,078 genes were increased and the expression of 7,103 genes were decreased in only LEDs irradiation group. For arachidonic acid-treated with LEDs irradiation group, the expression of 6,815 genes were increased, while the expression of 8,031 genes were decreased comparing with only arachidonic acid-treated group. IL-13alpha2 receptor was the most expressed gene in LEDs irradiation group comparing with control, followed by MMP3. Genes which the most down regulated was BIRC3 in LEDs irradiation group. PLAB genes was the most up-regulated in arachidonic acid treated with LEDs irradation group, followed by ranked RARRES1. Considering the classification by cell function, genes associated with signal transduction were the most affected by LEDs irradiation, followed by the genes associated with nucleoside, nucleotide and nucleic acid metabolism. In arachidonic acid treated with LEDs irradiation, genes associated with signal transduction and protein metabolism were affected. Taken together, LEDs irradiation could affect various biological process and could identify many genes related to LEDs irradiation, which could be used for clinical application.

곰피추출물의 CCD-986sk cell line monolayer (human fibroblast, KCBL-21947)에 대한 피부세포 생리활성효과를 측정하고, 또한 곰피추출물의 Clone M-3 mouse melano-cyte cell line에 대한 melanin formation 저해효과를 측정하기 위해 in vitro레벨에서 실험을 실시하였다. 곰피는 다년생 갈조류의 일종으로 이 종은 한국 연안해역에서 중요한 1차생산자의 역할을 담당하고 있는

Many researchers are interested in wound healing in the t reatment of burns, prevention of post surgical adhesions and cosmetic s urgery by excess collagen production and scar formatlOn Synthetic epidermal substi tutes with cultured epi thelial cells seem to be an attractive strategy since keratinocytes have been demonstrated to modulate fibroblast growth and collagen synthesis. Bioa bsorbable and biocompatible chitosan structurally mimics hyaluronic acid. Recently, a bio compatible synthesi zecl ch itosa n-PVP(polyvinyl pyrrolidone) hydrogels demonstrated in vitro biocompat ibi li ty for bio medical applications . However. there is no re port on this hydrogeJ"s ability to modulate human gingival fibroblast growth. The purpose of this study were to investigate different growth modulation between human gingival fibroblast and normal human oral keratinocyte by chitosan- PVP hydrogel, and to apply this biocompatible synthetic polymer to oral and maxillofacial wound healing. We have synthesized a hydrogel from chitosan-PVP and examined its effect on human gingival fibroblast growth modulation in vitro. Non-toxic and biocompatible hydrogel with human gingival fi broblasts and epithelial cells was tested by MTT assay. HGF showed a higher growth proliferation than that of NHOK after cell seeding. In MTT assay, 30% hydrogel leach out products showed a higher cellular viability in NHOK than that of any other products. In MTT assay, 30% hyclrogel leach out products showed relatively lower cellular viability of HGF ln growth profile, NHOK showed about 7 fo lcls higher than HGF after 1 day, while about 2 fo lds higher after 5 days. And also NHOK showed above about 70% cell ular via bility from 1 to 7 days. It suggested that Chitosan-PVP hydrogel would inhibit relatively the growth of HGF and s timulate the growth of NHOK_ This phenomenon may prove to be of use in wound management 0 1' oral and maxillofacial area as epitheli al substitutes.

Periodontitis is a chronic infectious disease that leads to periodontal destruction, and is one of the major causes of tooth loss in humans. The osteoclast differentiation factor (ODF), which is also known as the receptor activator of the NF-kB ligand (RANKL), is a surface-associated ligand on bone marrow stromal cells and osteoblasts. RANKL activates its cognate receptor, RANK, on osteoclast progenitor cells, which leads to the differentiation of mononucleated precursor cells. Osteoprotegerin (OPG) is a decoy receptor that is released from stromal cells and osteoblasts to inhibit the interaction between RANKL and RANK. Although the precise mechanism of bone loss in periodontitis is unknown, the differentiation and activation of osteoclasts by OPG-ODF-RANK signaling might play the role in periodontal bone destruction. The relationship between the concentration of sex hormones and the expression of ODF and OPG was examined by treating human gingival fibroblasts and periodontal ligament cells with the normal serum concentration of estrogen or progesterone during menstruation or at menopause. The ODF/OPG relative ratio was elevated at the concentration observed during ovulation in human gingival fibroblasts and at the concentration observed between ovulation and menstruation in periodontal ligament cells treated with estrogen. However, the ratio was <1 at all concentrations in both cells treated with progesterone. In the case of menopause simulated by estrogen depletion, the ratio was <1 in human gingival fibroblasts but >1 in periodontal ligament cells.

The process 0 1' wound healing needs the deposition of collagen and non-collagenous compounds, followed by the remodelling of extracellul ar matrix Recently, it has known that LEDs irradiation can help wound healing to accelerate the cell proliferation. But its mec hanism is not elucidated yot. The purpose of the present study is to observe the expression level of extracellular matrix by 635nm LEDs ir radiation. Human gingival fibroblasts were primary cultured, treated arac hidonic acid (때 and followed by LEDs irradiation. To observe the mRNA expression of extracellular matrix, cDNA mlcroarray was ca n‘ ied out 1n present study, 3 experimental groups were categorized into control, AA-treated group, and AA-treated with LEDs irradiation group. The differential expressions of MMP-1, -2, -3, - 10, - 11, -14, -16, - 17, -25 and TIMP-1, -2, -3. -4 were observed. Especially, mRNA expression of T1MP-3 was 10 fold decreased in arach idonic acid -treated with 635nm LEDs irradiation group. Finally, LEDs irradation can affect the expression level of MMPs and TIMPs, which lead to prolifer ation of gingival fibroblasts and result in would healing

It has been reported that light-emitting diodes(LED) can be used in the treatment of oral diseases. Although bio-stimulatory effects of LED irradiation such as promotes stimulation of wound healing have been well known, there are few reports about molecular mechanism associated with cell cycle by LED irradiation. The purpose of present study was to examine the molecular event in cell cycle of LED irradiation on primary human gingival fibroblast(hGF) in vitro. The source of light for irradiation was a continuous-wave LED emitting at a wavelength of 635nm, and manufactured that energy density was 5mW/cm2 on sample surface. The hGF were irradiated for 1 hour at 37℃ in 5% CO2 humidified chamber. Experimental samples were acquired at 0 (right after irradiation), 8 and 24 hour after irradiation. To investigate the molecular mechanisms associated with cell cycle, growth phase was determined by flow cytometry and mRNA expression of cyclin A, cyclin B, cyclin D1, cyclin E, cdc2, PCNA, p18, p27, p21, and p53 were determined by real time RT-PCR. Flow cytometric analysis demonstrated the percentage of cells in the G1 and S phase were decreased, but the G2 phase increased, which showed cells irradiated by LED were transitioned from S to G2 phase. For mRNA expression, cyclin B, cdc2, PCNA and p53 were increased at 0 hour after irradiation, and most of cell cycle molecules were increased at 8 hour after irradiation. At 24 hour after irradiation, cyclin A, cyclin E, PCNA and p18 were increased. Taken together, LED irradiation induced proliferation of hGF cells through transition from S to G2 phase.

Periodontitis is a chronic infectious disease that leads to the destruction, one of the major cause of tooth loss in human. Osteoclast Differentiation Factor(ODF), also called as Receptor activator of NF-xB ligand(RANKL), a surface-associated ligand on bone marrow stromal cells and osteoblasts, activates its cognate receptor RANK on osteoclast progenitor cells, which leads to differentiation of these mononucleated precursor cells. Osteoprotegerin(OPG), a decoy receptor, is released from stromal cells and osteoblasts to inhibit the interaction between RANKL and RANK. The experiment for the effect of pregnancy on gingival health showed greater gingival inflammation and edema during pregnancy, despite similar plaque index. There should be many factors affecting the periodontal health in pregnancy. In this experiment, we examined the direct effects of sex hormones(estrogen and progesterone) on the ODF/OPG expression in human gingival fibroblasts and periodontal ligament cells at the serum concentration of pregnancy. The ratio was high in the 1st trimester of pregnancy by estrogen and in the late 2nd trimester by progesterone. Therefore, the local periodontal destruction might be accelerated by these hormonal effect on the periodontal cells.

The aim of this study was to investigate the cytotoxicity of dental casting gold alloys. Recently, "biocompatability" is considered the most important requirement of dental materials. Dental metals and alloys were estimated by quantity of released ions, which had influenced to living tissues. The requirement of using normal human cells for cytoxicity strudy were abruptly increased. We used the cultured normal human gingival fibroblasts to estimate the cytotoxicity of dental casting gold alloys. The product of S company(Korea, AIGIS-SOFT, AIGIS-PLUS, AIGIS-A, AIGIS-PT, experimental group) and D company's (German, Biocclus inlay, Biolor SG, Stabilor NF Ⅳ, Degulor B, control group) dental casting gold alloys were used. The morphological investigation, hemolysis test, MTT assay and SRB assay were done in vitro. In vivo, inflammatory reaction in rat was examined for 2 weeks. 1. In the result of cytotoxicity assay, there were some differences but was no significancy among the results between two group's hemolysis, MTT and SRB assay. 2. The gingival fibroblasts attached to the surface of dental casting gold alloy showed various features and increased in number as the time had passed. 3. In vivo, chronic inflammatory cell infiltration was prominent from 3 days to 1 week and inflammation was reduced as time had gone. From the aboving results, there were no significant differences in cytoxicity depending on the ratio of gold content, but showed differences depending on the ratio of total precious and non-precious metal content between two groups. In vitro study showed few differences in inflamation reaction.

In order to investigate whether or not CCD-980sk cell line can be affected by Korean Citrus junos and medicinal herbs, we examined the MTT assay when we treated Korean Citrus junos and medicinal herbs in CCD-986sk human fibroblast cell line. The samples t