• Title/Summary/Keyword: In-vitro study

Search Result 8,888, Processing Time 0.052 seconds

The Effect of 1,25-Dihydroxyvitamin D3 on the Viability of Periodontal Ligament Cells and the Experimental Tooth Movement in Rats (1,25-Dihydroxyvitamin D3가 치주인대세포활성 및 실험적 치아이동에 미치는 영향에 관한 연구)

  • Kim, Sung-Woo;Park, Dong-Kwon;Kim, Sang-Cheol
    • The korean journal of orthodontics
    • /
    • v.27 no.2
    • /
    • pp.335-347
    • /
    • 1997
  • Vitamin D is known to exert its action by activating DNA and RBA within target cells to produce proteins and enzymes that can be used in bone resorption process. Particularly, the active form of vitmain D, 1,25-dihydroxycholecalciferol $[1,25-(OH)_2D_3]$, is considered to be one of the most potent stimulators of osteoclatic acitivity in vitro. The purpose of this study was to evaluate the effect of 1,25-Dihydroxyvitamin $D_3$ on the avtivity of periodotal ligament cells and, the experimental tooth movement. Human periodontal ligament cells were collected from the first premolar tooth extracted for the orthodontic treatment, and were incubated in the environment of $37^{\circ}C$, 5% $CO_2$ and 95% humidity. Microtitration(MIT) assay was done at 10, 25, 50 and 100ng/ml of 1,25-Dihydroxyvitamin $D_3$. 21 Sprague-Daft rats were divided into a control gmup(3), and experimental groups(18) where 100g of force from helical spring was applied across the maxillary incisors 1,25-Dihydroxyvitamin $D_3$ was injected into periodontal ligament at the mesial or distal surface of maxillary incisors so that we can compare the control side and the experimental side. Expreimental groups were sac rifled at 12, 24, 36, 48, 72hours and 7 days after force application, respectively. And the obtained tissues were evaluated histologically. The observed results were as follows. 1. The activity of periodontal ligament cells in l0ng/ml or 25ng/ml of 1,25-Dihydroxyvitamin $D_3$ 1,25-Dihydroxyvitamin $D_3$ was not significantly different to the control at the cultivation of 1, 2 and 3 days. 2. The activity of periodontal ligament cells was significantly increased at 3 days in 50 ng/ml of 1,25-Dihydroxyvitamin $D_3$ and 2, 3 days in 100g/ml of 1,25-Dihydroxyvitamin $D_3$. 3. Up to 7 days after force application, there was no difference in osteoblastic activity, tearing of periodontal ligament and proliferation of capillary at tension side between 1,25-Dihydroxyvitamin $D_3$ injection side and the control side. 4. The osteoclastic activity and the resorption of alveolar bone was greater in 1,25-Dihydroxyvitamin $D_3$ injection side than the control side at 36 hours after force application.

  • PDF

The Ability of Cervus Elaphus Sibiricus Herbal Acupuncture to Inhibit the Generation of Inflammatory Enzymes on Collagen-induced Arthritis Mice (녹용약침(鹿茸藥鍼)이 CIA 모델 생쥐의 염증인자 생성억제에 미치는 영향)

  • Hwang, Jong-Soon;Hwang, Ji-Hye;Lee, Hyun-Jin;Lee, Dong-Gun;Kang, Min-Joo;Back, Song-Ook;Cho, Hyun-Seok;Kim, Kyung-Ho;Kim, Kap-Sung
    • Journal of Acupuncture Research
    • /
    • v.24 no.6
    • /
    • pp.1-14
    • /
    • 2007
  • Backgrounds : Rheumatoid Arthritis(RA) is known as the chronic inflammatory diseasethat induces persistent inflammation in the joint cavity. The destruction of cartilage occurs as the result of bones destoyed by pannus, several influential cytokines induced by the synovial capsulitis, varieties of proteinases, $O_2$ radicals, and the secondary degenerative changes of articular cartilage. The type 2 collagen-induced arthritis model is used in recent experimental research on rheumatoid arthritis. Cervus elaphus sibiricus (Nockyong) has the effect of relieving pain by nourishing the muscles, joints, and bones. It is also known to be efficacious in promoting and enhancing the immune system. The objective of this study was to investigate the effect of Cervus elaphus sibiricus herbal acupuncture to inhibit the generation of proinflammatory enzyme on type 2 collagen-induced arthritis. I investigated the inhibition of mRNA transcription of MIF(macrophage migration inhibitory factor), $TNF-{\alpha}$(Tumor necrosis $factor-{\alpha}$) and MMP-9 (matrix metalloproteinase-9) of Cervus elaphus sibiricus herbal acupuncture using an in vitro test. Also investigated was the inhibition of differentiation of Th 1 cells and activation of cytokines(MIF, $TNF-{\alpha}$, IL-6, MMP-9), which are known to cause initial RA ,and are also related to the morphology of the synovial membranes of the joint capsule, by an in vivo test, using CIA(collagen induced arthritis) model mice. Materials & methods : The laboratory animals used in this experiment were 4 week-old DBA female mice, weighing approximately 20 grams, and adjusted to the laboratory environment. The experiment was divided into the normal group(NOR)-no treated group, control group(CON)-CIA induced group, and sample group(SAM)-Cervus elaphus sibiricus herbal acupuncture treated group. RA was induced in the mice via injection of $50{\mu}{\ell}$ C II mixed CFA. The Cervus elaphus sibiricus herbal acupuncture solution was applied on $GB_{35}$(陽陵泉) for 26 days from the 3rd day of RA inducement. The concentration of the solution was determined via a MTT assay. To research the effect on the expression of MIF, $TNF-{\alpha}$ and MMP-9 mRNA, RT-PCR was performed on synovial membrane cells from the knee joint of CIA mice. C II induced RA knee joint's histo-chemical synovial membrane was observed using a specimen model via the Hematoxilin and Eosin dying technique. Results : The expression of mRNA of RA-related cytokines such as MIF, $TNF-{\alpha}$, and MMP-9 dosedependently decreased in the cell from the synovial membranes of the joint, which is treated with Cervus elaphus sibiricus herbal acupuncture solution. In mice treated with Cervus elaphus sibiricusherbal acupuncture, the damage of synovial membranes of the joint was lessened, and differentiation of Th 1 cells was suppressed. The activation of RA-related cytokines such as MIF was suppressed, and the generation of $TNF-{\alpha}$ and MMP-9 showed a statistically significant decreas. Conclusions : It is speculated that Cervus elaphus sibiricus herbal acupuncture has the therapeutic effect of palliating the damage of the tissue impaired by RA by inhibition of the initial RA progression and by regulating excessive differentiation of Th 1 cell as it suppresses the generation of RA-related cytokines during the highest stage of RA by acting on pro-inflammatory enzymes.

  • PDF

Protective Effects of Perilla frutescens Britt var. japonica Extracts from Oxidative Stress in Human HaCaT Keratinocytes (HaCaT 피부각질세포에서 들깻잎 추출물의 산화적 스트레스에 대한 항산화 효과)

  • Ji, Na;Song, Jia-Le;Kil, Jeung-Ha;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.42 no.2
    • /
    • pp.161-167
    • /
    • 2013
  • The aim of this study was to investigate the protective effects of methanolic extract from perilla (Perilla frutescens Britt var. japonica) leaves (PLME) on oxidative injury from hydrogen peroxide ($H_2O_2$) in human HaCaT keratinoctyes. Cells were co-incubated with various concentrations (0~200 ${\mu}g/mL$) of PLME for 24 hr, and then exposed to $H_2O_2$ (500 ${\mu}M$) for 4 hr. $H_2O_2$ significantly decreased cell viability (p<0.05). However, PLME provided protection from $H_2O_2$-induced HaCaT cell oxidation in a dose-dependent manner. To further investigate the protective effects of PLME on $H_2O_2$-induced oxidative stress in HaCaT cells, the cellular levels of lipid peroxidation, and antioxidant enzymes (including superoxide dismutase (SOD), glutathione peroxidase (GSH-px) and catalase (CAT)) were measured. PLME decreased cellular levels of lipid peroxidation, and also increased the activities of antioxidant enzymes. In addition, the antioxidant activities of PLME were also determined by DPPH and hydroxyl (${\cdot}OH$) radical scavenging assay, and major antioxidant compounds of PLME were measured by colorimetric methods. DPPH and ${\cdot}OH$ radical scavenging activities of PLME increased in a dose dependent manner and was similar to the DPPH scavenging activity of ascorbic acid at 50 ${\mu}g/mL$; however PLME activities were stronger than ascorbic acid (50 ${\mu}g/mL$) in the ${\cdot}OH$ scavenging assay. The amounts of antioxidant compounds, including total polyphenolics, total flavonoids, and total ascorbic acid from PLME were $52.2{\pm}1.1$ mg gallic acid (GAE)/g, $33.7{\pm}4.7$ mg rutin (RUE)/g, and $17.0{\pm}0.5$ mg ascorbic acid (AA)/g, respectively. These results suggest that PLME has a strong free radical-scavenging activity and a protective effect against $H_2O_2$-induced oxidative stress in the keratinocytes.

Effects of Herbal Complex on Blood Glucose in Streptozotocin-induced Diabetic Rats and in Mice Model of Metabolic Syndrome (생약복합제의 Streptozotocin 유발 당뇨 및 대사성증후군 모델 동물에서의 혈당에 미치는 효과)

  • Park, Han-Seok;Lee, Yeon-Sil;Choi, Se-Jin;Kim, Jin-Kyu;Lee, Yun-Lyul;Kim, Hyun-Gwen;Koo, Sam-Hoi;Ku, Dae-Hoy;Ki, Seung-Il;Lim, Soon-Sung
    • Korean Journal of Pharmacognosy
    • /
    • v.40 no.3
    • /
    • pp.196-204
    • /
    • 2009
  • This study was carried out to investigate the in vivo and in vitro inhibitory effect of a traditional herbal complex (HC) extract prepared from a mixture of four oriental herbs (Dioscorea Rhizoma, Glycine soja Sieb. et Zucc, Bombycis corpus, Fermented Glycine soja) that have been widely used for the treatment and prevention of diabetes mellitus on hyperglycemia. The water extract of HC showed potent inhibitory effect on $\alpha$-glucosidase with $IC_{50}$ value of 1.24 mg/mL. Additionally, the ethanol extract of HC was also found to exhibit significant inhibitory effect against protein tyrosine phosphatase $1{\beta}$ ($PTP1{\beta}$), which is known as a major regulator of both insulin and leptin signaling. In the $PTP1{\beta}$ inhibitory assay, the most active n-hexane fraction obtained from the ethanol extract of HC, was identified as a mixture of fatty acid derivatives by gas chromatography-mass spectrometry (GC-MS). In high-fat diet-low dose streptozotocin (STZ)-induced diabetic rat, the water extract of HC improved the oral glucose intolerance as compared with rosiglitazone. HC also caused a marked decrease of body weight and fasting blood glucose and a significant improvement on glucose tolerance in metabolic syndrome mice model. These findings support that this traditional HC may be useful in the control of blood glucose in diabetes mellitus and metabolic syndrome.

Distinct Spatio-temporal Expression Patterns of Patatin Promoter-GUS Gene Fusion in Transgenic Potato Microtubers (형질전환 감자 소괴경의 발달단계에 따른 Patatin Promoter-GUS 유전자의 발현 분석)

  • Youm, Jung-Won;Kim, Mi-Sun;Lee, Byoung-Chan;Kang, Won-Jin;Jeon, Jae-Heung;Joung, Hyouk;Kim, Hyun-Soon
    • Journal of Plant Biotechnology
    • /
    • v.30 no.1
    • /
    • pp.13-18
    • /
    • 2003
  • This study was carried out to investigate the expression patterns of foreign gene that controlled by tuber-specific patatin promoter in transgenic potatoes. Potato leaf disc cultured in vitro were transformed by the Agrobacterium strain LBA4404 containing pBl121 or pATGUS from potato cv. Desiree. In order to select the transgenic lines, gene-specific primers deduced from the NPTII were synthesized and used for polymerase chain reaction. The down part of the putative transgenic potatoes was transplanted weekly onto sucrose-enriched medium to accelerate the microtuber formation. RNA gel blot analysis was performed on the total RNAs obtained from tuber that had been harvested at a week interval. Also, histochemical assay was observed in the explants transformed with either pBI121 or pATGUS. Results showed that the transgenic plant containing pATGUS expressed GUS transcripts mainly at the tuber, not in stem, with the highest expression level in 5 weeks-grown microtubers. In contrast to pATGUS plants, the transformed plants with pBI121 showed an equal expression pattern throughout the whole developing stages. Consistent with RNA gel blot analysis, histochemical GUS staining and enzyme activity exhibited pATGUS transcripts were at the highest level in 5 weeks cultures. From these results, we suggest that the best stage to analyze the foreign gene introduced by patatin promoter into potato plants is at 5 weeks cultures after tuber formation.

TENSILE STRENGTHS OF PRE-LIGATURED BUTTON WITH SEVERAL TYPES OF CONTAMINATION IN DIRECT BONDING PROCEDURE WHICH CAN HAPPEN DURING THE SURGICAL EXPOSURE OF UNERUPTED TEETH (치아의 견인을 위한 버튼 접착시 오염이 인장강동에 미치는 영향)

  • Kim, Seong-Oh;Choi, Byung-Jai;Lee, Jae-Ho;Sohn, Heung-Kyu
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.25 no.2
    • /
    • pp.400-420
    • /
    • 1998
  • We already know that it is very difficult to obtain an "isolated field" for direct bonding during the surgical exposure of unerupted teeth. The aim of this in-vitro study is to simulate the clinical situation of forced eruption and to evaluate the tensile strengths of preligatured button with several types of contamination which can happen during the surgical exposure of unerupted teeth. Four orthodontic direct bonding systems were used. ($Ortho-One^{TM}$, $Rely-a-Bond^{(R)}$, $Ortho-Two^{TM}$, Phase $II^{(R)}$) Each material was divided into four groups(n=20) : Group 1. (Control, no contamination), Group 2. (Rinse etching agent with saline instead of water), Group 3. (Blood contamination of etched surface for 30 seconds), Group 4. (Blood contamination of primed surface for 30 seconds) 320 bovine anterior permanent teeth were divided into the above mentioned 16 groups. Enamel surface was flattened and ground under water coolant. Pre-ligatured buttons were prepared to the same form. (Cut 0.25 ligature wire 10 cm in length. Twist the ligature wire 30 times clockwise. Mark the wire 15mm and 35mm points from button. Make a loop sticking two points together and twist the loop 6 times counterclockwise.) The bonded specimens were stored at $37^{\circ}C$ saline solution for 3 days. Then the tensile strength of each sample was measured with Instron universal testing machine, crosshead speed of 0.5mm/min. The following results were obtained: 1. As compared to control groups (Group 1) of each material, Rely-a-Bond had a significantly lower mean tensile strengths than other material. (p<0.01) 2. In Group 2. of Ortho-One and Rely-a-Bond, the mean tensile strengths decreased about 7.7% and 11.1%, respectively with statistical significances. (p<0.05) 3. In Group 2. of Ortho-Two and Phase II, the mean tensile strengths did not decrease. 4. In Group 3. of Ortho-One, Rely-a-Bond, Ortho-Two, and Phase II, the mean tensile strengths decreased about 60.8%, 56.1%, 60.2%, and 46.0%, respectively with statistical significances. (p<0.01) 5. In Group 4. of Ortho-One and Rely-a-Bond, the mean tensile strengths did not decrease. 6. In Group 4. of Ortho-Two and Phase II, the mean tensile strengths were decreased about 20.95% and 22.28%, respectively with statistical significances. (p<0.01) There were formations of a hump shaped mass from bonding resin under blood contamination which disturbed direct bonding procedure. According to Reynolds, the proper bond strength for clinical manipulation should be at least 45N or about 4.5Kg.F. According to these results, it can be concluded that Ortho-One could be used during surgical exposure of unerupted teeth. In any case, blood contamination of the etched surface should be avoided, but the blood contamination of primed surface of Ortho-One may not decrease bond strength. Just 'blowing-out' is enough to remove blood from primed surface of Ortho-One. You can verify the clean surface of the primer of Ortho-One after blowing out the blood contamination.

  • PDF

Lipase-Inhibitory and Anti-Oxidative Activity of the Methanol Extract and the Powder of Phellinus linteus (상황버섯 자실체 메탄올 추출물과 분말의 지방소화효소 억제 및 항산화 활성)

  • Kim, Ji-Hyun;Son, In-Suk;Kim, Jong-Sang;Kim, Ki-Hoon;Kwon, Chong-Suk
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.37 no.2
    • /
    • pp.154-161
    • /
    • 2008
  • Phellinus linteus (PL) has been known to exhibit potent biological activity. The present study was designed to investigate lipase-inhibitory and anti-oxidative activity of the methanol extract and the powder of PL fruiting body. The methanol extract of PL appeared to have the inhibitory activity against pancreatic lipase with an $IC_{50}$ value of $36.3\;{\mu}g/mL$, and the scavenging activity of DPPH radical with an $IC_{50}$ value of $20.1\;{\mu}g/mL$, which was similar to that of vitamin C ($IC_{50}\;18.3\;{\mu}g/mL$). To investigate the lipase-inhibitory and anti-oxidative effect of PL on animal, Sprague-Dawley rats were fed with high-fat diet supplemented with either 2% or 5% PL powder for 8 weeks. Total food intake was significantly increased, but body weight was not changed by PL powder supplementation. However, fecal fat excretion of the experimental groups fed with the PL powder were higher than that of the control group. PL powder showed a decrease in the plasma total cholesterol, LDL-cholesterol, and the hepatic total cholesterol levels. The anti-oxidative enzyme activities were also affected by PL supplementation. Glutathione peroxidase (GSH-Px) in the plasma and liver were significantly increased by 98% and 46% in the 2% PL group, and 99% and 32% in the 5% PL group, respectively. Total superoxide dismutase (T-SOD) activity was not affected by PL supplementation. DNA damage was measured by the comet assay in the lymphocytes collected after 2 weeks, 4 weeks, and 8 weeks of feeding PL supplemented diet. Lymphocyte DNA damage was decreased in the PL supplemented group. Furthermore, PL feeding enhanced the resistance to lymphocyte DNA damage caused by an oxidant challenge with $H_2O_2$.

Association of β-Catenin with Fat Accumulation in 3T3-L1 Adipocytes and Human Population (β-catenin 유전자의 3T3-L1 지방세포 및 인체에서의 지방축적 연관성 연구)

  • Bae, Sung-Min;Lee, Hae-Yong;Chae, Soo-Ahn;Oh, Dong-Jin;Park, Suk-Won;Yoon, Yoo-Sik
    • Journal of Life Science
    • /
    • v.21 no.9
    • /
    • pp.1301-1309
    • /
    • 2011
  • The major function of adipocytes is to store fat in the form of triglycerides. One of the signaling pathways known to affect adipogenesis, i.e. fat formation, is the WNT/${\beta}$-catenin pathway which inhibits the expression and activity of key regulators of adipogenesis. The purpose of this research is to find genes among the WNT/${\beta}$-catenin pathway which regulate adipogenesis by using small interfering (si) RNA and to find the association of single nucleotide polymorphisms (SNPs) of the gene with serum triglyceride levels in the human population. To elucidate the effects of ${\beta}$-catenin siRNA on adipogenesis key factors, PPAR${\gamma}$ and C/EBP${\alpha}$, we performed real-time PCR and western blotting experiments for the analyses of mRNA and protein levels. It was found that the siRNA-mediated knockdown of ${\beta}$-catenin upregulates adipogenesis key factors. However, upstream regulators of the WNT/${\beta}$-catenin pathway, such as DVL2 and LRP6, had no significant effects compared to ${\beta}$-catenin. These results indicate that ${\beta}$-catenin is a candidate gene for human fat accumulation. In general, serum triglyceride level is a good indicator of fat accumulation in humans. According to statistical analyses of the association between serum triglyceride level and SNPs of ${\beta}$-catenin, -10,288 C>T SNP (rs7630377) in the promoter region was significantly associated with serum triglyceride levels (p<0.05) in 290 Korean subjects. On the other hand, serum cholesterol levels were not significantly associated with SNPs of the ${\beta}$-catenin gene. The results of this study showed that ${\beta}$-catenin is associated with fat accumulation both in vitro and in the human population.

A Study on the Antioxidative, Anti-inflammatory and Anti-thrombogenic Effects of Zanthoxylum piperitum DC. Extract (초피나무 추출물의 항산화, 항염증 및 항혈전 효능에 관한 연구)

  • Jang, Mi-Jin;Rhee, Soon-Jae;Cho, Sung-Hee;Woo, Mi-Hee;Choi, Jeong-Hwa
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.35 no.1
    • /
    • pp.21-27
    • /
    • 2006
  • Effects of root, stem and leaf extracts of Zanthoxylum piperitum on the inhibition of lipid peroxidation in the hepatic microsome of rat, DPPH radical scavenging activity, soybean lipoxygenase activity and activated partial thromboplastin times (APTT) were examined in vitro. The highest inhibition of hepatic microsomal lipid peroxidation was observed by ethyl acetate fraction of the root and stem extracts. The high inhibition of lipid peroxidation was observed in the leaf, the root and the stem in order. The DPPH radical scavenging activity of ethyl acetate fraction was higher than that of n-butanol fraction and it was similar to the root and the steam extract. It was similar to the inhibition of hepatic microsomal lipid peroxidation. The DPPH radical scavenging activity was the highest in 0.50 mg/mL of ethyl acetate fraction, and it was 4.4-fold higher than that of a-tocopherol, as an antioxidant standard. The DPPH radical scavenging activity was dependent on the extract concentration in the range of $0.12\~5.00$ mg/mL. The soybean lipoxygenase activity of ethyl acetate fraction was higher than that of n-butanol fraction and it was similar to the root and the stem extracts. The soybean lipoxygenase activity was the highest in 0.50 mg/mL of ethyl acetate fraction. The soybean lipoxygenase activity was dependent on the extract concentration in the range of $0.12\~5.00$ mg/mL. The leaf extract showed the highest antithrombogenic effect followed by the stem and then the root extract. The activated partial thromboplastin times were dependent on the extract concentration in the range of $0.10\~2.00$ mg/mL.

Effect of Hijikia fusiforme Fractions on Proliferation and Differentiation in Osteoblastic MC3T3-E1 Cells (톳 분획물이 조골세포의 증식 및 분화에 미치는 영향)

  • Jeon, Min-Hee;Kim, Mi-Hyang
    • Journal of Life Science
    • /
    • v.21 no.2
    • /
    • pp.300-308
    • /
    • 2011
  • Osteoporosis is a disease involving a decrease in bone mineral density and increased risk of fractures. Osteoblast and osteoclast activities are important for bone formation. The MC3T3-E1 osteoblastic cell line is a well-accepted model of osteogellsis in vitro. Hijikia fusiforme is a kind of edible brown seaweed that grows mainly in the Northwest Pacific region, including the countries of Korea, Japan and China, and it has been widely used as a medicinal and health food in Korea. In this study, by using osteoblasts, the effects of Hijikia fusiforme fractions on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and mineralization of cells were investigated. Hijikia fusiforme were subjected to fractionation by using hexane, methanol, butanol and aqueous. Proliferation of the MC3T3-E1 osteoblastic cells that were treated with Hijikia fusiforme fractions increased by approximately 120%. Regarding effects of Hijikia fusiforme fractions on ALP activity, 1 ${\mu}g$/ml butanol fraction showed the highest activity. The synthesis of collagen increased significantly in response to treatment with Hijikia fusiforme fractions, with the exception of the hexane fraction. Moreover, mineralization in the MC3T3-E1 cells that were treated with 100 ${\mu}g$/ml butanol fraction increased by 281%. Also, when 100 ${\mu}g$/ml aqueous fraction was added, mineralization increased by 240%. These results indicate that Hijikia fusiforme fractions have anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.