• Polymer(Korea)
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• v.34 no.5
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• pp.474-479
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• 2010

We developed laminated cylindrical scaffolds composed of poly(lactide-co-glycolide)(PLGA) and keratin, and investigated their potential for tissue engineering and disk regeneration. The scaffold was designed to have two parts, i.e. inner cylinder and outer disk, to mimic a natural disk. The outer disk was composed of PLGA and the inner cylinder was prepared using PLGA film or PLGA/keratin hybrid film. In this study, we investigated the effects of keratin on the growth and proliferation of annulus fibrous(AF) cells in the cylindrical scaffolds. Scaffolds containing PLGA/keratin films showed a significantly higher cell proliferation and expression of collagen I and II than the counterpart with PLGA films. Keratin containing scaffolds also exhibited an excellent mechanical strength, demonstrating that keratin influences the proliferation of annulus fibrous cells. The results provide valuable information on PLGA/keratin films for tissue engineered disk regeneration.

• The Journal of Korean Medicine
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• v.28 no.2 s.70
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• pp.93-113
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• 2007

Objective : To research Trends of studies about anti-inflammation and pain relief, obesity, and safety of herbal acupuncture therapy by analyzing domestic theses, published since 2001, about herbal acupuncture therapy. Methods : Domestic theses, published since 2001, mentioning anti-inflammation or pain relief, obesity, or safety of herbal acupuncture therapy were reviewed and analyzed. These theses were then classified by university, year, and subject. Results : The following results were obtained in this study. 1. Among published theses related to anti-inflammation effects of herbal-acupuncture, studies about arthritis comprised the most part, 52. In theses relating to causes of arthritis, 16 were about adjuvant, which was the most, followed by Type II collagen, LPS and carrageenan. Blood test, reactions of inflammation and revelation of cytokine and immune cellswere methods for evaluating anti-inflammation effect. The tendency of experimental methods was to focus on molecular biologic method. 2. In theses related to pain relief, many clinical attempts with herbal injection were carried out, and Carthami Flos and Scolopendrawere used most. Observing reduction of pain inducing factor and checking behavioral change were methods for evaluating pain relief. 3. In theses related to obesity, research focused on effects in association with spots on the body suitable for acupuncture. There were also attempts comparing effectiveness between single injections and complex injections. Astraball Radix, Angelica Gigantis Radix, Coicis Semen and Taeumjowetang were used. Evaluation of anti-obesity effects were by weight loss, food efficiency, blood lipid profile and evaluation of liver function. 4. In theses related to safety of herbal-acupuncture, Herba Chelidonii Chaenonelis Fructus, Clematis Florida Thunb, Corydalidis Tuber, Paeoniae Radix, and Carthami Flos which marked 2 theses each were most studied. Methods of evaluating safety were mostly by observing liver and kidney functions based on blood test, and by applying herbal injections to clinical treatment. Conclusion : Herbal acupuncture is being used in various ways associating with its anti-inflammation, pain relief and anti-obesity effect. Studies on efficacy and mechanism of herbal acupuncture are being conducted even at the molecular biology level.

• Journal of Periodontal and Implant Science
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• v.35 no.1
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• pp.99-109
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• 2005

It has been focused on the importance of the host inflammatory response in periodontal pathogenesis and progression, treatment has been introduced to control the host response and the method, which diminishes production and activity of MMP by doxycycline, has been used in periodontal field. MMP is a proteolytic enzyme which plays a major role in tissue destruction and MMP-1 is secreted in the periodontally healthy tissue, while MMP-8, 9, 13, etc in the inflammatory state. Among these, MMP-13 has been discovered lately and reported to degrade primarily type II collagen. Periodontal ligament (PDL) cell plays a role in destruction of periodontal tissue. This study was to evaluate the effect of doxycycline and mefenamic acid, non-steroidal antiinflammatory drug on MMP-13 mRNA expression in the rat PDL cell. Doxycycline concentration of $1{\sim}100\;{\mu}g/ml$ was added rat PDL cell and cell activity was measured by MIT assay at day 1 and 3. MMP-13 gene expression was evaluated by RT-PCR after PDL cells were pre-treated for 1hour with doxycycline (50 ${\mu}g/ml$) alone or with mefenamic acid ($10^{-6}M$), then added $IL-1{\beta}$(1.0 ng/ml) and incubated for 16-18 hours. The results are as follows: 1. Cell activity decreased Significantly at 24 and 72 hours in 100 ${\mu}g/ml$ (p<0.05). 2. Level of MMP-13 mRNA was in 20.2% increase by $IL-1{\beta}$ and in pre-treating doxycycline group, expression of $IL-1{\beta}$ induced MMP-13 mRNA was inhibited by 31% than $IL-1{\beta}$ treated only. 3. Mefenamic acid did not inhibit on the expression of $IL-1{\beta}$ induced MMP-13 mRNA, while mefenamic acid in combination with doxycycline inhibited the expression by 41% compared to only $IL-1{\beta}$ stimulation. These results suggest that doxycycline synergistically inhibit the expression of $IL-1{\beta}$ induced MMP-13 mRNA in combination with mefenamic acid.

• Archives of Plastic Surgery
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• v.33 no.1
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• pp.46-52
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• 2006

High-density micromass culture was needed to take three dimensions culture with ASCs(adipose derived stromal cells) and chondrogenesis. However, the synthetic polymer has hydrophobic character and low affinity to cells and other biomolecules. Therefore, the surface modification without changes of physical and chemical properties is necessary for more suitable condition to cells and biomolecules. This study was performed to investigate the effect of surface modification of poly (lactic-co-glycolic acid)(PLGA) scaffold by plasma treatment (P(+)) on the adhesion, proliferation and chondrogenesis of ASCs, and not plasma treatment (P(-)). ASCs were isolated from human subcutaneous adipose tissue obtained by lipectomy and liposuction. At 1 hour 30 minutes and 3days after cell seeding onto the P(-) group and the P(+) group, total DNA amount of attached and proliferated ASCs markedly increased in the P(+) group (p < 0.05). The changes of the actin under confocal microscope were done for evaluation of cellular affinity, at 1 hour 30 minutes, the shape of the cells was spherical form in all group. At 3rd day, the shape of the cells was fiber network form and finely arranged in P(+) group rather than in P(-) group. RT-PCR analysis of cartilage-specific type II collagen and link protein were expressed in 1, 2 weeks of induction. Amount of Glycoaminoglycan (GAG) markedly increased in P(+) group(p < 0.05). In a week, extracellular matrix was not observed in the Alcian blue and Safranin O staining. However in 2 weeks, it was observed that sulfated proteoglycan increased in P(+) group rather than in P(-) group. In conclusion, we recognized that plasma treatment of PLGA scaffold could increase the hydrophilic property of cells, and provide suitable environment for high-density micromass culture to chondrogenesis

• Journal of Ginseng Research
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• v.36 no.2
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• pp.161-168
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• 2012

The abnormal maturation and ossification of articular chondrocytes play a central role in the pathogenesis of osteoarthritis (OA). Inhibiting the enzymatic degradation of the extracellular matrix and maintaining the cellular phenotype are two of the major goals of interest in managing OA. Ginseng is frequently taken orally, as a crude substance, as a traditional medicine in Asian countries. Ginsenoside $Rb_1$, a major component of ginseng that contains an aglycone with a dammarane skeleton, has been reported to exhibit various biological activities, including anti-inflammatory and anti-tumor effects. However, a chondroprotective effect of ginsenoside $Rb_1$ related to OA has not yet been reported. The purpose of this study was to demonstrate the chondroprotective effect of ginsenoside $Rb_1$ on the regulation of pro-inflammatory factors and chondrogenic genes. Cultured rat articular chondrocytes were treated with 100 ${\mu}M$ ginsenoside $Rb_1$ and/or 500 ${\mu}M$ hydrogen peroxide ($H_2O_2$) and assessed for viability, reactive oxygen species production, nitric oxide (NO) release, and chondrogenic gene expression. Ginsenoside $Rb_1$ treatment resulted in reductions in the levels of pro-inflammatory cytokine and NO in $H_2O_2$-treated chondrocytes. The expression levels of chondrogenic genes, such as type II collagen and SOX9, were increased in the presence of ginsenoside $Rb_1$, whereas the expression levels of inflammatory genes related to chondrocytes, such as MMP1 and MMP13, were reduced by approximately 50%. These results suggest that ginsenoside $Rb_1$ has potential for use as a therapeutic agent in OA patients.

• Polymer(Korea)
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• v.38 no.3
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• pp.278-285
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• 2014

Demineralized bone particles (DBP) and alginate hybrid sponges were fabricated at 10, 20, 40 and 80% DBP/alginate hybrid ratios for seeding chondrocyte. Cell proliferation was measured via MTT assay. Morphological observation, histology, biological assay and RT-PCR were performed at each time point 1, 2 and 3 weeks. The cell viability was better in 20% DBP/alginate sponges than in other sponges. SEM results showed that more attached and more proliferated cells in the 20% DBP/alginate sponges with the lapse of time. Finally, histochemical assay results showed that the phenotype of chondrocyte was well maintained and both acidic mucopolysaccharide and type II collagen was well formed at 20% sponges. This study suggested that DBP/alginate sponge may serve as a potential cell delivery vehicle and a structural basis for tissue engineered articular cartilage.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.1
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• pp.30-39
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• 2014

HanPoong Leading (HPL)-04 were prepared with different oriental medicines (balk of Kalopanax pictus balk, Chaenomelis Fructus, Angelica gigas root, Zingiber officinale, Raphanus sativus Linne and Saururus chinensis Baill.) to investigate the protective effects of HPL-04 on cartilage degradation in knee osteoarthritis (OA). Rat articular chondrocytes incubated with rhIL-$1{\alpha}$ markedly increased matrix metalloproteinase (MMP)-2 and 9 activities, decreased cell viability and reduced chondrogenic gene expression. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, MMP-2 and 9 activities and real time RT-PCR indicated that HPL-04 counteracted these harmful effects in dose-dependent manner. In addition, for experimental OA in vivo, monosodium iodoacetate (MIA, 0.5 mg/50 ${\mu}L$) was injected into knee joints of rats and administered HPL-04 to rats for 4 consecutive weeks after MIA treatment. The experimental data showed that treatment with HPL-04 significantly prevented of MMP-2 and 9 activities in articular cartilage. Histopathological and micro-CT evaluations of the knee joints also revealed that HPL-04 effectively ameliorated MIA-induced degenerative OA. In conclusion, HPL-04 has potential applicability for the prevention and treatment of degenerative OA.

• Journal of Haehwa Medicine
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• v.9 no.1
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• pp.201-214
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• 2000

Precise and detailed clinical research and evaluation based on objective standards are imperative factors in securing reliability of a clinical test. Built on this principle, this clinical test has been conducted during the period between March 1999 and March 2000 dealing with 31 outpatients treated with ShiHo-GuizhiTang or ShiHo-GuizhiTang modify and ShiHo-GuizhiTang mixed prescriptions at the Sung-bo ORIENTAL MEDICAL CLINIC. A variety of information relating to those patients has been collected and analyzed under such criteria as precise diagnoses and their clinical effectiveness. The analysis was duly based on "The theory of cold syndrome". Then, the 31 patients' individual clinical information was compared one another by breaking down the results into sub-categories including gender, age, disease, main symptom, blood pressure, pulse beats, syndrome of abdomen, treatment period, modify and mixed prescription, and evaluation. The result of this clinical test can be summarized as follows: First, ShiHo-GuizhiTang appears to be more frequently prescribed in the case of female patients than in the case of male patients despite the fact that it does not necessarily need to be applied only to female. In addition, the numbers of prescriptions of ShiHo-GuizhiTang in the two age groups consisting of elementary school children and economically active adults respectively were the highest among other age groups. Second, ShiHo-GuizhiTang proved to be most effective in treatment for respiratory ailments and arthritis. Third, ShiHo-GuizhiTang brought down blood pressure of hypertension patients and at the same time benefited patients with normal or lower-than-normal blood pressure who were vulnerable to diseases due to low disease-resistance. Fourth, ShiHo-GuizhiTang was effective in the case of frequent pulse(rapid pulse) and thereby indicating the fact it carries Taiyang superficies syndrome. 지맥 arises from suppressed immune responses owing to adrenocortical hormones. ShiHo-GuizhiTang controls and revitalizes those suppressed immune systems which stem from slow pulse and, as a consequence of that process, helps them return to normal condition of pulse. Fifth, from the standpoint of syndrome of abdomen, feeling of obstruction in the epigastrium serves as an important standard in the process of diagnosing diseases and evaluating effectiveness of treatments. Sixth, according to the results of the clinical test with the 31 patients, a total of 81 percent of test subjects benefited from the treatment. The figure is the sum of 52 percent of I-class (both main symptom and accompanying symptoms had been eliminated) and 29 percent of II-class (part of main symptom and accompanying symptoms had been eliminated) respectively. All told it is safe to say that ShiHo-GuizhiTang can elect to be a viable clinical treatment. In conclusion, it is estimated that this clinical study has drawn up guidelines for objective diagnostic standards and evaluation on specific treatments' effectiveness. This will lead to more general application of ShiHo-GuizhiTang. On top of that, this study could also provide an opportunity to stress the significance of ShiHo-GuizhiTang and ShiHo-GuizhiTang modify and ShiHo-GuizhiTang mixed prescriptions as an alternative treatment for collagen disease which comes from environmental degradation and pollution.

• Journal of Periodontal and Implant Science
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• v.35 no.3
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• pp.549-561
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• 2005

이 연구의 목적은 새로이 제작된 chitosan-poly(L-lactic acid)(PLLA) 다층 다공성 차폐막의 생체적합성 및 골세포활성도 및 골재생능을 평가하는 것이다. 제작된 차폐막을 24 well에 넣고 clonal osteoblast-like cell line(MC3T3-E1)을 접종한 군을 실험군으로, 차폐막을 사용하지 않은 대조군으로 하였다. 배양 1일, 7일 및 14일째에 각 well에서 세포수를 측정하였다. 주사전자현미경을 이용하여 차폐막에 부착된 세포의 형태관찰을 시행하였다. RNA 추출 및 RT-PCR을 실시한 후, agarose gel상에서 전기영동하여 조골세포 표식자인 collagen type I(COL), osteopontin(OP) 및 osteocalcin(OC) mRNA의 발현을 관찰하였다. 제작된 매트릭스의 생체적합성 및 골재생능을 관찰하기 위하여 백서의 두개골에 직경 8mm의 원형 결손부를 형성한 후 차폐막을 이식한 군을 실험군으로, 아무 것도 넣지 않은 군을 대조군으로 하여 4주 경과 후 실험동물을 희생시킨 후 조직학적관찰을 시행하였다. 시간경과에 따른 부착세포수 관찰결과, 배양 14일까지 조골세포의 수가 지속적으로 증가하였고, 주사전자현미경으로 세포의 형태 관찰결과, 배양된 세포들은 중층의 형태로 성장하면서 시간경과에 따라 세포가 응집되는 양상을 나타내었다. 관찰 기간동안 COL, OP, 및 OC mRNA의 발현이 관찰되어 배양 전 기간동안 조골세포의 형질이 잘 유지되고 있음을 알 수 있었다. 백서 두개골 결손부에 이식된 차폐막은 염증반응 없이 주위 조직과 우수한 생체적합성을 나타내었으며, 차폐막을 이식하지 하지 않은 대조군에 비해 높은 신생골 형성을 나타내었다. 이상의 관찰결과로 새로이 제작된 chitosan-PLLA 차폐막은 우수한 생체적합성 및 골재생능을 나타냄을 알 수 있었으며, 향후 이를 골조직 재생 및 치주조직유도재생 분야에 응용될 수 있을 것으로 생각된다.

• Journal of Physiology & Pathology in Korean Medicine
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• v.33 no.2
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• pp.131-140
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• 2019

The objective of present study is to evaluate concentration-dependent in vitro anti-osteoarthritic (OA) effects of synergic mixed formula consisted of dried pomegranate juice concentrate powder, Eucommiae Cortex aqueous extract and Achyranthis Radix aqueous extract 5:4:1 (g/g) mixture on the primary cultured rat articular chondrocytes. First, any cytotoxic effect of mixture was observed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay. Next, cyto-protective effect of test substances was evaluated by using the recombinant human interleukin $(rhIL)-1{\alpha}$ induced chondrocytes. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2(PGE_2)$ productions and 5-lipoxygenase (LPO) activities, and inhibitory effects on matrix metalloproteinase (MMP)-2 and MMP-9 activities were observed on $rhIL-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions. No obvious cytotoxic effects of mixture were demonstrated. Inflammatory damages of chondrocytes and related ECM degradations induced by treatment of LPS or $rhIL-1{\alpha}$ were significantly and concentration-dependently inhibited by pretreatment of mixture from a concentration level of 0.001 mg/ml to 1 mg/ml. In addition, mixture showed $IC_{50}$ for $rhIL-1{\alpha}-induced$ MMP-2 and MMP-9 activities as 44.01 and $162.47{\mu}g/ml$, and also showed $EC_{50}$ for $rhIL-1{\alpha}-induced$ inhibition of collagen type II, SOX9 and aggrecan mRNA expression as 8.61, 10.79 and $4.47{\mu}g/ml$, respectively. It is observed that mixture showed concentration-dependent anti-inflammatory and cytoprotective ECM preserved effects on the primary cultured rat articular chondrocytes without cytotoxicity.