• Journal of Haehwa Medicine
• /
• v.24 no.2
• /
• pp.35-57
• /
• 2016

Objectives : The purpose of this study is to prove the effect and mechanism of Gamikyejakjimogawusul-tang(GKHA) herbal acupuncture on induced rheumatoid arthritis model of DBA/1 mice. Methods : We check effect of GKHA extract on the AST, ALT, Creatinine, BUN of serum and cell viability of GK extract in RAW 264.7 cells to test the stability of this study. In vitro, we measure total phenol contents, total flavonoid contents, DPPH free radical scavenging activity, ABTS cation radical scavenging activity of Gamikyejakjimogawusul-tang, effect of GK extract on ROS(Reactive Ooxygen Species) production to estimate a anti-oxidant capacity, and we also measure effect of GK extract on NO (Nitric Oxid), IL-$1{\beta}$, IL-6, IL-17, IL-21, TNF-${\alpha}$, MCP-1, GM-CSF production in RAW 264.7 cells to estimate a anti-inflammatory efficacy. In vivo, we compare a rheumatoid arthritis manifestation between control and experimental group and estimate a AI. Then we check effect of GKHA on the level of WBC, neutrophil, lympocyte, monocyte in the blood to see the effect of immune cells in blood. In addition we measure effect of GKHA on the level of hs-CRP, IgM, IgG, IL-$1{\beta}$, IL-6, IL-17, IL-21, TNF-${\alpha}$, MCP-1, GM-CSF in serum. We observe effects of GKHA on imaging of cartilage degeneration using micro CT-arthrography in paw hind. And we calculate effects of GKHA that reduced BV ratio, BS/BV ratio using 3D Micro-CT. Lastly we observe effects of GKHA histopathologic examination analysis. Results : 1. The toxicity on liver and kidney was disregardable and the cytotoxicity against RAW 264.7 cells was also disregardable. 1. Total phenol contents and total flavonoid contents in GK extract were in high level. 2. DPPH free radical scavenging activity and ABTS cation radical scavenging activity were increased according to concentration of GK extract 3. ROS production was significantly decreased in GK extract (at 10, $100{\mu}g/ml$). 4. NO, IL-6, TNF-${\alpha}$, MCP-1 production were significantly decreased in GK extract(at 10, $100{\mu}g/ml$). IL-17, GM-CSF production were significantly decreased in GK extract(at 1, 10, $100{\mu}g/ml$). IL-$1{\beta}$, IL-21 production were also decreased but there was no statistical significance. 5. 25x observation after H&E and M-T staining, infiltration of immune cells and subsidence of the cartilage and damage to the synovial cells were decreased. Conclusions : This study showed that GKHA extract had anti-oxidant capacity, anti-inflammatory efficacy. GKHA extract also had inhibiting effect on the process of rheumatoid arthritis and can protect joint and cartilage. So we expect that GKHA extract can be a meaningful treatment to rheumatoid arthritis patients.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.33 no.6
• /
• pp.459-466
• /
• 2011

Purpose: This study evaluated the capability of bone formation with silk fibroin/nano-hydroxyapatite/corn starch composite scaffold as a bone defect replacement matrix when grafted in a calvarial bone defect of rabbits $in$ $vivo$. Methods: Ten New Zealand white rabbits were used for this study and bilateral round-shaped defects were formed in the parietal bone (diameter: 8.0 mm). The silk fibroin 10% nano-hydroxyapatite/30% corn starch/60% composite scaffold was grafted into the right parietal bone (experimental group). The left side (control group) was grafted with a nano-hydroxyapatite (30%)/corn starch (70%) scaffold. The animals were sacrificed at 4 weeks and 8 weeks. A micro-computerized tomography (${\mu}CT$) of each specimen was taken. Subsequently, the specimens were decalcified and stained with Masson's trichrome for histological and histomorphometric analysis. Results: The average ${\mu}CT$ and histomorphometric measures of bone formation were higher in the control group than in the experimental group at 4 weeks and 8 weeks after surgery though not statistically significant ($P$ >0.05). Conclusion: The rabbit calvarial defect was not successfully repaired by silk fibroin/nano-hydroxyapatite/corn starch composite scaffold and may have been due to an inflammatory reaction caused by silk powder. In the future, the development of composite bone graft material based on various components should be performed with caution.

• Restorative Dentistry and Endodontics
• /
• v.33 no.6
• /
• pp.507-517
• /
• 2008

There are considerable in vitro and in vivo evidences for remineralization and demineralization occurring simultaneously in incipient enamel caries. In order to "heal" the incipient dental caries, many experiments have been carried out to determine the optimal conditions for remineralization. It was shown that remineralization is affected by different pH, lactic acid concentrations, chemical composition of the enamel, fluoride concentrations, etc. Eighty specimens from sound permanent teeth without demineralization or cracks, 0.15 mm in thickness, were immersed in lactic acid buffered demineralization solutions for 3 days. Dental caries with a surface zone and subsurface lesion were artificially produced. Groups of 10 specimens were immersed for 10 or 12 days in lactic acid buffered remineralization solutions consisting of pH 4.3 or pH 6.0, and 100, 50, 25, or 10 mM lactic acid. After demineralization and remineralization, images were taken by polarizing microscopy (${\times}100$) and micro-computed tomography. The results were obtained by observing images of the specimens and the density of the caries lesions was determined. 1.As the lactic acid concentration of the remineralization solutions with pH 4.3 was higher, the surface zone of the carious enamel increased and an isotropic zone of the subsurface lesion was found. However, the total decalcification depth increased at the same time. 2.In the remineralization solutions with pH 6.0, only the surface zone increased slightly but there was no significant change in the total decalcification depth and subsurface zone. In the lactic acid buffer solutions with the lower pH and higher lactic acid concentration, there were dynamic changes at the deep area of the dental carious lesion.

• Journal of Biomedical Engineering Research
• /
• v.38 no.4
• /
• pp.175-182
• /
• 2017

Microcurrent electrical stimulation(MES) has been used to accelerate recovery of atrophied skeletal muscle. However, convincing stimulation parameters for suppressing muscle atrophy due to injured sciatic nerve remains unclear. The objective of this study was to investigate the effective intensity of MES on restraining muscle atrophy with rat model underwent sciatic nerve injury(SNI). Twenty-5-week-old Sprague Dawley male rats were equally assigned to five groups : Control group(Control, CON, n = 4), Denervation group(Denervation, D, n = 4), Denervation with MES of $22{\mu}A$ group(Denervation + $22{\mu}A$, D+22, n = 4), Denervation with MES of $100{\mu}A$ group (Denervation + $100{\mu}A$, D+100 n = 4), Denervation with MES of $400{\mu}A$ group(Denervation + $400{\mu}A$, D+400, n = 4). To induce muscle atrophy, all rats in the D, D+22, D+100, and D+400 groups, were subjected to sciatic nerve injury on their right hindlimb and allowed to have 1 week of resting period. Following this period, rats underwent daily MES(60 min/ a day, 5times/1week) for 4 weeks. After that, we investigate morphological changes in muscle volume by using in vivo micro-computed tomography at week 0, 1, 3 and 5. After 5 weeks, the muscle volume had the highest value in D+400 group, and also noticeably increased in D+100 group compared to it in D group. The results of this study imply that MES with current intensities between $100-400{\mu}A$ can suppress muscle atrophy effectively.

• Proceedings of the Korean Society of Precision Engineering Conference
• /
• 2006.05a
• /
• pp.183-184
• /
• 2006

Some researchers proposed positive effects of whole body vibration (WBV) on osteoporotic trabecular bones of animals. In the present study, the correlation between the improvement of morphological characteristics and the effect of WBV was investigated and analyzed in OVX rats. The rats were randomly assigned to 5 groups: control, sham, WBV 17Hz, WBV 30Hz and WBV 45Hz. The WBV groups were exercised on a vibration platform (magnitude : $1mm_{peak-peak}$, frequency : 17Hz, 30Hz and 45Hz, 30 minutes/day for 5 days/week). The 4th lumbar on rats was scanned by In-vivo Micro-CT at the week 0(just before WBV) and the week 8(after WBV). Structural parameters of the 4th lumbar, based on two dimensional (2D) scan image data, were investigated and analyzed. The quantitative decrement rate of trabecular bone on WBV groups with 30Hz and 45Hz were lower than control and sham groups. The results showed the positive effect of WBV on osteoporotic bones of OVX rats.

• Proceedings of the Korean Society of Precision Engineering Conference
• /
• 2006.05a
• /
• pp.649-650
• /
• 2006

Some researchers proposed positive effects of whole body vibration (WBV) on osteoporotic trabecular bones of animals. In the present study, the correlation between the improvement of morphological characteristics and the effect of WBV was investigated and analyzed in OVX rats. The rats were randomly assigned to 5 groups: control, sham, WBV 17Hz, WBV 30Hz and WBV 45Hz. The WBV groups were exercised on a vibration platform (magnitude $1mm_{peak-peak}$, frequency : 17Hz, 30Hz and 45Hz, 30 minutes/day for 5 days/week). The 4th lumbar on rats was scanned by In-vivo Micro-CT at the week 0(iust before WBV) and the week 8(after WBV). Structural parameters of the 4th lumbar, based on two dimensional (2D) scan image data, were investigated and analyzed. The quantitative decrement rate of trabecular bone on WBV groups with 30Hz and 45Hz were lower than control and sham groups. The results showed the positive effect of WBV on osteoporotic bones of OVX rats.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.42 no.5
• /
• pp.697-704
• /
• 2013

The inhibitory effect of ethanol extracts from Curdrania tricuspidata leaves (CTL) on osteoarthritis was investigated in primary cultured rat cartilage cells and a monosodium-iodoacetate (MIA)-induced arthritis rat model. To identify the effects of CTL 80% ethanol extracts (CTL80) and CTL 10% ethanol extracts (CTL10) against $H_2O_2$ treatment in vitro, cell survival was measured by the MTT assay. Cell survival after $H_2O_2$ treatment increased with CTL80 and CTL10 close to normal up to $300{\mu}g/mL\;H_2O_2$. The mRNA expression of matrix metalloproteinases (MMPs) was determined MMP-7 and MMP-13 (known catabolic factors), were significantly inhibited by CTL 80 and CTL10; a $200{\mu}g/mL$ dose of CTL80 especially decreased MMP-13 expression. In vivo, osteoarthritis was induced by an intra-articular injection of MIA into the knee joints of rats, then CTL80 and CTL10 orally administered daily for 35 days. After the animals were sacrificed, histological evaluations of their knee joints revealed a reduction in polymorphonuclear cell infiltration and smooth synovial lining in the CTL80-500 group. Micro-CT analysis of hind paws from CTL80-500 and CTL10 showed a protection against osteophyte formation, soft tissue swelling, and bone resorption. In conclusion, CTL ethanol extracts are effective in ameliorating joint destruction and cartilage erosion in MIA-induced rats. CTL decreases and normalizes articular cartilage through preventing extracellular matrix degradation and chondrocyte injury, and could potentially serve as a therapeutic treatment for humans.

• Journal of Korean Medicine Rehabilitation
• /
• v.25 no.2
• /
• pp.15-35
• /
• 2015

Objectives This study was performed to investigate the effects of Bujasasim-tang ethanol extract (BST) on oxidative stress, inflammation and osteoarthritic rat model. Methods To ensure safety of BST, heavy metal levels were measured and cytotoxicity test was done. In vitro, To evaluate antioxidative effects of BST, total phenolic contents, 1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging activity, reactive oxygen species (ROS) levels were measured. Also, to evaluate anti-inflammatory effects of BST treated group, total nitric oxide (NO) and pro-inflammatory cytokines (IL-$1{\beta}$, IL-6, TNF-${\alpha}$) levels were measured in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. In vivo, We injected MIA $50{\mu}l$ (60 mg/ml) into knee joints of rats to induce osteoarthritis. Rats were divided into total 3 groups (normal, control, BST treated group, each n=7). Normal group was not treated at all without inducing osteoarthritis and taken normal diet. Control group was induced osteoarthritis by MIA and taken with 2 ml of distilled water once a day for 4 weeks. BST treated group was induced osteoarthritis by MIA and taken BST 2 ml (200 mg/kg/mouse) once a day for 4 weeks. We evaluated dynamic weight bearing with the Incapacitance Test Meter. At the end of experiment, the rats were sacrificed to observe the functions of liver and kidney, changes of WBC, neutrophil, lymphocyte, monocyte levels in blood, to evaluate the levels of pro-inflammatory cytokines, tissue inhibitor of metallopreteinases-1 (TIMP-1), matrix metalloproteinase-9 (MMP-9), prostaglandin $E_2$ ($PGE_2$), leukotriene $B_4$ ($LTB_4$) within serum. We observed change of articular structures by Hematoxylin & Eosin (H&E), safranin-O staining method and measured amount of cartilage by micro CT-arthrography. Statistical analysis was done by unpaired student's t-test with significance level at p<0.05 in SPSS 11.0 for windows. Results 1. Safety of the BST was identified. 2. AST, ALT, BUN, creatinine levels of BST treated group were within normal limit. In vitro, 1. DPPH and ABTS free radical scavenging activities of BST showed dose-dependent increase. 2. ROS production were significantly decreased. 3. Total nitric oxide (NO) and IL-$1{\beta}$ production were decreased. 4. IL-6 and TNF-${\alpha}$ production were significantly decreased. In vivo, 1. Weight bearing ability was significantly increased. 2. WBC, neutrophil, lymphocyte, monocyte levels in blood were decreased. 3. IL-$1{\beta}$ and TNF-${\alpha}$ levels in serum were significantly decreased. and the IL-6 level was decreased. 4. TIMP-1, MMP-9, $LTB_4$, $PGE_2$ levels in serum were significantly decreased. 5. Cartilage volume of BST treated group was significantly increased. Also changes of cartilage, synovial membrane, fibrous tissue were suppressed. Conclusions The results obtained in this study Bujasasim-tang have effects of antioxidative, anti-inflammatory, relieve pain and protection of cartilage. Therefore we expect that Bujasasim-tang is effective treatment for osteoarthritis.

• Transactions of the Korean Society of Mechanical Engineers B
• /
• v.36 no.11
• /
• pp.1081-1090
• /
• 2012

This study aims to track the longitudinal alteration pattern on the trabecular bone microarchitecture at tibial epiphysis induced by T-OA over time using in vivo micro computed tomography (${\mu}CT$). Ten SD rats were divided into control (n = 5) and T-OA (n = 5) groups. Anterior cruciate ligament transaction was performed for the T-OA group. The results showed that the alteration pattern on the trabecular bone microarchitecture at tibial epiphysis in the T.OA group was definitely different compared with that in the CON group from 0 to 8 weeks (approximately 4-16%, P > 0.05). In particular, a difference was observed in the bone formation and density distributions over time (from 0 or 4 to 8 weeks; approximately 5.15%, P < 0.05). An improved understanding of the alteration pattern on the trabecular bone microarchitecture at tibial epiphysis may assist in developing more targeted treatment interventions for T-OA.

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