• Biomedical Science Letters
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• v.14 no.3
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• pp.167-172
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• 2008

We compared the clinical efficacy of LED therapy using 880 nm and 630 nm LED to test collagen accumulations in subcutaneous tissue of mouse after LED irradiation by measuring the quantity of collagen. 880 nm and 630 nm LED was irradiated on the back of ICR mouse given at $10.8J/cm^2$ followed for 30 minutes everyday for 5 weeks. Histological observation was performed by Hematoxylin & Eosin staining and Masson's Trichrome collagen staining. We also used Sircol soluble collagen assay kit for measuring the amounts of collagen in the mouse skin tissue after 1, 3, and 5 weeks post LED irradiation, respectively. Collagen generation was found at subcutaneous tissue, and the quantity of collagen in 880 nm LED group had grown more than that of 630 nm LED group at 5 weeks follow-up later. About 75% more efficacies for collagen generation were found in the group of 5th week of 880nm LED irradiation. The efficacy of 880nm LED could be more useful than 630 nm LED for synthesizing collagens in mouse subcutaneous tissue as time followed.

• The Korea Journal of Herbology
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• v.31 no.4
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• pp.1-10
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• 2016

Objectives: In Korean medicine, Curculiginis Rhizoma was treated for arthritis in remedy. But efficacy of Curculiginis Rhizoma on collagen induced arthritis was not revealed.Methods: Anti inflammatory effect of Curculiginis Rhizoma was researched in vitro with RAW264.7 cell and cell toxicity, levels of proinflammatory cytokines (TNF-α, IL-1β, IL-6 and IL-12) and PGE2 were analyzed by ELISA assay. Inflammatory protein were analyzed by western blotting assay (JNK, ERK, COX-2, TNF-α and IL-1β). In vivo, collagen induced arthritis mice model was used to evaluate anti-inflammation effect through arthritis index, immune cell number and cytokine levels (TNF-α, IL-6 and IL-1β) in serum.Results: ECR(Extract of Curculiginis Rhizoma) has not shown cell toxicity in 200 ㎍/㎖ on RAW264.7 cell. ECR suppressed releases of NO, TNF-α, IL-1β, IL-6, IL-12 and PGE2 on RAW264.7 cell treated with lipopolysacharide (1 ㎍/㎖). And ECR inhibited regulation of TNF-α, IL-1β and IL-6 mRNA, reduced protein release of JNK, ERK, iNOS, COX-2, IL-1β and TNF-α. AI of group treated with ECR 200 ㎎/㎏ and 100 ㎎/㎏ were significantly decreased compared to vihicle arthritis mice, the number of immune cell in foot joint was increased on control mice but those of group treated with ECR 200 ㎎/㎏ and 100 ㎎/㎏ were significantly reduced. This results correspond with contens of cytokines (TNF-α, IL-1β and IL-6) in serum.Conclusions: Curculiginis Rhizoma has anti-inflammation effect on RAW264.7 cell in vitro and collagen induced arthritis in vivo. So it is necessary to research more mechanism for cascade imfact.

• Journal of Acupuncture Research
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• v.22 no.2
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• pp.191-201
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• 2005

Background & Objective: Articular cartilage is a potential target for drugs designed to inhibit the activity of matrix metalloproteinases (MMPs) to stop or slow the destruction of the proteoglycan and collagen in the cartilage extracellular matrix. The purpose of this study was to investigate the effects of Aralia cordata Thunb. in inhibiting the release of glycosaminoglycan (GAG), the degradation of collagen, and MMP activity in rabbit articular cartilage explants. Methods : The cartilage-protective effects of Aralia cordata Thunb. were evaluated by using glycosaminoglycan degradation assay, collagen degradation assay, colorimetric analysis of MMP activity, measurement of lactate dehydrogenase activity and histological analysis in rabbit cartilage explants culture. Results : Interleukin-la (IL-1a) rapidly induced GAG, but collagen was much less readily released from cartilage explants. Aralia cordata Thunb. significantly inhibited GAG and collagen release in a concentration-dependent manner. Aralia cordata Thunb. dose-dependently inhibited MMP-3 and MMP-13 expression and activities from IL-1a-treated cartilage explants cultures when tested at concentrations ranging from 0.02 to 0.2 mg/ml. Aralia cordata Thunb. had no harmful effect on chondrocytes viability or cartilage morphology in cartilage explants. Histological analysis indicated that Aralia cordata Thunb. reduced the degradation of the cartilage matrix compared with that of IL -1a-treated cartilage explants.

• Proceedings of the Materials Research Society of Korea Conference
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• 2011.05a
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• pp.54.2-54.2
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• 2011

DTBP (dimethyl 3,3`-dithiobispropionimidate) was applied to collagen and gelatin coating on BCP granules and a crosslinking agent. The DTBP crosslinking was done for decreasing the solubility of the coating and hence increasing the stability. The nanostructure of collagen and gelatin coating surfaces were observed by SEM technique. Based on the DSC thermograms and FT-IR spectrums, the crosslinkings were confirmed between collagen molecules and gelatin molecules. The compressive strength was measured before crosslinking and after that. In-vitro study was carried out by measuring cell viability and observing cell morphology after DTBP crosslinking. Moreover, the proliferation ability of MG-63 osteoblast-like cells on the crosslinked BCP granules was evaluated by Western blot assay. The BCP granules were implanted into rabbit femur for 4 weeks and 12 weeks. The bone tissue formation was analyzed with micro-computed tomography (micro-CT) and histological analysis was also carried out by hematoxylin and eosin (H&E) staining for visualization of cells.

• Proceedings of the SCSK Conference
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• 1996.07a
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• pp.52-67
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• 1996

새로운 노화방지 물질로 개발한 3-APPA가 노화에 의해 야기되는 여러 변화들, 특히 세포 증식, 유전자 수준 및 단백질 수준에서의 collagen의 생합성 변화, 면역조직화학염색을 이용한 collagen 생합성의 변화등을 세포배양 및 동물실험을 통하여 측정하였다. MTT assay를 이용한 인체 피부 섬유아세포의 증식 실험에서 3-APPA는 무처치군에 비교해서 최고 2배의 섬유아세포 증식 효능을 나타내었으며, $^3$[H]-proline incorporation 방법을 이용한 단층세포 배양 및 3차원 dermal equivalent 섬유아세포 배양에서 무처치군 및 vitamin C 처리군에 비해 최고 1.5배의 collagen 생합성 증가를 나타내었다. 그러나 type I alpha-procollagen mRNA expression에는 영향을 미치지 않는 것으로 나타났다. H&E 염색을 이용한 hairless mice의 피부에 대한 형태학적 변화 및 type I pM procollagen antibody를 이용한 면역조직화학염색에서, 3-APPA는 collagen 생합성을 증가시키는 것으로 나타났다. 이상의 결과에서 3-APPA는 섬유아세포 배양 및 hairless mouse를 이용한 실험에서 피부 섬유아세포 증식을 촉진시키며 collagen 생합성을 증가시켜 피부노화를 억제 할 수 있는 물질임을 밝혔다.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.4
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• pp.365-371
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• 2014

To identify new active anti-wrinkle ingredients, this study investigated the anti-wrinkle effects of Schizandra chinensis Baillon fermented with Lactobacillus plantarum (SCF) by assessment of cytotoxicity of human dermal fibroblast, collagen biosynthesis, matrix metalloproteinase-I (MMP-1) inhibition and elastase inhibition. S. chinensis was fermented with L. rhamnosus for 1 day at $37^{\circ}C$. The cytotoxicity of SCF was evaluated by a cytopathic effect reduction method. Effects on collagen biosynthesis and matrix metalloproteinase-I (MMP-1) of SCF were evaluated by previous reported method using procollagen type-IC peptide EIA kit and Matrix Metalloproteinase-1 Biotrack activity Assay Kit, respectively. Elastase inhibition assay was conducted by reaction of enzyme and substrate using N-Suc-$(Ala)_3$-nitroanilide as the substrate. As the results, SCF didn't show cytotoxicity against human dermal fibroblast at concentration of $100{\mu}g/mL$. Also, SCF was increased collagen synthesis and showed inhibitory effect of MMP-1 (p < 0.05). In the elastase inhibition assay, the $IC_{50}$ of SCF was $36.4{\mu}g/mL$. Therefore, our results indicated that SCF possesses anti-wrinkle effects and can be used practically for anti-wrinkle care of skin.

• Archives of Plastic Surgery
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• v.36 no.3
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• pp.247-253
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• 2009

Purpose: Hydroxyapatite(HA) has been widely used due to its chemical similarity to bone and good biocompatibility. HA is composed of macropores and micropores. Too much irregularities of the micropores are ineffective against the adhesion and proliferation of osteoblast. Many efforts have been tried to overcome these drawbacks. HA crystal coating on the irregular surface of HA scaffold, crystallized HA, is one of the method to improve cell adhesion. Meanwhile, the collagen has been incorporated with HA to create composite scaffold that chemically resembles the natural extracellular matrix components of bone. The authors proposed to examine the effect of collagen - coated crystallized HA on the adhesion and proliferation of osteoblast. Method: HA powder containing $10{\mu}m$ pore size was manufactured as 1 cm pellet size. For the making crystallized HA, 0.1 M EDTA solution was used to dissolve HA powder and heated $100^{\circ}C$ for 48 hours. Next, the crystallized HA pellets were coated with collagen (0.1, 0.5, and 1%). The osteoblasts were seeded into HA pellets and incubated for the various times (1, 5, and 9 days). After the indicating days, methylthiazol tetrazolium (MTT) assay was performed for cell proliferation and alkaline phosphatase (ALP) activty was measured for bone formation. Result: In SEM study, the surface of crystallized HA pellet was more regular than HA pellet. MTT assay showed that the proliferation of osteoblasts increased in a collagen dose - dependent and time - dependent manner and had a maximum effect at 1% collagen concentration. ALP activity also increased in a collagen dose - dependent manner and had a highest effect at 1% collagen concentration. Conclusion: These data showed that crystallization and collagen coating of HA was effective for osteoblast proliferation and ALP activity. Therefore, our results suggest that crystallized - HA scaffold with collagen coating is may be a good strategy for tissue engineering application for bone formation.

• The Journal of Internal Korean Medicine
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• v.32 no.4
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• pp.504-518
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• 2011

Objectives : This study was performed to investigate the anti-fibrogenic effect of greater celandine on cultured rat hepatic stellate cells. Materials and Methods : Hepatic stellate cells (HSC-T6) were treated with various concentrations of greater celandine extract for 24, 48, and 72 hours. The extraction was done with distilled water. After the treatment, cell viability, proliferation, mRNA of the ${\alpha}SMA$, TIMP-1, TIMP-2, collagen I ${\alpha}$ 1, MMP-2, IL-6, TGF-${\beta}1$, PDGFr-${\beta}1$, Bcl-2, Bax, Bcl-xl, caspase-3, caspase-9 and the activities of SOD and catalase were measured by using MTT assay, BrdU assay, real-time PCR, superoxide dismutase assay and catalase assay. Results : The viability, proliferation, mRNA expression and synthesis of collagen of the hepatic stellate cells were inhibited as the concentration increased, which indicates the herb has an inhibitory effect on fibrogenesis of the liver by regulating the fibrosis associated genes in transcription. Conclusions : These results suggest that greater celandine would be beneficial in the treatment of fibrotic patients as well as for patients with chronic hepatitis.

• Nutrition Research and Practice
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• v.1 no.4
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• pp.279-284
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• 2007

Solar ultraviolet (UV) irradiation leads to distinct changes in the skin connective tissues by degradation of collagen, which is a major structural component in the extracellular matrix. UV irradiation induces the production of matrix metalloproteinases (MMP) capable of attacking native fibrillar collagen and responsible for inhibiting the construction of collagenous extracellular matrix. In this study, we attempted to investigate the protective actions of Rubus coreanus ethanol extract (RCE) on the MMP production and the consequent procollagen/collagen degradation in UV-B-irradiated human dermal fibroblasts. The analytical data showed that Rubus coreanus ethanol extract was mostly comprised of cyanidin 3-rutinoside. Pre-treatment of fibroblasts with this extract inhibited UV-B-induced production of MMP-1, MMP-8 and MMP-13 in dose-dependent manners. In addition, Western blot analysis and immunocytochemical staining assay revealed that RCE markedly augmented the cellular levels of procollagen/collagen declined in UV-B-exposed dermal fibroblasts. These results demonstrate that RCE blocks UV-B-induced increase of the collagen degradation by inhibiting MMP production. Thus, RCE may act as an agent inhibiting excessive dermal collagen degradation leading to the skin photoaging.

• Archives of Plastic Surgery
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• v.32 no.3
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• pp.363-368
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• 2005

It was assumed that the effect of estrogen on wound healing would be variable according to patient's gender and age since estrogen is a sex steroid. This study was designed to determine the variability of the effect of estrogen on proliferation of human dermal fibroblasts and collagen synthesis which are most important in wound healing considering patient's gender and age. Fibroblasts were isolated from the dermis of female patients in premenstrual, menstrual, or postmenopausal age group and that of male patients. The isolated fibroblasts were cultivated in the presence of estrogen($1.0{\mu}g/ml$). The cells were seeded at $5.0{\times}10^3cell/well$ in Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient including 5% fetal bovine serum in 96-well plates. The cells were incubated for 3 days. For fibroblast proliferation MTT assay method was used. To measure the production of collagen, the collagen type I carboxy- terminal propeptide enzyme immunoassay was carried out. Estrogen stimulated the proliferation of fibroblasts in female patients, but not in male patients. The greatest cell proliferation and collagen synthesis was seen at women in menstrual and postmenopausal age. These results demonstrated that effects of estrogen on dermal fibroblast proliferation and collagen synthesis were variable with gender and age.