• Archives of Pharmacal Research
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• 제28권11호
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• pp.1311-1316
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• 2005

To better define the relationship between dermal regeneration and wound contraction and scar formation, the effects of epidermal growth factor (EGF) loaded in collagen sponge matrix on the fibroblast cell proliferation rate and the dermal mechanical strength were investigated. Collagen sponges with acid-soluble fraction of pig skin were prepared and incorporated with EGF at 0, 4, and 8 $\mu$g/1.7 $cm^{2}$. Dermal fibroblasts were cultured to 80$\%$ confluence using DMEM, treated with the samples submerged, and the cell viability was estimated using MTT assay. A deep, $2^{nd}$ degree- burn of diameter 1 cm was prepared on the rabbit ear and the tested dressings were applied twice during the 15-day, post burn period. The processes of re-epithelialization and dermal regeneration were investigated until the complete wound closure day and histological analysis was performed with H-E staining. EGF increased the fibroblast cell proliferation rate. The histology showed well developed, weave-like collagen bundles and fibroblasts in EGF-treated wounds while open wounds showed irregular collagen bundles and impaired fibroblast growth. The breaking strength (944.1 $\pm$ 35.6 vs. 411.5 $\pm$ 57.0 Fmax, $gmm^{-2}$) and skin resilience (11.3 $\pm$ 1.4 vs. 6.5 $\pm$ 0.6 mJ/$mm^{2}$) were significantly increased with EGF­treated wounds as compared with open wounds, suggesting that EGF enhanced the dermal matrix formation and improved the wound mechanical strength. In conclusion, EGF-improved dermal matrix formation is related with a lower wound contraction rate. The impaired dermal regeneration observed in the open wounds could contribute to the formation of wound contraction and scar tissue development. An extraneous supply of EGF in the collagen dressing on deep, $2^{nd}$ degree-burns enhanced the dermal matrix formation.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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• pp.80-80
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• 2003

Insulin-dependent or Type 1 diabetes mellitus (IDDM) has been associated with an increased severity of periodontal disease. Since periodontal ligament (PDL) cells play a significant role in maintenance and regeneration of mineralized tissue, the success of procedures, such as guided tissue regeneration, is directly related to the ability of these cells to augment mineralized tissue. In this study, we investigated the time- and dose-dependent effect of high glucose on the proliferation and collagen synthesis of human periodontal ligament (PDL) cells. PDL cells were treated with high glucose (22mM, 33mM, 44mM) for 1 or 2 days. High glucose significantly inhibited proliferation of PDL cells as a time- and dose-dependent manner as evidenced by MTT assay. PDL cells were cultured in high glucose media (22mM, 33mM, 44mM) for 24 h. The ratio of collagen content to total protein was evaluated, and the gene expression of type I collagen was assessed by RT - PCR. The high concentration of glucose inhibited collagen synthesis, a marker of bone formation activity. This study indicated high glucose concentration could alter the metabolism of periodontal ligament cell, leading to alveolar bone destruction.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 1999년도 IFSCC . ASCS 학술대회 발표 논문
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• pp.145-154
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• 1999

A novel retinol derivative, polyethoxylated retinamide (Medimin A) was synthesized, as an anti-aging agent. Collagen synthesis, skin permeation, stability, and toxicity of Medimin A were evaluated and compared with those of retinol and retinyl palmitate. In vitro collagen synthesis was evaluated by quantitative assay of [$^3H$]-proline incorporation into collagenase sensitive protein in fibroblast cultures. For in vitro skin permeation experiments, Franz diffusion cells (effective diffusion area: $1, 766{\;}\textrm{cm}^2$) and the excised skin of female hairless mouse aged 8 weeks were used The stabilities of retlnoids were evaluated at two different temperature ($25{\;}^{\circ}C$ and $40{\;}^{\circ}C$) and under UV in solubilized state and in OW emulsion. To estimate the safety, acute oral toxicity, acute dermal toxicity, primary skin irritation, acute eye irritation and human patch test were performed The effect of Medimin A on collagen synthesis was similar to that of retinol. The skin permeability of Medimin A was higher than those of retinol and retinyl palmitate. The Medimin A was more stable than retinol and retinyl palmitate. Medimin A was nontoxic in various toxicological tests. These results suggest that Medimin A would be a good anti-aging agent for enhancing bioavailability and stability.

• 생약학회지
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• 제38권2호통권149호
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• pp.101-107
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• 2007

This study was carried out to examine the in vitro effects of alginate (LVA) which is low viscosity alginic acid, on collagen type II synthesis of chondrocytes and the in vivo effect, orally administered, on cartilage degradation. Rabbit articular chondrocytes were cultured in 35 mm dishes and then LVA was treated. The effects of LVA of various viscosity (86.5 cP(LVA1),45.4 cP(LVA2), 21.2 cP(LVA3) and 9.6 cP(LVA4)) and various concentration (50, 100, 200 ${\mu}$g/ml of LVA4) on chondrocytes were determined by western blotting assay for the detection of collagen type II production. In western blotting assay, collagen type II production in chondrocytes were 1.00 in control,0.95 in LVA1, 1.41 in LVA2, 1.57 in LVA3 and 1.58 in LVA4. Collagen type II production of various concentration of LVA4 were 1.00 in control, 1.24 in 50 ${\mu}$g/ml of LVA4, 1.52 in 100 ${\mu}$g/ml of LVA4 and 1.86 in 200 ${\mu}$g/ml of LVA4. Osteoarthritis (OA) was induced in 24 rabbits by unilateral anterior cruciate ligament transection (ACLT) and randomly divided into 6 groups. The experimental group was given oral administration of 0.5 ml/kg of saline(control), 12.5 mg/kg of LVA (A12.5), 25 mg/kg of LVA (A25), 50 mg/kg of LVA (A5O), 75 mg/kg of LVA (A75) and 20 mg/kg of aceclofenac (AC) for 6 weeks after ACLT. All knees were harvested at 6 weeks after surgery and cartilage degradation was evaluated. Cartilage degradation in the control group was significantly more severe than that in the A25, A5O and A75 groups but AC group had no significant changes on the macroscopic grading scale.

• Archives of Plastic Surgery
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• 제32권4호
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• pp.529-532
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• 2005

Expanding cells ex-vivo is very important in tissue-engineering. Culture medium is usually supplemented with fetal bovine serum(FBS) in most of the experiments. However, cells grown in bovine serum media may posses the possibilities of disseminating bovine diseases and/or stimulating the patient's immune reactions. To overcome these problems, autologous or homologous serum should be used instead of the FBS. The purpose of this study is to compare cell proliferation and collagen synthesis depending on the kind of sera mixed on media and to provide a guideline on applying established experimental data to clinical cases. Human dermal fibroblasts were obtained from four patients. Five thousand cells per well in 96-well plates were incubated DMEM/F-12 Nutrient with varying serum mixture; 10% autologous serum, 10% homologous serum, and 10% FBS. Five days after incubation fibroblast proliferation and collagen production were determined by MTT assay and CICP enzyme immunoassay. The mean cell number were; $3.95{\times}10^4/well$, $2.97{\times}10^4/well$ and $2.30{\times}10^4/well$, respectively. The average amounts of collagen synthesized were; 238.13 ng/ml, 204.88 ng/ml, and 163.88 ng/ml in each. These results show that the use of human serum mixture may contribute to, not only preventing disseminated infection of bovine diseases. but also increase cell proliferation and collagen synthesis without simulating the patient's immune reactions.

• Journal of Applied Biological Chemistry
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• 제58권2호
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• pp.183-187
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• 2015

시호 추출물로부터 자외선에 의한 주름개선 효과를 확인하여 화장품 소재로서의 가능성을 검증하였다. 주름 활성 검증을 위하여 전자공여능, elastase, pro-collagen 생합성, Matrix metalloprotease-1 (MMP-1)의 활성을 측정하였다. 시호 추출물의 세포 독성을 측정하기 위하여 MTT assay를 하여 세포 독성이 100%에 가까운 농도인 5, 10, $50{\mu}g/mL$의 농도에서 pro-collagen 생합성과 MMP-1의 활성 검증하였다. 시호 추출물의 전자 공여능과 elastase 활성을 측정한 결과 $1,000{\mu}g/mL$의 농도에서 각각 80, 52%의 저해 활성을 가졌다. 또한 pro-collagen 생합성 역시 UVB를 조사한 후 시호 추출물을 농도 별로 처리 한결과 농도의존적으로 증가하는 것을 확인하였다. 주름생성에 관련되어져 있는 MMP-1의 발현을 알아본 결과 시호 추출물에 의해 total protein 양이 또한 감소되는 것을 확인 할 수 있었다. 따라서 시호는 주름활성을 개선시킬 수 있는 기능성 소재로 활용 될 수 있을것으로 사료된다.

• 생약학회지
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• 제31권4호
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• pp.373-382
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• 2000

This study was performed to investigate the antithrombotic activity and protective effect of hexane fraction of Kamihyulbuchukeotang (KHCTH) on brain injury by KCN and MCA occlusion a prescription of HCT added with Lumbricus and Notoginseng Radix. Experiemental parameters are brain ischemia by MCA occlusion assay, KCN-induced brain injury, pulmonary thrombosis and platelet aggregation assay. The results were summarized as follows; 1. KHCTH extracts significantly inhibited the duration of KCN-induced coma (67%) and mortality (80%). 2. KHCTH extracts significantly suppressed brain ischemic area and edema following MCA occlusion and protected neuron cells as compared with control data. 3. KHCTH extracts inhibited pulmonary thrombosis induced by collagen and epinephrine. 4. KHCTH extracts inhibited platelet aggregation induced by collagen, ADP as agonist up to 76.9% and 32.3% respectivey at 1 mg/ml more effective than water extract of KHCT These data suggested that KHCTH could be applied as the protector of brain ischemia and injury and antithrombotic agent.

• Restorative Dentistry and Endodontics
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• 제34권5호
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• pp.430-441
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• 2009

본 연구는 사람 치수세포 및 치주인대세포의 차이를 알아보고자 배양한 각각의 세포를 CDNA microarray assay를 통하여 유전자의 발현정도의 차이를 비교하였다. 그 결과를 바탕으로 각각의 세포에서 2배 이상의 유전자 발현의 차이를 보이는 유전자중 특징적인 3가지 유전자를 선택하여 RT-PCR로 검증한 결과 다음과 같은 결론을 얻었다; 1. Microarray assay 결과, 치주인대 세포에 비해 치수 세포에서 2배 이상 발현한 유전자 수는 총 51개가 나타났다. 2. RT-PCR의 결과, 치주인대세포에 비해 치수 세포에서 ITGA4, TGF-${\beta}2$ 등이 높게 나타났다. 3. Microarray assay결과, 치수 세포에서 비해 치주인대 세포에서 2배 이상 발현한 유전자 수는 총 19개가 나타났다. 4. RT-PCR의 결과, 치수 세포에 비해 치주인대세포에서 LUM, WISP1, MMP1 등이 높게 나타났다. 본 연구 결과로 치수세포에는 상아질 형성에 관여하는 특징적인 유전자가 치주인대세포에 비해 높게 발현되었으며, 치주인대세포에는 교원질 합성에 관여하는 특징적인 유전자가 치수세포에 비해 높게 발현되어, 치수세포와 치주인데 세포는 유전자 발현의 차이가 나타남을 알 수 있었다.

• Journal of Periodontal and Implant Science
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• 제29권1호
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• pp.31-40
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• 1999

Many factors may affect periodontal changes during the physiologic conditions of woman(e.g. puberty, menstrual cycle, pregnancy, menopause). Recently many research has focused on the immunological changes of host, but the exact mechanism is not clear. Collagen is a major constituent of periodontium, and collagenase specifically digests the collagen and plays a role in destruction of periodontal tissue. So, I suppose that it participates with the cytokines in the inflammation of gingiva and vascular response during the changes of female sex hormones. Because there are some evidences of the existence of the receptors of estrogen and progesterone in the gingiva, it may be a target tissue of female sex hormones. In this experiment, gingival fibroblast and periodontal ligament cell were cultured in the presence of various concentrations of estrogen or progesterone corresponding to the menstrual cycle and pregnancy. Collagenase activity of the supernatant of culture media was determined by Spectrophotometric collagenase assay. The enzyme activity was calculated by the % decrease of the coated collagen. 1. The estrogen at both concentrations had no effect on the activity of collagenase of the gingival fibroblast. 2. The progesterone had some effect on the collagenase activity of the gingival fibroblast at low and high concentration of menstrual cycle, and elevated the enzyme activity at all range of pregnancy concentrations. 3. In periodontal ligament cells, estrogen elevated the enzyme activity at the early pregnancy concentration and progesterone elevated at the concentration just before menstruation. In this experiment, pregesterone elevated the collagenase activity of gingival fibroblast and periodontal ligament cells. But the mechanism of the up-regulation of the enzyme activity was not confirmed. The more experiments of direct effect of progesterone on gingival at the molecular level(e.g. northern blot analysis) can reveal the exact mechanism.

• 대한한방내과학회지
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• 제17권1호
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• pp.34-50
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• 1996

The present experiment was desinged to investigate the effects of Tongdosan water extracts on the Cardiovascular System in the Experimental Animals. Thus, the changes of blood pressure and heart rate were measured after oral admini- stration. Measurment of Mortality rate was observed for measuring the effect of Tongdosan water extract. Tongdosan water extract against pulmonary thrombo- embolism induced by collagen the mixture(0.1ml/10g, 2mg/kg B.W) plus serotonin(5mg/kg B.W) in mouse. The effect of Tongdosan water extract was examined by observing the change of collagen-induced platelet aggregation, coagulation activity, ex vivo and in vitro fibrinolytic activity of euglobulin fraction in rats. The results were summarized as followings. 1. Tongdosan dropped the blood pressure in spontaneous hypertensive rat. 2. The drug increased the auricular blood flow in rabbit. 3. The drug relaxed the artery contraction by pretreated norepinephrine in rat. 4. The drug inhibited the death rate of mouse which was led to thromboembo- lism by serotonin and collagen. 5. The drug inhibited the platelet aggregation in rat. 6. The drug prolonged the prothrombin time and activated partial thromboplastin time on the test of plasma coagulation factor activity in rat, but was not valuable. 7. The drug reduced the fibrinogen lyses time of rat ex vivo assay and lyses area was increased. 8. Tongdosan reduced fibrinogen lyses time of rat in vitro assay. According to the above mentioned results, Tongdosan increased the blood flow and dropped the blood pressure by the dilation of blood vessel. And the drug presented the antithrombin acivity, inhibited the platelet aggregation.