• Archives of Pharmacal Research
• /
• v.28 no.11
• /
• pp.1311-1316
• /
• 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.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 2003.11a
• /
• pp.80-80
• /
• 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.

• Proceedings of the SCSK Conference
• /
• 1999.10a
• /
• pp.145-154
• /
• 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.

• Korean Journal of Pharmacognosy
• /
• v.38 no.2 s.149
• /
• pp.101-107
• /
• 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
• /
• v.32 no.4
• /
• pp.529-532
• /
• 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
• /
• v.58 no.2
• /
• pp.183-187
• /
• 2015

The electron donating ability, elastase inhibitory, procollagen synthesis and Matrix metalloprotease-1 (MMP-1) activities were measured in order to verify the anti-wrinkle properties of extracts from Buplerum falcatum as a functional ingredient for cosmetic products. Electron donating ability and elastase inhibition activities were 80 and 52% at a dose of $1,000{\mu}g/mL$ of B. falcatum 70% ethanol extract. Pro-collagen synthesis was increased with the increase concentration of B. falcatum extract on CCD-986sk in addition to decrease the amount of protein of MMP-1. The results suggested that B. falcatum extract can be used to reduced electron donating ability, elastase, pro-collagen synthesis and MMP-1 activity and is a potential candidate for cosmedical materials.

• Korean Journal of Pharmacognosy
• /
• v.31 no.4
• /
• pp.373-382
• /
• 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
• /
• v.34 no.5
• /
• pp.430-441
• /
• 2009

The purpose of this study was to characterize functional distinction between human dental pulp cells(PC) and periodontal ligament cells(PDLC) using cDNA micro array assay and to confirm the results of the microarray assay using RT-PCR. 3 genes out of 51 genes which were found to be more expressed(>2 fold) in PC were selected, and 3 genes out of 19 genes which were found to be more expressed(>2 fold) in PDLC were selected for RT-PCR as well. According to this study, the results were as follows: 1. From the micro array assay, 51 genes were more expressed (2 fold) from PC than PDLC. 2. RT-PCR confirmed that ITGA4 and TGF ${\beta}2$ were more expressed in PC than in PDLC 3. From the micro array assay, 19 genes were more expressed (2 fold) from PDLC than PC. 4. RT-PCR confirmed that LUM, WISP1. and MMP1 were more expressed in PDLC than in PC. From the present study, different expression of the genes between the PC and PDLC were characterized to show the genes which play an important role in dentinogenesis were more expressed from PC than PDLC, while the genes which were related with collagen synthesis were more expressed from PDLC than PC.

• Journal of Periodontal and Implant Science
• /
• v.29 no.1
• /
• pp.31-40
• /
• 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.

• The Journal of Internal Korean Medicine
• /
• v.17 no.1
• /
• pp.34-50
• /
• 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.