• Journal of Periodontal and Implant Science
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• v.28 no.1
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• pp.17-35
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• 1998

Chitosan, with a chemical structure similar to hyaluronic acid, has been implicated as a wound healing agent. The purpose of this research was to evaluate the effects of chitosan on the characteristics of periodontal ligament cells, calvaria cells and gingival fibroblasts and to define the effects of chitosan on bone formation in vitro. In control group, the cells were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% Fetal bovine serum, 100unit/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, chitosan($40{\mu}g/ml$) is added into the above culture condition. And then each group was characterized by examining the cell proliferation at 1,3,5,7,9,12,15 day, the amount of total protein synthesis, alkaline phosphatase activity at 3, 7 day and the ability to produce mineralized nodules of rat calvaria cell at 11 day. The results were as follows : 1. At early time both periodontal ligament cells and calvaria cells in chitosan-treated group proliferated more rapidly than in non-treated control group, but chitosan-treated group of periodontal ligament cells at 9 days and calvaria cells at 12days showed lower growth rate than control group. Gingival fibroblast in chitosan-treated group had lower growth rate than in control group but the difference was not statistically significant (P< 0.01).2. Both periodontal ligament cells and calvaria cells in chitosan-treated group showed much protein synthesis than in control group at 3 days, but showed fewer than in control group at 7 days. Amount of total protein synthesis of gingival fibroblast didn't have statistically significant difference among the two groups(P< 0.01). 3. At 3 and 7 days, alkaline phosphatase activity of periodontal ligament cells and calvaria cells was increased in chitosan-treated group, but at 7 days there was not statistically significant difference among the two groups of calvaria cells (P< 0.01). Alkaline phosphatase activity of gingival fibroblast didn't have statistically significant difference among the two groups(P<0.01). 4. Mineralized nodules in chitosan-treated group of rat calvaria cells were more than in control group. In summery, chitosan had an effect on the proliferation, protein systhesis, alkaline phosphatase activity of periodontal ligament cells and calvaria cells, and facilitated the formation of bone. It is thought that these effects can be used clinically in periodontal regeneration therapy.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.41 no.4
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• pp.303-313
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• 2015

Titrated extract of Centella asiatica (TECA), which is poorly soluble in water is well known for wound healing and anti wrinkle agent. This study was conducted to find the optimum condition for the preparation of water in oil in water ($W_1/O/W_2$) emulsion containing TECA. Solubility of TECA were measured by UV spectrophotometer. 2.55 g of TECA was dissolved in solution composed of dipropylene glycol (40.0 g), ethanol (20.0 g), and water (10.0 g). Factors affecting stability of the emulsions ($W_1/O$, $W_1/O/W_2$) was investigated. The optimum conditions for the preparation of $W_1/O$ emulsion was composed of dipropylene glycol : ethanol : water : TECA in a weight ratio of 40.0 : 20.0 : 10.0 : 2.5 for water phase and squalane : cetyl PEG/PPG-10/1 dimethicone : cetearyl alcohol in a weight ratio of 22.5 : 4.0 : 2.5 for oil phase. The optimum conditions for the preparation of $W_1/O/W_2$ multiple emulsion was composed of water : $W_1/O$ emulsion : polysorbate 80 : carbomer : triethanolamine in a weight ratio of 55.8 : 40.0 : 4.0 : 0.1 : 0.1.

• Journal of Nutrition and Health
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• v.55 no.2
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• pp.240-249
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• 2022

Purpose: Glabridin (GD) is a bio-available isoflavane isolated from the root extract of licorice (Glycyrrhiza glabra L.). It exhibits a variety of pharmacological activities such as anti-inflammatory and anti-oxidant activities. However, extracellular vesicles (EVs) secretion and the anti-cancer mechanism of action remains largely unknown. The present study investigates the anticancer effects of GD by determining the inhibition of EVs secretion in the human breast cancer cell line, MDA-MB-231. Methods: Cell viability, reactive oxygen species (ROS) production, migration, invasion rate, and vascular endothelial growth factor (VEGF) concentration were assessed in MDA-MB-231 cells treated with increasing concentrations of GD (0.1, 1, 5, 10, 20 µM). Subsequently, EV secretion and exosomal DEL-1 protein expression were evaluated to determine the anticancer effects of GD. Results: The results showed that GD significantly inhibited the cell proliferation of MDA-MB-231 cells in a dose- or time-dependent manner. Also, ROS production and apoptosis marker protein cleaved caspase-3 were significantly increased in GD-treated MDA-MB-231, compared to control. Furthermore, GD exposure resulted in significantly decreased not only migration and invasion rates but also the VEGF concentration, thereby contributing to a reduction in angiogenesis. Interestingly, the concentration and number of EVs as well as EV marker proteins, such as CD63 and TSG101, were decreased in GD-treated MDA-MB-231 cells. Markedly, extracellular matrix protein DEL-1 as angiogenesis factor was decreased in EVs from GD-treated MDA-MB-231 cells. Conclusion: This study identifies that the anti-cancer molecular mechanism of GD is exerted via inhibition of angiogenesis and EVs secretion, indicating the potential of GD as a chemotherapeutic agent for breast cancer.

• Journal of Periodontal and Implant Science
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• v.29 no.3
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• pp.561-578
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• 1999

The goal of periodontal therapy is the periodontal regeneration by the removal of microorganisms and their toxic products from the periodontally diseased root surface. To achieve periodontal regeneration, root conditioning as an adjunct to root planing has been done. There are low pH etchants such as citric acid, tetracycline-HCl, and EDTA solution which is a neutral chelating agent. The purpose of present study was to examine the effect of root conditioning by citric acid, tetracycline HCl, and EDTA. Total 35 root specimens(6${\times}$3${\times}$2mm) were prepared from the periodontally diseased teeth, scaled and root planed. The specimens were treated with normal saline for 1 minute, saturated citric acid(pH 1) for 3 minutes, 50mg/ml tetracycline-HCl(pH 2) for 5 minutes, 15% EDTA(pH 7) for 5 minutes using rubbing technique. The specimens were examined under scanning electron microscopy at 1000, and 3000 magnification. On the microphotographs taken at 1000 magnification, the numbers of opened and patent dentinal tubules per unit area(10,640${\mu}m^2$) were counted. And the diameters of opened dentinal tubules per unit are (10,640${\mu}m^2$) were measured. The differences of number and diameter among all groups were statistically analyzed by Kruskal Wallis Test. The results were as follows; 1. In the specimens applied with normal saline(control group), the root surface was finely cracked, and was covered by irregular smear layer. Neither exposed dentinal tubules nor any patent dentinal tubules could be seen. 2. In the specimens applied with saturated citric acid(experimental 1 group), the globular collagen fibers were exposed around the peritubular space, and many dentinal tubules were revealed. 3. In the specimens applied with tetracycline-HCl(experimental 2 group), the process-like collagen fibers were exposed around the peritubular space, and some dentinal tubules were revealed. 4. In the specimens applied with 15% EDTA(experimental 3 group), the root surface was covered by the collagenous fibrillar network, and many dentinal tubules were revealed. 5. The numbers of opened and patent dentinal tubules were significantly more in exp. 1 group and exp. 3 group than in exp. 2 group(P<0.05). But there was no significant difference between exp. 1 group and exp. 3 group. In control group, the number of opened and patent dentinal tubules could not be counted because any dentinal tubules couldn't be seen. 6 . The diameter of opened dentinal tubules was significantly smaller in exp. 1 group and exp. 3 group than in exp. 2 group(P<0.05). But there was no significant difference between exp. 1 group and exp. 3 group. In control group, the diameter of opened dentinal tubules could not be measured because any dentinal tubules couldn't be seen. The results demonstrate that root conditioning with citric acid, tetracycline- HCl, and EDTA is more effective in periodontal healing than only root planing, and 15% EDTA solution can replace low pH etching agents such as citric acid, tetracycline-HCl for root conditioning.

• Journal of Periodontal and Implant Science
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• v.24 no.2
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• pp.219-237
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• 1994

The ultimate goal of clinical periodontal therapy is to achieve regeneration of a healthy connective tissue reattachment. Conventional therapy including scaling, root planing, gingival curettage, gingivectomy and flap procedures of various types results primarily in repair rather than regeneration of the periodontium. In order for periodontal regeneration to occur, progenitor periodontal ligament cells must migrate to the denuded root surface, attach to it, proliferate and mature into an organized and functional fibrous attachment apparatus. Polypeptide growth factors belong to a class of potent biologic mediators which regulate cell differentiation, proliferation, migration and metabolism. Insulin-like growth factor-I (IGF- I ) of these factors appear to have an important role in periodontal wound healing and bone formation. The purpose of this study is to evaluate the effects of IGF- I on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were obtained from periodontal tissue explants culture of the first premolar tooth extracted for the orthodontic treatment. Cells were cultured in Dulbecco's modified Eagle medium(DMEM) with 10% fetal bovine serum. Fourth to seventh passage cells were plated in 24 well tissue culture plates and medium changed to serum-free medium prior to addition of growth factors. Cell proliferation was measured by the incorporation of $[^3H]-thymidine$ into DNA, Protein synthesis was determined by measurement of $[^3H]-proline$ incorporation into collagenase-digestible protein(CDP) and noncollagenous protein(NCP) according to the method of Peterkofsky and Diegelmann (1971), And alkaline phosphatase activity was measured as one parameter of osteoblastic differentiation. The results were as follows : The DNA synthetic activity was increased in a dose-dependent manner with IGF- I except for 0.1ng/ml concentration of IGF- I At the concentration of 10, 100ng/ml, IGF- I significantly increased the DNA synthetic activity(P<0.05) The total protein, collagen and noncollagen synthesis was increased in a dose-dependent manner with IGF- I except for 0.1ng/ml concentration of IGF- I. At the concentration of 1, 10, 100ng/ml, IGF- I significantly increased the total protein, collagen and noncollagen synthesis activity(P<0.95, P<0.001). The % of collagen was not effected according to the concentration of IGF- I. The alkaline phosphatase activity was increased in a dose-, time-dependent manner with IGF- I (10, 100ng/ml). In conclusions, the present study shows that IGF- I has a potentiality to enhance the DNA synthesis of periodontal ligament cells with including the increase of the total protein and collagen synthetic activity. The use of IGF- I to mediate biological stimulation of periodontal ligament cells shows promise for future therapeutic applications.

• Journal of Periodontal and Implant Science
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• v.24 no.2
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• pp.303-320
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• 1994

Current acceptable methods For promotin gperiodontal regeneration are base on removal of diseased soft tissue, root treatment, guided tissue regeneration, inteoduction of new graft materials and biological mediators. Platelet-derived growth factor(PDGF) is one of polypeptide growth factor. PDGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purposes of this study is to evaluate the effects of PDGF-AA, BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/10% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Author measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis and alkaline phosphatase activity according to the concentration of PDGF-AA and BB(0, 0.1, 1, 10, 100ng/ml). The results were as follows : The DNA synthetic activity was increased dose dependently by PDGF-AA and BB. The maximum mitogenic effect was at the 100ng/ml of PDGF-AA and 10ng/ml of PDGF-BB. The total protein, collagen and noncollagen systhesis was increased dose dependently by PDGF-AA and BB. The % of collagen was slightly decresed according to the concentration of PDGF-AA and BB. The effect of PDGF-AA and BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. The effect of PDGF-AA and BB on alkaline phosphatase activity did not show any significant, meanwhile the alkaline phosphatase activity of 14 days group showed significnat increase. In conclusion, PDGF-AA and BB may have important roles in stimulation of DNA synthesis in human periodontal ligament cells, which means an increase in collagen-synthesizing cells, and may be useful for clinical application in periodontal regenerative procedures.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.44 no.3
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• pp.349-362
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• 2018

An annual plant, Eclipta prostrata (Linn) is a member of the Asteraceae plant family and inhabited in tropical or subtropical regions of the world. Through many previous researches, E. prostrata has been extensively studied for its hepatoprotective effect, antivenom potential against viper venom, antioxidant, hair-growth, wound-healing efficacy and so on. In this study, for better understanding of the potential of E. prostrata as skin protectant, we conducted the experiments evaluating the antioxidant and antiaging efficacy. To this end, 50% ethanolic extract of E. prostrata and its ethyl acetate fraction were prepared and investigated. For the evaluation of antioxidant capacity of the samples, $FSC_{50}$ and $OSC_{50}$ were estimated. As a result, $OSC_{50}$ of ethyl acetate fraction was 2.7 times superior to $OSC_{50}$ of L-ascorbic acid, a well known antioxidant agent. Futhermore E. prostrata showed notable reactive oxygen species (ROS) scavenging effect and protective effect against $H_2O_2$ in the celluar level as well. Especially, in the $^1O_2$ induced hemolysis test, $64{\mu}g/mL$ of ethyl acetate fraction showed greater than 6 times increased retardation effect compare to control which means E. prostrata has remarkable antioxidant capacity. To validate the antiaging effect of the samples, we conducted elastase inhibition assay using elastase solution extracted from human skin fibroblasts, Hs68. As a result, $16{\mu}g/mL$ of each sample showed 6.8% and 14.0% of elastase inhibition respectively. Finally, antimicrobial activity of E. prostrata was assessed to validate the possibility as alternative preservative. From the result, ethyl acetate fraction showed oustanding antimicrobial activity as of methyl paraben, a well known chemical preservative. In conclusion, these results suggest that E. prostrata can be used as natural skin protectant or preservative as natural ingredient in food or cosmetics industry.

• Journal of Periodontal and Implant Science
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• v.27 no.4
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• pp.785-804
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• 1997

It was well known that calcium sulfate was biocompatible, resorbed rapidly in the body, had potential as a good barrier membrane. Platelet-derived growth factor(PDGF) was one of polypeptide growth factor that had been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purpose of this study was to evaluate the effects of a combination of calcium sulfate and PDGF on periodontal ligament cells in vitro to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the premolar tooth extracted for the orthodontic treatment. Cells were cultured in ${\alpha}-MEM$ contained with 20% FBS, at the $37^{\circ}C$, 100% of humidity, 5% $Co_2$ incubator. Cells were inoculated and cultured into 96 well culture plate with $1{\times}10^4cells/well$ of ${\alpha}-MEM$ for 1 day. After discarding the medium, those cells were cultured in ${\alpha}-MEM$ contained with 10% FBS alone(control group), in calcium sulfate(calcium sulfate group), in calcium sulfate treated with 15ng/ml of PDGF-BB(calcium sulfate+PDGF group), in ${\alpha}-MEM$ contained with 10% FBS treated with 15ng/ml of PDGF-BB(PDGF group) for 1, 2, 3 day respectively. And then each group was characterized by examining of the cell counting, MTT assay, collagen synthesis. The results were as follows. 1. In the analysis of cell proliferation by cell counting, both calcium sulfate group and calcium sulfate plus PDGF group showed no stastically significant difference compared to control group, but there was stastically significant difference between PDGF group and calcium sulfate group at 1, 2 day(P<0.05). 2. In the analysis of cell proliferation by MTT assay in calcium sulfate extracts, both calcium sulfate group and calcium sulfate plus PDGF group showed no stastically significant difference compared to control group, but there was stastically significant difference between PDGF group and calcium sulfate group at 2, 3 day, and between calcium sulfate plus PDGF group and calcium sulfate group at 2 day(P

• Journal of Veterinary Clinics
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• v.18 no.3
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• pp.257-264
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• 2001

This study was performed to investigate the effects of chitosan-trimer (CT) on the prevention of postoperative adhesion formation in the rate model. All animals divided into PBS (control), 1% CT, 3% CT, and chitin treated group. The mean adhesion score in 1% CT group (1.03$\pm$0.63), 3% CT group (0.64$\pm$0.53) and chitin group (1.67$\pm$0.71) was found to be lower than that in control group (2.07$\pm$0.81). More favorable adhesion prevention was achieved in 3% CT group (0.64$\pm$0.53) in comparison with the control group, 1% CT group, and chitin group without any hemorrhagic complications. A statistically significant difference was observed in adhesion formation between control group and 3% CT group (p<0.001). In control group, 44 of 45 sites (97.7%) formed adhesions between the intestine defects. In contrast, 3% CT was effective in reducing the incidence of adhesion formation to 17 to 45 sites (62.2%) (p<0.05). The locations of adhesions were observed in serosa-serosa (60%), serosa-mesentery (13.3%), serosa-connective tissue of testis (10%), omentum-liver (10%), serosa-omentum (3.3%), serosa-cecum (3.3%), and serosa-incision (0%). On the results of histological analysis, grade of inflammation and fibrosis at the sites of postoperative peritoneal adhesion formation were not significantly different in all groups. But, 3% CT showed the lowest score of inflammation and fibrosis. In 3% CT group, the rate of increase of plasma fibrinogen was significantly lower compared with that in control group from pre-operation to 10 days later (p<0.05). There were no appreciable difference in the CBC, leukocyte differential counts and total protein concentrations among four groups. In conclusion, our data suggested that CT should be effective on reducing adhesion formation in experimental rat models. The results also showed that 3% CT does not adversely affect normal wound healing and healthy recovery after operation.

• Journal of Periodontal and Implant Science
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• v.23 no.3
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• pp.535-563
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• 1993

New techniques for regenerating the destructed periodontal tissue have been studied for many years. Current acceptable methods of promoting periodontal regeneration alre basis of removal of diseased soft tissue, root treatment, guided tissue regeneration, graft materials, biological mediators. Platelet-derived growth factor (PDGF) is one of polypeptide growth factor. PDGF have been reported as a biological mediator which regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the possibility of using the PDGF as a regeneration promoting agent for furcation involvement defect. Eight adult mongrel dogs were used in this experiment. The dogs were anesthetized with Pentobarbital Sodium (25-30 mg/kg of body weight, Tokyo chemical Co., Japan) and conventional periodontal prophylaxis were performed with ultrasonic scaler. With intrasulcular and crestal incision, mucoperiosteal flap was elevated. Following decortication with 1/2 high speed round bur, degree III furcation defect was made on mandibular second(P2) and fourth(P4) premolar. For the basic treatment of root surface, fully saturated citric acid was applied on the exposed root surface for 3 minutes. On the right P4 20ug of human recombinant PDGF-BB dissolved in acetic acid was applied with polypropylene autopipette. On the left P2 and right P2 PDGF-BB was applied after insertion of ${\beta}-Tricalcium$ phosphate(TCP) and collagen (Collatape) respectively. Left mandibular P4 was used as control. Systemic antibiotics (Penicillin-G benzathine and penicillin-G procaine, 1 ml per 10-25 1bs body weight) were administrated intramuscular for 2 weeks after surgery. Irrigation with 0.1% Chlorhexidine Gluconate around operated sites was performed during the whole experimental period except one day immediate after surgery. Soft diets were fed through the whole experiment period. After 2, 4, 8, 12 weeks, the animals were sacrificed by perfusion technique. Tissue block was excised including the tooth and prepared for light microscope with H-E staining. At 2 weeks after surgery, therer were rapid osteogenesis phenomenon on the defected area of the PDGF only treated group and early trabeculation pattern was made with new osteoid tissue produced by activated osteoblast. Bone formation was almost completed to the fornix of furcation by 8 weeks after surgery. New cementum fromation was observed from 2 weeks after surgery, and the thickness was increased until 8 weeks with typical Sharpey’s fibers reembedded into new bone and cementum. In both PDGF-BB with TCP group and PDGF-BB with Collagen group, regeneration process including new bone and new cementum formation and the group especially in the early weeks. It might be thought that the migration of actively proliferating cells was prohibited by the graft materials. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.