• Journal of Periodontal and Implant Science
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• v.26 no.2
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• pp.396-413
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• 1996

Current acceptable methods for promoting periodontal regeneration are based on removal of diseased soft tissue. root treatment, guided tissue regeneration, introduction of new graft materials and biological mediators. Insulin-like growth factor-I(IGF-I) and Platelet-derived growth factor-BB(PDGF-BB), the members of the polypeptuyde growth factor family have been reported as the biological mediators which regulate a variety cellular matrix biologic activities of wound healing process including the cell proliferation, migration and extracellular matrix synthesis.The purposes of this study is to evaluate the combination effects of IGF-I and PDGF-BB on the cellular activity of the periodontal ligament cells to act 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 containing 10% FBS at the $37^{\circ}C$, 5% CO2 incubator. Author measured the DNA synthetic activity, and total protein, collagen and noncollagenous protein synthetic activities according to the concentration of 10,100ng/ml IGF-I and1,10 ng/ml PDGF-BB in combination. The results were as follows: Significantly increased in the 1 ng/ml PDGF-BB alone compared to the 10 ng/ml PDGF-BB alone(P<0.01) and in the 1 ng/ml PDGF-BB and 10, 100ng/ml IGF-I in combination compared to the 1 ng/ml PDGF-BB alone(P<0.05, P<0.0l). The synthetic activity of the total protein and collagen is significantly increased like to the synthetic activity of the DNA(P<0.05). The synthetic activity of the noncollagenous protein is increased according to the concentration of IGF_I, but not statistically statistically significant(P>0.05). The percent of the collagen is significantly in the 1ng/ml PDGF-BB and 10ng/ml IGF-I in combination compared to the 1ng/ml PDGF-BB alone(P<0.05) and in the 10ng/ml IGF-I in combination compared to the 10ng/ml PDGF-BB alone(P<0.05). The synthetic activity of the DNA is In conclusions, the percent study shows that PDGF-BB and IGF-I in combination have a potentiality to enhance the DNA synthesis and the total protein and collagen synthesis of The periodontal ligament cells, especially it is more significant in the low concentration of PDGF-BB compared to the high one. Thus, the PDGF-BB and IGF-I in combination may have important roles in promotion of periodontal litgment healing, and consequently, may useful for clinical application in periodontal regenerative procedures.

• Journal of Periodontal and Implant Science
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• v.27 no.3
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• pp.603-619
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• 1997

The use of laser in the treatment of soft tissue minimizes hemorrhage, provides better view of the operating field, and thereby minimizes operating time. Also, there will be far less post-operative swelling, pain and scar formation, and sterilizing effect are shown in some portions of the wound site. All these advantages of laser therapy contribute to its widespread use in the field of medicine and dentistry. Regarding such facts, we used CO2 laser of different watts in gingivectomy for white rats to compare initial healing process. For the control group, the least amount of output in performing gingivectomy(4watts) was offered, and for the experimental group, 6watts was given. Animals were sacrificed on the second, third days, 1 weeks, 2 weeks, and 3 weeks after operation, and their specimens were histologically analyzed. The following results were obtained: 1. Blood clot of small size was observed in both the control and experimental groups after two days, and no more thereafter. 2. In both the control and experimental groups, the inflammation zone size was the greatest after two days, and it decreased gradually to become almost invisble by the second week. The experimental group showed larger size of inflammation zone during second and third days: however, there was no difference after one week. 3. Granulation tissue in both the control and experimental groups showed gradual maturation with time, and by the second week, it was almost replaced by normal connective tissue. By the third week, complete healing pattern was observed. The experimental group showed larger granulation tissue than the control group until the third day, but there was no significant difference after one week. 4. In both the control and experimental groups, gingival epithelialization began on the second day. After one week, regeneration of rete peg and partial formation of junctional epithelium were observed: by the second week, keratinization of oral sulcular epithelium began, and it was completed by the third week.

• Journal of Periodontal and Implant Science
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• v.33 no.3
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• pp.457-474
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• 2003

The ultimate objective of periodontal treatment is to get rid of an on-going periodontal disease and further regenerate the supporting tissue, which is already destroyed, functionally. Currently, the bone grafting operation using various kinds of bone grafting materials and the operation for induced regeneration of periodontal tissue using the blocking membrane are performed for regeneration of the destroyed periodontal tissue. However, there are respective limitations Galenical preparations, which are used for regeneration of periodontal of tissue, has less risk of rejective reaction or toxicity that may be incidental to degradation and their effect is sustainable. Thus, in case they are applicable to a clinic, they can he used economically. Chitosan has such compatibility, biological actions including antibacterial activity, acceleration of wound treatment, etc., and excellent mechanical characteristics, which has recently aroused more interest in it. Also, it has been reported that it promotes osteogenesis directly or indirectly by functioning as a matrix to promote migration and differentiation of a specific precussor cell (for example, osteoblast) and further inhibiting the function of such a cell as fibroblast to prevent osteogenesis. In this study, the pure chitosan solution, which was obtained by purifying chitosan, was used. However, since this chitosan is of a liquiform, it is difficult to sustain it in a defective region. It is, therefore, essential to use a carrier for delivering chitosan to, and sustaining it gradually in the defective region. In the calvarial defect model of the Sprague-Dawley rat, it is relatively easy to maintain a space. Therefore, in this study, the chitosan solution with which ACS was wetted was grafted onto the defective region, For an experimental model, a calvarial defect of rat m s selected, and a critical size of the defective region was a circular defect with a diameter of 8 mm. A group in which no treatment was conducted for the calvarial defect was set as a negative control group. Another group in which treatment was conducted with ACS only was set as a positive control group (ACS group). And another group in which treatment was conducted was conducted with by grafting the pure chitosan solution onto the defective region through ACS which was wetted with the chitosan solution was set an experimental group (Chitosan/ACS group). Chitosan was applied to the Sprague-Dawley rat's calvarial bone by applying ACS which was wetted with the chitosan solution, and each Sprague-Dawley rat was sacrificed respectively 2 weeks and 8 weeks after the operation for such application. Then, the treatment results were compared and observed histologically and his tometrically. Thereby, the following conclusions were obtained. 1. In the experimental group, a pattern was shown that from 2 weeks after the operation, vascular proliferation proceeded and osteogenesis proceeded through osteoblast infiltration, and at 8 week after the operation, ACS was almost absorbed, the amount of osteogensis was increased and many osteoid tissue layers were observed. 2. At 2 weeks after the operation, each amount of osteogenesis appeared to be 8.70.8 %, 13.62.3 % and 4.80.7 % respectively in the experimental group, the positive control group and the negative control group. Accordingly, it appeared to be higher in the Experimental group and the positive control group than in the negative control group, but there was no significant difference statistically (p<0.01). 3. At 8 weeks after the operation, each amount of osteogenesis appeared to be 62.26.1%, 17.42.5 % and 8.21.4 % respectively in the experimental group, the positive control group and the negative control group. Accordingly, it appeared to be substantially higher in the experimental group than in the positive control group and the negative control group, and there was a significant difference statistically (p<0.01). As a result of conducting the experiment, when ACS was used as a carrier for chitosan, chitosan showed effective osteogenesis in the perforated defective region of the Sprague-Dawley rat's calvarial bone.

• Journal of Periodontal and Implant Science
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• v.31 no.1
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• pp.1-23
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• 2001

Chitosan is biodegradable natural polymer that has been demonstrated its ability to improve wound healing, and calcium metaphosphate(CMP) is a unique class of phosphate minerals having a polymeric structure. In this study, chitosan/CMP and platelet derived growth factor(PDGF-BB) loaded chitosan/CMP sponges were developed, and the effect of the sponges on bone regeneration and their possibility as scaffolds for bone formation by three-dimensional osteoblast culture were examined. PDGF-BB loaded chitosan/CMP sponges were prepared by freeze-drying of a mixture of chitosan solution and CMP powder, and soaking in a PDGF-BB solution. Fabricated sponge retained its 3-dimensional porous structure with $100-200\;{\mu}m$ pores. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with $^{125}I-labeled$ PDGF-BB. In order to examine their possibility as scaffolds for bone formation, fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 28 days. Cell proliferation, alkaline phosphatase activity were measured at 1, 7, 14 and 28 days, and histologic examination was performed. In order to examine the effect on the healing of bone defect, the sponges were implanted into rat calvarial defects. Rats were sacrificed 2 and 4 weeks after implantation and histologic and histomorphometrical examination were performed. An effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. PDGF-BB loaded chitosan/CMP sponges supported the proliferation of seeded osteoblastic cells as well as their differentiation as indicated by high alkaline phosphatase activities. Histologic findings indicated that seeded osteoblastic cells well attached to sponge matrices and proliferated in a multi-layer fashion. In the experiments of implantation in rat calvarial defects, histologic and histomorphometric examination revealed that chitosan/CMP sponge promoted osseous healing as compared to controls. PDGF-BB loaded chitosan/CMP sponge further echanced bone regeneration. These results suggested that PDGF-BB loaded chitosan/CMP sponge was a feasable scaffolding material to grow osteoblast in a three-dimentional structure for transplantation into a site for bone regeneration.

• 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.

• Journal of Periodontal and Implant Science
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• v.47 no.3
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• pp.143-153
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• 2017

Purpose: The aim of the present exploratory study was to evaluate extraction socket healing at sites with a history of periodontal and endodontic pathology. Methods: The mandibular 4th premolar teeth in 5 adult beagle dogs served as experimental units. Periodontal and endodontic lesions were induced in 1 premolar site in each animal using wire ligatures and pulpal exposure over 3 months (diseased sites). The contralateral premolar sites served as healthy controls. The mandibular 4th premolar teeth were then extracted with minimal trauma, followed by careful wound debridement. The animals were sacrificed at days 1, 7, 30, 60, and 90 post-extraction for analysis, and the healing patterns at the healthy and diseased extraction sites were compared using radiography, scanning electron microscopy, histology, and histometry. Results: During the first 7 days of healing, a significant presence of inflammatory granulation tissue was noted at the diseased sites (day 1), along with a slightly accelerated rate of fibrin clot resolution on day 7. On day 30, the diseased extraction sites showed a greater percentage of persistent fibrous connective tissue, and an absence of bone marrow formation. In contrast, healthy sites showed initial signs of bone marrow formation on day 30, and subsequently a significantly greater proportion of mature bone marrow formation on both days 60 and 90. Radiographs exhibited sclerotic changes adjoining apical endodontic lesions, with scanning electron microscopy showing collapsed Volkmann canals protruding from these regions in the diseased sites. Furthermore, periodontal ligament fibers exhibited a parallel orientation to the alveolar walls of the diseased sites, in contrast to a perpendicular arrangement in the healthy sites. Conclusions: Within the limitations of this study, it appears that a history of periodontal and endodontic pathology may critically affect bone formation and maturation, leading to delayed and compromised extraction socket healing.

• Journal of Periodontal and Implant Science
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• v.28 no.1
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• pp.133-143
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• 1998

Diabetes mellitus is a systemic disease with profound effects on oral health and periodontal wound healing. Uncontrolled diabetes adversely affects surgical wound healing and is often associated with abnormal proliferation of fibroblasts. Human gingival fibroblasts and PDL cells were chosen because they are intimately involved in periodontal therapy and are important for the success of surgical procedure such as guided tissue regeneration. The aim of the present study was to elucidate whether cellular activity and collagen synthesis by glucose pre-treated human gingival fibroblasts and PDL cells are influenced by insulin, and whether healthy cells differ from glucose treated cells. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidified incubator. To evaluate the effect of glucose on gingival fibroblasts and periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4\;cells/well$ culture plates and treated with 20 and 50mM of glucose for 5 days. Then MTT assay was carried out. To evaluate the effect of insulin on glucose-pretreated cells, the cells were seeded at a cell density of $1{\times}10^4\;cells/well$ culture plates and treated with 20 and 50mM of glucose for 5 days. After incubation, $10^3$, $10^4$ and $10^5mU/l$ of insulin were also added to the each well and incubated for 2 days, respectively. Then, MTT assay and collagen synthesis assay were carried out. The results indicate that cellular activity of gingival fibroblasts significantly increased by glucose while periodontal ligament cells were unaffected and cellular activity of gingival fibroblasts and periodontal ligament cells were unaffected by insulin. Collagen synthesis of gingival fibroblast with 20mM glucose and insulin unaffected, but 50mM glucose and insulin increased than control. Collagen synthesis of periodontal ligament cell with 20mM glucose and $10^5mU/l$ insulin significantly increased than other groups and 50mM glucose pretreated PDL cells significantly increased at $10^3mU/l$ insulin but decreased at $10^4mU/l$ insulin. Our findings indicated that these cell types differed in their growth response to glucose, and the increase in collagen synthesis was significantly raised at insulin level of $10^3mU/l$ in gingival fibroblasts and periodontal ligament cells except 20mM glucose pretreated periodontal ligament cells.

• Journal of Periodontal and Implant Science
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• v.32 no.2
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• pp.403-414
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• 2002

I. Purpose of Study Zea Mays L. has been known to be effective for improving periodontal health and Magnoliae cortex to have effective antibacterial and antimicrobial activity against periodontal pathogens. The purpose of this study was to examine the biologic effects of Zea Mays L. and Magnoliae cortex extract mixtures on healing of rat calvarial bone defects. II. Materials & Methods 8mm circular defects were prepared on rat calvaria during surgical procedures of 180 Sprague-Dawley rats. The ethanolic extracts of Magnoliae cortex and Zea Mays L. and these two natural extract 1:1 and 2:1 (Magnoliae: Zea Mays L.) ratio mixtures were oral administrated by oral zondes once a day at two different dose of 94.5mg/kg, 189mg/kg body weight. There are nine groups of rats in this study: control group (no sample loading), Magnoliae cortex extract loading groups (I,II)(94.5mg/kg,189mg/kg respectively), Zea Mays L. extract loading groups (I,II), M:Z(1:1) loading groups (I,II), M:Z(2:1) loading groups(I,II). Rats were sacrificed at 4 weeks and 6 weeks after surgery. New bone formations around calvarial defects were radiographically and histologically measured by computerassisted histomorphometry. Each data was statistically analyzed by One-way ANOVA test. III. Results There were statistical significances between negative control group and the other test groups on radiographical and histological quantitative assessments. Among test groups, mixture groups showed statistical significances, especially, M:Z (2:1) groups (I and II) were highly significant.(p<0.05) These results implicated that the mixture of Magnoliae and Zea Mays L. (2:1 mixing ratio) with 94.5mg/kg concentration might be highly effective on the wound healing of bony defected site and have potential possibilities as a useful drug to promote bone tissue regeneration.

• Journal of Life Science
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• v.28 no.6
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• pp.681-687
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• 2018

Recently, several researchers have been developing cosmetics from natural ingredients for skin whitening and anti-aging products. The red sea cucumber (RSC), Apostichopus japonicas, is a species of sea cucumber in the family stichopodiae, which is widely distributed in China, Japan, and Korea. To use Red Sea Cucumber as a cosmetic ingredient, its inhibitory effects on melanogenesis and the anti-aging effects of RSC extracts were investigated. First, a tyrosinase activity assay was performed, which showed that RSC inhibited tyrosinase activity at a concentration of $200{\mu}g/ml$. An MTT assay was carried out to evaluate cell toxicity, and the results showed that RSC extract has no cytotoxicity in HaCaT cells. Furthermore, the mRNA expression levels of tyrosinase, tyrosinase related protein 1 (TRP-1), tyrosinase related protein 2 (TRP-2), microphthalmia-associated transcription factor (MITF), and matrix metalloproteinase (MMPs) genes treated with RSC extract in B16F10 and HaCaT cells decreased. Moreover, a wound-healing assay was performed to identify the cell regeneration effect of RSC extracts. Also, a skin turnover effect was confirmed by creating a three-dimensional cell culture with HaCaT and human fibroblasts. Altogether, the results suggested that Red Sea Cucumber may possess a high ability to induce whitening and anti-wrinkle effects as a cosmeceutical ingredient.

• Journal of Periodontal and Implant Science
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• v.33 no.2
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• pp.259-269
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• 2003

The ideal goal of periodontal therapy is the regeneration of periodontal tissue repair of function. Although is very difficult to attain the goal, recent advances in periodontal wound healing concepts encourage hope reaching it. Recently many efforts are concentrated on the regeneration potential of material used in traditional Korean medicine. Phlomidis Radix has been used for the treatment of blood stasis, bone fracture and osteoporosis in traditional Korean medicine. The purpose of this study is to examine effects of dichloromethane fraction Phlomidis Radix on Bone Formation in Human Fetal Osteoblasts. Human fetal osteoblastic cell line(hFOB1 1.19 ;American Type Culture Collection, Manassas, VA) were used and cells were cultured containing DMEM and dichloromethane fraction Phlomidis Radix(100 ng/ml , 1 ${\mu}$/ml, 10 ${\mu}$/ml) at 34$^{\circ}C$ with 5% $CO_2$ in 100% humidity. MTT was performed to examine the viability of the cell, and alkaline phosphatase activity was analyzed to examine the mineralization. Also bone calcification nodules were evaluated. The cellular activity of hFOB1 was increased in 100 ng/ml, 1 ${\mu}$/ml , 10 ${\mu}$/ml of dichloromethane fraction of Phlomidis Radix and especially significant increation was showed in 100 ng/ml of dichloromethane fraction of Phlomidis Radix at 6days (p <0.05). ALP level of hFOB1 was significantly increased in 100 ng/ml , 1 ${\mu}$/ml, 10 ${\mu}$/ml of dichloromethane fraction of Phlomidis Radix and especially more increation was showed in 10 ${\mu}$/ml of dichloromethane fraction of Phlomidis Radix (p <0,05). Calcification nodules of hFOB1 significantly increased in 10 ${\mu]$/ml of dichloromethane fraction of Phlomidis Radix at 21 days of incubation(p<0.05). The results indicate that dicholoromethane fraction of Phlomidis Radix has excellent effects on mineralization of hFOB1.