Regeneration of the periodontal tissue destroyed by periodontal disease is one of the final goals of periodontal therapy. In the past few years, periodontists have used various alloplastic grafting materials in an attempt to regenerate bone lost from periodontal disease. These materials have used widely because they have shown to be nontoxic, biologically compatible with surrounding host tissue and chemically similar to bone. The purpose of this study was to investigate the effect of Porous Resorbable Calcium Carbonate and Porous Replamineform Hydroxyapatite on the regeneration of the alveolar bone and the healing of roots transplanted into the periodontally diseased extraction sockets of dogs. The experimental chronic periodontitis was induced by elastic ligatures on the 2nd and 3rd mandibular premolars of 2 adult dogs for 8weeks after surgically creating periodontal defect. The extracted root were split in half along the long-axis, and the extend of plaque exposure was marked on the root surfaces with burs. The roots were inserted in extraction sockets with Porous Resorbable Calcium Carbonate(PRCC) in left side and with Porous Replaminefrom Hydroxyapatite(PRH) in right side. The flaps were sutured to cover the sockets completely. The animals were sacrificed after 12 weeks of healing, and the specimens were examined histologically. The results were as follows: 1. No inflammatory reactions were observed in either groups. 2. Hoot resorption was observed in both groups while the general outline of the roots were maintained. 3. PRCC was almost completely resorbed and replaced with new bone, while R.H.A. was not resorbed & remained encased in newly-formed C-T and alveolar bone. 4. PRH was encapsulated with alveolar bone which has been deposited from apical & lateral area of the sockets, while the coronal portion of the sockets were filled with C-T. 5. In both groups, the resorbed portions of the roots were replaced with new bone. These results suggest that either PRCC or PRH may not interfere with bone formation or healing in extraction sockets, and in some degree, retard the root resorption. Because the roots maintained in anatomy, we think that graft materials prevent the root resorption.

The present study was performed to evaluate the amount of bony regeneration following the guided tissue regeneration(GTR). Re-entry procedure has been performed at 1 year following the GTR with Gore-tex membranes on the furcal defects and the amount of bony regeneration was measured. Sites treatedwith open flap procedures were used as controls. The results reveated that significant amount of bone could be regenerated through the GTR procedures compared with convention flap procedures.

The present study was done to evaluate the antibacterial effects of $Minoclin^R$ which was localally delivered on the $Gore-tex^R$ barrier membrane in the guided tissue regeneration(GTR) therapy for treatment of human furcal defects. Beneath the membranes. the antibiotics were applied for 1 week and then changed with new one. The $Minoclin^R$ was removed out one week later. 6 weeks after the GTR therapy. No systemic antibiotics were administered except for oral mouthrinses with chlorhexidines. 2 weeks and 6 weeks following the membrane therapy, the bacterial samples were examined for periodontopathic microorganisms. The results indicated that the locally delivered $Minoclin^R$ successfully inhibited the growth of periodontopathic organisms. This results might be further applied in the subgingival plaque control regimen in the GTR procedure, especialy in patients who is contraindicated for oral administration of systemic antibiotics

5 serotypes(a, b, c, d, e) of Actinobacillus actinomycetemcomitans showed distinct hybridization patterns(DNA fingerprinting patterns) when the bacterial DNA were hybridized with randomly cloned 4.7-Kb sized DNA probe. The sizes of hybridized bands in each serotypes were different among serotypes and represented unique patterns of hybridization with the probe used. The serotype a showed two bands of fingerprinting patterns: 23.1 kb and 2.5 kb respectively. Serotype b and c showed single band: 6.6 kb and 9.5 kb, respectively. Serotype d and e showed two bands of hybridization: 23.1 kb and 2.8 kb, and 23.7 kb and 2.1 kb, respectively. The results indicate that this standard fingerpriting patterns of DNA hybridization with 4.7 kb probe can be further used for genotyping clinical isolates of Actinobacillus 8ctinomycetemcomitansand its relevance with periodontal disease activity.

The neuropeptide substance P(SP) has been recognized to modulate immune systems, with close proximity between peptidergic sensory nerve endings and immune cells. These include the macrophage and neutrophil activation, IL-2 production in T cell, augmentation of Ig synthesis, mast cell degranulation, $PGE_2$ and collagenase secretion in synoviocytes. In this study I examined SP-induced various biological activities such as antimicrobial action, cytokine production, and mast cell degranulation in the presence or absence of other inflammatory cell activators. Antimicrobial studies showed that undifferentiated HL-60 cells were not affected by SP. However, SP significantly enhanced antimicrobial action of TPA-treated or dbcAMP-treated HL-60 cells which had been differentiated into PMN or macrophage/monocyte. I could not find synergistic relationship between SP and LPS in parallel experiments of the above. SP did not induce IL-l production from murine macrophage cell line RAW264.7 whether costimulated with LPS or not. Mast cell degranulation was occured only when stimulated with high dose ($10^{-5}M$) of SP and the degree of this activation was slightly reduced by simultaneous application of $MIP-1{\alpha}$. In addition, CGRP which is known to be a common coexisting neuropeptide with SP within specific fibers did not augment the function of SP on mast cell degranulation. These results suggest that immunoregulatory activities of SP could be mediated through direct upregulation of various functions of immune cells and also upregulation of responsiveness of immune cells to other immune activators.

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.

The use of basic fibroblast growth factor which function as potent biologic mediators regulating numerous activities of wound healing has been suggested for the promotion of periodontal regeneration. The mitogenic effects of basic fibroblast growth factor on human periodontal ligament cells and human gingival fibroblasts were evaluated by determining the incorporation of 5-Bromo-2'deoxy-uridine into DNA of the cells in a dose -dependent manner. The cells which were prepared were the primary cultured gingival fibroblasts and periodontal ligament cells from human the fourth or sixth subpassages were used in the experiments. The cells which were seeded DMEM contain 10% FBS. The added concentrations of basic fibroblast growth factor were 0.1, 1, 10, 50, $l00{\eta}g/ml$ and basic fibroblast growth factor were added to the quiescent cells for 24 hours, 48 hours and 72 hours. They were labeled with $10{\mu}l/200{\mu}l$ 5Bromo-2'-deoxy-uridine for the last 6 hours of each culture. The results of the five determinants were presented as mean and S.D.. The results were as follows. : The DNA synthetic activity of human gingival fibroblasts was increased dose dependently by basic fibroblast growth factor at 24 hours, 48 hours and 72 hours. The similar mitogenic effects were at the 24 and 48 hours of basic fibroblast growth factor, but the DNA synthetic activity of human gingival fibroblasts generally decreased at 72 hours. The DNA synthetic activity of human periodontal ligament cells was increased dose dependently to $50{\eta}g/ml$ by basic fibroblast growth factor at 24, 48 and 72 hours, but the DNA synthetic activity decreased at $l00{\eta}g/ml$ of each hour. Generally the maximum mitogenic effects were at the 48 hours application of basic fibroblast growth factor. The DNA synthetic activity of human periodontal ligament cells generally decreased lower at 72 hours than at 24, 48 hours the application of basic fibroblast growth factor. In the comparison of DNA synthetic activity between human gingival fibroblasts and human periodontal ligament cells, human periodontal ligament cells had slightly higher proliferation activity than human gingival fibroblasts for a longer time at the high dosage of the basic fibroblast growth factor.In conclusion, basic fibroblast growth factor have important roles in the stimulation of DNA synthesis in human periodontal ligament cells and human gingival fibroblasts, and thus may be useful for clinical applications in periodontal regenerative procedures.

Transforming growth factor $-{\beta}$ is one of the polypeptide growth factors that mediate the activity of mesenchymal cells and regulate wound healing process via cell proliferation, migration and extracellular matrix formation. The purposes of this study is to evaluate the effects of transforming growth factor $-{\beta}$ on the protein synthetic activity of human periodontal ligament cells and human gingival fibroblasts. The cells which were prepared were primary cultured gingival fibroblasts and periodontal ligament cells from humans, and the fourth or sixth subpassage were used in the experiments. Cells were seeded and at a confluent state, 0, 0.5, I, 2.5, 5, 10 ng/ml $TGF-{\beta}$ and $2{\mu]Ci/ml\;[^3H]$ proline were added to the cells and cultured for 24 hours. Then, 1 and 5 ng/ml concentrations were selected and added to confluent cells and cultured for 24 and 48 hours. They were labeled with $2{\mu}Ci/ml\;[^3H]$ proline for 24 hours and a collagen assay was done by the Peterkofsky and Diegelman method. The results were presented as the mean disintegration per minute (dpm) per well and S.D. of four determinations, The results were as follows. : The total protein, collagen and noncollagenous protein synthesis in periodontal ligament cells and gingival fibroblasts were increased dose- dependently by transforming growth factor-p to 2.5-5 ng/ml concentration and decreased at 10 ng/ml concentration. The percent of collagen was slightly changed according to the concentration of transforming growth factor-po The effect of transforming growth $factor-{\beta}$ was not specific for collagen synthesis since it increased the total, noncollagenous and collagenous protein, simultaneously. In the comparison of protein synthetic activity between the human periodontal ligament cells and human gingival fibroblasts, the human gingival fibroblasts had higher activities than the human periodontal ligament cells at all times and concentrations of $TGF-{\beta}$. In the comparison of protein synthetic activity between the 24 hour effect and the 48 hour effect of $TGF-{\beta}$, the 48 hour cultured cells' synthetic activity decreased more than the 24 hour cultured cells at human periodontal ligament cells and human gingival fibroblasts. In conclusion, $TGF-{\beta}$ has important roles in the stimulation of protein synthesis in human periodontal ligament cells and human gingival fibroblasts. Thus, it may be useful for clinical application in periodontal regenerative procedures.

To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.

The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.

This study was performed to evaluate the effects of platelet-derived growth factor (PDGF) and epidermal growth factor (EGF) on the characteristics of beagle dog's periodontal ligament (BPD) cells and bone marrow (BBM) cells which have the important role on the early stage of periodontal tissue regeneration in vitro. In control group, the cells ($1.5{\times}10^5$cells/ml) were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum, $50{\mu]g/ml$ ascorbic acid, and 10mM/ml ${\beta}-glycerophosphate$. In experimental groups, growth factors, PDGF or EGF(10ng/ml), were added into the above culture condition. And then each group was characterized by examining the cell proliferation rate, amount of total protein synthesis, alkaline phosphatase activity at 1, 5, 9, 13, 17th day after seeding of cells into the culture wells. The results were as follows: 1. Both BPD and BBM cells in PDGF-treated group proliferated more rapidly than non-treated cells. This finding also was observed in EGF-treated group but it was not as prominent as that of PDGF-treated group. The proliferation rates of both cells showed the time-dependent pattern during experimental periods in all three groups. 2. Amount of total protein synthesis was more increased in PDGF-treated group than in control group. But no significant difference between EGF-treated group and control group was observed throughout experimental periods even though the tendency of amount of protein synthesis was time-dependent pattern. 3. Alkaline phosphatase activity also more increased in PDGF-treated group than control group. But slight decrease tendency was seen in both cells of EGF-treated group. From the above results, PDGF appeared to enhance the proliferation and cellular activities including amount of total protein synthesis and alkaline phosphatase activity of BPD and BBM cell, but EGF did not show notable effects. The optimal application of these growth factors was thought to be useful as the adjunctive means in periodontal regeneration procedures.

Periodontal pocket is one of the most frequently developed clinical feature on the teeth with periodontal disease. In order to determine the gingival curettage effect of intrapocket irradiation of a pulsed Nd : YAG laser on periodontally involved teeth, bilateral 60 teeth with $4{\sim}6mm$ in probing pocket depth 1 week after supragingival scaling were selected. On half of them the intrapocket irradiation($300{\mu}m$ fiber optic, 1.5W power, for 2min.) of a pulsed-Nd : YAG laser(EL.EN.EN06O, Italy) was applied as the lased group. On the contralateral 30 teeth the subgingival curettage was accomplished by Gracey curettes as the curattage group. The periodontal pocket tissues were surgically excised by the modified Widman flap technique immediately after the intrapocket irradiation or subgingival curettage, subsequently fixed with 10% neutral formalin, sectioned in $4{\sim}6{\mu}m$ thickness, and stained with hematoxylin-eosin. Surface characteristics and incomplete removal of the pocket epithelium were evaluated under light microscope. And the difference between the lased group and the curettage group was statistically analyzed by Chi-square test in Microstat program. The results were as follows ; 1. The plane surface was observed more frequently in the curettage group(73.3%) than in the lased group(23.3%), and the rough surface was observed more frequently in the lased grOoup(63.3%) than in the curettage group(6.7%)(p<0.05). 2. The rate of incomplete removal of the pocket epithelium was relatively high in both the lased group(76.6%) and the curettage group(86.6%), and there was no significant difference between the lased group and the curettage group(p>0.l). The results suggest that the further studies including various power control of laser should be succeeded in order to obtain more favorable results by the intrapocket irradiation of a pulsed Nd:YAG laser than the subgingival curettage with Gracey curettes.

Most dentists are very interested in laser therapy on the intraoral soft tissue lesions because they want to accomplish the analgesic and aseptic surgery with little or no bleeding. In order to determine the difference of pain threshold according to different gingival tissues with or without inflammation, 25 patients with inflammatory periodontal disease and 10 volunteers with good general and oral health were selected as the inflamed group and the normal group, respectively. Interdental papilla, marginal gingiva, attached gingiva, and alveolar mucosa were irradiated by the contact delivery($300{\mu]m$ fiber optic, for 5 seconds) of a pulsed Nd:YAG laser(EN.EL.EN06O, Italy). And the laser power was gradually increased from 0.5W by the increment of 0.1W. The highest laser power was recorded as the first painful power when the painful gesture was recognized at first. The difference of the first painful power of laser according to different gingival tissues with or without inflammation was statistically analyzed by paired t-test in MICROSTAT program. Following results were obtained: 1. In the comparison related with the inflammation, the first painful power was significantly lower in the inflamed group than in the normal group, regardless of interdental papilla and marginal gingiva(p<0.05). 2. In the comparison related with the tissue structure, the first painful. power was significantly lower in alveolar mucosa than in attached gingiva(p<0.05). The results suggest that, for the painless therapy by a pulsed-Nd:YAG laser irradiation, the laser surgery over 2.0W of power should be necessarily accomplished under the local anethesia, and the local anesthesia should be considered according to the degree of inflammation, the tissue structure, and the purpose of laser therapy.

Periodontal disease is characterized by destruction of supporting tissues caused by invasion of plaque bacteria and defense mechanism of host. Many dentists are very interested in laser therapy on various intraoral soft tissue lesions including inflammatory periodontal pocket. In order to determine the therapeutic effect of intrapocket irradiation of a pulsed- Nd : YAG laser on the inflammatory periodontal pockets, bilateral 60 teeth with 4-6mm in probing pocket depth and gingival inflammation were selected and evaluated by sulcus bleeding index(SBI), and plaque index(pI) for baseline record. Intrapocket irradiation($300{\mu}m$ fiber optic, I.5W power, for 2 min.) of a pulsed-Nd : YAG laser(EL.EN.EN060, Italy) was applied on half of them. As the control group, the same procedure except power-off was repeated on the contralateral 30 teeth. At 1-, 2-, 3-, and 4-week after intrapocket manipulation, every tooth was reevaluated by the same clinical indices. And the difference between the lased group and control group was statistically analyzed by paired t-test and Chi-square test in Microstat program. Following results were obtained: 1. Until I-week and 2-week after intrapocket manipulation, SBI was lowered in both lased group and control group, compared to baseline SBI, but from 3-week after, the recovering tendency toward baseline was noted, and at only 2-week after, the number of teeth showing lowered SBI was significantly more in lased group than in control group(p<0.05). 2. PI of both lased group and control group was lowered through whole experimental period from I-week to 4-week after, compared to baseline PI(p<0.05), but there was no significant difference between lased group and control group(p>0.1). The results suggest that intrapocket irradiation of a pulsed-Nd:YAG laser may lead somewhat remission of gingival inflammation, but for more favorable therapeutic result the thorough root planing should be necessarily accompanied with gingival curettage.

Previous studies have shown that Magnoliae cortex and Ginkgo biloba extracts were showed on the antimicrobial and anti-inflammatory action, in vitro. The purpose of this study was to evaluate on the effect of antimicrobial and anti-inflammatory activity of Magnoliae cortex and Ginkgo biloba extracts containing dentifrice in gingivitis. 70 subjects with gingivitis were divided into an experimental group which performed normal oral hygiene procedure with Magnoliae cortex and Ginkgo biloba extracts containing dentifrice and a control group which also performed normal oral hygiene procedure with the same dentifrice without the natural extracts and completed a doubleblind, cross-over study. At baseline and 3 weeks, subjects were assayed for clinical study by plaque index, gingival index, pocket depth, GCF rate, and microbiological study by subgingival dental plaque bacterial morphotypes by phase contrast microscopy, total anaerobes, total aerobes, Black pigmented bacteroides, A.actionomycetemcomitans, A.viscosus, C.rectus, Ssenguis; P.gingivalis, P.intennedia by bacterial culture and immunofluorescence microscopy. After 3 weeks using their respective dentifrices, reductions in the clinical indices of subjects were similar between the experimental dentifrice group and a control dentifrice group except for statistically significant much reductions in PI, GI, and GCF rate in the experimental dentifrice group as compared to control dentifrice group. Also statistically significant reductions in the motile rods and Spirochetes were found in both experimental group to compare with control group, however statistically much reduction in total anaerobes, Black pigmented bacteroides, and P.gingivalis, P.intennedia were found in the experimental dentifrice group as compared to control dentifrice group. This results indicates that Magnoliae cortex and Ginkgo biloba extracts containing dentifrice might be useful for elimination of gingival inflammation.

It is known that some natural extracts from plants have a various range of antimicrobial and antiinflammatory activity. There are lots of clinical trials to develop toothpastes containing natural extracts for prevention of dental caries and gingival inflammation. The purpose of this study was to evaluate antimicrobial and antiinflammatory activity of magnolol containing toothpastes and other commercial toothpastes. Eleven kinds oftoothpastes were used. They include magnolol, sanguinarine, Myrrha, Mori radicis cortex,Cimicifugae rhizoma, sodium fluoride, aminocaprolactic acid etc. Six strains of bacteria were used for this test, ego Porphylomonas gingivalis, Prevotellain-termedia, Actinobacillus actinomy cetemcomitans, Streptococcus mutans, Stretococcus sanguis, and Actinomyces species. Antimicrobial activity was determined by an agar dillution method and a broth microdillution method. Antiinflammatory activity was assessed by the inhibition of $PGE_2$ production from gingival fibroblast with the addition of rHIL-1 and centrifuged solution of toothpastes. Control group was only rHIL-1 additive sample. $PGE_2$ enzyme immunoassay systemfAmersham, In. Buckinghamshire, U.K). $PGE_2$ level was measured by ELISA reader with 450 nm, The results from the study revealed that toothpastes containing natural extracts generally had high antimicrobial and antiinflammatory activity. Especially magnolol containing toothpaste showed higher antimicrobial activity than other toothpastes, and sanguinarine containing toothpaste showed particularly high antimicrobial activity in A. actinomicetemcomitans and A. viscosus. In some degree all toothpastes inhibited $PGE_2$ production, but magnolol containing toothpaste was potent inhibitor of $PGE_2$. Sodium chloride containing toothpaste had also effective result. The results suggested that toothpastes containing natural extracts were promising in plaque control and prevention of dental caries and gingivitis.