• Journal of Periodontal and Implant Science
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• v.37 no.1
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• pp.45-51
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• 2007

This study evaluated the possibility of the 3-dimensional attachment of human periodontal ligament fibroblasts to a periodntally involved root surface after an EDTA treatment in vitro. The human PDL fibroblasts were isolated from the middle third of the root of periodontally healthy teeth extracted for orthodontic reasons. The cells were cultured in a medium containing Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at $37^{\circ}C$ in humidified air containing 5% $CO_2$. Eight single-rooted teeth were obtained from patients diagnosed with periodotitis. After scaling and root planing, four teeth were etched with 24% ethylenediaminetetracetic acid (EDTA) for two minutes (Experimental group). The other four teeth were not treated with EDTA and were used as the control group. The human PDL fibroblasts were placed in the total root surface and cultured for 4 weeks. The teeth were fixed in 2.5% glutaraldehyde in PBS before preparation for the scanning electron microscopy (SEM) examination. The human PDL fibroblasts showed a healthy morphology on the root surfaces treated with EDTA (Experimental group) and a relatively unhealthy appearance on the treated root surfaces (Control group). This suggests that EDTA favorably affects the 3-dimensional attachment of human PDL fibroblasts cultured on the root surfaces. which may play an important role in periodontal healing and regeneration.

• The Journal of Korean Orthopaedic Ultrasound Society
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• v.1 no.2
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• pp.117-121
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• 2008

Prolotherapy can be defined as the injection of growth factors or growth factor production stimulants to grow normal cells or tissue. Even though it has been a controversal procedure for decades, it is currently gaining in popularity among many physicians. The term prolotherapy was coined by Hacket in the 1950s to imply proliferation of normal tissue at ligamentous and tendinous entheses. The procedure has been described by other terms, such as sclerotherapy, regenerative injection therapy, and stimulated ligament repair. Incomplete healing from sprains or strains is common and can lead to chronic pain, joint instability and laxity, and is a risk factor for the development of osteoarthritis. Prolotherapy is commonly used for these musculoskeletal conditions which are refractory to usual care therapies. The proliferant solution and technique varies according to physicial training and preferance. Commonly reported proliferants include 10% to 25% dextrose, P2G and sodium morrhuate. High resolution ultrasound imaging of musculoskeletal tissue is increasing in popularity because of patient tolerability, low cost, ability to visualize tissue in real time motion and superior resolution of highly organized tissue such as a tendon. This procedure can be introduced by ultrasound imaging and tissue growth and repair after this procedure in a tendon or a ligament can be documented with ultrasound.

• Journal of Periodontal and Implant Science
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• v.47 no.6
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• pp.352-362
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• 2017

Purpose: Furcation involvement in the molars is difficult to treat, and has been recognized as a risk factor for tooth loss. Although periodontal regenerative therapies, including guided tissue regeneration and various types of bone grafts, have been applied to furcation defects, the effects of these treatments are limited, especially in large class III furcation defects. The purpose of this pilot study was to investigate the effect of reciprocal autologous root transplantation on periodontal wound healing and regeneration in class III furcation defects in dogs. Methods: Furcation defects (7 mm wide and 6 mm high) were surgically created after root separation of the unilateral third and fourth premolars in 4 dogs. Eight furcation defects were randomized to receive either reciprocal autologous root transplantation (test) or no further treatment (control). In the test group, the mesial and distal roots were transplanted into the distal and mesial extraction sockets, respectively. The animals were sacrificed 10 weeks after surgery for histologic evaluation. Results: The healing pattern in the control group was characterized by extensive collapse of the flap and limited periodontal regeneration. New bone formation in the test group ($3.56{\pm}0.57mm$) was significantly greater than in the control group ($0.62{\pm}0.21mm$). Dense collagen fibers inserting into the residual cementum on the transplanted root surfaces were observed in the test group. Slight ankylosis was observed in 2 of the 4 specimens in the test group on the mesiodistal sides where the root-planed surfaces faced the existing bone. Root resorption (RR) was detected in both the control and test groups. Conclusions: Within the limits of this study, it can be concluded that reciprocal autologous root transplantation was effective for bone regeneration in class III furcation defects in dogs. However, further studies are required to standardize the approach in order to prevent unwanted RR prior to clinical application.

• Journal of Periodontal and Implant Science
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• v.23 no.1
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• pp.16-26
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• 1993

In order to investigate the healing effect on inflammation and bone regeneration of low power density laser radiation in dogs, experimental periodontitis was made in dog mandibular 3rd, 4th premolars. All teeth were classified with four groups of two experimental group and control groups. The second group were irradiated on periodontitis site and the first group were control. The fourth group were irradiated on periodontitis site flap operation and the third group were flap control. Experimental groups were irradiated with GaAs low power density laser of pulse wave and continuous wave of 904nm every day by five days respectively and then control group and experimental groups were evaluated by histo-pathological study. The results were as follows : 1. Experimental periodontits site of dog were irradiated with GaAs low power laser results in reducing of pseudoepitheliomatous proliferation and inflammation at light microscope. 2. After irradiation with low power density laser, experimental groups were revealed that PDL forming activity were increased and newly formed collagen deposition were observed. 3. Low power density lsaer irradiation on experimental periodontits site after flap operation showed that decreasing of inflammation, reducing of osteoclast activity. Capillary proliferation, reduction of pseudoepitheliomatous proliferation. 4. After irradiation with low power density laser on flap experimental site, experimental groups were revealed that newly formed collagen in periodontal ligament and alveolar bone were detected on MT staining.

• Journal of Periodontal and Implant Science
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• v.26 no.3
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• pp.591-604
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• 1996

The present study evaluates the effects of dura mater barrier membranes In class III furcation defects on the regeneration of periodontal tissues in dogs. Experimental class III furcation defects were created surgically by removing alveolar bone horizontally down to 4mm from CEJ in mandibular premolars of adult dogs. Dura mater barrier membranes were applied bucco-lingually in the test group, and flap surgery only with no membranes in the control group. The healing was evaluated clinically and histologically after 8weeks. Clinically, the test group showed slight exposures of the membranes, while the control group showed no furcation exposure, The test specimens showed new bone formation coronal to the notch, while the control specimens had new bone formation up to the level of the notch. New cementum was observed in both groups. The test specimens showed functional arrangements of connective tissue fibers between new bone and new cenentum, while irregular arrangements were observed in the controls. No root resorption or ankylosis were observed in either groups,These results suggest that dura mater resorbable barrier membranes on class III furcation defects may be effective in regeneration of alveolar bone and peridontal ligament.

• Journal of the korean academy of Pediatric Dentistry
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• v.36 no.4
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• pp.640-646
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• 2009

In case of luxation injuries, loss of tooth vitality is common. And in case of trauma in the immature permanent teeth, precise diagnosis of pulp necrosis is very difficult. That is because limitation in distinguishing between normal dental papilla in immature permanent teeth, transient apical breakdown(TAB), which is part of normal healing process, and apical radiolucency in pulp necrosis. Especially in non-vital immature permanent tooth, the treatment is complex and requires long time. This clinical case report shows that severely infected immature teeth with periradicular periodontitis can undergo healing and apexogenesis or maturogenesis with no definative treatment or after conservative treatment. In the cases reported, we emphasize the considerable power of regeneration of the tooth, probably due to its large number of undifferentiated mesenchymal cells in the dental papilla, pulp tissue, periodontal ligament tissues. Thus, when endodontic treatment in immature permanent teeth, over instrumentation is not recommend for preserve the apical vital stem cells.

• Journal of Periodontal and Implant Science
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• v.27 no.3
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• pp.443-457
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• 1997

The purpose of this study was to evaluate the effects of ethanolic extracts of Scutellaria Radix on the alveolar bone formation in the extract socket of rat. Thirty-six Sprague-dawley rats were used in this study. Mean body weight of rat was $130{\pm}5g$. Experimental animal were administered 0.4% ,${\beta}-aminopropionitril$(Sigma, USA) with the solid commercial food for 5 days. All the maxillary 1st molar of the rats were extracted by using of the tissue forcep under the general anesthesia with Pentobarbital sodium(Tokyo Chemical Co, Japan) injection into intraperitoneal space. All the extracted rats were divided into two group, experimental group which were feeded the solid food mixed ethanolic extracts of Scutellaria Radix, and control group which were feeded same food without reagent. At 1, 3, 5, 7, 9 and 14th days after tooth extraction, rats in both groups were serially sacrificed respectively. All the specimen were treated as usual method and prepared Hematoxylin-eosin stain for the light microscopic observation. The results were as follows : 1. Bone formation of extracted socket starts from the area on remained periodontal ligament than other area. 2. In the case of administration of the extracted Scutellaria Radix showed rapid healing process of connective tissue than non-administrated group. 3. In the case of administration of the extracted Scutellaria Radixshowed rapid osteogenesis than non-administrated With above results, it was concluded that ethanolic extracts of Scutellaria Radix may play a favorable role on the healing process of exatraction socket after extraction in rats. It was suggested that further study to evaluate the different concentration and administration method of ethanolic extracts of the Scutellaria Radix into same experimental model.

• 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.27 no.4
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• pp.867-882
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• 1997

Safflower(Carthamus tinctorius $Linn\acute{e}$ has been traditionally used for the treatment of blood stasis, and Dipsasi Radix has been used as a drug for fracture in Chinese medicine. The purpose of present study was to examine the biologic effects of safflower extract and Disasi radix extracts on the periodontal. ligament cells and osteoblastic cells and on the wound healing of rat calvarial defect. The ethanolic extract of safflower blossom, safflower seed and Dipsasi Radix(125, 250, and 500 ${\mu}g/ml$) were prepared as test group, and PDGF-BB(lOng/ml) and unsafonifiable fraction of Zea Mays L.(125, 250, and 500 ${\mu}g/ml$) were employed as positive control. The effects of each agents on the growth and survival, ALPase activity, expression of PDGF-BB receptor, chemotactic response of PDL cell and ATCC human osteosarcoma MG63 cells in vitro were examined. The tissue regenerative effect of each extracts was evaluated by histomorphometric measuring of newly formed bone on the 8mm defect in rat calvaria after oral administration of 3 different dosages groups : 0.02, 0.1 and 0.35g/kg, per day. It was also employed the same dosages of unsaponifiable fraction of Zea Mays L. as positive controls. Safflower blossom extract, safflower seed extract, and Dipsasi Radix extract stimulate the cellular activity of MG63 cells in concentration range of $125-500{\mu}g/ml$, and safflower bolssom extract and safflower seed extract stimulate also the cellular activity of periodontal ligament cells in concentration range of $250-500{\mu}g/ml$. In activity of ALPase, $250-500{\mu}g/ml$ of safflower blossom extracts showed significant stimulating effects on MG63 cells, and the same concentration range of safflower seed extracts showed significant effect on periodontal ligament cells. In the recovery on PDGF-BB receptor expression which was depressed by $IL-1{\beta}$, $125-250{\mu}g/ml$ of safflower blossom extracts and $250-500{\mu}g/ml$ of safflower seed extracts showed significant increasing effect on MG63 cells, and $500{\mu}g/ml$ of safflower blossom extract and $250-500{\mu}g/ml$ of safflower seed extracts showed significant effect on periodontal ligament cells. In chemotactic response, among all tested group, safflower seed extracts only were chemotactic to MG63 cells and periodontal ligament cells in concentration range of $125-500{\mu}g/ml$. Also in the view of bone regeneration in rat calvarial defect model, the only group that was orally administrated 0.35g/kg, day of safflower seed extract showed significant new bone formation. These results suggested that safflower extracts might have a potential possibilities as an useful drug for adjunct to treatment for regeneration of periodontal defect.

• Journal of Periodontal and Implant Science
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• v.26 no.2
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• pp.469-490
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• 1996

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 $TGF-{\beta}1$.