• Journal of Periodontal and Implant Science
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• v.25 no.1
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• pp.24-34
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• 1995

The purpose of this study was to compare effects of demineralization of citric acid and tetracycline HCI on periodontally involved root surface. Twelve periodontally involved single rooted teeth were used. After scaling and root planing, root conditioning with citric acid and tetracycline HCI were carried and the teeth were processed scanning electron microscopic observation.The results were as follows: The scaled root surface was covered by much debris and calculus. The effect of demineralization of citric acid and tetracycline HCI was more reduced on scaling group than root planing group, because of hypermineralization of cementum surface and demineralization effect on root surface of tetracycline HCI showed tendency to reduction. The root planed group displayed more smooth root surface than scaling group, the surface was covered by smear layer, thus no exposure of dentinal tubule opening and collagen fiber, especially after root planing, citric acid and tetracycline HCl treated group showed exposure of dentinal tubule and collagen fiber, thus it was thought that new connective tissue attachment could be acquired.

• Journal of Periodontal and Implant Science
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• v.27 no.1
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• pp.79-90
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• 1997

The purpose of this study was to evaluate the substantivity of experimentally developed gel type tetracycline HCl and a mixture of tetracycline-citric acid gel, and compare to those of solution type tetracycline HCl. 11 extracted anterior teeth were subjected to this study. After scaling and root planing teeth were randomly divided into 3 treatments groups : group 1; 3 teeth were irrigated with tetracycline HCl(50mg/ml) solution , group 2; tetracycline gel (5%) was inserted in the periodontal pockets of 3 teeth, group 3; a mixture of tetracycline and citric acid gel was inserted in the pockets of 3 teeth. And 2 teeth treated in 0.9 % sterile saline served as controls. After 5-minute exposure, each tooth immediately extracted and incubated at room temperature for 22 days in tris-buffered saline as a desorption media. The total volume of TBS was removed and replaced with fresh TBS, at 24-h intervals. Removed desorption media transferred to a sterile vial and stored at -70 oC. This procedure was repeated every 24 h throughout the 22-day desorption period. Using Porphyromonas gingivalis as an indicator organism, a microtiter assay was used to evaluate antimicrobial activity desorbed from the teeth. 1. 50mg/ml tetracycline HCl solution exhibited the longest antimicrobial activity. Compared to saline treated group, it showed significant difference on the day 1 and day 2 desorption period. 2. The ODs of 5% tetracycline gel and a mixture of tetracycline-citric acid gel were significantly different during the first 24 hour only. 3. There was no statistically significant difference after the day 3 between the groups.(p<0.05). Despite our expectation a mixture of tetracycline-citric acid gel did not show longer antimicrobial activities than those of tetracycline gel, and the solution type exhibited the longest activities. Because the gel type agents may stay in the subgingival environment longer than the solution, if the teeth were not extracted immediately after the delivery of the agent, the result could be different. hus this result suggests the possibilities of practical use of these kind of gel type agents.

• Journal of Periodontal and Implant Science
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• v.23 no.2
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• pp.283-299
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• 1993

The purpose of this study was to observe early connective tissue attachment on dentin surface treated with citric acid, tetracycline, and fibrin sealants and compare their conditioning effects on dentin surface. Experimental dentin blocks conditioned with citric acid, tetracycline or fibrin sealant, and only root planned control block were surgically implanted in the pouch under buccal mucoperiosteal flaps of left mandible, right maxilla, left maxilla, right mandible of 18 male rabbits. Rabbits were sacrificed after 1 and 6 hours, 1, 3, 7 and 14 days after implantation and then specimens including dentin block and surrounding soft tissue were obtained, and prepared for light and transmission electron microscopic examination. 1 and 6 hours after dentin block implantation, there was plasma proteins adsorption followed by fibrin clot formation and no differences among specimens. At the 1-day observation interval, delicate fibrin network was observed in the all groups, and there were proliferative fibroblasts, angiogenesis and macrophage in the all 3-day specimens. Cellular aggregates and abundant connective tissue adhered dentin surface and tetracycline or citric acid treated group showed much proliferative fibroblast and abundant collagen fibers at 1 week. But at 2 week, citric acid treated group showed much proliferative fibroblast and abundant collagen fibers. These observations suggested that new connective tissue attachment to dentin was initiated by the adsorption of plasma proteins to the dentin surface and followed by fibrin clot formation. Tetracycline and citric acid seemed to make dentin surface more biologically favorable for the connective tissue attachment.

• Journal of Periodontal and Implant Science
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• v.26 no.4
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• pp.889-905
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• 1996

It is known that growth factors function as potent biologic mediators regulating numerous activities of wound healing via cell proliferation, migration and extracellular matrix formation and they also promote periodontal regeneration. But, method of growth factor application is controversial yet. So purpose of this study is to evaluate the effect of demineralized root surface as one of method of growth factor application. The ginigival fibroblasts were primary cultured and fifth or sixth subpassages were used in these experiments. In first experiment, root surface blocks demineralized with 100mg/ml tetracycline for 5 minutes and pH 1 citric acid for 3 minutes(experimental groups) and nonteminerilized root surface blocks (control groups) were placed in 100ng/ml PDGF-BB for 5 minutes. Then the cells were seeded on each root surface blocks and cultured for 6, 24, 48, 72 hours. In second experiment, root surface blocks deminerilized with tetracycline and citric acid and nondemineralized root surface blocks were placed in 200ng/ml PDGF-BB for 5 minutes and another non-demineralized root surfcae blocks were placed in DMEM without PDGF-BB. At 1, 2, 4, 6, 8 days, the cells were seeded in 24-well plate and using of each eluent, cultured for 72 hours. The results of the four determinants were presented as mean and S.D.. The results were as follows : The attachment and proliferation of human gingival fibroblast on root surface were more increased when PDGF-BB was applicated on root surfrace demineralized with tetracycline or citric acid than non-demineralized root surface. And, in comparision tetracycline with citric acid, there were more attachment and proliferation of human gingival fibroblast on root surface demineralized with tetracycline than citric acid, and proliferation of human gingival fibroblast on demineralized root surface was increased time dependently 1 day to 3 days. In second experiment using eluent, proliferation of human gingival fibroblast was more increased to 6 days when human gingival fibroblast was cultured in eluent that PDGF-BB was applicated on demineralized root surface than two control groups, and degree of proliferation was decreased time dependently 1 day to 6 days. Proliferation of human gingival fibroblast cultured in eluent without PDGF-BB was constant 1 day to 6 days. After 6 days, degree of proliferation of human gingival fibroblast was similar in four groups. This means that release duration of PDGF-BB from demineralized root surface is 6 days. And in comparision tetracycline with citric acid, there was more proliferation of human gingival fibroblast in tetracycline-treated group than citric acid. In conclusion, demineralized root surface as primary site for PDGF-BB application, especially demineralized with tetracycline has important roles in attachment and proliferation of human gingival fibroblast, and may be useful clinical applications in periodontal regenerative procedures.

• Journal of Periodontal and Implant Science
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• v.24 no.1
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• pp.109-119
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• 1994

An essential fact in the regeneration of new periodontal tissue after periodontal therapy is the reattachment of collagen fibers to the tooth. Two phenomena play a fundamental role in preventing new connective tissue attachment to the exposed root surface ; 1) The apical migration of the junctional epithelium 2) The contamination of cementum by toxic substances, especially endotoxins. Authors have used rat submucosal implantation of root sections to study the connective tissue healing to periodontally diseaed root, previously planed and demineralized with citric acid and tetracycline- HCl. The results were obtained as follows. 1. The connective tissue attachment was increased in tetracycline, citric acid, non disease, scaling and root planing order and inflammatory reaction was seen in the rat teeth, no treatment group. 2. Collagen fiber attachment at the dentin surface was more increased than cementum surface 3. In 2 week of citric acid and tetracycline-HCl specimens, osteoid was seen near the fibrotic band. 4. In the MT view, collagen fiber formation was increased with time and the numerous collagen fiber and connective tissue was more densly attached to the tooth surfaces in the tetracycline-HCl group than the citric acid group.

• Journal of Periodontal and Implant Science
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• v.23 no.2
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• pp.219-227
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• 1993

The purpose of the present study was to evaluate the effects of 5% tetracycline(Tc) with or without citric acid on periodontally diseased root surfaces. Six single-rooted teeth extracted from one patient was selected and received thorough scaling and root planning, followed by saline irrigation. Each one tooth was divided into eight fragments with a thin separating disc. Total 48 fragments were prepared and setted into 4 groups for this study. Group I (control group)were treated with saline. Group II were treated with 5% Tc gel. Group III were treated with 33% citric acid-5% Tc gel. Group IV were treated with Tc solution. All the specimens are evaluated under Scanning Electron Microscope(SEM). Group I showed large amount of debris in spite of thorough scaling and root planing, but Group II, III & IV showed clean and soft root surface texture. In higher magnification(x3, 000), Group II, III & IV showed nunmerous dentinal tubules, especially Group IV showed collagen fibrils. In the present study, Tc gel and Tc gel with citric showed clinically successful result when treated on periodontally diseased root surface, in vitro.

• 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.31 no.1
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• pp.21-41
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• 2001

The goal of Periodontal treatment is predictable periodontal regeneration. But until now, many products including GTR materials and growth factors are beyond of complete regeneration. BMP can induce ectopic bone formation when implanted into sites such as rat muscle and can greatly enhance healing of bony defects when applied exogenously. BMP can promote periodontal regeneration by their ability to stimulate new bone and new cementum formation. But little is known about optimal conditions required for the application. Root conditioning is used for bioacive root change so altered root surface provides a substrate that promotes chemotaxis, migration and attachment of peridontal cells encouraging connective attachment to the denuded root surface. The aim of this study is to investigate whether the acid conditioning change effect of rhBMP-2 on human periodontal ligament cell and osteoblast cell line. 288 periodontally involved root dentin slices are divided into 6 groups, each 48, 1)control, 2)treated with BMP, 3)treated with citric acid 4)treated with citric acid+BMP 5)treated with tetracycline 6)treated with TC+BMP. Each group was devided half, so 12 root dentin slices were seeded with periodontal ligament cells and 12 were seeded with osteoblasts. At day 2 and 7, cell number, protein assay, ALP activitiy was measured. To investigate morphology of cultured cells, SEM was employed. Statistical analysis was performed with SPSS 8.0 either t-test or ANOVA test. The results are ; Protein assay and cell number was slightly decreased in CA+BMP group compared to Ca group but it was not statistically significant and ALP activity was much more increased in CA+BMP group compared to CA group so there was no statistically significance between BMP and CA+BMP group and statistically significant compared to control group. Cell number and protein assay was slightly increased in TC group and ALP activity was much less the BMP group and CA group. Cell number and protein and ALP activity was not much increased in TC+BMP group. TC group and TC+BMP group showed cell morphology change in SEM. This results suggested that application of root surface with citric acid before BMP treatment might give better result in periodontal regeneration.

• Journal of Periodontal and Implant Science
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• v.24 no.1
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• pp.87-97
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• 1994

The ultimate goal of periodontal therapy is to fully reconstruct the periodontal attachment apparatus. Commonly used techniques for treatment of infrabony defects include a combination of root planing, curettage and root treatment. To prevent the apical migration of epithelial cells, the technique of guided tissue regeneration is used. The aim of this study is to compare the effects of root treatment with Citric acid & Tetracycline and Guided tissue regeneration in dogs. Experimental periodontitis was induced by the ligation of orthodontic elastic threads in the upper right and left premolars 3, 4 of five adult dogs for 6 weeks. 4 types of procedures were performed as follows; 1) Control graup : Mucoperiosteal flap 2) Experinental I : GTR used Gore-tex(R) membrane 3) Experinental II : Root treatment with citric acid (PHl) 4) Experinental III : Root treatment with tetracycline HCl (50mg/ml) There after, dogs were serially sacrificed at the 1, 2, 4, 5, 8 weeks, and the specimens were prepared, and stained with hematoxylin-eosin for the light microscopic evaluation. The results of this study were as follows; 1. Junctional epithelium reached to the notch through the furcation area in control group at 8 weeks. 2. In the aspects of the inflammatory cell infiltration, control group showed severe aggregation than experimental group I, II, III through the experimental period 3. New cementum was observed over the notch from 5 weeks in experimental group II 4. In the aspects of the amount of new bone formation, experimental group was better than control group, but there was not significant differences among the experimental group, I, II, III

• Journal of Periodontal and Implant Science
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• v.35 no.3
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• pp.719-730
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• 2005

The purpose of this study was to compare the effect of tetracycline HCL, Citric acid and PrefGel applied on the root surfaces that planed with periodontal curret with Roto bur. In this study, 20 extracted teeth with advanced periodontal disease were used. The teeth were root planing with periodontal curette and Roto bur. Following root planing, each agents was burnished on the prepared root surface for 3 minutes to find opened dentinal tubules. And then, each specimens were investigated using scanning electron microscope. Amount of remained cementum by loss of tooth substance index and the number of opened dentinal tubules were evaluated to each specimens The results were as follows. 1. Groups treated with periodontal curette were almost seemed no removed. Other groups treated with Roto bur showed partially opened dentinal tubule orifices. 2. Loss of tooth substance index were compared between groups. There was no statistically difference between periodontal curette groups. Between Roto bur groups was alike. But there were statistically differences between periodontal curette and Roto bur groups. 3. At comparing with various root conditioning agents, Tetracycline HCL group took statistically higher than Citric acid and PrefGel in opened dentinal tubules. On the other hand, there was no statistically difference between Citric acid group and PrefGel group. As a result of this study, groups treated with Roto bur showed more cementum removed than groups treated with periodontal curette. In a treatment for regeneration of periodontal tissue, it was regarded that Roto bur should be used and that Tetracycline HCL would be more effective as chemical root conditioning agent.