An ultimate goal of periodontal therapy is to stop the disease process and to regenerate a functionally-oriented periodontium destroyed as a result of periodontal disease. The purpose of this study was to observe the effect of grafting granulation tissue obtained from extraction socket on the regeneration of horizontal furcation defect. Six dogs were used in this study. All mandibular first and third premolars were extracted. At 2, 3, and 5 days after extraction, tissues were obtained from extraction socket of 1 mongrel dog and examined by light microscope. Granulation tissue obtained at 5 days after extraction was chosen as the graft material. Five days later, horizontal furcation defects were created surgically at mandibular second and fourth premolars in the right and left side of the 5 beagle dogs. The entrance area of the artificially prepared "key hole" defects were about $3\;4mm^2$. By random selections, 2 exposed furcation defects were grafted with granulation tissue obtained from extraction socket as experimental group and 1 furcation defect was as control. The flaps were replaced to their original position and sutured with 4-0 chromic cat-gut. Three dogs were sacrificed 4 weeks and two dogs 8 weeks after surgery, and the prepared specimens were examined by light microscope. At 4 weeks, furcations were filled with epithelial lining and fibrous connective tissue infiltrated with chronic inflammatory cells. New bone formation was observed in all groups. Only experimental group showed new cementum formation. At 8 weeks, new cementum, functional arrangement of new PDL fiber, root resorption, and some ankylotic union of newly formed alveolar bone and root surface were observed in all groups. Experimental group showed that epithelial downgrowth was inhibited and new bone formation was more active compared to control. The success rate of the furcation defect healing was higher in experimental group than control. These results suggested that grafting of granulation tissue obtained from extraction socket which combined with reconstructive periodontal flap surgery may promote periodontal regeneration of horizontal furcation defect.
Purpose: Avulsed tooth can be completely recovered, if sound periodontal ligament (PDL) of tooth is maintained. Although a lot of storage solutions have been explored for the better storage of avulsed tooth, there is a shortcoming that the preservation time is much short. On the other hand, there has been studies that (-)-epigallocatechin-3-gallate (EGCG), the most abundant polyphenol in green tea, which is related to the anti inflammatory, antioxygenic, and antibacterial effects, allows the successful preservations of tissues and cells. This study evaluated the effect of EGCG on avulsed-teeth preservation of Beagle dogs for a period of time. Methods: The atraumatically extracted teeth of Beagle dogs were washed and preserved with 0/10/$100\;{\mu}M$ of EGCG at the time of immediate, period 1 (4 days in EGCG-contained media and additional 1 day in EGCG-free media), period 2 (8 days in EGCG-contained media and additional 2 days in EGCG-free media) and period 3 (12 days in EGCG-contained media and additional 2 days in EGCG-free media). Then, the cell viabilities of preserved teeth was calculated by dividing optical density (OD) of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay with OD of eosin assay to eliminate the measurement errors caused by the different tissue volumes. Results: From the results, the immediately analyzed group presented the highest cell viability, and the rate of living cells on teeth surface decreased dependent on the preservation period. However, the $100\;{\mu}M$ of EGCG-treated group showed statistically significant positive cell activity than EGCG-free groups throughout preservation periods. Conclusions: Our findings showed that $100\;{\mu}M$ EGCG could maintain PDL cell viability of extracted tooth. These results suggest that although EGCG could not be a perfect additive for tooth preservation, it is able to postpone the period of tooth storage. However, further in-depth studies are required for more plausible use of EGCG.
Purpose: The aim of this study was to compare osteoblast behavior on zirconia and titanium under conditions cultured with bone morphogenetic protein-2. Methods: MC3T3-E1 cells were cultured on sandblasted zirconia and sandblasted/etched titanium discs. At 24 hours after seeding MC3T3-E1, the demineralized bone matrix (DBM) gel alone and the DBM gel with bone morphogenetic protein-2 (BMP-2) were added to the culture medium. The surface topography was examined by confocal laser scanning microscopy. Cellular proliferation was measured at 1, 4, and 7 days after gel loading. Alkaline phosphatase activity was measured at 7 days after gel loading. The mRNA expression of ALPase, bone sialoprotein, type I collagen, runt-related transcription factor 2 (Runx-2), osteocalcin, and osterix were evaluated by real-time polymerase chain reaction at 4 days and 7 days. Results: At 1, 4, and 7 days after loading the DBM gel alone and the DBM gel with BMP-2, cellular proliferation on the zirconia and titanium discs was similar and that of the groups cultured with the DBM gel alone and the DBM gel with BMP-2 was not significantly different, except for titanium with BMP-2 gel. ALPase activity was higher in the cells cultured with BMP-2 than in the other groups, but there was no difference between the zirconia and titanium. In ALPase, bone sialoprotein, osteocalcin, Runx-2 and osterix gene expression, that of cells on zirconia or titanium with BMP-2 gel was much more highly increased than titanium without gel at day 7. The gene expression level of cells cultured on zirconia with BMP-2 was higher than that on titanium with BMP-2 at day 7. Conclusions: The data in this study demonstrate that the osteoblastic cell attachment and proliferation of zirconia were comparable to those of titanium. With the stimulation of BMP-2, zirconia has a more pronounced effect on the proliferation and differentiation of the osteoblastic cells compared with titanium.
Purpose: Recently, interest in chitosan has increased due to its excellent biological properties such as biocompatibility, antibacterial effect, and rapid healing capacity. On the other hand, hydroxyapatite is used as a bone substitute in the fields of orthopedics and dentistry. The hydroxyapatite-chitosan (HA-CS) complex containing hydroxyapatite nanoparticles was developed for synergy of both biomaterials. The objective of this study was to evaluate the effect of hydroxyapatite (HA)-chitosan (CS) membrane on bone regeneration in the rat calvarial defect. Methods: Eight-millimeter critical-sized calvarial defects were created in 70 male Sprague-Dawley rats. The animals were divided into 7 groups of 10 animals and received either 1) chitosan (CS) 100% membrane, 2) hydroxyapatite (HA) 30%/CS 70% membrane, 3) HA 30%/CS 70%, pressed membrane, 4) HA 40%/CS 60% membrane, 5) HA 50%/CS 50% membrane, 6) HA 50%/CS 50%, pressed membrane, or 7) a sham . surgery control. The amount of newly formed bone from the surface of the rat calvarial defects was measured using histomorphometry, following 2- or 8- week healing intervals. Results: Surgical implantation of the HA - CS membrane resulted in enhanced local bone formation at both 2 and 8 weeks compared to the control group. The HA - CS membrane would be significantly more effective than the chitosan membrane in early bone formation. Conclusions: Concerning the advantages of biomaterials, the HA-CS membrane would be an effective biomaterial for regeneration of periodontal bone. Further studies will be required to improve the mechanical properties to develop a more rigid scaffold for the HA-CS membrane.
Purpose: The purpose of this study was to retrospectively evaluate the survival of periodontally hopeless teeth that were intentionally extracted and replanted after a delay and to compare the radiographic characteristics of the survival group with those of the failure group. Methods: The clinical and radiographic data from patients who underwent delayed intentional replantation between March 2000 and July 2010 were reviewed. Twenty-seven periodontally hopeless teeth were extracted and preserved in medium supplemented with antibiotics for 10-14 days. The teeth were then repositioned in the partially healed extraction socket and followed for 3 to 21 months. The radiographic parameters were analyzed using a paired t test and the cumulative survival rate was analyzed using Kaplan-Meier analysis. Results: Seven replanted teeth failed and the overall cumulative survival rate was 66.4%. In the survival group, the amount of bone loss was reduced from 68.45% to 34.66% three months after replantation. There was radiologic and clinical evidence of ankylosis with 5 teeth. However, no root resorption was found throughout the follow-up period. In the failure group, bone formation occurred from the bottom of the socket. However, a remarkable radiolucent line along the root of a replanted tooth existed. The line lengthened and thickened as time passed. Finally, in each case of failure, the tooth was extracted due to signs of inflammation and increased mobility. Conclusions: Delayed intentional replantation has many advantages compared to immediate intentional replantation and could serve as an alternative treatment for periodontally involved hopeless teeth. However, techniques for maintaining the vitality of periodontal structures on the tooth surface should be developed for improved and predictable results.
The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of periodontal ligament cells. Primary human periodontal ligament cells were cultured in dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells of 4th to 7th passage were inoculated in the multiwell plates coated with chitosan in concentration of 0.22, 0.2, and $2mg/m{\ell}$. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized modules was evaluated after 21 days of culture. The results were as follows : 1. The morphology of periodontal ligament cells on the chitosan coating was round or spheric. Round cells were aggregated after 6 hours of culture. Aggregated cells on the chitosan coated surface showed nodule-like appearance after 24 hours of culture and not achieved confluency at 7 days. 2. During early period of culture, the attachment of periodontal ligament cells were inhibited by chitosan coating. Inhibition of cell attachment tended to increase with the concentration of chitosan. 3. At the chitosan concentration of 0.02 and $0.2mg/m{\ell}$, periodontal ligament cells were more rapidly proliferated at 7 days, compared to the control group. At the concentration of $2mg/m{\ell}$, the proliferation of periodontal ligament cells was inhibitied(p<0.01). 4. Alkaline phosphatase activity of periodontal ligament cells was increased in chitosan coated group, especially at the concentration of $0.02mg/m{\ell}$after 4 days of culture.5. Periodontal ligament cells produced mineralized nodules on chitosan coated wells without the addition of mineralized nodule forming materials (ascorbic acid, ${\beta}-glycerophosphat$, dexamethasone). With the addition of mineralized nodule forming materials, periodontal ligament cells produced more mineralized nodules at the concentration of $0.02mg/m{\ell}$, compared to the control. In summary, the attachment, proliferation, cell activity, and alkaline phosphatase activity of periodontal ligament cells depended on the concentration of coated chitosan. Chitosan stimulated mineralized nodule formation by periodontal ligament cells. At the appropriate concentration($0.02mg/m{\ell}$), chitosan could increase alkaline phosphatase activity and stimulate the formation of mineralized nodule by periodontal ligament cells. These results suggest that chitosan can be used as an adjunct for bone graft material, and the matrix of tissue engineering for periodontal regeneration, especially bone regeneration.
The present study was performed to compare effects of demineralized freeze-dried bone allograft(DFDBA) with deproteinized bovine bone mineral(DBBM) on periodontal fenestration defect in rats. Twelve adult male rats weighing 500 to 540 grams were used in this study. Periodontal fenestration defects were surgically created with tapered fissure bur(${\Phi}1mm$) at the left side of buccal surface of the mandible. The defect size was from anterior border of the first molar to anterior of the ascending ramus mesiodistally and from just below the alveolar crest to apically 1.5-2mm area apicocoronally with 2mm in depth. Rats were divided into control group, test group I and II. Four defects were assigned to the test group I grafted with DBBM and other 4 defects were assigned to the test group II grafted with DFDBA. The rest of defects were the negative control group. At 10 days and 35 days after surgery, 12 rats were sacrificed through intracardiac perfusion and specimens were obtained prepared with Hematoxylin-Eosin stain for light microscopic evaluation. The results of this study were as follows : 1. In the control group, new bone, osteoid, dense connective tissue were observed in the defects at 10 days. new bone formation was not found but loose connective tissue was formed in the defect and fibrous encapsulation of graft materials was shown in two test groups at 10 days. 2. In all groups, new bone formation was shown in the defect at 35 days. And in the control group, bone formation increased at 35 days than at 10 days. 3. In the test group I and II at 35 days, graft materials were combined with new bone and joined host bone. There was very close contact between new bone, graft materials, and host bone with no gaps. 4. In the test group I and II, new bone formation was similar to that in the control group but not exeeded. In conclusion, in the test group I new bone formation was similar to that in the test group II at 35 days, but there was infiltration of inflammatory cells at 10 days. DFDBA and DBBM were considered as the biocompatible graft materials and effective in the regeneration of new bone.
치과에서 수복물제작에는 지난 50년간 금속을 이용한 금속도재관이 많이 사용되어 왔다. 그러다 보니 심미적이고 생체 친화적이며 금속 알러지등의 문제로 인한 환자들의 사고의 변화에 따라서 'metal free restoration'에 대한 관심이 점점 높아지고 있다. 특히 그 동안 많이 사용되어왔던 귀금속 가격이 급등하여 더 이상 구강 내 수복 물로써 제 역할을 하는 것은 불가능하게 되었으며, PFM수복은 술자로 하여금 chipping과 파절로 인한 문제점을 안고 있다. 따라서 구치부 임플란트 수복물로써 보다 심미적이고 강도가 높은 재료의 요구로 인하여 그 어느 때보다 CAD/CAM이 주목을 받고 있다. Full zirconia 수복을 위해서 고려해야 할 사항은 1. 강도 2. 콤비네이션 작업은? 3. 빛 투과성은? 4. crack이 발생한 경우의 처치는? 5. 블럭의 색조 재현상은? 6. 대합치 마모도 7. 저온 열화 현상 등을 들 수가 있는데, 본 연구에서는 블럭의 색조 재현상에 대해서 정리하고자 한다. Full zirconia 수복을 꺼려하는 가장 커다란 이유 중에 하나가 바로 기존의 PFM 수복과 비교했을 때 자연스러운 색조를 재현하기가 어렵다는 점이며, 교합 조정 후에 컬러링한 표면이 삭제된 후 나타나는 보기 싫은 블럭의 노출로 인하여 많은 임상가들이 꺼려하고 있다. 지르코잔의 블럭을 약 4년 여 동안 사용하면서 이러한 점들은 어느 정도 극복되어질 수 있는 문제라고 여겨지며, 어떻게 하면 그 가급적 주변 보철물 또는 자연 치아와의 조화 이룰 수 있는 수복물을 만들 수 있을 것인가에 관해 정리였다.
The present study was to evaluate the healing patterns of guided tissue regeneration( GTR) using resorbable $Vicryl^{(R)}$(polyglactin 910) mesh and nonresorbable expanded polytetrafluoroethylene(ePTFE) membrane with or without bone grafting using autogeneous bone and demineralized freeze-dried bone allograft(DFDBA) in the grade II furcation defects. Mucoperiosteal flaps were reflected buccally in the mandibular 2nd, 3rd and 4th premolar areas and furcation defects were created surgically by removing $5{\times}6mm$ alveolar bone in 4 dogs. Root surfaces were thoroughly debrided of periodontal ligament and cementum, and notches were placed on root surface at the most apical bone level. In the right and left mandibular quadrant, each tooth was received $Vicryl^{(R)}$ mesh(ACE Surgical Supply Co., USA) only, $Vicryl^{(R)}$ mesh with DFDBA, $Vicryl^{(R)}$ mesh with autogeneous bone grafts, ePTFE membrane($Core-tex^{(R)}$ membrane, W.L. Gore & Associates Inc., USA) only, ePTFE membrane with DFDBA or ePTFE membrane with autogeneous bone grafts. For the fluorescent microscopic examination, fluorescent agents were injected at 2, 4 and 8 weeks after surgery. Four weeks after surgery, 2 dogs were sacrificed and ePTFE membranes were removed from remaining 2 dogs, which were sacrificed at 12 weeks after surgery. Undecalcified tissues were embedded in methylmethacrylate and $10{\mu}m$ thick sections were cut in a buccolingual direction. These sections were stained with hematoxylin-eosin stain and Masson's trichrome stain, and evaluated by descriptive histology and linear measurements. The results were as follows : 1) $Vicryl^{(R)}$ mesh group showed less connective tissue attachment than ePTFE membrane group. 2) The combination of GTR using $Vicryl^{(R)}$ mesh and osseous grafts resulted in new attachment and new bone formation more than GTR using $Vicryl^{(R)}$ mesh only. 3) GTR using ePTFE membrane, with or without osseous grafts, enhanced periodontal regeneration. 4) Root resorption and dentoalveolar ankylosis were observed in the areas treated with the combination of GTR and DFDBA. It was suggested that the effect of adjunctive bone grafting in GTR procedure depends on the materials and the physical properties of barrier membranes. $Vicryl^{(R)}$ mesh performed a barrier function and the use of adjunctive bone grafting may enhance the periodontal regeneration.
The purpose of this study was to compare effects of various bone grafts on periodontal regeneration of alveolar bone defects in dogs. Seven adult dogs aged 12 to 18 months were used in this study. Experimental alveolar bone defects were created surgically with a #1/2 round bur at the furcation area of the buccal surface of the mandibular 3rd, 4th premolars and 1st molar. Each experimental alveolar bone defects were grafted with dense hydroxyapatite, natural coral, and decalcified freeze-dried bone, and respectively divided into DHA, NC, DFDB group. An area without bone graft was divided into control group. At 1,2,4,6, and 12 weeks, dogs were serially sacrificed and specimens were prepared with Hematoxylin-Eosin stain and Mallory stain for light microscopic evaluation. The results of this study were as follows : 1. In control group, the matrix change of granulation tissue was observed at 1 week. And in experimental groups, the appearance of connective tissue around graft materials was loosely formed at 1 week, but densely formed at 2 weeks. 2. In every group, the slight formation of new trabecular bone was seen from remaining bone at 1 week. 3. The DHA and NC particles were gradually encapsulated by new trabecular bone from remaining bone, and the osteoid tissue was directly induced from DFDB particles. 4. The presence of osteoblasts was first observed at 1 week in control group and at 2 weeks in NC group, but at 6 weeks in DHA group. 5. In DHA group, the resorption of particles was not observed until 12 weeks. But in NC and DFDB group, the particles were resorbed at 6 weeks and replaced by new bone. And the amount and size of particles were reduced, and their border represented irregular form. In summary, in three experimental groups the inflammatory or foreign body reaction were slight, but the regeneration of new osteoid tissue and the matrix change of dense connective tissue fiber were observed. Especially, NC and DFDB materials were considered as the biocompatible graft materials which were effective in the regenertion of new bone.
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