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.
Journal of the Korean Academy of Esthetic Dentistry
/
v.22
no.1
/
pp.30-46
/
2013
Porcelain fused to metal crown has been used mostly over the last 50 years for restorations in dentistry. However, the patients' awareness of aesthetic aspect, biocompatibility and the problems such as an allergy to metals led to the growing interest in the 'metal free restoration'. In particular, the price of the precious metals that have been mainly used to date has risen drastically, which made them impossible to play their role as oral restorative materials anymore, and in addition, the PFM restoration has intrinsic problems of chipping and fracture. Therefore, the CAD/CAM has been drawing more attention than ever due to the popular needs for the material that is more aesthetic and stronger for restoration of the molar implant. Considerations in carrying out the full zirconia restoration are as follows: 1) strength, 2) combination work, 3) light penetrability, 4) treatment of cracks, 5) the color reproducibility of the block, 6) the abrasivity of antagonistic tooth, 7) low temperature degradation. In this presentation, the color reproducibility of the block will be discussed. One of the biggest reasons for avoiding the full zirconia restoration is that it is difficult to reproduce the natural color compared to the conventional PFM restoration. Thus, many clinicians show reluctance due to the exposure of the ugly block when the coloring on the surface is removed after occlusal adjustment. From the experience of using blocks by Zirkonzahn for more than 4 years, it is considered that these problems can be addressed to some degrees. Accordingly, how to make restorations that are well in harmony with surrounding prosthesis or natural teeth will be discussed.
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.
The purpose of this study was to study of the effects of the bioglass and the natural coral on healing process of the alveolar bone defects. Three adult dogs aged 1 to 2 years were used in this study. Experimental alveolar bone defects were created surgically with surgical bur and bone chisel at the furcation area of the buccal surface of the right and left mandibular 3rd, 4th premolars. Twelve experimental alveolar bone defects were devided into four groups according to the type of graft materials. The groups were as follows : 1. flap operation with root planing & curettage(Negative control group) 2. flap operation with autogenous bone(Positive control group) 3. flap operation with bioglass(BG group) 4. flap operation with natural coral(NC group) At 2, 4, and 8 weeks, the dogs were serially sacrificed and specimens were prepared with Hematoxylin-Eosin stain for light microscopic evaluation. The results of this study were as follows : 1. The defect areas were filled with granulation tissue at two weeks in negative control group. But in other groups, the appearance of connective tissues around graft materials were formed more densely and the response of inflammation by graft materials itself was not found. 2. In every control and experimental groups at two weeks, there was seen the accumulation of the formation of new bone trabeculae at the bottom of defects and gradually expanded toward the graft materials and in autogenous group there was slightly seen the formation of new cementum. 3. There was seen the erosion of central portion of bioglass particles at two weeks in BG group, and the erosion of the central portion was developed more progressively and was filled with bone-like tissues at eight weeks. 4. The natural coral particles were encapsulated by densely connective tissues and seen the formation of new bone tissues at four weeks and developed more new bone and cementum formation at eight weeks. From the results of this study, the bioglass and the natural coral may be biocompatible and have a weak adverse reaction to the periodontal tissues.
Kim, Seong-Ho;Park, Kui-Woon;Yoo, Hyung-Keun;Shin, Hyung-Shik
Journal of Periodontal and Implant Science
/
v.25
no.3
/
pp.539-556
/
1995
Periodontal therapeutic modalities should be re-establishing and regenerating the periodontal tissue previously lost to the disease. To achieve periodontal regeneration, periodontal ligament cells must selective migrate to the deneded root surface, attached and proliferated it. Local pH concentration is one of the most factors that periodontal regeneration. The aims of this study were to examine on biological effects of pH to the human periodontal ligament cells in vitro, especially on the cell morphology, attachment, activity, vitality and viability. Human periodontal ligament cells were cultured from extracted tooth for non-periodontal reason. Immediately after extraction, any soft tissue adhering to the cervical parts of the roots was carefully removed with a sterile curette. To produce different pH levels in the media, Eagle's MEM was adjusted from pH 6.6 to 8.2 in 0.2 intervals with 1 M NaOH and 1 N HCl. After cultivation, Then, Periodontal ligament cells were cultured at pH ranging from 6.6-8.2. attachment assay was done at 1, 2 day incubation and activity assay was done at 1, 2, 3 day incubation. The experiments were evaluated by scaning electron microscopic techniques (HITACHIX-650 Scaning Electron Microanalyzer, Tokyo, Japan), MTT assay, and the cultured periodontal ligament cells were fixed in neutral formalin for 24 hours and immunohistochemically processed by PCNA for proliferating ability. The surviving cells in the medium showed slightly increased volume and widening intercellular distances at low concentration of pH than control group (pH 7.4), and apparently shrinkage at high concentration of pH than control group (pH 7.4). The results of the statistical analysis from the experiment on attachment, vitality and viability were as follows. Attachment of periodontal ligament cells at 1st and 2nd day, similar attachment rate of low concentration pH compared with control value (pH 7.4). But above pH 8.0, attachment rate were statistically significant decrease from control value(P<0.05). Periodontal ligament cell's activities were maximum at pH 7.6 by MTT assay. Similar with control value at low concentration of pH. But, the activities were statistically significant decrease at high concentraration of pH(P<0.05). Cellular proliferating rate (PCNA index) were statistically significant decrease from control value at low and high concentration of pH(p<0.05). This results suggested that hjgh concentration pH, in other words, alkali pH was cytotoxic effects on human periodontal ligament cells in vitro.
The purpose of this study was to observe the effects of the periodontal ligament on the healing and the formation of alveolar bone in the extraction socket, when this ligament had artificially remained in the socket during the tooth removal. Twenty rats aged 4 weeks were used and devided into the control groups (10) and the experimental groups (10) in this study. The maxillary right and left first molars were extracted in both groups. In the experimental groups the periodontal ligament was remained in the extraction sockets using 0.4% ${\beta}-aminopropionitrile$, and in the control the periodontal ligament was completely removed by curettage. At 1, 3, 5, 7 and 14 days after the tooth extraction, rats in both groups were serially sacrificed. And the specimens were prepared with Hematoxylin-Eosin stain for the light microscopic evaluation. The results of this study were as follows ; 1. On 1 day, the periodontal ligament was only found in the extraction socket walls of the experimental groups, and there was not the distinguishable difference between the control and the experimental groups. 2. On 3 days, there were more collagen fibers and the appearance of higher cellular density in the experimental groups than in the control. And the cells and collagen of the periodontal ligament were so actively proliferated and synthesized that invaded into the connective tissue of the extraction sockets in the experimental groups. 3. In the experimental groups, the trabecular bone was formed on the basal and lateral bone surface on 5 days. However, there was not the new bone forming appearance in the control groups at this time. 4. On 7 days, the trabecular bone was formed in the control groups. 5. On 14 days, the extraction sockets were almost entirely filled with the bony trabeculae in both groups. But, compared to the control group, the experimental groups showed the prominent differences in the amount & the density of the new bone formed. In conclusion, it was suggested that the residual periodontal ligament tissue in the extraction socket will play a major role as the important cell source in the healing and the new bone formation of the extraction socket.
Since laser therapy has been applied to dentistry, many dental practitioners are very interested in laser therapy on various intraoral soft tissue lesions including gingival hyperplasia and aphthous ulcer. The purpose of the present study was to determine the therapeutic effect and the harmful effect of a pulsed-Nd:YAG laser irradiation on human gingival tissue. In twenty periodontal patients with gingival enlargement, the facial gingival surface of maxillary anterior teeth was randomly irradiated at various power of 1.0W(100mJ, 10Hz), 3.0W(100mJ, 30Hz) and 6.0W(l50mJ, 40Hz) for 60 seconds by contact delivery of a pulsed-Nd:YAG laser(EN.EL.EN060, Italy). Immediately after laser irradiation, the gingival tissues were surgically excised and prepared in size of 1mm3. Subsequently the specimens were processed for prefixation and postfixation, embedded with epon mixture, sectioned in $1{\mu}$ thickness, stained with uranyl acetate and lead citrate, and observed under transmission electron microscope(JEM 100 CXII). Following findings were observed; l. In the gingival specimens irradiated with l.OW power, widening of intercelluar space and minute vesicle formation along the widened intercellular space were noted at the epithelial cells adjacent to irradiated area. 2. In the gingival specimens irradiated with 3.0W power, the disruption of cellular membrane, aggregation of cytoplasm, and loss of intercellular space were observed at the epithelial cells adjacent to irradiated area. 3. In the gingival specimens irradiated with 6.0W power, the disruption of nuclear and cellular membrane was observed at the epithelial cells adjacent to irradiated area. The ultrastructural findings of this study suggest that surgical application of a pulsed-Nd:YAG laser on human gingival tissue may lead somewhat delayed wound healing due to damage of epithelial cells adjacent to irradiated area.
The purpose of this study was to evaluate histomorphometrically a toothash - plaster of Paris mixture associated with collagen membrane ($Bio-Gide^{(R)}$), regarding new bone formation in the peri-implantitis defects in dogs. Three mandibular molars were removed from 1-year-old mongrel dogs. After 2 months of healing, 2 titanium implants with sandblasted with large grit and acid etched (SLA) surface were installed in each side of the mandible. Experimental peri-implantitis was induced with ligatures after successful osseointegration. Ligatures were removed after identification of bone defect beneath the level of 5th thread of fixture on radiographic image. The mucoperiosteal flaps were elevated and the contaminated fixtures were treated with chlorhexidine and saline. The bone defects were assigned to one of the following treatments: no guided bone regeneration (GBR) procedure (group 1), GBR with Bio-$Oss^{(R)}$ and Bio-$Gide^{(R)}$ (group 2), or GBR with toothash - plaster of Paris mixture (TPM) and Bio-$Gide^{(R)}$ (group 3). The dogs were sacrificed after 8 or 16 months. The mean percentages of new bone formation within the limits of the 5 most coronal threads were $17.83{\pm}10.69$ (8 weeks) and $20.13{\pm}13.65$ (16 weeks) in group 1, $34.25{\pm}13.32$ (8 weeks) and $36.33{\pm}14.21$ (16 weeks) in group 2, and $46.33{\pm}18.39$ (8 weeks) and $48.00{\pm}17.78$ (16 weeks) in group 3, respectively. The present study confirmed statistically considerable new bone formation within the threads in group 3 compared with group 1 at 8 and 16 weeks (P<0.05). Although, data analysis did not reveal significant differences between group 2 and 3, the latter showed better results during the period of 8 or 16 weeks. Our findings support the effectiveness of TPM as a GBR material in the treatment of peri-implantitis bone defect.
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