Removal Of Periodontal Ligament Research Articles

Does periodontal ligament removal using natural proteolytic enzymes alter the surface structure and microhardness of cementum of avulsed tooth? An in vitro analysis

Introduction: Surface treatment of avulsed tooth must be directed towards the debridement of necrotic periodontal fibers while having no effect on the cementum microhardness. The role of proteolytic enzymes on the periodontal ligament (PDL) surface and hardness of cementum has not been investigated in the literature. Aim: The present study aimed to evaluate the effect of 5.25% sodium hypochlorite (NaOCl), 10% bromelain, 10% papain and 10% panzyme on root surface topography and microhardness of avulsed teeth. Materials and Methods: Forty healthy premolars were freshly extracted and randomly allocated into five treatment groups based on the surface agent employed; Group 1: 50 ml saline, Group 2:20 ml of 5.25% NaOCl, Group 3: 20 ml of 10% bromelain enzyme, Group 4: 20 ml of 10% papain enzyme, and Group 5: 20 ml of 10% mixed panzyme enzyme. The specimens were then subjected to confocal laser scanning microscopy and scanning electron microscopic examination. Supplemental set of freshly extracted forty premolars were decoronated longitudinally and one-half from each specimen was submitted for Vickers microhardness testing before and after application of the above-mentioned agents. Statistical Analysis: Intragroup comparison of mean percentage hardness reduction was assessed using paired t-test. Results: Treatment with NaOCl led to complete removal of PDL fibers with the presence of cracks and crazes on the root surface along with a significant reduction in microhardness. Bromelain and papain revealed regular and even distribution, while few PDL fibers were evident with papain. Panzyme revealed a shiny surface with minute cracks and a significant reduction in microhardness. Conclusion: Ten percentage bromelain for 10 min was effective in the removal of necrotic PDL fibers and preserved the cementum integrity better.

Effect of periodontal ligament removal with gauze prior to delayed replantation in rabbit incisors on rate of replacement resorption.

Delayed (dry storage > 60minutes) replantation results in ankylosis and replacement resorption. It has been suggested to remove the non-viable periodontal ligament before replantation to possibly reduce the rate of replacement resorption. However there has been no study on the rate of replacement resorption after such measures. The aim of this study was to investigate if there was any difference in the rate of replacement resorption by either removing the periodontal ligament (PDL) with gauze or not removing PDL in teeth subjected to delayed replantation followed by healing for 2 or 6weeks. Maxillary central incisors were extracted in 8 rabbits. In the right central incisors, the necrotic PDL was removed by dry gauze over the root surface. In the left eight extracted teeth PDL was left on the root surface. All extracted teeth were left to dry for 60minutes. Extra-oral root canal treatment was performed before replantation. The rabbits were sacrificed after 2weeks and 6weeks respectively. Histologic processing and evaluation was done. In the 2weeks group, all teeth showed ankylosis. The cementum was intact, and fusion of the bone and root was generally seen without resorption of the root, whereas in the 6weeks group regardless of whether PDL had been kept or not, ankylosis and osseous replacement of the dentin was seen. There was no evidence of inflammatory infiltrate in the sections examined. Removal of PDL prior to delayed replantation may result in some initial protection of the cementum during the first few weeks. However, over longer times there seems to be neither protection of the dentin from ankylosis and osseous replacement, nor any influence on the rate of replacement resorption.

An experimental model for studies on delayed tooth replantation and ankylosis in rabbits.

To develop an experimental in vivo model to study dentoalveolar ankylosis and osseous replacement resorption after delayed replantation. The maxillary right incisors of eight rabbits were extracted and replanted, while the maxillary left incisors served as controls. A special technique for extraction was used, luxating the incisor laterally prior to extraction. Extra-oral root canal treatment was performed. In four of the eight replanted teeth, periodontal ligament was removed using gauze. All teeth were replanted after one-hour dry storage. A horizontal line was drilled on the labial enamel surface of the incisors to enable registration of possible subsequent infraposition. The rabbits were sacrificed after 1 week or 4 weeks, respectively. Percussion test of the teeth was carried out and infraposition was registered. Histological processing and evaluation were performed. Percussion sound was normal after 1 week and high in the 4 week group; infraposition was noted in all replanted teeth in the 4-week group. Fusion between the bone and dentin was seen in some areas on the root already after 1 week, but there was little or no osseous replacement. Larger areas of ankylosis were seen in the 4-week group although not significant, but deeper progressive osseous replacement was noted in this group. There was no significant difference in ankylosis with regard to periodontal ligament removal or not. Pulp and periodontal tissues were normal in all control teeth. The rabbit tooth model may be useful for experimental in-vivo studies on delayed replantation, subsequent ankylosis, osseous replacement and infraposition.

The effect of propolis solution on root surface treatment in replanted teeth

Objective: The objective of this study was to assess the effect of propolis solution on root surface treatment in cases of delayed tooth replantation following chemical or mechanical removal of the periodontal ligament. Material and methods: Maxillary right central incisors of 36 male rats (Wistar) were extracted, kept in a dry environment for 15 minutes and then distributed into six groups, as follows: mechanical-propolis and mechanical-fluoride (mechanical removal followed by immersion in 6% propolis and 2% sodium fluoride, respectively), mechanical (mechanical removal only), chemical-propolis and chemical-fluoride (chemical removal followed byimmersion in 6% propolis and 2% sodium fluoride, respectively), and chemical (chemical removal only). Following periodontal ligament removal, root canals were filled with calcium hydroxide paste and replanted. After 60 days, histological analysis with light microscopy was carried out to detect areas of dentoalveolar ankylosis and root resorption. Results: Results revealed that chemical removal of the periodontal ligament was associated with slightly better tissue repair findings when compared with mechanical removal. Conclusions: The use of 6% propolis solution encouraged tissue repair, while the 2% sodium fluoride solution combined with chemical removal resulted in a better-organized healing process, with less destruction of dentine when compared with the 6% propolis solution.

Delayed tooth replantation: MTA as root canal filling.

MTA has been investigated as a root-end filling material. Its mechanism of action has some similarities to that of Ca(OH)2. The purpose of this study was to evaluate the repair process taking place in the delayed replantation of monkey teeth using calcium hydroxide and MTA as root canal filling materials. Five monkeys had their lateral incisors extracted and bench-dried for 60 minutes. After root canal preparation, the teeth were assigned to two groups according to root canal filling material: I, calcium hydroxide; and II, MTA. The same treatment sequence was followed for both groups: coronal seal, periodontal ligament removal, immersion of the tooth in 2% acidulated-phosphate sodium fluoride, irrigation of the socket with saline and replantation. Both groups exhibited replacement resorption, areas of ankylosis and absence of inflammatory root resorption. Statistically similar results (p > 0.05) were observed for both groups regarding replacement root resorption, but the groups differed significantly (p < 0.05) regarding the occurrence of ankylosis. MTA may be a viable clinical option for filling teeth submitted to delayed replantation, and is an acceptable option for treating replanted permanent teeth in order to prevent tooth resorption, particularly when dressing changes are not possible.

Expression of OPG, RANK, and RANKL Proteins in Tooth Repair Processes After Immediate and Delayed Tooth

This study evaluated the immunohistochemical expression of OPG, RANK, and RANKL proteins in the repair after immediate and delayed replantation of rat teeth. Fifty-six Wistar rats (Rattus norvegicus albinus) had their maxillary right lateral incisor extracted and then replanted, according to the following conditions: group I (control; n = 8), teeth were not extracted; group II (n = 16), immediate replantation; group III (n = 16), delayed replantation without treatment; and group IV (n = 16), delayed replantation after root surface treatment (periodontal ligament removal and immersion in 2% acidulated-phosphate sodium fluoride) and calcium hydroxide intracanal dressing. Rats in group I were euthanized on the first day of the experiment, while the animals in the other groups were euthanized 10 and 60 days after replantation (n = 8/period). Hematoxylin and eosin-stained sections were obtained for histological analysis. The immunohistochemical analysis revealed expression of OPG and RANKL proteins in all groups and both postreplantation times, except for group II at 60 days. In the experimental groups, RANK expression was observed only at 10 days. In conclusion, there was strong immunostaining for the OPG-RANK-RANKL system at the earlier postreplantation time, suggesting a more effective participation of these proteins at the start of the healing process, as their expression decreased at 60 days.

Mechanical removal of necrotic periodontal ligament by either Robinson bristle brush with pumice or scalpel blade. Histomorphometric analysis and scanning electron microscopy

One of the important factors accounting for successful delayed replantation of avulsed teeth is seemingly the type of root surface treatment. Removal of necrotic cemental periodontal ligament remnants may prevent the occurrence of external root resorption, which is the major cause of loss of teeth replanted in such conditions. The purpose of this study was to compare the efficacy of two mechanical techniques for removal of root-adhered periodontal ligament. Preservation or removal of the cementum layer concomitantly with these procedures was also assessed. Forty-five roots of healthy premolars extracted for orthodontic purposes were selected. After extraction, the teeth were kept dry at room temperature for 1 h and then immersed in saline for rehydration for an additional 10 min. Thereafter, the roots were assigned to three groups, as follows: group 1 (control)--the cemental periodontal ligament was preserved; group 2--removal of the periodontal ligament by scraping root surface with a scalpel blade (SBS); group 3--periodontal ligament remnants were removed using a Robinson bristle brush at low-speed with pumice/water slurry (RBP). The specimens were analysed histomorphometrically and examined by scanning electron microscopy. The quantitative and qualitative analyses of the results showed that the RBP technique was significantly more effective than the SBS technique for removal of the periodontal ligament remnants adhered to root surface. Both techniques preserved the cementum layer.

Delayed tooth replantation after root surface treatment with sodium hypochlorite and sodium fluoride: histomorphometric analysis in rats

In cases of delayed tooth replantation, non-vital periodontal ligament remnants have been removed with sodium hypochlorite in an attempt to control root resorption. Nevertheless, reports of its irritating potential in contact with the alveolar connective tissue have been described. Therefore, this study evaluated the healing process on delayed replantation of rat teeth, after periodontal ligament removal by different treatment modalities. Twenty-four rats, assigned to 3 groups (n=8), had their upper right incisor extracted and left on the workbench for desiccation during 60 min. Afterwards, the teeth in group I were immersed in saline for 2 min. In group II, root surfaces were scrubbed with gauze soaked in saline for 2 min; and in group III, scrubbing was done with gauze soaked in 1% sodium hypochlorite solution. Thereafter, root surfaces were etched with 37% phosphoric acid and immersed in 2% acidulate-phosphate sodium fluoride solution, at pH 5.5. Root canals were filled with a calcium hydroxide-based paste and the teeth were replanted. The animals were sacrificed 60 days postoperatively and the pieces containing the replanted teeth were processed and paraffin-embedded. Semi-serial transversally sections were obtained from the middle third of the root and stained with hematoxylin and eosin for histomorphometric analysis. Data were analyzed statistically using Kruskal-Wallis and Dunn's tests. The results showed that root structure and cementum extension were more affected by resorption in group III (p<0.05). All groups were affected by root resorption but the treatment performed in group III was the least effective for its control. The treatment accomplished in groups I and II yielded similar results to each other.

Relationship of cellular proliferation to expression of osteopontin and bone sialoprotein in regenerating rat periodontium.

Cellular repopulation was studied in a model in which adjacent mineralising and soft connective tissue matrices are regenerated. Window wounds were created through alveolar bone, with either preservation or removal of periodontal ligament, in 30 male Wistar rats. Three animals per time period were killed on days 1, 3, 7, 14, and 28 after surgery for each wound type. Cellular proliferation in alveolar bone and periodontal ligament was assessed by 3H-thymidine labelling 1 h before death, followed by radioautographic analysis. Cellular differentiation was determined by the temporal expression of the bone-related markers osteopontin and bone sialoprotein, using immunohistochemical methods. In regenerating periodontium, osteopontin was expressed earlier than bone sialoprotein, which was restricted to alveolar bone. After wounding, transient expression of osteopontin was detected in the periodontal ligament at days 1 and 3. In general, wounding induced fivefold higher proliferation and clonal growth of periodontal ligament cells compared to the unwounded (control) side. Combined immunostaining and radioautography demonstrated colocalisation of osteopontin in sites with high numbers of labelled cells in both nascent periodontal ligament and regenerating alveolar bone at days 3 and 7. In contrast, bone sialoprotein, which appeared in regenerating alveolar bone on days 14-28 after wounding, was expressed much later than the peak of cellular proliferation. We conclude that (1) the intact periodontal ligament influences cell proliferation and osteopontin expression; (2) osteopontin is an early marker of periodontal tissue regeneration that is temporally and spatially associated with intensive cell proliferation and migration in osteogenic and periodontal ligament cell populations; and (3) bone sialoprotein is expressed after proliferation at sites of mineralising bone formation.

Periodontal healing after replantation and autotransplantation of incisors in monkeys

The effect of injury to the cemental or alveolar part of the periodontal ligament upon periodontal healing after replantation and autotransplantation of teeth was studied in green Vervet monkeys. Sixty-six maxillary central incisors were replanted or autotransplanted after extra-alveolar periods of 18 min. saline storage or 120 min, dry storage. The animals were sacrificed after 8 weeks and the teeth examined histometrically. Identical healing was found in the following experimental groups: replantation or auto transplantation after 18 min. and replantation with or without removal of periodontal ligament in the alveolus. A significant increase in replacement resorption (ankylosis) compared to the 18 min. replantation group was found in the following groups: replantation and autotransplantation after 120 min., replantation after removal of periodontal ligament on the root surface and autotransplantation after 120 min. to a socket where the tooth was extracted 18 min. earlier. Autotransplantation after 18 min. to a socket where a tooth had been extracted 120 min, earlier led to a significant increase in the amount of replacement resorption compared to transplantation to an 18 min. socket. Based on these findings, it is concluded that the presence of an intact and viable periodontal ligament on the root surface is the most important factor in assuring healing without root resorption. The length of the extra-alveolar period also seems to exert some influence upon the socket, enhancing the development of ankylosis.