Evaluation of the Viability of Rat Periodontal Ligament Cells after Storing at 0℃/2 MPa Condition up to One Week: In Vivo MTT Method

PURPOSE: The aim of this study was to evaluate the rat periodontal ligament cell viability under 0℃/2 MPa condition up to one week using in vivo 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay. MATERIALS AND METHODS: As soon as 110 upper molar teeth of rats were extracted, they were stored in Hartman's solution under 0℃/2 MPa condition for 1, 2, 3, 4, and 7 days each. All specimens were treated with in vivo MTT assay and the value of optical density was measured by ELISA reader. These values were statistically analyzed by one-way ANOVA. RESULT: There was no statistical difference on MTT value between immediate and 1 day storage group. There were statistically significant differences between 1 day and 2 days tsorage, 2 and 3 days storage groups, respectively. Teeth of 3,4, and 7 days storage groups showed significantly lower MTT valuesc ompared with shorter period storage groups. CONCLUSION: When the MTT values were substituted in standard curve, 1 day storage group at 0℃/2 MPa condition showed 68% cell viability when compared with immediate group. It dropped to 13% at 2 days, and to less than 5% at 3 days or more.

Delayed intentional replantation of periodontally hopeless teeth: a retrospective study

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.

Vital tooth with periapical lesion: spontaneous healing after conservative treatment

It is often presumed that apical periodontitis follows total pulp necrosis, and consequently root canal treatment is commonly performed. Periapical lesion development is usually caused by bacteria and its byproduct which irritate pulp, develop pulpitis, and result in necrosis through an irreversible process. Afterwards, apical periodontitis occurs. This phenomenon is observed as an apical radiolucency in radiographic view. However, this unusual case presents a spontaneous healing of periapical lesion, which has developed without pulp necrosis in a vital tooth, through conservative treatment.

Pulp necrosis following luxated injury to teeth in a patient with uncontrolled type II diabetes mellitus: a case report

Patients with diabetes mellitus show delayed wound healing and increased susceptibility to infection. Therefore, the effects of diabetes on pulpal and periodontal healing should be taken into consideration when treating diabetic dental traumatized patients. This case presents the treatment for dental traumatized 20 yr old female with uncontrolled type II diabetes. The traumatized upper central incisors had showed pulpal healing in early days. However, 7 mon after the trauma, the teeth had been diagnosed with pulp necrosis with apical abscess. Eventually, non surgical root canal treatment on the teeth had been performed.

Changes in periodontium after extraction of a periodontally-involved tooth in rats

PURPOSE: Recent interest has focused on intentional replantation to restore an original tooth. Some studies have shown successful results with intentional replantation for periodontally involved teeth. For long-term success of replantation, a healthy periodontal status of the recipient site is required so that delayed replantation is more suitable for periodontally involved teeth. To reveal the ideal timing for delayed replantation of periodontally involved teeth, the healing process of extraction sockets after extraction of periodontitis-induced teeth in rats was evaluated. METHODS: Twenty-eight rats were randomly divided into two groups: a control group (n=8) and test group (n=20). In the test group, periodontitis was induced by a ligature around the cervix of the mandibular first molar of all of the rats. Two weeks later, the mandibular first molars were extracted in all of the animals. The animals were sacrificed on days 0, 3, 7, and 10 after extraction and histological and immunohistochemical analysis was performed. RESULTS: In histological analysis of the test group, inflammatory cell infiltrate was found abundantly in the remaining periodontium 3 days after tooth extraction and decreased gradually at later time points. In immunohistochemical analysis of the test group, both interleukin-6 (IL-6) and, tumor necrosis factor-alpha (TNF-alpha) were numerous in the furcation area at each postextraction day. IL-6 was stained more heavily between 3 and 7 days after extraction; at day 10 after extraction, little staining was observed. TNF-alpha staining was more intense at 3 days after extraction and gradually weakened at later points in time. CONCLUSIONS: Within the limits of this study, it takes at least 10 days to resolve periodontal inflammation in rat extraction sockets.

Hypoesthesia after IAN block anesthesia with lidocaine: management of mild to moderate nerve injury

Hypoesthesia after an inferior alveolar nerve (IAN) block does not commonly occur, but some cases are reported. The causes of hypoesthesia include a needle injury or toxicity of local anesthetic agents, and the incidence itself can cause stress to both dentists and patients. This case presents a hypoesthesia on mental nerve area followed by IAN block anesthesia with 2% lidocaine. Prescription of steroids for a week was performed and periodic follow up was done. After 1 wk, the symptoms got much better and after 4 mon, hypoesthesia completely disappeared. During this healing period, only early steroid medication was prescribed. In most cases, hypoesthesia is resolved within 6 mon, but being aware of etiology and the treatment options of hypoesthesia is important. Because the hypoesthesia caused by IAN block anesthesia is a mild to moderate nerve injury, early detection of symptom and prescription of steroids could be helpful for improvement of the hypoesthesia.

Minimizing the extra-oral time in autogeneous tooth transplantation: use of computer-aided rapid prototyping (CARP) as a duplicate model tooth

OBJECTIVES: The maintenance of the healthy periodontal ligament cells of the root surface of donor tooth and intimate surface contact between the donor tooth and the recipient bone are the key factors for successful tooth transplantation. In order to achieve these purposes, a duplicated donor tooth model can be utilized to reduce the extra-oral time using the computer-aided rapid prototyping (CARP) technique. MATERIALS AND METHODS: Briefly, a three-dimensional digital imaging and communication in medicine (DICOM) image with the real dimensions of the donor tooth was obtained from a computed tomography (CT), and a life-sized resin tooth model was fabricated. Dimensional errors between real tooth, 3D CT image model and CARP model were calculated. And extra-oral time was recorded during the autotransplantation of the teeth. RESULTS: The average extra-oral time was 7 min 25 sec with the range of immediate to 25 min in cases which extra-oral root canal treatments were not performed while it was 9 min 15 sec when extra-oral root canal treatments were performed. The average radiographic distance between the root surface and the alveolar bone was 1.17 mm and 1.35 mm at mesial cervix and apex; they were 0.98 mm and 1.26 mm at the distal cervix and apex. When the dimensional errors between real tooth, 3D CT image model and CARP model were measured in cadavers, the average of absolute error was 0.291 mm between real teeth and CARP model. CONCLUSIONS: These data indicate that CARP may be of value in minimizing the extra-oral time and the gap between the donor tooth and the recipient alveolar bone in tooth transplantation.

Patients' perception and satisfaction with apicoectomy / 대한치과보존학회지

OBJECTIVES: This study was aimed to examine the patients' perception and satisfaction with the results of endodontic microsurgery which was apicoectomy with retrofilling. MATERIALS AND METHODS: A questionnaire was given to 109 patients, who were recalled after a minimum of 3 months upon endodontic microsurgery in the Department of Conservative Dentistry, Yonsei University. A contingency table and correlation analysis were used to determine if there were any correlations between age/gender and the patients' responses (p = 0.05). RESULTS: Approximately 60% of respondents answered they had never heard of surgical endodontic procedures. 63.3% of respondents chose the surgical option because they wanted to keep their natural teeth. If the patient required the same procedure on another tooth later, 100 out of 109 respondents answered they would choose microsurgery instead of extraction. Most patients (82.57%) appeared to be satisfied with the surgical procedure. CONCLUSIONS: Endodontic microsurgery consisting of apicoectomy and retrofilling seems to appeal to majority of patients as a satisfactory and valuable treatment choice.

Partial pulp necrosis caused by excessive orthodontic force / 대한치과보존학회지

As the dental pulp is encased with a rigid, noncompliant shell, changes in pulpal blood flow or vascular tissue pressure can have serious implication for the health of pulp. Numerous studies have demonstrated that orthodontic force application may influence both blood flow and cellular metabolism, leading degenerative and/or inflammatory responses in the dental pulp. The aim of this case report is to present a case about tooth with chronic periapical abscess which showed normal vital responses. Excessive orthodontic force is thought to be the prime cause of partial pulp necrosis. Owing to remaining vital tissue, wrong dianosis can be made, and tooth falsely diagnosed as vital may be left untreated, causing the necrotic tissue to destroy the supporting tissuses. Clinician should be able to utilize various diagnostic tools for the precise diagnosis, and be aware of the endodontic-orthodontic inter-relationship.

Histology of dental pulp healing after tooth replantation in rats / 대한치과보존학회지

The objective of this study was to observe the histology of dental pulp healing after tooth replantation in rats. The maxillary right first molars of 4-week-old rat were extracted, and then the teeth were repositioned in the original socket. At 3 days after replantation, there was localized inflammatory reaction. But, pulp revasculization and healing had already begun in the root area. At 5 days after replantation, odontoblast-like cells were observed. Tertiary dentin deposition was observed beneath the pulp-dentin border from 1 week after replantation. And tertiary dentin was increased at 2 weeks after replantation. The presence of odontoblast-like cells and the formation of tertiary dentin were continued to 4 weeks after replantation. At 4 weeks after replantation, the deposition of bone-like tissues and cementum-like tissues was observed. This results show that there is a possibility of pulp healing after tooth replantation in rats and the mineralization of tooth can progress. The mineralization of tooth after replantation was initially occurred by the deposition of tertiary dentin, but as time passed, the deposition of bone-like tissues and cementum-like tissues was begun and increased.

The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure / 대한치과보존학회지

The purpose of this study was to evaluate the viability of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure by means of MTT assay, WST-1 assay. 12 teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both side of the first and second maxillary molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group 1 (Immediate extraction), group 2 (Slow freezing under pressure of 3 MPa), group 3 (Slow freezing under pressure of 2 MPa), group 4 (Slow freezing under no additional pressure), group 5 (Rapid freezing in liquid nitrogen under pressure of 2 MPa), group 6 (Rapid freezing in liquid nitrogen under no additional pressure), group 7 (low-temperature preservation at 0degrees C under pressure of 2 MPa), group 8 (low-temperature preservation at 0degrees C under no additional pressure), group 9 (low-temperature preservation at -5degrees C under pressure of 90 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in 37degrees C water bath, then MTT assay, WST-1 assay were processed. One way ANOVA and Tukey HSD method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 7 (0degrees C/2 MPa) showed higher viability of periodontal ligament cells than other group (2-6, 8) and this was statistically significant (p < 0.05), but showed lower viability than group 1, immediate extraction group (no statistical significance). By the results of this study, low-temperature preservation at 0degrees C under pressure of 2 MPa suggest the possibility for long term preservation of teeth.

In vivo evaluation of accuracy and consistency of two electronic apex locators / 대한치과보존학회지

OBJECTIVES: To evaluate the accuracy and consistency of two different apex locators at both the Apex and 0.5 marks. MATERIALS AND METHODS: Twenty-six root canals was scheduled for extraction for periodontal or prosthodontic reasons. Thirteen canals were measured using Root ZX and the rest by i-ROOT. The root canal length was measured both the at 0.5 mark and the Apex mark. The file was then fixed to the tooth, and the distance from the file tip to the major foramen of each canal was measured after removing the root dentin under the microscope so that the major foramen and the file tip were seen. RESULTS: 1. When the Apex mark was used, 100% of both the Root ZX and i-ROOT groups were within 0.5 mm of the major foramen. 2. When 0.5 mark was used, 100% of the Root ZX group and 77% of the i-ROOT group were within 0.5 mm of the major foramen. 3. In terms of standard deviation and quartile value, the Apex mark was more consistent than 0.5 mark in the Root ZX group, and 0.5 mark was more consistent in the i-ROOT group, but there was no statistically significant difference when compared with t-test. 4. The root canal length difference between the Apex mark and 0.5 mark was 0.22 mm and 0.46 mm in the Root ZX and i-ROOT groups, respectively. CONCLUSIONS: In this study, the Apex mark was the more consistent mark. Therefore, it is recommended to subtract 0.5 mm, which is the average length between the apex and apical constriction, from the root canal length at the Apex mark to obtain the working length clinically.

Evaluation of vitrification for cryopreservation of teeth

PURPOSE: The aim of this study was to investigate whether vitrification in the cryopreservation of periodontal ligament (PDL) cells could be useful for tooth banking. METHODS: In step 1, primary cultured human PDL cells were cryopreserved in 100% conventional cryopreservation media and 100% vitrification media (ESF40 media) in different temperatures for 2 weeks. In step 2, a series of modified vitrification formulae named T1 (75% vitrification media + 25% F media), T2 (50% vitrification media + 50% F media) and T3 (25% vitrification media + 75% F media) were used to store PDL cells for 2 weeks and 4 weeks in liquid nitrogen. MTT assay was performed to examine the viability of PDL cells. RESULTS: Maximum cell viability was achieved in cells stored in 100% conventional cryopreservation media at -196degrees C (positive control group) in step 1. Compared to the positive control group, viability of the cells stored in 100% vitrification media was very low as 10% in all test conditions. In step 2, as the percentage of vitrification media decreased, the cell viability increased in cells stored for 2 weeks. In 4-week storage of cells in step 2, higher cell viability was observed in the T2 group than the other vitrification formulae while the positive control group had the highest viability. There was no statistically significant difference in the cell viability of 2-week and 4-week stored cells in the T2 group. CONCLUSIONS: These observations indicate 100% vitrification media is not successful in PDL cell cryopreservation. Conventional cryopreservation media is currently the most appropriate media type for this purpose while T2 media would be interesting to test for long-term storage of PDL cells.

The efficacy of programmed cryo-preservation under pressure in rat periodontal ligament cells / 대한치과보존학회지

The purpose of this study was to evaluate the viability of periodontal ligament cells in rat teeth using slow cryo-preservation method under pressure by means of MTT assay and WST-1 assay. Eighteen teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both sides of the first and second maxillary molars were extracted as atraumatically as possible under Tiletamine anesthesia. The experimental groups were group 1 (Immediate control), group 2 (Cold preservation at 4degrees C for 1 week), group 3 (Slow freezing), group 4 (Slow freezing under pressure of 3 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in 37degrees C water bath, then MTT assay and WST-1 assay were processed. One way ANOVA and Tukey method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 4 showed significantly higher viability of periodontal ligament cells than group 2 and 3 (p < 0.05), but showed lower viability than immediate control group. By the results of this study, slow cryo-preservation method under pressure suggests the possibility for long term cryo-preservation of the teeth.

Comparison of viability of oral epithelial cells stored by different freezing methods / 대한치과보존학회지

This study examined the influence of the storage methods on the viability of oral epithelial cells using conventional cell freezing storage, slow freezing preservation, rapid freezing preservation, and slow freezing preservation with a pressure of 2 Mpa or 3 Mpa. The cell viability was evaluated by cell counting, WST-1 and the clonogenic capacity after 6 days of freezing storage. After 6 days, the frozen cells were thawed rapidly, and the cell counting, WST-1, and clonogenic capacity values were measured and compared. 1. The results from cell counting demonstrated that conventional cryopreservation, slow freezing under a 2 Mpa pressure and slow freezing under a 3 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p<0.05). 2. The results from the optical density by WST-1 demonstrated that slow freezing under a 2 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p<0.05). 3. The clonogenic capacity demonstrated that slow freezing under a 2 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p<0.05).

In vitro evaluation of the consistency of two electronic apex locators / 대한치과보존학회지

The purpose of this study was to evaluate the consistency of two electronic apex locators in vitro model. Materials consisted of fifty two extracted premolars and two electronic apex locators; Root ZX (J. Morita, Osaka, Japan) and E-Magic Finder Deluxe (S-Denti, Cheonan, Korea). After access preparation, the teeth were embedded in a saline-mixed alginate model. Canal lengths of each tooth were measured at "0.5" and "Apex" mark of the apex locators, respectively so that each tooth had two measurements from 0.5 and Apex points. The file was fixed at final measurement using a glass ionomer cement. The apical 4 mm from the apex was exposed to measure the distance from the file tip to the major apical foramen of each tooth. Average distances and standard deviations were used to evaluate the consistency. Results showed that all measurements of both Root ZX and E-Magic Finder located the major foramen the range of +/- 0.5 mm level. Both apex locators showed better consistency at Apex mark than at 0.5 mark. The average distance of file tip-major foramen was - 0.18 mm at 0.5 mark and - 0.07 mm at Apex mark in Root ZX, - 0.25 mm at 0.5 mark and - 0.02 mm at Apex mark in E-Magic Finder. Standard deviation was 0.21 at 0.5 mark and 0.12 at Apex mark in Root ZX, 0.12 at 0.5 mark and 0.09 at Apex mark in E-Magic Finder.

Evaluation of the viability of periodontal ligament cell in rat teeth using slow cryopreservation method with magnetic field / 대한치과보존학회지

The purpose of this study was to evaluate the viability of periodontal ligament cell in rat teeth using slow cryopreservation method with magnetic field through MTT assay and TUNEL test. For each group, 12 teeth of 4 weeks old white female Sprague-Dawley rat were used for MTT assay, and 6 teeth in TUNEL test. The Maxillary left and right, first and second molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group1 (immediately extraction), group 2 (cold preservation at 4degrees C for 1 week), group 3 (rapid cryopreservation in liquid nitrogen), group 4 (slow cryopreservation with magnetic field of 1 G), and group 5 (slow cryopreservation). F medium was used as preservation medium and 10% DMSO as cryoprotectant. After preservation and thawing, the MTT assay and TUNEL test were processed. One way ANOVA and Scheffe method were performed at the 95% level of confidence. The value of optical density obtained after MTT analysis was divided by the value of eosin staining for tissue volume standardization. In both MTT assay and TUNEL test, it had showed no significant difference among group 3, 4, and 5. And group 3 had showed higher viability of periodontal ligament cell than group 2. From this study, slow cryopreservation method with magnetic field can be used as one of cryopreservation methods.

Dual Plane Method with Orbital Fat Repositioning in Lower Blepharoplasty

Blepharoplasty is a commonly performed aesthetic surgery, but there is still no commonly used standard method for the correction of orbital fat bulging in lower blepharoplasty. We performed dual plane method by elevating skin flap and muscle flap separately, and then repositioned orbital fat with preserving orbital septum for correction of orbital fat bulging in lower blepharoplasty. Between July, 1998 and June, 2005, one hundred and fifty six patients underwent lower blepharoplasty by author's method. The results were satisfactory in all of the patients. Complications were six cases of hematoma and ten cases of visible scar in lateral canthal area. There was no skin flap necrosis, hyperpigmentation, skin irregularity or ectropion. In lower blepharoplasty for the sufficient excision of skin and operation of orbicularis oculi, we performed precise correction of skin and muscle independently by dual plane approach, by separate elevation of skin flap and muscle flap, and were able to reposition orbital fat easily and safely with preservation of orbital septum.

Evaluation of periodontal ligament cell viability in rat teeth after frozen preservation using in-vivo MTT assay / 대한치과보존학회지

The purpose of this study was to examine the viability of PDL cells in rat molars by using in vivo MTT assay, which was used to compare fast cryopreservation group by liquid nitrogen (-196degrees C) with 4degrees C cold preservation group. A total of 74 Sprague-Dawley white female rats of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted as atraumatically as possible under ketamine anesthesia. Ten teeth of each group were divided as six experimental groups depending upon the preservation. Cryopreservation groups were Group 1 (5% DMSO 6% HES in F medium), Group 2 (10% DMSO in F medium), Group 3 (5% DMSO 6% HES in Viaspan(R)), Group 4 (10% DMSO in Viaspan(R)) which were cryopreserved for 1 week and cold preservation groups were Group 5 (F medium), Group 6 (Viaspan(R)) at 4degrees C for 1 week. Immediate extraction group was used as a control. After preservation and thawing, the in vivo MTT assay was processed. Two way ANOVA and Duncan's Multiple Range Test was performed at the 95% level of confidence. Another 2 teeth of each group were treated as the same manner and frozen sections 10 microm thick for microscopic observation. The value of optical density obtained after in vivo MTT analysis was divided by the value of eosin staining for tissue volume standardization. Group 1, 2 had significantly higher optical density than Group 3 and 4 which had the lowest OD value. Group 6 had higher OD value than in Group 5 (P < 0.05). Histological findings of periodontal ligament cell, after being stained with MTT solution were consistent with the in vivo MTT assay results. In this study, the groups which were frozen with DMSO as a cryoprotectant and the groups with F medium showed the best results.

In vitro evaluation of accuracy and consistency of four different electronic apex locators / 대한치과보존학회지

The purpose of this study was to evaluate the accuracy and the consistency of four different electronic apex locators in an in vitro model. Fourty extracted premolars were used for the study. Four electronic apex locators (EAL) were Root ZX, SmarPex, Elements Diagnostic Unit (EDU), and E-Magic Finder Deluxe (EMF). After access preparation, the teeth were embedded in an alginate model and the length measurements were carried out at "0.5"and "Apex"mark using four EALs. The file was cemented at the location of the manufacturers'instruction (Root ZX, EDU, EMF: 0.5 mark, SmarPex: Apex mark). The apical 4mm of the apex was exposed and the distance from the file tip to the major foramen was measured by Image ProPlus (x 100). The distance from the file tip to the major foramen was calculated at 0.5 and Apex mark and the consistency of 0.5 and Apex mark was compared by SD and Quartile of Box plots. In this study, Root ZX and EMF located the apical constriction accurately within +/- 0.5 mm in 100%, whereas SmarPex and EDU located in 90% and in 70% respectively. For Root ZX and EMF, there was no significant difference between the consistency of 0.5 and Apex mark. However, for the EDU and SmarPex, Apex mark was more consistent than 0.5 mark. From the evaluation of the consistency in this study, for Root ZX and EMF, both 0.5 and Apex mark can be used as a standard mark. And for EDU and SmarPex, the Apex mark can be recommended to be used as a standard mark.