Endodontic Microsurgical Treatment of a Three-rooted Mandibular First Molar with Separate Distolingual Root: Report of One Case.

The separate distolingual (DL) roots of three-rooted mandibular first molars are thought to be too difficult for performing apical surgery. This article represents microsurgical treatment of a three-rooted mandibular first molar with a separate DL root. The procedure includes incision and flap retraction, osteotomy, apicoectomy, retropreparation and retrofilling of the root canal, using micro instruments, ultrasonic retrotips and mineral trioxide aggregate (MTA) under a dental operating microscope. Two mm in length of apical root resection, 2 mm in depth of root canal retropreparation with a personalised ultrasonic retrotip, and 2 mm in length of retrofilling with MTA are the key points for accomplishment of apical surgery on separate DL roots. The case was followed up for 15 months after surgery. Clinical and radiographic examinations revealed complete healing of periapical tissue. Separate DL roots of three-rooted mandibular first molars can be treated by endodontic microsurgery with modifications from standard protocol.

Healing after root-end microsurgery by using mineral trioxide aggregate and a new calcium silicate-based bioceramic material as root-end filling materials in dogs.

INTRODUCTION: The purpose of this study was to compare healing after root-end surgery by using grey mineral trioxide aggregate (MTA) and EndoSequence Root Repair Material (RRM) as root-end filling material in an animal model. METHODS: Apical periodontitis was induced in 55 mandibular premolars of 4 healthy beagle dogs. After 6 weeks, root-end surgeries were performed by using modern microsurgical techniques. Two different root-end filling materials were used, grey MTA and RRM. Six months after surgery, healing of the periapical area was assessed by periapical radiographs, cone-beam computed tomography (CBCT), micro computed tomography (CT), and histology. RESULTS: Minimal or no inflammatory response was observed in the majority of periapical areas regardless of the material. The degree of inflammatory infiltration and cortical plate healing were not significantly different between the 2 materials. However, a significantly greater root-end surface area was covered by cementum-like, periodontal ligament-like tissue, and bone in RRM group than in MTA group. When evaluating with periapical radiographs, complete healing rate in RRM and MTA groups was 92.6% and 75%, respectively, and the difference was not statistically significant (P = .073). However, on CBCT and micro CT images, RRM group demonstrated significantly superior healing on the resected root-end surface and in the periapical area (P = .000 to .027). CONCLUSIONS: Like MTA, RRM is a biocompatible material with good sealing ability. However, in this animal model RRM achieved a better tissue healing response adjacent to the resected root-end surface histologically. The superior healing tendency associated with RRM could be detected by CBCT and micro CT but not periapical radiography.

The effects of human platelet lysate on dental pulp stem cells derived from impacted human third molars.

Human platelet lysate (PL) has been suggested as a substitute for fetal bovine serum (FBS) in the large-scale expansion of dental pulp stem cells (DPSCs). However, the biological effects and the optimal concentrations of PL for the proliferation and differentiation of human DPSCs remain unexplored. We isolated and expanded stem cells from the dental pulp of extracted third molars and evaluated the effects of PL on the cells' proliferative capacity and differentiation potential in vitro and in vivo. Before testing, immunocytochemical staining and flow cytometry-based cell sorting showed that the cells derived from human dental pulp contained mesenchymal stem cell populations. Cells were grown on tissue culture plastic or on hydroxyapatite-tricalcium phosphate (HA/TCP) biomaterials and were incubated with either normal or odontogenic/osteogenic media in the presence or absence of various concentrations of human PL for further investigation. The proliferation of DPSCs was significantly increased when the cells were cultured in 5% PL under all testing conditions (P < 0.05). However, this enhancement was inconsistent when the cells were cultured in 1% PL or in 10% PL; 10% PL significantly inhibited cell proliferation and was therefore excluded from further differentiation testing. Culture medium containing 5% PL also significantly promoted the mineralized differentiation of DPSCs, as indicated by the measurement of alkaline phosphatase activity and calcium deposition under mineral-conditioned media (P < 0.05). Scanning electron microscopy and modified Ponceau trichrome staining showed that the cells treated with 5% PL and mineralizing media were highly capable of integrating with the HA/TCP biomaterials and had fully covered the surface of the scaffold with an extensive sheet-like structure 14 d after seeding. In addition, 5% PL showed significantly positive effects on tissue regeneration in two in vivo transplantation models. We conclude that the appropriate concentration of PL enhances the proliferation and mineralized differentiation of human DPSCs both in vitro and in vivo, which supports the use of PL as an alternative to FBS or a nonzoonotic adjuvant for cell culture in future clinical trials. However, the elucidation of the molecular complexity of PL products and the identification of both the essential growth factors that determine the fate of a specific stem cell and the criteria to establish dosing require further investigation.

Root canal morphology of permanent maxillary teeth in the Han nationality in Chinese Guanzhong area: a new modified root canal staining technique.

INTRODUCTION: The purpose of this study was to investigate the canal morphology of 504 maxillary permanent teeth of subjects of Han nationality in Chinese Guanzhong area. METHODS: Maxillary permanent teeth were randomly collected in Guanzhong area. After regular preparation, the teeth were immersed into ink without preparing access cavities and then put into hyperbaric oxygen chamber (0.6 Mpa) for 2 hours to let the ink penetrate into root canal from apical foramen, apical deltas and foramen of lateral canals under stable positive pressure. After demineralization and clearing, the following observations were made: (1) number of root canals, (2) root canal configuration by using Vertucci's classification, (3) presence of lateral canals, and (4) frequency of apical deltas. RESULTS: All the teeth were well-stained, and the fine details were well-revealed. Apical deltas (12.2%-83.3%) and lateral canals (13.7%-68.8%) could be frequently found in all types of maxillary teeth. Most of central incisors (95.8%), lateral incisors (91.4%), and canines (75.4%) displayed type I canal configuration, whereas most of first premolars (87.3%) and second premolars (72.3%) possessed 2 canals with type II, IV, or VI canal configuration. The majority of distobuccal roots and palatal roots of first molars (88.9%, 97.8%), second molars (92.0%, 94.0%), and third molars (87.5%, 91.6%) possessed type I canal configuration. The prevalence of mesiobuccal roots with type I configuration was 66.7% in maxillary first molars, 82% in second molars, and 62.5% in third molars. CONCLUSIONS: The modified technique of canal staining can effectively reveal detailed root canal system. The canal configuration of maxillary teeth in subjects of Han nationality in Chinese Guanzhong area is consistent with previous reports in other races.

[Establishment and application of an in vitro model for apatite crystal mineralization].

OBJECTIVE: To establish an in vitro model for the apatite crystal mineralization. To evaluate the influences of bovine serum albumin (BSA) and fluoride to the mineralization of apatite crystal. METHODS: The model was constructed using cation selective membrane (CMV) and dialysis membrane. Double distilled water (DDW), BSA, 5, 20, 100 mg x L(-1) fluoride were added into the reaction space of the model. Reaction was carried out at 37 degrees C for 3 days under gentle stirring. The crystals were identified by scanning electron microscope (SEM) and X-ray diffraction (XRD). RESULTS: The model was established successfully. When DDW and BSA were added respectively, the main component of the deposit was octacalcium phosphate (OCP), but the shape and size of the crystals differs from each other. When fluoride with different concentration were added, the main component of the crystal turned to rod-like and prism-like fluoroapatite (FAP) crystal. The size and crystallinity of the FAP increased with the increase of the fluoride concentration. CONCLUSION: It is an effective way to evaluate the influence factors of the apatite crystal mineralization by using the in vitro model.

[Tissue engineering of dentin-pulp complex-like structures by human dental mesenchymal cells].

OBJECTIVE: To establish three-dimensional culture model of human dental mesenchymal cells and bioengineer in vivo with ceramic bovine bone (CBB) and Collagraft as scaffolds. METHODS: Human dental mesenchymal cells induced upon stimulation of bFGF and IGF-1 or TGF-beta(1) were implanted onto CBB and Collagraft containing the same kinds of growth factors respectively. Then cell/scaffold constructs were transplanted into nude mice to establish in vivo culture model of dental mesenchymal cells. Control groups were set up at the same time. After 4 weeks or 10 weeks, the implants were taken out for histological and immunohistochemical analysis. RESULTS: Within 10-week implant tissues, typical dentin-pulp complex-like structures were generated in scaffolds containing growth factors. Human dentin sialoprotein (DSP) was expressed in the newly formed dentin. This phenomenon wasn't observed in control groups and 4-week implants. CONCLUSIONS: Dentin-pulp complex-like structures could be bioengineered successfully with human dental mesenchymal cells and CBB or Collagrafts containing growth factors in nude mice.

[Immortalized human odontoblast-like cell line expressed dentin extracellular matrix in vitro].

OBJECTIVE: To determine whether dentin matrix proteins were expressed by the human odontoblast-like cell line hTERT-hOd-1 in vitro. METHODS: Collagen type I, bone sialoprotein (BSP), dentin matrix protein 1 (DMP1) and the marker for odontoblast, dentin sialophosphoprotein (DSPP) and dentin sialoprotein (DSP) were detected in these cells by immunohistochemistry, RT-PCR and in situ hybridization. During being cultured in mineralizing medium for 5 weeks, the secretion of OC and activity of ALP were measured once a week. RESULTS: DSPP, DMP1, BSP and collagen type I were expressed in hTERT-hOd-1 either at mRNA or protein level. Under the induction of mineralizing medium, the cells showed higher activity of ALP and increased secretion of OC. CONCLUSION: hTERT-hOd-1 expressed dentin extracellular matrix in vitro, which means the cell line has the potential of mineralization.