Axin2-CreERT2 mediated knockout of bone morphogenetic protein receptor type 1A caused abnormal secondary dentine and altered cell fate of Axin2-expressing odontogenic cells.

AIM: Inducing odontogenic differentiation and tubular dentine formation is extremely important in dentine repair and tooth regeneration. Bone morphogenic proteins (BMPs) signalling plays a critical role in dentine development and tertiary dentine formation, whilst how BMPR1A-mediated signalling affects odontoblastic differentiation of Axin2-expressing (Axin2+ ) odontogenic cells and tubular dentine formation remains largely unknown. This study aims to reveal the cellular and molecular mechanisms involved in the formation of secondary dentine. METHODOLOGY: Axin2lacZ/+ mice harvested at post-natal 21 (P21) were used to map Axin2+ mesenchymal cells. Axin2CreERT2/+ ; R26RtdTomato/+ mice and Axin2CreERT2/+ ; R26RDTA/+ ; R26RtdTomato/+ mice were generated to observe the tempo-spatial distribution pattern of Axin2-lineage cells and the effect of ablation of Axin2+ cells on dentinogenesis, respectively. A loss-of-function model was established with Axin2CreERT2/+ ; Bmpr1afl/fl ; R26RtdTomato/+ (cKO) mice to study the role of BMP signalling in regulating Axin2+ cells. Micro-computed tomography, histologic and immunostainings, and other approaches were used to examine biological functions, including dentine formation, mineralization and cell differentiation in cKO mice. RESULTS: The results showed rich expression of Axin2 in odontoblasts at P21. Lineage tracing assay confirmed the wide distribution of Axin2 lineage cells in odontoblast layer and dental pulp during secondary dentine formation (P23 to P56), suggesting that Axin2+ cells are important cell source of primary odontoblasts. Ablation of Axin2+ cells (DTA mice) significantly impaired secondary dentine formation characterized with notably reduced dentine thickness (Mean of control: 54.11 µm, Mean of DTA: 27.79 µm, p = .0101). Furthermore, malformed osteo-dentine replaced the tubular secondary dentine in the absence of Bmpr1a with irregular cell morphology, abnormal cellular process formation and lack of cell-cell tight conjunction. Remarkably increased expression of osteogenic markers like Runx2 and DMP1 was detected, whilst DSP expression was observed in a dispersed manner, indicating an impaired odontogenic cell fate and failure in producing tubular dentine in cKO mice. CONCLUSIONS: Axin2+ cells are a critical population of primary odontoblasts which contribute to tubular secondary dentine formation, and BMP signalling pathway plays a vital role in maintaining the odontogenic fate of Axin2+ cells.

Loss of IκBζ Drives Dentin Formation via Altered H3K4me3 Status.

Enforced enrichment of the active promoter marks trimethylation of histone H3 lysine 4 (H3K4me3) and acetylation of histone H3 lysine 27 (H3K27ac) by inhibiting histone demethylases and deacetylases is positively associated with hard tissue formation through the induction of osteo/odontogenic differentiation. However, the key endogenous epigenetic modulator of odontoblasts to regulate the expression of genes coding dentin extracellular matrix (ECM) proteins has not been identified. We focused on nuclear factor (NF)-κB inhibitor ζ (IκBζ), which was originally identified as the transcriptional regulator of NF-κB and recently regarded as the NF-κB-independent epigenetic modulator, and found that IκBζ null mice exhibit a thicker dentin width and narrower pulp chamber, with aged mice having more marked phenotypes. At 6 mo of age, dentin fluorescent labeling revealed significantly accelerated dentin synthesis in the incisors of IκBζ null mice. In the molars of IκBζ null mice, marked tertiary dentin formation adjacent to the pulp horn was observed. Mechanistically, the expression of COL1A2 and COL1A1 collagen genes increased more in the odontoblast-rich fraction of IκBζ null mice than in wild type in vivo, similar to human odontoblast-like cells transfected with small interfering RNA for IκBζ compared with cells transfected with control siRNA in vitro. Furthermore, the direct binding of IκBζ to the COL1A2 promoter suppressed COL1A2 expression and the local active chromatin status marked by H3K4me3. Based on whole-genome identification of H3K4me3 enrichment, ECM and ECM organization-related gene loci were selectively activated by the knockdown of IκBζ, which consistently resulted in the upregulation of these genes. Collectively, this study suggested that IκBζ is the key negative regulator of dentin formation in odontoblasts by inhibiting dentin ECM- and ECM organization-related gene expression through an altered local chromatin status marked by H3K4me3. Therefore, IκBζ is a potential target for epigenetically improving the clinical outcomes of dentin regeneration therapies such as pulp capping.

Use of 0.4-Tesla static magnetic field to promote reparative dentine formation of dental pulp stem cells through activation of p38 MAPK signalling pathway.

AIM: To investigate whether static magnetic fields (SMFs) have a positive effect on the migration and dentinogenesis of dental pulp stem cells (DPSCs) to promote reparative dentine formation. METHODOLOGY: In vitro scratch assays and a traumatic pulp exposure model were performed to evaluate the effect of 0.4-Tesla (T) SMF on DPSC migration. The cytoskeletons of the DPSCs were identified by fluorescence immunostaining and compared with those of a sham-exposed group. Dentinogenic evaluation was performed by analysing the expressions of DMP-1 and DSPP marker genes using a quantitative real-time polymerase chain reaction (qRT-PCR) process. Furthermore, the formation of calcified deposits was examined by staining the dentinogenic DPSCs with Alizarin Red S dye. Finally, the role played by the p38 MAPK signalling pathway in the migration and dentinogenesis of DPSCs under 0.4-T SMF was investigated by incorporating p38 inhibitor (SB203580) into the in vitro DPSC experiments. The Student's t-test and the Kruskal-Wallis test followed by Dunn's post hoc test with a significance level of P < 0.05 were used for statistical analysis. RESULTS: The scratch assay results revealed that the application of 0.4-T SMF enhanced DPSCs migration towards the scratch wound (P < 0.05). The cytoskeletons of the SMF-treated DPSCs were found to be aligned perpendicular to the scratch wound. After 20 days of culture, the SMF-treated group had a greater number of out-grown cells than the sham-exposed group (nonmagnetized control). For the SMF-treated group, the DMP-1 (P < 0.05) and DSPP genes (P < 0.05), analysed by qRT-PCR, exhibited a higher expression. The distribution of calcified nodules was also found to be denser in the SMF-treated group when stained with Alizarin Red S dye (P < 0.05). Given the incorporation of p38 inhibitor SB203580 into the DPSCs, cell migration and dentinogenesis were suppressed. No difference was found between the SMF-treated and sham-exposed cells (P > 0.05). CONCLUSION: 0.4-T SMF enhanced DPSC migration and dentinogenesis through the activation of the p38 MAPK-related pathway.

Expression and localization of CRAMP in rat tooth germ and during reparative dentin formation.

OBJECTIVES: Cathelicidin-related antimicrobial peptide (CRAMP) is an antimicrobial peptide in mice and rats homologous to LL-37 in humans. In addition to its antibacterial activity, CRAMP has various physiological functions by binding to formyl peptide receptor 2 (FPR2). However, the role of these peptides in teeth is unknown. Therefore, we investigated the role of CRAMP and FPR2 in tooth development, reparative dentin formation, and defense response. MATERIAL AND METHODS: First, we examined the localization of CRAMP and FPR2 during tooth development by immunohistochemical analysis. Next, we investigated the localization of CRAMP, FPR2, and CD68-positive macrophages by immunohistochemical analysis during pulp inflammation and reparative dentin formation after cavity preparation. Finally, we analyzed the effect of lipopolysaccharide (LPS) on the expression of CRAMP and FPR2 in dental pulp cells by real-time reverse transcription PCR. RESULTS: At the late bell stage in tooth development, CRAMP was detected in odontoblasts, and FPR2 was observed in the sub-odontoblastic layer. In mature teeth, CRAMP was not detected, but FPR2 continued to be localized in the sub-odontoblastic layer. After cavity preparation, CRAMP-positive cells and macrophages were found in dental pulp tissues below the cavity at an early stage of repair. At subsequent stages of reparative dentin formation, CRAMP was observed in odontoblast-like cells that contacted reparative dentin. FPR2 immunoreactivity was also detected in odontoblast-like cells and neighboring cells. LPS stimulated the expression of CRAMP mRNA in dental pulp cells in vitro. CONCLUSIONS: Localization of CRAMP and its receptor FPR2-positive cells were observed during physiological and reparative dentin formation. CLINICAL RELEVANCE: CRAMP/LL-37 has a possibility that induce reparative dentin formation.

GaAlAs laser-induced pulp mineralization involves dentin matrix protein 1 and osteopontin expression.

OBJECTIVES: GaAlAs lasers induce pulp mineralization by promoting reparative dentinogenesis. This study analyzed the expression of dentin matrix protein 1 (DMP1) and osteopontin in GaAlAs laser-irradiated rat molars, to examine the hypothesis that these proteins play a role in the laser-induced reparative dentinogenic process. MATERIALS AND METHODS: The mesial surfaces of the upper first molars of 8-week-old Wistar rats were irradiated with a pulsed GaAlAs laser. After 1-14 days, mRNA expression of DMP1 and osteopontin in the coronal pulp was analyzed using real-time PCR. DMP1, osteopontin, and heat shock protein 25 (HSP25) were immunolocalized at 1-21 days. RESULTS: The pulp exhibited a degenerative zone in its mesial portion on days 1-3, and progressive formation of reparative dentin lined with HSP25-immunoreactive odontoblast-like cells, from day 7 onwards. DMP1 and osteopontin mRNA expression were significantly upregulated on days 1-7 and 3-7, respectively. From day 7 onwards, DMP1 and osteopontin immunoreactivity colocalized along the boundary between the primary and reparative dentin. CONCLUSION: GaAlAs laser irradiation of rat molars induced upregulated DMP1 and osteopontin mRNA expression in the coronal pulp, followed by the formation of reparative dentin and the colocalization of DMP1 and osteopontin immunoreactivity at the site at which this tissue first appeared.

Induction of reparative dentin formation on exposed dental pulp by dentin phosphophoryn/collagen composite.

The ultimate goal of vital pulp therapy is to regenerate rapidly dentin possessing an excellent quality using a biocompatible, bioactive agent. Dentin phosphophoryn (DPP), the most abundant noncollagenous polyanionic protein in dentin, cross-linked to atelocollagen fibrils was applied to direct pulp capping in rats. After 1, 2, and 3 weeks, the teeth applied were examined on the induction of reparative dentin formation and the response of pulp tissue, compared to calcium hydroxide-based agent conventionally used. The reparative dentin formation induced by DPP/collagen composite was more rapid than by calcium hydroxide. In the morphometrical analysis, the formation rate of reparative dentin by DPP/collagen composite was approximately the same as that by calcium hydroxide at 3 weeks. Nevertheless, the compactness of reparative dentin formed by DPP/collagen composite was much superior to what resulted from calcium hydroxide. Also, DPP/collagen composite showed high covering ability of exposed pulp. Moreover, DPP/collagen composite led only to slight pulp inflammation at the beginning whereas calcium hydroxide formed necrotic layer adjacent to the material and induced severe inflammation in pulp tissue at 1 week. The present study demonstrates a potential for DPP/collagen composite as a rapid biocompatible inducer for the formation of reparative dentin of excellent quality in rats.

Effects of industrial noise on circumpulpar dentin--a field emission scanning electron microscopy and energy dispersive spectroscopy analysis.

Chronic exposure to Industrial Noise (IN), rich in Low Frequency Noise (LFN), causes systemic fibrotic transformation and sustained stress. Dental wear, significantly increased with exposure to LFN, affects the teeth particularly through the circumpulpar dentin. Our goal is to understand the consequences of IN exposure on the circumpulpar dentin of Wistar rats. 10 Wistar rats were exposed to IN for 4 months, according to an occupationally simulated time schedule and 10 animals were used as age-matched controls. The first and the second upper and lower molars of each animal were processed for observation by Field Emission Scanning Electron Microscopy (FESEM) and Energy Dispersive Spectroscopy (EDS) analysis was performed. In exposed animals FESEM showed a 2.0 to 6.0 µm-dense mineral band between dentin and the pulp with no regular continuity with the tubules. This structure had a few tubules where the odontoblasts processes could be observed embedded within the band and collagen fibers were trapped inside. EDS analysis revealed that it was hydroxyapatite similar to dentin, with a higher carbon content. FESEM results show that the band may be tertiary reparative dentin formed by odontoblast-like cells, but the increased amount of carbon (EDS) could mean that it is sclerotic dentin. IN should be acknowledge as a strong stimulus, able to cause an injury to odontoblasts and to the formation of reparative tertiary dentin, in a process that may accelerate the aging of the teeth, either by direct impact of acoustic pressure pulsations or by increased stress and dental wear.

Biodentine used as a pulp-capping agent in primary pig teeth.

PURPOSE: The purpose of this study was to assess and compare, in primary pig teeth, the pulp response after a pulpotomy using either Biodentine (a new tricalcium-silicate cement), white mineral trioxide aggregate (WMTA), or formocresol (FC) and repeat the same after direct pulp capping using either Biodentine, WMTA, or calcium hydroxide. METHODS: A total of 180 primary teeth from 9 healthy 4-month-old female pigs were divided into 3 experimental periods (7, 28, and 90 days) for each material used for the pulpotomy and direct pulp capping treatments. Seven, 28, and 90 days later, the animals were euthanized and the specimens were prepared for histological examination and evaluation. The data collected from the histological examinations were statistically analyzed using Kruskal-Wallis and Dunn multiple comparison tests. RESULTS: In pulpotomy groups, there was a significant difference between Biodentine and WMTA vs FC in terms of inflammatory cell response and hard tissue formation. In direct pulp capping groups, there was only a significant difference between Biodentine and calcium hydroxide in terms of hard tissue formation in a 7-day period. CONCLUSIONS: Biodentine and white mineral trioxide aggregate are both suitable, biocompatible materials for pulp capping in primary teeth of pigs.

The levels of vascular endothelial growth factor and bone morphogenetic protein 2 in dental pulp tissue of healthy and diabetic patients.

INTRODUCTION: Vascular endothelial growth factor (VEGF) and bone morphogenetic protein 2 (BMP 2) are growth factors (GFs) identified within the dentine-pulp complex and involved into the cellular events connected to the pulp-healing response. It is well established that the expression of these GFs is increased in different tissues in diabetes mellitus. Because there are no data concerning the levels of VEGF and BMP 2 in human dental pulp, the aim of present study was to quantify VEGF and BMP 2 levels in intact dental pulp and dental pulp that underwent reactive dentinogenesis in healthy and diabetic human subjects. METHODS: The study was conducted on 28 healthy and 28 subjects with controlled diabetes type II who underwent pulp extirpation as a part of prosthetic rehabilitation. Pulp were collected from intact teeth and teeth treated by indirect pulp capping. The levels of VEGF and BMP 2 were determined in the pulp tissue lysates with enzyme-linked immunosorbent assay. RESULTS: The levels of VEGF and BMP 2 were significantly higher in intact teeth pulp of diabetic than in healthy subjects. The concentrations of these GFs were significantly lowered in teeth with indirect pulp capping both in healthy and diabetic persons. Furthermore, VEGF and BMP 2 levels were in strong positive correlation. CONCLUSIONS: Similar changes in the levels of VEGF and BMP 2 in intact and treated teeth of healthy and diabetic patients could be suggestive of associated roles of these GFs in responses of healthy and diabetic dental pulp.

Changes in proliferation and osteogenic differentiation of stem cells from deep caries in vitro.

INTRODUCTION: It is suggested that dental pulp stem cells are involved in tooth regeneration and play an important role in maintaining pulp homeostasis. Previously, normal dental pulps were more widely used for experimental models than carious dental pulps. The aim of this study was to isolate and culture the dental pulp stem cells from carious and normal teeth and to evaluate stem cell parameters. METHODS: Pulp tissues were obtained and dissociated from normal and carious teeth. Single-cell suspensions were seeded into 6-well plates and purified by collecting multiple colonies. Normal dental pulp stem cells (DPSCs) and carious dental pulp stem cells (CDPSCs) were compared for morphologic appearance and for their capacity to differentiate into 3 lineages. Colony-forming and MTT assays, cell cycle analysis, gene expression, and alkaline phosphatase activity were also evaluated. RESULTS: Stem cells were cultured successfully from normal and carious dental pulps. CDPSCs displayed increased proliferation ability compared with DPSCs. CDPSCs also showed enhanced ALP activity, mineralization ability, and expression of osteogenesis/dentinogenesis-related genes. All cultures differentiated into 3 cell types. CONCLUSIONS: Our data suggest that caries as a local microenvironment should be taken into account when DPSCs are intended to be used for investigations and application. Furthermore, the mechanism of the underlying changes in cell properties requires further study.

Quantitative determination of high-temperature requirement protein A1 and its possible associated molecules during induced reparative dentin formation.

INTRODUCTION: The equilibrium of inhibitors and inducers plays an important role in the mineralization of dentin during dentinogenesis. High-temperature requirement protein A1 (HtrA1) is a novel bone mineralization inhibitor involved in physiological and pathological bone formation. However, the expression of HtrA1 in tooth mineralization is unknown. The purpose of the present study was to investigate the localization and quantity of HtrA1 and its possible related molecules during induced reparative dentin formation. METHODS: Rats were randomly sacrificed after direct pulp capping on days 0, 7, 14, and 21. Maxillary segments were obtained and routinely prepared for histological analysis, immunohistochemistry, quantum dots-based double immunofluorescence, and CRi's Nuance imaging system (CRI, Woburn, MA)-based quantitative determination. RESULTS: The square measure values of reparative dentin significantly increased on day 7 and continued to increase until day 21. HtrA1, matrix Gla protein (MGP), nestin, and bone sialoprotein were positively stained and colocalized in the odontoblasts and/or odontoblast-like cells zone and reparative dentin during induced reparative dentin formation. The expressions of HtrA1 and MGP were significantly enhanced after direct pulp capping on day 7 and did not significantly change between days 7, 14, and day 21. Both expressions of HtrA1 and MGP were positively correlated with the square measure values of reparative dentin; however, no correlation was found between the expressions of HtrA1 and MGP. CONCLUSIONS: HtrA1 could be observed and might possibly be involved in the process of reparative dentin formation associated with MGP.

Role of CTGF/CCN2 in reparative dentinogenesis in human dental pulp.

Connective tissue growth factor/CCN family 2 (CTGF/CCN2) has been considered to participate in tooth development. To date, the expression and role of CTGF/CCN2 in reparative dentinogenesis have been unclear. Our previous study revealed that matrix metalloproteinase-3 (MMP-3) stimulates cell migration via CTGF/CCN2 expression and secretion in human dental pulp cells, and that this is dependent on dynamin-related endocytosis and independent of protease activity. The objective of the present study was to determine the expression of CTGF/CCN2 in reparative dentin in human carious teeth and to examine the effect of CTGF/CCN2 on mineralization in cultured human dental pulp cells. Minimal expression of CTGF/CCN2 was evident in odontoblasts subjacent to the dentin-pulp junction in healthy teeth, whereas strong expression was detected in odontoblast-like cells lining the reparative dentin subjacent to dental caries. In human dental pulp cells, CTGF/CCN2 promoted mineralization but failed to induce proliferation, suggesting that this molecule has the ability to induce the differentiation of human dental pulp cells. Taken together, the data suggest that CTGF/CCN2 is likely involved in reparative dentinogenesis through formation of hard tissue in human carious teeth.

Immunohistochemical analysis of pulpal regeneration by nestin expression in replanted teeth.

AIM: To investigate dental pulp healing after tooth replantation in rats using nestin as an odontoblastic marker for immunohistochemical analysis. METHODOLOGY: Twenty-five maxillary right first molars from 25 female Sprague-Dawley rats, aged 4 weeks post-natally, were extracted and immediately repositioned in the original socket within 5 s. Five rats each were later killed on days 3, 5 and weeks 1, 2 and 4. The maxillae were removed en bloc and the tissue samples containing the maxillary right first molars were decalcified, sectioned, mounted and stained with anti-nestin antibody to be observed under a light microscope. RESULTS: At 3 days after replantation, there was a localized inflammatory reaction, but pulp revascularization and healing had begun in the root area. At 5 days after replantation, odontoblast-like cells were observed. Reparative dentine deposition was observed beneath the pulp-dentine border from 1 week after replantation, and gradually increased until 2 weeks after replantation. The presence of odontoblast-like cells and the formation of reparative dentine continued from the first week throughout the experimental period. At week four, deposition of osteodentine and cementum-like tissues were observed. CONCLUSIONS: Pulpal mineralization after replantation initially occurred via the deposition of reparative dentine, followed by the deposition of osteodentine and cementum-like tissues in rat teeth.

Evaluation of mineral trioxide aggregate (MTA) versus calcium hydroxide cement (Dycal(®) ) in the formation of a dentine bridge: a randomised controlled trial.

AIM: To assess the effectiveness of mineral trioxide aggregate (MTA) used as an indirect pulp-capping material in human molar and premolar teeth. METHODOLOGY: We conducted a clinical evaluation of 60 teeth, which underwent an indirect pulp-capping procedure with either MTA or calcium hydroxide cement (Dycal(®) ). Calcium hydroxide was compared with MTA and the thickness of the newly formed dentine was measured at regular time intervals. The follow-up was at 3 and 6 months, and dentine formation was monitored by radiological measurements on digitised images using Mesurim Pro(®) software. RESULTS: At 3 months, the clinical success rates of MTA and calcium hydroxide were 93% and 73%, respectively (P = 0.02). At 6 months, the success rate was 89.6% with MTA, and remained steady at 73% with calcium hydroxide (P = 0.63). The mean initial residual dentine thickness was 0.23 mm, and increased by 0.121 mm with MTA and by 0.136 mm with calcium hydroxide at 3 months. At 6 months, there was an increase of 0.235 mm with MTA and of 0.221 mm with calcium hydroxide. CONCLUSIONS: A higher success rate was observed in the MTA group relative to the Dycal(®) group after 3 months, which was statistically significant. After 6 months, no statistically significant difference was found in the dentine thickness between the two groups. Additional histological investigations are needed to support these findings.

Odontoblast-targeted Bcl-2 overexpression promotes dentine damage repair.

OBJECTIVE: Bcl-2 is widely expressed in a developing tooth organ and regulates tooth morphogenesis. However, whether Bcl-2 is related to tooth damage repair is unknown yet. Using an odontoblast-targeted Bcl-2 overexpression transgenic mouse (Col2.3Bcl-2) and artificial cavity preparation as a model system, the relationship between Bcl-2 and reparative dentinogenesis is investigated in this study. METHODS: The odontoblastic-like cell cultures derived from mouse molar pulps were established. The expression of transgenic human Bcl-2 (hBcl-2) and endogenous mouse Bcl-2 (mBcl-2) and mouse Bax (mBax, a Bcl-2 antagonist) was detected in vivo and in vitro by Western blot and immunocytochemistry, respectively. Basal level and artificial cavity-induced odontoblast apoptosis was detected by the Deoxynucleotidyl Transferase (TdT) dUTP Nick End labelling (TUNEL) technique. Reparative dentine formation induced by artificial cavity drilled to a half dentine thickness on mesial cervical region of mandibular first molars 2, 4, and 6 weeks post-op was evaluated histologically and via micro-CT. RESULTS: The transgenic hBcl-2 was stably expressed in odontoblasts of the transgenic animals without interference with the expression of mBcl-2 and mBax. Basal level as well as artificial cavity- induced odontoblast apoptosis was prevented by the transgene. Compared to the wild type, the transgenic animals produced reparative dentine with significantly higher mineral density 6 weeks after the operation. CONCLUSIONS: Bcl-2 overexpression prevents odontoblast apoptosis and promotes dentine damage repair, indicating that genetic manipulation of Bcl-2 may be a novel strategy to maintain the vitality and function of dentine-pulp complex under detrimental mechanical stimuli.

Evaluation of dentin formed in autogenous tooth transplantation in the dog: a comparison between one- and two-stage surgical techniques.

This study was designed to compare the thickness of dentin formed associated with autogenous tooth transplantation in dogs, using either one- or two-stage surgical techniques. The study consisted of three Beagles, older than 5 months, in which six incisors and six premolars were transplanted to mechanically prepared recipient sockets. One group was transplanted using a one-stage method to recipient beds prepared immediately before transplantation. The second groups of teeth were transplanted using a two-stage method in which the recipient beds were prepared and left to heal for 7 days before transplantation. Dogs were injected with xylenol orange, calcein and oxytetracycline at 2 days before, 3 and 9 weeks after transplantation, respectively, for vital staining. Clinical examinations were carried out every week, and the animals were euthanized 9 weeks later. The jaws were resected, fixed in formaldehyde and embedded in resin. Undemineralized sections were cut and examined by fluorescent microscopy. The thickness of dentin formed in the third week after transplantation and 9 weeks was evaluated by undertaking histomorphometric analysis and analysed using the Mann-Whitney U test (P = 0.05). All the transplanted teeth in both groups survived, and the dentin was formed. No statistically significant difference was found in the thickness of dentin formed in the third week and formed in the third to ninth week between the treatment groups (P = 0.999 and P = 0.998, respectively). This study demonstrated that there was no difference between the two surgical techniques in terms of the thickness of dentin formed in transplanted teeth.

Influence of etching ability of one-step self-etch adhesives on bonding to sound and non-carious cervical sclerotic dentin.

The purpose of this study was to evaluate influence of etching ability of one-step self-etch adhesives on bonding to non-carious cervical sclerotic lesion. Forty-eight human maxillary premolars with and without natural non-carious cervical sclerotic lesions at buccal surfaces were used in this study. Artificial wedge-shaped lesions were prepared in sound premolars. All lesions were bonded with Bond Force (BF; pH=2.3) or G-Bond Plus (GBA; pH=1.5) according to the manufacturers' instructions. After water storage for 24 hours at 37°C, bonded specimens were subjected to microtensile bond test to measure bond strength to the gingival wall in each lesion. The µTBS of BF to the natural lesion was significantly lower than that of artificial lesion (p<0.05), while for GBA, there is no significant difference (p>0.05). There was no difference in µTBS to artificial lesion between these two adhesives, however in case of non-carious cervical sclerotic lesion, the µTBS of GBA was statistically higher than that of BF.

Histologic response and tenascin and fibronectin expression after pulp capping in pig primary teeth with mineral trioxide aggregate or calcium hydroxide.

This study evaluated the histological response and the expression of tenascin (TN) and fibronectin (FN) after pulp capping with mineral trioxide aggregate (MTA) or calcium hydroxide (CH). Class V cavities and pulp exposure were performed in 40 primary pig teeth. The pulps were capped with either MTA or CH, and the cavities were sealed with resin-modified glass ionomer cement. CH was used as a control. Seven and 70 days posttreatment, the animals were sacrificed and teeth were prepared for histological evaluation. TN and FN were detected by immunostaining. A severe inflammatory response was observed after 7 days in the CH group (p<0.043), while in the MTA group, a mild response was observed. Similar reparative dentin deposition was observed after 70 days for both groups (p<0.005). The expression of FN and TN was similar for both groups in the two periods evaluated. TN and FN were expressed during pulp reparative events, independently of the capping material.

GaAlAs laser irradiation induces active tertiary dentin formation after pulpal apoptosis and cell proliferation in rat molars.

INTRODUCTION: This study aimed to clarify pulpal responses to gallium-aluminum-arsenide (GaAlAs) laser irradiation. METHODS: Maxillary first molars of 8-week-old rats were irradiated at an output power of 0.5 or 1.5 W for 180 seconds, and the samples were collected at intervals of 0 to 14 days. The demineralized paraffin sections were processed for immunohistochemistry for heat-shock protein (HSP)-25 and nestin in addition to cell proliferation assay using bromodeoxyuridine (BrdU) labeling and apoptosis assay using deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL). RESULTS: Intense HSP-25 and nestin immunoreactivities in the odontoblast layer were weakened immediately after 0.5-W irradiation and recovered on day 1, resulting in slight tertiary dentin formation by day 14. On the contrary, 1.5-W irradiation immediately induced the loss of HSP-25 and nestin-immunoreactivities in the odontoblast layer. On day 1, numerous TUNEL-positive cells appeared in a degenerative zone that was surrounded by intense HSP-25 immunoreactivity. BrdU-positive cells occurred within the intensely HSP-25-immunopositive areas during days 2 through 5, whereas TUNEL-positive cells gradually decreased in number by day 5. HSP-25- and nestin-positive odontoblast-like cells were arranged along the pulp-dentin border by day 7, resulting in remarkable tertiary dentin formation on day 14. CONCLUSIONS: The output energy determined pulpal healing patterns after GaAlAs laser irradiation; the higher energy induced the apoptosis in the affected dental pulp including odontoblasts followed by active cell proliferation in the intense HSP-25-immunoreactive areas surrounding the degenerative tissue, resulting in abundant tertiary dentin formation. Thus, the optimal GaAlAs laser irradiation elicited intentional tertiary dentin formation in the dental pulp.

Biomimetic approach to perforation repair using dental pulp stem cells and dentin matrix protein 1.

INTRODUCTION: Dentin regeneration could be an ideal treatment option to restore tissue function. This study was conducted to evaluate the ability of dental pulp stem cells (DPSCs) and dentin matrix protein 1 (DMP1) impregnated within a collagen scaffold to regenerate dentin. METHODS: Simulated perforations were created in 18 dentin wafers made from freshly extracted human molars. Six groups were established. They were (1) empty wafers, (2) mineral trioxide aggregate, (3) collagen scaffold, (4) scaffold with DMP1, (5) scaffold with DPSCs, and (6) scaffold with DPSCs and DMP1. One sample was placed subcutaneously in each mouse with three mice in each group. After 12 weeks, the samples were subjected to radiographic, histological, and immunohistochemical evaluations. RESULTS: DPSCs impregnated within a collagen scaffold differentiated into odontoblast-like cells forming a highly cellular, vascular, and mineralized matrix in the presence of DMP1. CONCLUSIONS: A triad consisting of DPSCs, DMP1, and a collagen scaffold promotes dentin regeneration in a simulated perforation repair model.