Allogeneic Bone Marrow Mesenchymal Stromal Cell Transplantation Induces Dentin Pulp Complex-like Formation in Immature Teeth with Pulp Necrosis and Apical Periodontitis.

INTRODUCTION: Dental pulp regeneration is challenging in endodontics. Cellular therapy is an alternative approach to induce dental pulp regeneration. Mesenchymal stromal cells (MSCs) have the capacity to induce dental pulp-like tissue formation. In this study, we evaluated the capacity of allogeneic bone marrow MSCs (BM-MSCs) to regenerate pulp following necrosis and apical periodontitis in children's permanent immature apex teeth. METHODS: Patients aged 8 to 12 years with pulp necrosis and apical periodontitis were evaluated. The study included 15 teeth (13 incisors and 2 molars) from 14 patients (8 boys and 6 girls). Radiographic evaluation showed periapical radiolucency and immature apex teeth. There was no response to cold or electric pulp testing. The root canal of each tooth was cleaned, shaped, and Ca(OH)2 used as an interappointment medication. Cryopreserved allogeneic BM-MSCs were thawed, expanded, incorporated into preclotted platelet-rich plasma, and implanted into the tooth's pulp cavity. They were sealed with bioceramic cement and composite. Sensibility, apical foramen, calcium deposits within the root canal, and resolution of periapical lesions were evaluated in each tooth over the following 12 months. RESULTS: Based on 9 variables established for dental pulp-like tissue regeneration, all MSC-treated teeth showed evidence of successful regeneration. Clinical and radiographic evaluation of the treated teeth showed periapical lesion healing, sensitivity to cold and electricity, decreased width of the apical foramen, and mineralization within the canal space. CONCLUSIONS: Transplantation of allogeneic MSCs induces the formation of dental pulp-like tissue in permanent immature apex teeth with pulp necrosis and apical periodontitis. Implant of MSCs constitutes a potential therapy in regenerative endodontics in pediatric dentistry. Future studies incorporating a larger sample size may confirm these results.

[Recognition on dentin dysplasia type â ¡].

Dentin dysplasia type Ⅱ (DD-Ⅱ) is a subtype of hereditary dentin disorders. The dentin sialophosphoprotein (DSPP) gene has been revealed to be the causative gene, whose mutations could affect the normal tooth development process. The lesions involve both deciduous and permanent dentition, mainly manifested as tooth discoloration, attrition and even the subsequent malocclusion. If not treated in time, it will significantly affect the physical and psychological health of patients. The disease is difficult to be diagnosed in clinic accurately as its low incidence and hidden manifestations. The present article aims to discuss the clinical and radiographic characteristics, diagnosis, treatment of DD-Ⅱ, in order to improve the overall understanding on DD-Ⅱ for clinicians.

Iron level participates in the pathological damages of dental caries in infant rats by affecting enamel mineralization.

Iron deficiency anemia (IDA) is a common nutritional disease associated with early childhood caries. This study aimed to explore the role of iron levels in pathological changes of dental caries in childhood. Rats were divided into four groups based on their iron content: IDA, positive control (PC), high iron (HI), and negative control (NC). Except for the rats in the NC group, rats in the other groups were inoculated with Streptococcus mutans and fed cariogenic high-sugar fodder to induce caries. Three months later, the caries status of the molars was evaluated at both the smooth and sulcal surfaces according to Keyes scores. Scanning electron microscopy (SEM) was performed to reveal microstructural changes in caries. Energy-dispersive spectroscopy (EDS) was used to determine the elemental composition of the enamel and dentin. In addition, the histopathology of the salivary gland was detected using hematoxylin and eosin (HE) staining.The results showed that rats in the PC group exhibited obvious carious lesions. The carious score was significantly higher in the IDA group than in the PC group but was lower in the HI group. SEM revealed complete destruction of the enamel and damage to the middle dentin in the IDA group. In contrast, the molars in the HI group exhibited some degree of enamel demineralization, but the underlying dentin was almost intact. In addition, the elemental compositions of the enamel and dentin were similar among the four groups, and iron was detected only in the HI group. No differences were observed in the morphological structures of the salivary glands of rats from the different groups. In conclusion, ID enhanced the pathological damage of caries, whereas HI weakened it. Iron may participate in the pathological damage caused by childhood caries by affecting enamel mineralization.

Identification of DSPP novel variants and phenotype analysis in dentinogenesis dysplasia Shields type II patients.

OBJECTIVES: To investigate the genetic causes and teeth characteristics of dentin dysplasia Shields type II(DD-II) in three Chinese families. MATERIALS AND METHODS: Data from three Chinese families affected with DD-II were collected. Whole-exome sequencing (WES) and whole-genome sequencing (WGS) were conducted to screen for variations, and Sanger sequencing was used to verify mutation sites. The physical and chemical characteristics of the affected teeth including tooth structure, hardness, mineral content, and ultrastructure were investigated. RESULTS: A novel frameshift deletion mutation c.1871_1874del(p.Ser624fs) in DSPP was found in families A and B, while no pathogenic mutation was found in family C. The affected teeth's pulp cavities were obliterated, and the root canals were smaller than normal teeth and irregularly distributed comprising a network. The patients' teeth also had reduced dentin hardness and highly irregular dentinal tubules. The Mg content of the teeth was significantly lower than that of the controls, but the Na content was obviously higher than that of the controls. CONCLUSIONS: A novel frameshift deletion mutation, c.1871_1874del (p.Ser624fs), in the DPP region of the DSPP gene causes DD-II. The DD-II teeth demonstrated compromised mechanical properties and changed ultrastructure, suggesting an impaired function of DPP. Our findings expand the mutational spectrum of the DSPP gene and strengthen the understanding of clinical phenotypes related to the frameshift deletion in the DPP region of the DSPP gene. CLINICAL RELEVANCE: A DSPP mutation can alter the characteristics of the affected teeth, including tooth structure, hardness, mineral content, and ultrastructure.

[Mutation of dentin sialophosphoprotein and hereditary malformations of dentin].

The classification as well as the clinical manifestations of hereditary malformations of dentin are of great concern and have been deeply elucidated. The understanding of its genetic basis also increases progressively. Dentin sialophosphoprotein (DSPP) is the pathogenic gene of dentinogenesis imperfecta type Ⅱ, dentinogenesis imperfecta type Ⅲ and dentin dysplasia type Ⅱ. In this article, the classification of DSPP mutations as well as the resultant dysfunction of the mutant DSPP are summarized respectively and the corresponding clinical manifestations are analyzed. This work will provide a reference for the diagnosis and treatment of hereditary malformations of dentin.

Novel strategy for dental caries by physiologic dentin regeneration with CPNE7 peptide.

OBJECTIVE: CPNE7-derived functional peptide (CPNE7-DP) has been introduced as a bioactive therapeutics for dentin diseases. CPNE7-DP regenerates tubular dentin on the pulpal side and occlude dentinal tubules. CPNE7-DP was capable to treat dentin hypersensitivity typically associated with dentinal wear at the neck of the tooth. However, the role of CPNE7-DP in another common dentin disease, dental caries, remains uninvestigated. In this study, we evaluated the potential application of CPNE7-DP in dentin caries using an experimental dentin caries model in rats. DESIGN: The stability of CPNE7-DP in caries-like environments including pathologic bacteria of caries or low pH was tested. We established a nutrition-time/hyposalivation-based dental caries rat model by inoculating caries-inducing bacteria and diet for sufficient time. Glycopyrrolate has been treated to induce reversible hyposalivation for accelerating caries progression. Then the tubular dentin regeneration was investigated with histologic methods. Also, modulation of inflammation or autophagy by CPNE7-DP was investigated with marker gene expression in human dental pulp cells (hDPCs) and immunohistochemistry. RESULTS: CPNE7-DP was stable with caries-inducing bacteria and low pH. Establishment of dentin caries was confirmed with radiographic and histologic evaluation. CPNE7-DP regenerated a substantial amount of tubular tertiary dentin and alleviated the pulp inflammation of dentin caries. Under inflammatory conditions, CPNE7-DP reduced the expression of inflammatory cytokines. These phenomena could be the consequence of the modulation of autophagy by CPNE7-DP, which reactivates inflamed odontoblasts. CONCLUSIONS: Overall, CPNE7-DP, which repairs caries through physiological dentin regeneration, might help overcoming the limitations of current restorative caries treatments.

Monitoring silver diamine fluoride application with optical coherence tomography and thermal imaging: An in vitro proof of concept study.

OBJECTIVES: The purpose of this study was to show that optical coherence tomography (OCT) and thermal imaging can be used to monitor changes in the structure and activity of caries lesions over time after treatment with silver diamine fluoride (SDF). METHODS: Artificial caries lesions were formed on enamel and dentin bovine blocks. Each block was partitioned into five windows with the central three windows exposed to a demineralization solution to create lesions: one sound window served as a sound control (SC), one sound window was exposed to SDF to serve as a test control (SCT), one lesion window served as a lesion control (LC), one lesion window received one application of SDF (L1), while the other lesion window received two applications of SDF (L2). Each window was scanned using OCT before SDF application, and every week subsequently, for 12 weeks after initial SDF treatment. Changes in the mean intensity and the width of the peak of increased reflectivity due to the lesion and SDF along with the intensity at a depth of 180 µm from the surface representing optical penetration through the lesion were monitored. Changes in the heat lost, ΔQ (temperature integrated over time) of each window during drying with air were also monitored using a thermal imaging camera. Transverse microradiography (TMR), and high-resolution microscopy were also used for the analysis of selected samples. RESULTS: The reflectivity and optical penetration of sound and lesion areas of enamel and dentin manifested significant changes in OCT images after SDF application. Thermal imaging showed significant differences in ΔQ indicative of permeability changes in the sound and lesion areas of enamel and dentin after SDF application.

Protective effect of titanium tetrafluoride and silver diamine fluoride on radiation-induced dentin caries in vitro.

This in vitro study evaluated the protective effect of titanium tetrafluoride (TiF4) varnish and silver diamine fluoride (SDF) solution on the radiation-induced dentin caries. Bovine root dentin samples were irradiated (70 Gy) and treated as follows: (6 h): 4% TiF4 varnish; 5.42% NaF varnish; 30% SDF solution; placebo varnish; or untreated (negative control). Microcosm biofilm was produced from human dental biofilm (from patients with head-neck cancer) mixed with McBain saliva for the first 8 h. After 16 h and from day 2 to day 5, McBain saliva (0.2% sucrose) was replaced daily (37 °C, 5% CO2) (biological triplicate). Demineralization was quantified by transverse microradiography (TMR), while biofilm was analyzed by using viability, colony-forming units (CFU) counting and lactic acid production assays. The data were statistically analyzed by ANOVA (p < 0.05). TiF4 and SDF were able to reduce mineral loss compared to placebo and the negative control. TiF4 and SDF significantly reduced the biofilm viability compared to negative control. TiF4 significantly reduced the CFU count of total microorganism, while only SDF affected total streptococci and mutans streptococci counts. The varnishes induced a reduction in lactic acid production compared to the negative control. TiF4 and SDF may be good alternatives to control the development of radiation-induced dentin caries.

Is it necessary to pre-treat Dentine before GIC Restorations? Evidence from an in Vitro Study / É necessário pré tratar a dentina antes das restaurações de CIV? Evidência de um estudo in vitro

ABSTRACT The aim of this study was to assess the influence of differentpretreatmentapproaches on glass ionomer cement (GIC)bond strength (BS) to dentine. Sixty bovine incisors wereallocated into six groups according to substrate - sound orcariesaffecteddentine; and substrate pretreatmentapproach -no conditioning (control), polyacrylic acid for 10 s andphosphoric acid for 7 s. Teeth in the cariesaffecteddentinegroup were previously submitted to cariogenic pHcyclingchallenge. After dentine pretreatment,according to experi mentalgroups, polyethylene tubes were placed on flat dentinesurfaces and filled with GIC. Teeth were stored in distilledwater for 24 h at 37 °C and then submitted to microshear test(0.5 mm/min). Failure pattern analysis was performed understereomicroscope (400x). Data were analysed using twowayANOVA and Tukey's test (α=5%). Statistically significantdifferences were found for the pretreatmentapproach,regardless of substrate (p<0.001). The polyacrylic acid groupand control group had similar BS values, and were both betterthan the phosphoric acid group. In general, GIC had betterbonding performance in sound dentine than in cariesaffecteddentine. In conclusion, dentine pretreatmentwith polyacrylicacid did not improve the performance of GIC restoration onclinically relevant substrates.

RESUMO O objetivo deste estudo foi avaliar a influência de diferentesprétratamentosna resistência de união (RU) de cimentos deionômero de vidro (CIV) a dentina. Sessenta incisivos bovinosforam alocados em 6 grupos de acordo com o substrato -hígido ou cariado; e com a abordagem de prétratamento-sem condicionamento (controle), ácido poliacrílico por 10 s, eácido fosfórico por 7 s. Os dentes pertencentes aos grupos dedentina cariada foram previamente submetidos ao desafiocariogênico por meio da ciclagem de pH. Após o prétratamentoda dentina, de acordo com os grupos experimentais,tubos de polietileno foram colocados sobre superfícies planas dedentina e preenchidos com CIV. Os dentes foram armazenadosem água destilada por 24 h a 37°C e então submetidos ao testede microcisalhamento (0,5 mm/min). A análise do padrão defratura foi realizada em estereomicroscópio (400x). Os dadosobtidos foram analisados usando ANOVA de dois fatores e testede Tukey (α=5%). Diferença estatisticamente significante foiencontrada para as diferentes abordagens de prétratamento,independente do substrato (pp<0,001). Aplicação de ácidopoliacrílico resultou em valores de RU similares aos do grupocontrole. Entretanto, ambos os grupos mostraram um melhordesempenho quando comparado a aplicação de ácidofosfórico. De forma geral, CIV apresentou melho5 desempenhoadesivo em dentina sadia quando comparada a dentinacariada. Em conclusão, o prétratamentoem dentina comácido poliacrílico não melhora o desempenho das restauraçõesde CIV em substratos clinicamente relevantes.

[Gene mutations and disorders of dental hard tissues].

Developmental disorders of dental hard tissues are important components of non-carious diseases, which mainly include amelogenesis imperfecta and hereditary dentin disorders with various subtypes. In the absence of effective intervention, these disorders would lead to tooth sensitivity, defects of tooth structure or even loss of tooth, affecting the masticatory function and facial aesthetic configuration. At present, many dental clinicians may not have sufficient understanding of the diseases, and it is urgent to pay attention to the diseases per se and the patients affected. Based on the summary of the current research progresses, this article focuses on the clinical classification, the disease phenotype and the pathogenesis of gene mutations, in order to provide reference and help for the dental clinicians as well as the patients.

Biodentine Induce Proteínas Apoptóticas Caspasa 3 y PARP-1 en Células de la Pulpa Dental Humana / Biodentine Induces Apoptotic Proteins Caspase 3 and PARP - 1 on Human Dental Pulp Cells

RESUMEN: Las patologías pulpares han sido un verdadero reto para la odontología principalmente por su tratamiento. Actualmente, existen numerosos biomateriales en el mercado que reportan tener propiedades inherentes en los tejidos dentarios. Sin embargo, diferentes estudios sobre múltiples líneas celulares expuestas a estos biomateriales demuestran resultados controversiales como biocompatiblidad y citotoxicidad celular. Biodentine, es un cemento endodóntico en base a silicatos cálcico de múltiples aplicaciones, que prestaría propiedades de biocompatibilidad como bioactividad celular, características que le permitirían incluso ser utilizado en contacto directo con la pulpa dental. El objetivo de este estudio es la evaluación in-vitro de Biodentine, sobre cultivos de células de la pulpa dental humana (CCPDH). Se prepararon discos de cemento de Biodentine™ de 2 x 6 mm, los que se expusieron a cultivos de células aisladas de la pulpa dental humana. Luego de 24, 48 y 72 horas de exposición, se realizaron ensayos de viabilidad celular utilizando el método colorimétrico MTT. También se realizaron ensayos de expresión proteica de dos proteínas involucradas en la vía de señalización de la apoptosis celular: Caspasa - 3 clivada y Poli (ADP-Ribosa) Polimerasa, PARP - 1. Existen diferencias estadísticamente significativas (p<0,05) en los ensayos de viabilidad celular entre las células expuestas a Biodentine y el grupo control, como también a medida que aumenta el tiempo de exposición (p<0,05). Por otra parte, también existen diferencias significativas (p<0,05) en la expresión de PARP- 1 en los grupos sometidos a Biodentine. Los resultados obtenidos en este estudio demuestran que Biodentine genera citotoxicidad celular en cultivos celulares de pulpa dental humana, por disminución de la viabilidad celular como por la expresión de proteínas apoptóticas. Es por esto que la utilización de este biomaterial debería ser estudiado y considerarse en cada caso clínico, especialmente como recubridor pulpar directo.

ABSTRACT: Oral pathologies have been a real challenge for dentistry, mainly due to its treatment. Currently, there are numerous biomaterials on the market that may present inherent properties in dental tissues. However, studies on multiple cell lines are based on biocompatible results such as biocompatibility and cellular cytotoxicity. Biodentine is endodontic cement based on calcium silicates of multiple applications, which would provide biocompatibility properties as cellular bioactivity, characteristics that will allow it to be used in direct contact with the dental pulp. The objective of this study is the in vitro evaluation of Biodentine, on cultures of cells of the human dental pulp (HDPC). Biodentine cement disks of 2 x 6 mm were prepared, and HDPC culture plates were introduced. After 24, 48 and 72 hours of exposure, cell viability tests were performed using the MTT colorimetric method. On the other hand, protein expression assays of two proteins involved in the signaling pathway of cell apoptosis Caspase-3 cleaved (cas-3 clv) and PARP-1 are carried out. There are statistically significant differences (p <0,05) in the cell viability tests between Biodentine and control group, as well as the exposure time increases (p <0,05). Otherwise, there are also significant differences (p <0,05) in the expression of PARP-1 in the groups, sometimes a Biodentine. The results in this study that Biodentine generates a cellular cytotoxicity in HDPC cultures, therefore, cell viability as the expression of apoptotic proteins. This is why the use of this biomaterial should be studied for each particular clinical case, especially as a direct pulp capping agent.

Characteristics and severity of tooth wear in 2 to 5-year-old kindergarten children in Medellin / Facetas de desgaste y características de severidad en niños de 2 a 5 años de un jardín infantil, Medellín

The aim of the present study was to identify the characteristics and severity of tooth wear in children aged 2 to 5 years attending a kindergarten in the city of Medellin. The study population comprised 92 children aged 2 to 5 years attending a kindergarten in Medellin; the final sample included 86 children selected using a non-probabilistic sampling method. All assessments were performed using the index devised by Hansson and Nilner (1989). Prevalence of tooth wear in the studied population was 100%. The observed results confirm previous findings showing that the severity of tooth wear is proportionally related with increase in age, and is mainly due to the physiological wear of teeth. No significant differences in tooth wear facets were observed among the different types of teeth (incisors, canines, molars). It is necessary to develop validated, precise, and comparable indices that allow establishing the etiology and orienting treatment of non-physiological tooth wear.

El objetivo del estudio fue identificar las facetas de desgaste y las características de severidad en niños de 2 a 5 años de un jardín infantil de la ciudad de Medellín. El estudio incluyó 92 niños entre los 2 y 5 años de un jardín infantil de la ciudad de Medellín, con una muestra final de 86 seleccionados a partir de un muestreo no probabilístico. Se utilizó como instrumento el índice propuesto por Hansson y Nilner (1989). El 100% (86) de los niños evaluados presentaron facetas de desgaste. Los resultados encontrados confirman datos previos en cuanto a que las severidades de las facetas de desgaste dentario están relacionadas proporcionalmente con el aumento de edad y se deben principalmente al desgaste fisiológico de los dientes. No se encontraron diferencias significativas en la prevalencia o severidad de desgaste entre los grupos anatómicos dentarios (incisivos, caninos y molares). Es necesario establecer índices validados, precisos y comparables para evaluar la etiología y orientar hacia la solución de las causas no fisiológicas.

In vivo fate of bone marrow mesenchymal stem cells implanted into rat pulpotomized molars.

In our previous work, we established an in vivo coronal pulp regeneration model in which biodegradable hydrogel-made scaffolds carrying rat bone marrow mesenchymal stem cells (BM-MSCs) were implanted in the coronal pulp chamber of pulpotomized rat maxillary first molars. In this study, we investigated the in vivo fate of LacZ-labeled BM-MSCs in our coronal pulp regeneration model. BM-MSCs were nucleofected with pVectOZ-LacZ plasmid encoding ß-galactosidase 1 day before implantation, and the LacZ-transfected BM-MSCs were implanted into the pulpotomized pulp chamber with biodegradable preformed scaffold-hydrogel constructs. Empty vector was used as a control. After 3 and 14 days, the molars were retrieved and subjected to ß-galactosidase staining. At 3 days, ß-galactosidase-expressing cells with a round profile were located mainly around the scaffold. At 14 days, when the pulp-like tissue had been generated, the majority of ß-galactosidase-expressing cells were detected under the newly formed dentin bridge-like structure, where nestin-expressing odontoblast-like cells were arranged. Immunoreactivity for dentin sialoprotein, a marker of mature odontoblasts, was strongly detected under the original dentin. No ß-galactosidase staining was observed in the control group. Thus, we demonstrated that BM-MSCs survived for 2 weeks after implantation and colonized within the site of potential cytodifferentiation. Our findings indicated that BM-MSCs could differentiate into cells involved in mineralized tissue formation in the functionally relevant region.

Shaping ability of BioRace, ProTaper NEXT and Genius nickel-titanium instruments in curved canals of mandibular molars: a MicroCT study.

AIM: To evaluate and compare the canal shaping ability of BioRace, ProTaper NEXT and Genius engine-driven nickel-titanium (NiTi) file systems in extracted mandibular first molars using micro-computed tomography (MCT). METHODOLOGY: Sixty mesial root canals of mandibular first molars were randomly divided into three equal groups, according to the instrument system used for root canal preparation (n = 20): BioRace (BR), ProTaper NEXT (PTN) or Genius (GN). Root canals were prepared to the full WL using a crown-down technique up to size 35, .04 taper instruments for BR and GN groups and size 30, .07 taper instruments for the PTN group. MCT was used to scan the specimens before and after canal instrumentation. Changes in dentine volume, the percentage of uninstrumented canal surface and degree of canal transportation were evaluated in the coronal, middle and apical thirds of canals. Data were analysed statistically using one-way analysis of variance and Tuckey's post hoc tests with the significance level set at 5%. RESULTS: There were no significant differences between the three groups in the terms of dentine removed after preparation and determination of the root canal volume, or percentage of uninstrumented canal surface (P > 0.05). No significant differences were found between the systems for canal transportation in any canal third (P > 0.05). CONCLUSIONS: The shaping ability of the BR, PTN and GN NiTi file systems was equally effective. All instrumentation systems prepared curved root canal systems with no evidence of undesirable changes in 3D parameters or significant shaping errors.

Root Canal Shaping Using Nickel Titanium, M-Wire, and Gold Wire: A Micro-computed Tomographic Comparative Study of One Shape, ProTaper Next, and WaveOne Gold Instruments in Maxillary First Molars.

INTRODUCTION: The aim of this study was to evaluate the root canal shaping effect of instruments manufactured from nickel titanium, M-Wire, and Gold wire with different glide path preparation techniques. METHODS: One hundred thirty-five mesiobuccal canals of extracted human maxillary molars were randomly divided into 3 equal groups (N = 45) for glide path preparation with K-files (KF) (Dentsply Sirona, Ballaigues, Switzerland), One G (OG) files (Micro-Mega, Besançon, France), and ProGlider (PG) files (Dentsply Sirona). Specimens of each glide path group were further divided equally into 3 groups for instrumentation with ProTaper Next (PTN, Dentsply Sirona), One Shape (OS, Micro-Mega), and WaveOne Gold (WOG, Dentsply Sirona) systems (n = 15). Micro-computed tomographic imaging was used to scan teeth before instrumentation and after shaping to compare centering ratio and canal transportation values at the apical, midroot, and coronal levels and the overall changes in canal volume. Data sets were statistically analyzed (analysis of variance and Kruskal-Wallis H tests). RESULTS: The centering ratios for all groups were statistically similar at all levels. Apical canal transportation was significantly high for K/OS and K/PTN (P = .003). Midroot canal transportation was significantly high for K/PTN, K/OS, and OG/OS (P = .0003). Coronal canal transportation was significantly high for K/PTN and K/OS (P = .011). The highest change in canal volume was observed with all PTN groups and the lowest with PG/WOG (P = .06). CONCLUSIONS: WOG manufactured from Gold wire combined with PG showed better root canal shaping ability and removed less dentin from the canal walls. The nickel-titanium (OS) and M-Wire (PTN) instruments used in combination with KF significantly transported more canals. PTN removed the most dentin from the canal walls regardless of the GPP technique.

Ideal Concentration of Advanced-Platelet Rich Fibrin (A-PRF) Conditioned Media for Human Dental Pulp Stem Cells Differentiation

Abstract Objective: To discover the ideal concentration of Advanced Platelet Rich Fibrin (A-PRF) as modification of PRF, for human Dental Pulp Stem Cells (hDPSCs) differentiation. Material and Methods: hDPSCs were devided into five experimental groups: Group I (control group) consist of hDPSCs cultured in 10% FBS, Group II consist of hDPSCs cultured in 1% A-PRF, Group III consist of hDPSCs cultured in 5% A-PRF, Group IV consist of hDPSCs cultured in 10% A-PRF and Group V consist of hDPSCs cultured in 25% A-APRF. All group have been observed for 7 and 14 days and each group had three biological replicates (triplo). Formation of the mineralized nodules was detected after 7 days by Alizarin red-based assay and Dentin Sialophosphoprotein (DSPP) expression after 7 and 14 days quantified by ELISA reader. Statistical analysis was proven with Kruskal-Wallis and post hoc Mann-Whitney test Results: The differentiation of hDPSCs in all A-PRF groups was significantly different on day-7 (p<0.05) compare to control group (Group I). There were no significant differences between all groups on day-14 (p>0.05). Significantly differences between Group II (1% A-PRF) and Group I (control), Group II (1% A-PRF) and Group III (5% A-PRF), also Group II (1% A-PRF) and Group V (25% A-PRF) was found from post hoc test analysis Conclusion: The ideal conditioned media concentration for differentiation of human dental pulp stem cells was on 1% up to 5% A-PRF group.

Microdontia after chemotherapy in a patient treated for neuroblastoma: Histopathological findings.

Microdontia is one of the late effects of antineoplastic therapy in children. This study is based on the comparative histological examination of abnormal, peg-shaped premolars, erupted in a patient treated for neuroblastoma, and of non-affected teeth, extracted in a healthy child. Apart from the size, the teeth vary in tissue morphology. The number of dentinal tubules, dependent on the number of odontoblasts, is smaller in the microdontal sample when observation in the same-sized field of view is conducted. Moreover, the youngest, more than 100-micrometer-thick layer of the microdontal dentin seems to be the secondary dentin, with crispy-shaped tubules and empty spaces between them. No irregular dentin is deposited in the samples of physiologically developed teeth. The structure of cementum is different as well. Unlike regularly shaped premolars, in which typical 2-layer tissue is seen, in sections of microdontal teeth, only acellular tissue with cementoblasts overlying its surface is present. Thorough analysis of drug administration effects, which are visible in microscopic sections, and of time of anticancer treatment could provide insight into the developmental mechanisms of tooth germ formation.

Periapical Microsurgery: Do Root Canal-retreated Teeth Have More Dentinal Defects?

INTRODUCTION: This microsurgical clinical study evaluated if teeth that have undergone endodontic retreatment are associated with more dentinal defects than primary root canal-treated teeth. METHODS: One hundred fifty-five patients who underwent periapical microsurgery treatment in a private practice setting were evaluated. The root ends were resected, and the roots were inspected for the presence of dentinal defects through the surgical operating microscope with the help of a 0.8-mm-diameter light-emitting diode probe light and methylene blue dye. The root canal treatment history (primary vs retreatment) of the teeth was documented and related to the presence or absence of dentinal defects. Bivariate analysis was performed using the chi-square test, and a multivariate analysis was performed using logistic regression to evaluate possible confounding effects of patient age, sex, and tooth location on the association between treatment and the presence of dentinal defects. RESULTS: Of the 155 treated teeth, 33 were excluded (3 fractured and 30 missing treatment history). Of the remaining 122 included teeth, 73 (59.8%) had undergone primary root canal treatment and 49 (40.2%) retreatment. Sixteen teeth (22.5%) of the primary root canal group versus 33 (64.7%) of the retreatment group had dentinal defects. The proportion of retreated teeth with dentinal defects compared with primary treatment was statistically significant (P < .001) with a higher proportion of retreated teeth having dentinal defects. In the multivariate analysis, only the type of treatment was statistically significant (P < .001). CONCLUSIONS: This clinical study showed that root canal-retreated teeth are associated with more dentinal defects than primary root canal-treated teeth.

Evaluation of the effects of the solution used for electrochemical dissolution of nickel-titanium endodontic files on dentine structure, microhardness and cell viability.

AIM: To evaluate the effects of the [NaF 12 g L-1  + NaCl 1 g L-1 ] solution used in the electrochemical dissolution process of fractured endodontic files, as well as its NiTi-containing product, on dentine hardness, topography and human fibroblast viability. METHODOLOGY: Sixty single-rooted human teeth were evaluated for dentine microhardness using the Vickers hardness test and the area and number of dentinal tubules by scanning electron microscopy. The samples were divided according to the dentine surface treatment: distilled water; 17% EDTA; [NaF 12 g L-1  + NaCl 1 g L-1 ]; and 17% EDTA + [NaF 12 g L-1  + NaCl 1 g L-1 ]. Thirty-six single-rooted human teeth were divided according to the irrigation protocol: Dulbecco's Modified Eagle's Medium + 10% foetal bovine serum; 5.25% NaOCl; [NaF 12 g L-1  + NaCl 1 g L-1 ]; and [NaF 12 g L-1  + NaCl 1 g L-1  + NiTi]. The extracts in contact with the apical foramen were used in the MTT assay to evaluate human fibroblast viability, with dilutions of 100%, 50%, 25% and 12.5%. Statistical tests used were paired t-tests, one-way anova, Tukey's test, Kruskal-Wallis test and Dunn's post-test. RESULTS: The [NaF 12 g L-1  + NaCl 1 g L-1 ] solution did not modify dentine microhardness or the average dentinal tubule area. However, EDTA induced changes in dentine structure and microhardness (P < 0.05). The [NaF 12 g L-1  + NaCl 1 g L-1 ] solution, and its NiTi-containing product had lower cytotoxicity than NaOCl at dilutions of 25% and 50% (P < 0.01). CONCLUSIONS: The [NaF 12 g L-1  + NaCl 1 g L-1 ] solution did not alter dentine microhardness or damage the dentine structure. It also demonstrated lower cytotoxicity than NaOCl.

Dentinogenesis imperfecta type II- genotype and phenotype analyses in three Danish families.

BACKGROUND: Dentinogenesis imperfecta (DI) is a rare debilitating hereditary disorder affecting dentin formation and causing loss of the overlying enamel. Clinically, DI sufferers have a discolored and weakened dentition with an increased risk of fracture. The aims of this study were to assess genotype-phenotype findings in three families with DI-II with special reference to mutations in the DSPP gene and clinical, histological, and imaging manifestations. METHODS: Nine patients participated in the study (two from family A, four from family B, and three from family C). Buccal swab samples were collected from all participants and extracted for genomic DNA. Clinical and radiographic examinations had been performed longitudinally, and the dental status was documented using photographic images. Four extracted and decalcified tooth samples were prepared for histological analysis to assess dysplastic manifestations in the dentin. Optical coherence tomography (OCT) was applied to study the health of enamel tissue from in vivo images and the effect of the mutation on the function and structure of the DSPP gene was analyzed using bioinformatics software programs. RESULTS: The direct DNA sequence analysis revealed three distinct mutations, one of which was a novel finding. The mutations caused dominant phenotypes presumably by interference with signal peptide processing and protein secretion. The clinical and radiographic disturbances in the permanent dentition indicated interfamilial variability in DI-II manifestations, however, no significant intrafamilial variability was observed. CONCLUSION: The different mutations in the DSPP gene were accompanied by distinct phenotypes. Enamel defects suggested deficit in preameloblast function during the early stages of amelogenesis.