Biocompatibility of a resin-modified glass-ionomer cement applied as pulp capping in human teeth

Purpose: to evaluate the human pulp response following pulp capping with calcium hydroxide (CI-I, Group 1), and the resin-modified glass-ionomer Vitrebond (VIT, Group 2). Materials and Methods: Intact teeth with no cavity preparation were used as control Group (ICG, Group 3). Buccal Class V cavities were prepared in 34 sound human premolars. After exposing the pulps, the pulp capping materials were applied and the cavities were Filled using Clearfil Liner Bond 2 bonding agent and Z100 resin-based composite. The teeth were extracted after 5, 30, and from 120 to 300 days, fixed in 10% buffered formalin solution, and prepared according to routine histological techniques. 6-mu m sections were stained with hematoxylin and eosin, Masson's trichrome, or Brown gr Brenn technique for bacterial observation. Results: At 5 days, CH caused a large zone of coagulation necrosis, the mononuclear inflammatory reaction underneath the necrotic zone was slight to moderate. VIT caused a moderate to intense inflammatory pulp response with a large necrotic zone. A number of congested venules associated with plasma extravasation and neutrophilic infiltration was observed. Over time, only CH allowed pulp repair and complete dentin bridging around the pulp exposure site. VIT components displaced into the pulp tissue triggered a persistent inflammatory reaction which appeared to be associated with a lack of dentin bridge formation. After 30 days a few histological sections showed a number of bacteria on the lateral dentin walls. In these samples the pulp response was similar to those samples with no microleakage. VIT was more irritating to pulp tissue than CH, which allowed pulp repair associated with dentin bridge formation. These results suggested that VIT is not an appropriate dental material to be used in direct pulp capping for mechanically exposed human pulps.

Human pulpal response to direct pulp capping with an adhesive system

Purpose: To evaluate clinically and microscopically the human pulp response when directly capped with an adhesive system or calcium hydroxide over short (9-12 days) and long (53-204 days) experimental periods. Materials and Methods: Fifty-one sound human premolars scheduled for orthodontic extraction, had their pulp horns gently exposed with a diamond point. Debris in the pulp wound was washed out with a sterile saline solution. The pulps were then capped with either an adhesive system (Scotchbond Multi-Purpose Plus) or calcium hydroxide. All teeth were subsequently restored with resin-based composite (Z-100) according to the manufacturer's instructions. After the experimental periods, the teeth were extracted and processed for light microscopic examination. Results: Short-term: the pulp tissue capped with SBMP-P exhibited dilated and congested blood vessels associated with a mo;derate inflammatory response and blanching of pulp cell nuclei. Long-term: no evidence of healing and bridge formation was observed. A persistent mild inflammatory pulp response was present. Micro-abscesses were detected in three cases associated with bacterial infiltration. Calcium hydroxide stimulated early pulp repair and dentin bridging which extended into the longest period.

Human pulp response to resin cements used to bond inlay restorations

Objective. The aim of this study was to evaluate the pulp response following cementation of inlays using two different resin cements.Methods. Deep Class V cavities were prepared on the buccal surface of 34 sound human premolars. impressions were taken and inlays were prepared which were cemented with the following luting materials-Group 1: Rely X(TM) Unicem. (3M ESPE); Group 2: Variolink(R) II (Ivoclar Vivadent). in Group 3 (control), after lining the cavity floor with Dycal(R) (Dentsply Caulk) the inlays were cemented with Rely X(TM) Unicem. Four additional teeth were used as an intact control group. For Variolink(R) II, the adhesive system Excite was used as part of the cementation procedure. After 7 or 60 days, the teeth were extracted and processed for histological assessment.Results. At 7 days, Rely X(TM) Unicern and Variolink(R) II system triggered in two samples a mild and moderate inflammatory response, respectively. At 60 days, the pulpal response decreased for both groups. A discrete persistent inflammatory response occurred in Group 2 in which displacement of resin components across the dentin tubules was observed. In the control group, normal histological characteristics were observed. The inflammatory response and tissue disorganization were related to the remaining dentin thickness between the cavity floor and the pulp tissue.Significance. Techniques for inlay cementation using distinct luting cements may cause specific pulpal damage. Variolink(R) II associated with the adhesive system Excite cause more aggressive effects to the pulp-dentin complex than Rely X(TM) Unicern cement when both are used to cement inlay restorations. (C) 2005 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Direct pulp capping with a self-etching adhesive system: Histopathologic evaluation in dogs' teeth

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Response of human dental pulp to dentin adhesive systems

Many in vivo studies have stated that the response of the dentin/pulp complex does not depend on the dental material used as the liner or pulp-capping agent. However, several in vitro studies have reported the metabolic cytotoxic effects of resin components applied to fibroblast and odontoblast cell lines. The aim of this study was to evaluate the human pulp response following direct pulp capping with current bonding agents and calcium hydroxide (CH). Sound premolars scheduled for orthodontic extraction had their pulp tissue mechanically exposed. After hemorrhage control and total acid conditioning, the experimental bonding agents, including All Bond 2, Scotchbond MP-Plus, Clearfil Liner Bond 2, and Prime & Bond 2.1 were applied on the pulp exposure site. CH saline paste was used as the control pulp-capping agent. All cavities were restored with Z-100 resin composite according to the manufacturer's instructions. Following extractions, the teeth were processed for microscopic evaluation. In the short term, the bonding agents elicited a moderate inflammatory pulp response with associated dilated and congested blood vessels adjacent to the pulp exposure site. A mild inflammatory pulp response was observed when Clearfil Liner Bond 2 or CH was applied on the pulp exposures. With time, macrophages and giant cells engulfing globules and components of all experimental bonding agents displaced into the pulp space were seen. This chronic inflammatory response did not allow complete pulp repair, which interfered with the dentin bridge formation. Pulp exposures capped with CH exhibited an initial organization of elongated pulp cells underneath the coagulation necrosis. CH stimulated early pulp repair and dentin bridging that extended into the longest period. The bonding agents evaluated in the present study cannot be recommended for pulp therapy on sound human teeth.

Biocompatibility of an adhesive system applied to exposed human dental pulp

Human pulp tissue was directly capped with All Bond 2, or calcium hydroxide and evaluated 7, 30, or 60 days after the procedures. Histological analysis was performed to assess the inflammatory cell response, tissue disorganization, dentin bridging, and the presence of bacteria. At 7 days, with All Bond 2 capping, there was a large area of neutrophilic infiltrate underlying the pulp capping material, and the death of adjacent odontoblasts, was observed. However, with time, the neutrophilic reaction was replaced by fibroblastic proliferation with macrophages and giant cells surrounding globules of resin scattered in the coronal pulp tissue. The persistent inflammatory reaction and hyaline alteration of extracellular matrix inhibited complete pulp repair or dentin bridging. In contrast, at 7 days, the pulp tissue capped with calcium hydroxide exhibited odontoblast-like cells organized underneath coagulation necrosis. Pulp repair evolved into apparent complete dentin bridge formation at 60 days. All Bond 2 did not appear to allow any pulp repair and does not appear to be indicated for direct pulp capping of human teeth. Copyright © 1999 by The American Association of Endodontists.

Evaluation of two mineral trioxide aggregate compounds as pulp-capping agents in human teeth

Aim: The present randomized, controlled prospective study evaluated the histomorphological response of human dental pulps capped with two grey mineral trioxide aggregate (MTA) compounds. Methodology: Pulp exposures were performed on the occlusal floor of 40 human permanent pre-molars. The pulp was capped either with ProRoot (Dentsply) or MTA-Angelus (Angelus) and restored with zinc oxide eugenol cement. After 30 and 60 days, teeth were extracted and processed for histological examination and the effects on the pulp were scored. The data were subjected to Kruskal-Wallis and Conover tests (α = 0.05). Results: In five out of the 40 teeth bacteria were present in pulp tissue. No significant difference was observed between the two materials (P > 0.05) in terms of overall histological features (hard tissue bridge, inflammatory response, giant cells and particles of capping materials). Overall, 94% and 88% of the specimens capped with MTA-Angelus and ProRoot, respectively, showed either total or partial hard tissue bridge formation (P > 0.05). Conclusions: Both commercial materials ProRoot (Dentsply) and MTA-Angelus (Angelus) produced similar responses in the pulp when used for pulp capping in intact, caries-free teeth. © 2009 International Endodontic Journal.

Response of human dental pulp to calcium hydroxide paste preceded by a corticosteroid/antibiotic dressing agent

Aim: To evaluate the treatment with corticosteroid/antibiotic dressing in pulpotomy with calcium hydroxide. Methods: Forty-six premolars were pulpotomized and randomly assigned into 3 groups. In Group I pulpal wound was directly capped with calcium hydroxide, and Group II and Group III received corticosteroid/antibiotic dressing for 10 min or 48 h, respectively, before pulp capping. Teeth were processed for histological analysis after 7, 30 or 60 days to determine inflammatory cell response, tissue disorganization, dentin bridge formation and presence of bacteria. Attributed scores were analyzed by Kruskal-Wallis and Mann-Whitney tests (α=0.05). Results: On the 7th day, all groups exhibited dilated and congested blood vessels in the tissue adjacent to pulpal wound. The inflammatory cell response was significantly greater in Group III (p<0.05). On the 30th day, in all groups, a thin dentin matrix layer was deposited adjacent to the pulpal wound and a continuous odontoblast-like cell layer underlying the dentin matrix was observed. On the 60th day, all groups presented a thick hard barrier characterized by an outer zone of dystrophic calcification and an inner zone of tubular dentin matrix underlined by a defined odontoblast-like cell layer. Conclusions: Within the limitations of present study, considering that the treatment was performed in healthy teeth, it may be concluded that the use of a corticosteroid/antibiotic dressing before remaining tissue protection with calcium hydroxide had no influence on pulp tissue healing.

Tissue-engineering-based Strategies for Regenerative Endodontics

Stemming from in vitro and in vivo pre-clinical and human models, tissue-engineering-based strategies continue to demonstrate great potential for the regeneration of the pulp-dentin complex, particularly in necrotic, immature permanent teeth. Nanofibrous scaffolds, which closely resemble the native extracellular matrix, have been successfully synthesized by various techniques, including but not limited to electrospinning. A common goal in scaffold synthesis has been the notion of promoting cell guidance through the careful design and use of a collection of biochemical and physical cues capable of governing and stimulating specific events at the cellular and tissue levels. The latest advances in processing technologies allow for the fabrication of scaffolds where selected bioactive molecules can be delivered locally, thus increasing the possibilities for clinical success. Though electrospun scaffolds have not yet been tested in vivo in either human or animal pulpless models in immature permanent teeth, recent studies have highlighted their regenerative potential both from an in vitro and in vivo (i.e., subcutaneous model) standpoint. Possible applications for these bioactive scaffolds continue to evolve, with significant prospects related to the regeneration of both dentin and pulp tissue and, more recently, to root canal disinfection. Nonetheless, no single implantable scaffold can consistently guide the coordinated growth and development of the multiple tissue types involved in the functional regeneration of the pulp-dentin complex. The purpose of this review is to provide a comprehensive perspective on the latest discoveries related to the use of scaffolds and/or stem cells in regenerative endodontics. The authors focused this review on bioactive nanofibrous scaffolds, injectable scaffolds and stem cells, and pre-clinical findings using stem-cell-based strategies. These topics are discussed in detail in an attempt to provide future direction and to shed light on their potential translation to clinical settings.

Immediate and late analysis of dental pulp stem cells viability after indirect exposition to alternative in-office bleaching strategies

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Immediate human pulp response to ethanol-wet bonding technique

To evaluate the short-term response of human pulps to ethanol-wet bonding technique. Methods Deep class V cavities were prepared on 17 sound premolars and divided into three groups. After acid-etching, the cavities from groups 1 (G1) and 2 (G2) were filled with 100% ethanol or distilled water, respectively, for 60 s before the application of Single Bond 2. In group 3 (G3, control), the cavity floor was lined with calcium hydroxide before etching and bonding. All cavities were restored with resin composite. Two teeth were used as intact control. The teeth were extracted 48 h after the clinical procedures. From each tooth serial sections were obtained and stained with haematoxylin and eosin (H/E) and Masson's trichrome. Bacteria microleakage was assessed using Brown & Brenn. All sections were blindly evaluated for five histological features. Results Mean remaining dentine thickness was 463 ± 65 μm (G1); 425 ± 184 μm (G2); and 348 ± 194 μm (G3). Similar pulp reactions followed ethanol- or water-wet bonding techniques. Slight inflammatory responses and disruption of the odontoblast layer related to the cavity floor were seen in all groups. Stained bacteria were not detected in any cavities. Normal pulp tissue was observed in G3 except for one case. Conclusions After 48 h, ethanol-wet bonding does not increase pulpal damage compared to water-wet bonding technique. Clinical significance Ethanol-wet bonding may increase resin-dentine bond durability. This study reported the in vivo response of human pulp tissue when 100% ethanol was applied previously to an etch-and-rinse simplified adhesive system.

Cytotoxicity of resin-based luting cements to pulp cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diabetes triggers the loss of tooth structure associated to radiographical and histological dental changes and its evolution to progressive pulp and periapical lesions in rats

The aim of this study was to evaluate the putative influence of diabetes without metabolic control in the loss of tooth structure as well as histological changes in dentin and pulp tissue in rats. Diabetes was induced in Wistar rats (n=25) by intravenous administration of alloxan (42mg/kg). Diabetic and non-diabetic control rats were evaluated at 1, 3, 6, 9 and 12 months of follow-up. In order to evaluate the presence and progression of dental caries and periapical lesions, hemimandibles were removed and submitted to radiographical, histological, and morphometrical procedures. Dental caries were detected after radiographical and histological evaluations in diabetic group from the third month of diabetes onset, increasing gradually in frequency and severity in periods. Diabetic rats dental pulps also presented significant reduction in volume density of collagen fibers and fibroblasts at third month, parallel with a trend towards the increase in inflammatory cells volume density. Diabetic rats presented a generalized pulp tissue necrosis after 6 months of diabetes induction. Moreover, periapical lesions were not detected in control group, while these lesions were observed in all rats after 3, 6, 9, and 12 months of diabetes induction. Uncontrolled diabetes seems to trigger the loss of tooth structure, associated to histological dental changes and mediates its evolution to progressive severe pulp and periapical lesions in rats. Therefore, diabetes may be considered a very important risk factor regarding alterations in dental pulp, development of dental caries, and periapical lesions.

Differential inflammatory response of dental pulp explants and fibroblasts to saliva

Abstract AIM: To investigate the inflammatory response of dental pulp fibroblasts and the respective explants to whole saliva. METHODOLOGY: Explants from human and porcine dental pulp tissue and isolated dental pulp fibroblasts were used to investigate the inflammatory response to sterile saliva. Cytokine and chemokine expression was assessed by RT-PCR. Western blot analysis and pharmacologic inhibitors were used to determine the involvement of signalling pathways. RESULTS: Dental pulp explants of human and porcine origin exposed to human saliva exhibited no major changes of IL-6 and IL-8 mRNA expression (P > 0.05). In contrast, isolated porcine and human dental pulp fibroblasts, when stimulated with human saliva, exhibited a vastly increased expression of IL-6 and IL-8 mRNA (P < 0.05). In pulp fibroblasts, saliva also increased the expression of other cytokines and chemokines via activation of NFkappaB, ERK and p38 signalling. Notably, a significantly reduced inflammatory response was elicited when pulp fibroblasts were transiently exposed to saliva. CONCLUSIONS: Saliva has a potential impact on inflammation of dental pulp fibroblasts in vitro but not when cells are embedded in the intrinsic extracellular matrix of the explant tissue.

Effects of Prolyl Hydroxylase Inhibitor L-mimosine on Dental Pulp in the Presence of Advanced Glycation End Products

INTRODUCTION Proangiogenic prolyl hydroxylase (PHD) inhibitors represent a novel approach to stimulate tissue regeneration. Diabetes mellitus involves the accumulation of advanced glycation end products (AGEs). Here we evaluated the impact of AGEs on the response of human pulp tissue to the PHD inhibitor L-mimosine (L-MIM) in monolayer cultures of dental pulp-derived cells (DPCs) and tooth slice organ cultures. METHODS In monolayer cultures, DPCs were incubated with L-MIM and AGEs. Viability was assessed based on formazan formation, live-dead staining, annexin V/propidium iodide, and trypan blue exclusion assay. Vascular endothelial growth factor (VEGF), interleukin (IL)-6, and IL-8 production was evaluated by quantitative polymerase chain reaction and immunoassays. Furthermore, expression levels of odontoblast markers were assessed, and alizarin red staining was performed. Tooth slice organ cultures were performed, and VEGF, IL-6, and IL8 levels in their supernatants were measured by immunoassays. Pulp tissue vitality and morphology were assessed by MTT assay and histology. RESULTS In monolayer cultures of DPCs, L-MIM at nontoxic concentrations increased the production of VEGF and IL-8 in the presence of AGEs. Stimulation with L-MIM decreased alkaline phosphatase levels and matrix mineralization also in the presence of AGEs, whereas no significant changes in dentin matrix protein 1 and dentin sialophosphoprotein expression were observed. In tooth slice organ cultures, L-MIM increased VEGF but not IL-6 and IL-8 production in the presence of AGEs. The pulp tissue was vital, and no signs of apoptosis or necrosis were observed. CONCLUSIONS Overall, in the presence of AGEs, L-MIM increases the proangiogenic capacity, but decreases alkaline phosphatase expression and matrix mineralization.