Scaling-Up of Dental Pulp Stem Cells Isolated from Multiple Niches

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

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.

Current status of pulp capping with dentin adhesive systems: a review

Several studies have assessed the morphology and thickness of hybrid layer, the dentin bend strengths as well as sealing ability of dentin adhesive systems. However, few in vivo studies have evaluated the biocompatibility of the adhesive systems following application to deep dentin or directly to the pulp of human teeth. Many studies performed in non-human primate teeth or teeth of rats have reported pulp healing and dentin bridging following pulp capping with bonding agents. In addition, a few clinical and radiographical reports of the success of resin pulp capping have been described in the dental literature.Objectives: the aim of this review was to evaluate the literature on pulp responses following total acid etching and application of adhesive resins on deep cavities or pulp exposures. In addition, the clinical/radiographical evidence for the apparent success of vital pulp therapy and results obtained from animal and human studies were compared and discussed.Significance and conclusions: the self-etching adhesive systems may be useful and safe when applied on dentin, In contrast, persistent inflammatory reactions as well as delay in pulpal healing and failure of dentin bridging were seen in human pulps capped with bonding agents. The results observed in animal teeth cannot be directly extrapolated to human clinical conditions. Consequently, vital pulp therapy using acidic agents and adhesive resins seems to be contraindicated. (C) 2000 Academy of Dental Materials. Published by Elsevier B.V. Ltd. All rights reserved.

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.

Scanning electron microscopy evaluation of the hard tissue barrier after pulp capping with calcium hydroxide, mineral trioxide aggregate (MTA) or ProRoot MTA

The aim of this study was to investigate the morphology and localisation of calcium hydroxide- and mineral trioxide aggregate (MTA)-induced hard tissue barriers after pulpotomy in dogs' teeth. Pulpotomies were performed on maxillary and mandibular premolars of five dogs. The teeth were assigned into three groups according to the pulp-capping agent used. The pulpal wounds were capped with calcium hydroxide (Ca(OH)(2) - control), MTA or ProRoot MTA, and the cavities were restored with amalgam. After a 90-day follow-up period, the dogs were euthanised and the teeth were examined under scanning electron microscopy (SEM). An image-processing and analysis software was used to delimit the perimeters of the root canal area and the hard tissue barrier to determine the percentage of root canal obliteration. SEM data were used to assess the morphology, localisation and extension of the reparative hard tissue barriers. ProRoot MTA was statistically different from MTA and Ca(OH)(2) (P < 0.05) regarding tissue barrier morphology. Localisation data showed that ProRoot MTA was significantly different from Ca(OH)(2) (P < 0.05) and similar to MTA (P > 0.01; P > 0.05). No statistically significant difference (P > 0.01; P > 0.05) was observed between MTA and Ca(OH)(2). A larger number of complete (centroperipheral) hard tissue barriers with predominance of dentinal tubules was observed to the ProRoot MTA when compared with the Ca(OH)(2) group.

Er : YAG laser: Antimicrobial effects in the root canals of dogs' teeth with pulp necrosis and chronic periapical lesions

Objective: Our goal in this study was to evaluate the antimicrobial effect of Er:YAG laser applied after biomechanical preparation of the root canals of dog's teeth with apical periodontitis. Background Data: Various in vitro studies have reported effective bacterial reduction in infected root canals using Er:YAG laser. However, there is no in vivo research to support these results. Methods: Forty root canals of dogs' premolar teeth with pulp necrosis and chronic periapical lesions were used. An initial microbiological sample was taken, and after biomechanical preparation was carried out, a second microbiological sample was taken. The teeth were divided into two groups: Group I-biomechanical preparation was taken of root canals without Er:YAG laser application; Group II-biomechanical preparation was taken of root canals with Er:YAG laser application using 140-mj input, 63-mJ output/15 Hz. After coronal sealing, the root canals were left empty for 7 days at which time a third microbiological sample was taken. The collected material was removed from the root canal with a #40 K file and placed in transport media. It was serially diluted and seeded on culture dishes selective for anaerobes, aerobes, and total streptococci. Colony-forming units per milliliter (CFU/mL) were counted. Results: Groups I and II showed an increase of CFU/mL for all microorganisms 7 days after treatment, being statistically significant for anaerobes in Group I and for anaerobes and total streptococci in Group II. When comparing CFU/mL of Groups I and II, there was a statistically significant increase after 7 d for total streptococci in Group II. Conclusion: Er:YAG laser applied after biomechanical preparation did not reduce microorganisms in the root canal system.

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

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Behavioral Response of the Leafcutter Ant, Atta sexdens rubropilosa (Hymenoptera, Formicidae), to Components of Citrus Pulp

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ontogenesis of the fruit pulp layer of Hymenaea stigonocarpa (Fabaceae : Caesalpinioideae)

Hymenaea, a genus of major economic importance, has been the subject of several botanical studies. However, there is disagreement over the origin of the edible fruit pulp of Hymenaea, as there are no ontogenetic studies on this organ. According to some authors, the edible layer results from transformations of the mesocarp and endocarp, while according to others, it is considered a seed aril. There are still others who regard this layer as originating from an undefined region of the pericarp. To understand the nature and origin of the pulp layer, Hymenaea stigonocarpa Mart. ex Hayne ovaries and fruit were processed according to standard techniques. The production of the fruit pulp layer starts immediately after anthesis. During anthesis, the inner epidermal cells of the ovary show periclinal division and form a new layer of cells towards the mesocarp; this remains meristematic and initiates cell production by predominantly periclinal divisions, producing a compact tissue towards the locule. This tissue will become the fruit pulp layer, the inner endocarp. The seed coat shows typical testal structure without evidence of aril formation. This allows us to conclude that the fruit pulp layers are exclusively made from part of the endocarp. We also observed resin cavities on outer mesocarp and outer endocarp.

A microsatellite-based, gene-rich linkage map for the AA genome of Arachis (Fabaceae)

Cultivated peanut (Arachis hypogaea) is an important crop, widely grown in tropical and subtropical regions of the world. It is highly susceptible to several biotic and abiotic stresses to which wild species are resistant. As a first step towards the introgression of these resistance genes into cultivated peanut, a linkage map based on microsatellite markers was constructed, using an F-2 population obtained from a cross between two diploid wild species with AA genome (A. duranensis and A. stenosperma). A total of 271 new microsatellite markers were developed in the present study from SSR-enriched genomic libraries, expressed sequence tags (ESTs), and by data-mining sequences available in GenBank. of these, 66 were polymorphic for cultivated peanut. The 271 new markers plus another 162 published for peanut were screened against both progenitors and 204 of these (47.1%) were polymorphic, with 170 codominant and 34 dominant markers. The 80 codominant markers segregating 1:2:1 (P < 0.05) were initially used to establish the linkage groups. Distorted and dominant markers were subsequently included in the map. The resulting linkage map consists of 11 linkage groups covering 1,230.89 cM of total map distance, with an average distance of 7.24 cM between markers. This is the first microsatellite-based map published for Arachis, and the first map based on sequences that are all currently publicly available. Because most markers used were derived from ESTs and genomic libraries made using methylation-sensitive restriction enzymes, about one-third of the mapped markers are genic. Linkage group ordering is being validated in other mapping populations, with the aim of constructing a transferable reference map for Arachis.

Genetic relationships among Arachis hypogaea L. (AABB) and diploid Arachis species with AA and BB genomes

The cultivated peanut (Arachis hypogaea L.) is an allotetraploid, with two types of genomes, classified as AA and BB, according to cytogenetic characters. Similar genomes to those of A. hypogaea are found in the wild diploid species of section Arachis, which is one of the nine Arachis sections. The wild species have resistances to pests and diseases that affect the cultivated peanut and are a potential source of genes to increase the resistance levels in peanut. The aim of this study was to analyze the genetic variability within AA and BB genome species and to evaluate how they are related to each other and to A. hypogaea, using RAPD markers. Eighty-seven polymorphic bands amplified by ten 10-mer primers were analyzed. The species were divided into two major groups, and the AA and the BB genome species were, in general, separated from each other. The results showed that high variation is available within species that have genomes similar to the AA and the BB genomes of A. hypogaea.

Testing the quick meal hypothesis: the effect of pulp on hoarding and seed predation of Hymenaea courbaril by red-rumped agoutis (Dasyprocta leporina)

Red-rumped agoutis (Dasyprocta leporina) are important seed dispersers/predators of Neotropical large-seeded plants. Several species of seeds cached by agoutis have an edible reward, in contrast to temperate rodent-dispersed diaspores. The quick meal hypothesis states that the presence of a reward such as edible pulp will enhance the efficiency of rodents as seed disperses by satiating the animal and, consequently, reducing seed predation and enhancing hoarding. In this study, this hypothesis was tested using as the reference system the pulp and seeds of Hymenaea courbaril. Seeds with and without pulp were offered to agoutis and the behaviour of each individual was recorded. Since the probability of predation and hoarding were complementary, we used the probability of predation. The proportion of agoutis that preyed on at least one seed was similar for seeds with (42.8% of individuals) and without (40.0% of individuals) pulp. In agoutis that preyed upon at least one seed, the probability that they killed a seed did not differ between seeds with (0.17 +/- 0.03) and without (0.20 +/- 0.08) pulp. Hence, these results do not support the 'quick meal hypothesis'.

Does attraction to frugivores or defense against pathogens shape fruit pulp composition?

Fruit traits evolve in response to an evolutionary triad between plants, seed dispersers, and antagonists that consume fruits but do not disperse seeds. The defense trade-off hypothesis predicts that the composition of nutrients and of secondary compounds in fruit pulp is shaped by a trade-off between defense against antagonists and attraction to seed dispersers. The removal rate model of this hypothesis predicts a negative relationship between nutrients and secondary compounds, whereas the toxin-titration model predicts a positive relationship. To test these alternative models, we evaluated whether the contents of nutrients and secondary compounds can be used to predict fruit removal by mutualists and pathogens in 14 bird-dispersed plants on a subtropical island in São Paulo state, southeastern Brazil. We selected eight to ten individuals of each species and prevented fruit removal by covering four branches with a net and left fruits on four other branches available to both, vertebrate fruit consumers and pathogens. The persistence of ripe fruits was drastically different among species for bagged and open fruits, and all fruit species persisted longer when protected against seed dispersers. We found that those fruits that are quickly removed by vertebrates are nutrient-rich, but although the attack rate of pathogens is also high, these fruits have low contents of quantitative defenses such as tannins and phenols. Thus, we suggest that the fruit removal rate by seed dispersers is the primary factor selecting the levels of fruit defense. Likewise, nutrient-poor fruits have low removal of seed dispersers and low probability of attack by pathogens. These species retain ripe fruits in an intact condition for a prolonged period because they are highly defended by secondary compounds, which reduce overall attractiveness. However, this strategy might be advantageous for plants that depend on rare or unreliable dispersers.

Effect of pectinolitic enzymes on the physical properties of caja-manga (Spondias cytherea Sonn.) pulp

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