Adaptive amelogenesis during unimpeded eruption of rat mandibular incisor.

The aim of the study was to see whether the length of the enamel secretion zone in unimpeded rat incisors, measured precisely, is in agreement with the observed decrease in enamel thickness. Unimpeded eruption of mandibular incisors of five experimental and two control rats was induced by cutting off the erupted part of the incisors three times per week for 5 weeks. The length of the zone of enamel secretion in unimpeded and impeded control incisors was measured on longitudinal and serial transverse histological sections of fixed, demineralised and embedded hemimandibles. Impeded contralateral incisors were also included in the study. The length of the zone of enamel secretion in unimpeded incisors showed an increase to 8,398 ± 558 µm, that is 161% of the length in control incisors (5,213 ± 95 µm). The contralateral incisor showed a reduction in eruption rate, in length of the secretion zone, and the whole tooth was shifted somewhat apically. The measured length of the secretion zone is in agreement with the observed thickness of enamel (98 µm) in unimpeded incisors. The reduced eruption rate and the apical shift of the contralateral incisor are probably due to an increased occlusal load.

On the method of revealing enamel structure by acid etching. Aspects of optimization and interpretation.

The study aimed at finding an optimal combination of acid concentration and etching time when nitric acid is used as etchant for the study of the finer details of human dental enamel structure. Four hundred 2-3-mm-thick segments of facio-lingually sectioned human third molar crowns were assigned to 20 groups with 20 specimens in each group, each group differing with respect to acid concentration (0.1, 1, 2.5, and 5%) and etching time (15, 30, 45, 90, and 180 s). After etching and preparation, specimens were observed in the scanning electron microscope (SEM). Surface roughness/topography increased with increasing acid concentration and increasing etching time, but not in a linear fashion; generally, prisms tended to go from flat-surfaced to cone-shaped and prism sheaths from fissure-like to wedge-shaped. Intragroup variations and intergroup similarities were considerable. The two major enamel factors determining the etch effect are crystal orientation and prism sheath properties. Other factors, such as distribution of porosities and crystal quality, also contribute probably. Slight to moderate topography is best for observing the finer enamel structure, for example, etching with concentrations in the range 0.1-1% and with etching times in the range 15-90 s, the stronger the acid, the shorter the time. The depth effect of nitric acid is judged to be relatively small. Considerable variations in expression of prism cross-striations were observed. SEM observations of acid-etched enamel in carefully selected planes are a powerful method for the study of enamel structure, bearing in mind the artifactual aspects of the observed surface.

Scanning Electron Microscopy (SEM) Methods for Dental Enamel.

Scanning electron microscopy (SEM) is exceptionally well suited for the study of the structure of dental enamel, due to its ability to create high-resolution images of hard surfaces. Continuous attention on how to arrive at the observation stage with a clean and dry specimen is one main aspect of specimen preparation. Other main aspects are choice of whether the specimen should be embedded or not, choice of plane of section, and choice of acid-etching regime. Special attention is given to the preparation of small specimens and how to prepare and observe more than one plane or aspect in the same specimen.

Tooth identification puzzle: A method of teaching and learning tooth morphology.

INTRODUCTION: The present report outlines a method of teaching/learning tooth morphology by tooth identification puzzle. MATERIALS AND METHODS: Students are presented with sets of extracted human teeth comprising complete dentitions except deciduous incisors and canines. The task is to place the teeth in correct positions in a schematic dentition diagram. The course, including 2-3 introductory lectures and a final test of one hour, has a time frame of 14-16 hours. A total of 506 2nd year students from several years participated. RESULTS: The course is much appreciated by the students who experience a marked progress in skills. In the final test, 51.8% of the students had no faults, whilst 3% failed (more than 12 faults). The average number of faults per student was 2.3. Of the 20 240 positioned teeth 5.7% were misplaced. The most frequently misplaced teeth were mandibular central incisors, maxillary second premolars and mandibular first premolars. The most common type of fault was inside determination. DISCUSSION: The course is cost-effective and facilitates learning through its multifaceted activity with involvement of many senses. An important asset is the appreciation of variations in tooth morphology. The course provides an arena for close and positive interaction between students and teachers.

Obtaining scratch-free specimens of dental enamel prepared by sectioning, grinding, polishing, and acid etching for scanning electron microscopy.

The aim of the present study was to identify the stage during acid etching of dental enamel where scratches introduced by sectioning and grinding procedures were eliminated. Nitric acid was used as etchant. Four hundred 2-3 mm-thick longitudinal sections of human third molar crowns were assigned to 80 groups, each group differing with respect to grinding procedure (silicon carbide paper grit 600; 1,000; 1,200; and 1,200 + polishing), acid concentration (0.1%, 1%, 2.5%, and 5%) and etching time (15, 30, 45, 90, and 180 s). Observation of the specimens with scanning electron microscopy (SEM) showed that elimination of scratches was facilitated by increasing fineness of grinding paper and polishing, and by increasing acid concentration and increasing etching time. An acid concentration of 0.1% did not remove scratches totally irrespective of grinding procedure or etching time, 1% acid removed most scratches after 45-90 s, while 2.5% and 5% acids removed most scratches after 30-45 s. It is proposed that the scratches are eliminated/obscured through a combined effect of the acid and the inherent structure of the enamel. The results of the present study may serve as a guide in choosing the best procedures for obtaining scratch-free enamel surfaces through acid etching with the least possible loss of enamel substance. Research highlights Sectioning and grinding produce scratches in dental enamel. Scratches are eliminated by acid etching prior to SEM studies of enamel structure. Scratch elimination increases with increasing acid strength and etching time and is facilitated by finer grit grinding paper and polishing.

Aspects of the final phase of enamel formation as evidenced by observations of superficial enamel of human third molars using scanning electron microscopy.

OBJECTIVE: Enamel structure reflects ameloblast function. By studying the structure of the superficial enamel, information about ameloblast function toward the end of the secretory stage may be obtained. DESIGN: The superficial enamel in midcoronal areas of acid-etched facio-lingual sections from human third molars was studied in the scanning electron microscope (SEM). RESULTS: A great variation was observed in occurrence of prism-free enamel. Prism-free enamel dominated in 40% (mandibular) and 47% (maxillary) of observed areas and had a mean thickness of about 30µm. Striations in the prism-free enamel had an interstriae distance of about 3.3-3.8µm. The angle between prisms and enamel surface was about 60°, between prisms and Retzius lines about 45° and between Retzius lines and enamel surface about 15°. The distances between regularly occurring Retzius lines and between striations in the prism-free enamel both tended to decrease toward the enamel surface. Prisms could change direction as they approached the enamel surface, mostly in cervical direction. Where Retzius lines curved and converged occlusally, prisms tended to deviate in an occlusal direction. CONCLUSIONS: Judged from the incremental lines and occurrence of prism-free enamel, ameloblasts slow down and tend to lose their Tomes' process as they approach the end of secretion. The crystals of prism-free enamel belong to the same system as the interprism crystals of prismatic enamel. A method, based on the disposition of fine incremental lines, is suggested for evaluation of ameloblast dynamics in the last stage of enamel secretion.

Microscopic structure of dental hard tissues in primary and permanent teeth from individuals with Prader-Willi syndrome.

OBJECTIVE: severe tooth wear, in terms of both erosive wear and attrition, is a significant problem in individuals with Prader-Willi syndrome (PWS). The purpose of the present study was to describe the structure of enamel and dentine in primary and permanent teeth from individuals with PWS. DESIGN: thirty-two primary and 10 permanent teeth representing 16 individuals with PWS were investigated in the study. The enamel surface was studied using scanning electron microscopy (SEM). The microscopic structure of enamel and dentine was studied using SEM, microradiography and light microscopy. RESULTS: the microscopic structure of enamel and dentine was found to be normal with the exception of a slight increase of interglobular dentine (IGD). Severe erosive defects were observed in primary teeth and also in permanent teeth with long exposure to the oral environment. CONCLUSION: the erosive enamel defects in individuals with PWS seem more related to the factors in the oral environment than to enamel structure which appeared normal. The occurrence of IGD indicate deficient mineralization but is probably of minor clinical significance. Gastro-oesophageal reflux is worthy of further investigation in individuals with Prader-Willi syndrome.

The effect of hypoxia on the formation of mouse incisor enamel.

OBJECTIVE: The permanently growing mouse incisors exhibit all stages of tooth development along their inciso-apical axis at any time. Any disturbance or injury of the ameloblasts during enamel formation or maturation may result in permanent defects in the finished enamel since the enamel does not undergo repair or remodeling after formation. In order to increase our understanding of how hypoxia affects enamel formation, we induced severe acute hypoxia in adult mice and observed its effects on the enamel in incisors. DESIGN: Incisors from hypoxic mice were obtained 5 and 49 days after the hypoxic insult. Hypoxic and control incisors were dissected out and observed by scanning electron microscopy (SEM). Incisors were subsequently ground longitudinally or transversely, etched, and observed again by SEM. The nature and position of defects were considered in relation to the configuration and dynamics of the incisors. RESULTS: The effect of hypoxia varied considerably, among mice, among incisors, and among ameloblasts. Affected enamel showed hypoplasia with hypomineralization or hypomineralization without hypoplasia. Vascular endothelial growth factor (VEGF) showed considerably stronger labeling in hypoxic compared to control ameloblasts. CONCLUSIONS: The present study demonstrates quantitative and qualitative defects in the enamel reflecting the vulnerability of ameloblasts toward severe acute hypoxia in mouse incisors.

Incremental lines in mouse molar enamel.

OBJECTIVE: The purpose of the present study was to investigate the occurrence and periodicity of enamel incremental lines in mouse molars in an attempt to draw attention to some key questions about the rhythm in the activity of the secreting ameloblasts during formation of mouse molar enamel. METHODS: The mouse molars were ground, etched, and studied using scanning electron microscopy. RESULTS: Lines interpreted as incremental lines generally appeared as grooves of variable distinctness, and were only observed cervically, in the region about 50-250µm from the enamel-cementum junction. The lines were most readily observable in the outer enamel and in the superficial prism-free layer, and were difficult to identify in the deeper parts of enamel, i.e. in the inner enamel with prism decussation. However, in areas where the enamel tended to be hypomineralized the incremental lines were observed as clearly continuous from outer into inner enamel. The incremental lines in mouse molar enamel exhibited an average periodicity of about 4µm, and the distance between the lines decreased towards the enamel surface. CONCLUSIONS: We conclude that incremental lines are to some extent visible in mouse molar enamel. Together with data from the literature and theoretical considerations, we suggest that they probably represent a daily rhythm in enamel formation. This study witnesses the layered apposition of mouse molar enamel and supports the theory that circadian clock probably regulates enamel development.

Expression of Clu and Tgfb1 during murine tooth development: effects of in-vivo transfection with anti-miR-214.

Expression of clusterin (Clu) in the murine first molar tooth germ was markedly increased at postnatal developmental stages. The time-course of expression of this gene paralleled those of other genes encoding proteins involved during the secretory phase of odontogenesis, as described previously. Immunohistochemical studies of clusterin in murine molar tooth germs suggested this protein to be located in outer enamel epithelium, regressing enamel organ, secretory ameloblasts, and the dental epithelium connecting the tooth to the oral epithelium at an early eruptive stage. Immunolabelling of transforming growth factor beta-1 (TGF-ß1) revealed it to be located close to clusterin. The levels of expression of Clu and Tgfb1 were markedly decreased following in-vivo transfection with anti-miR-214. In contrast, the expression of several genes associated with regulation of growth and development were increased by this treatment. We suggest that clusterin has functions during secretory odontogenesis and the early eruptive phase. Bioinformatic analysis after treatment with anti-miR-214 suggested that, whilst cellular activities associated with tooth mineralization and eruption were inhibited, activities associated with an alternative developmental activity (i.e. biosynthesis of contractile proteins) appeared to be stimulated. These changes probably occur through regulation mediated by a common cluster of transcription factors and support suggestions that microRNAs (miRNAs) are highly significant as regulators of differentiation during odontogenesis.

Expression of delta-like 1 homologue and insulin-like growth factor 2 through epigenetic regulation of the genes during development of mouse molar.

Delta-like 1 homolog (Dlk1) and insulin-like growth factor 2 (Igf2) are two of six well-studied mouse imprinted gene clusters that are paternally expressed. Their expression is also linked to their maternally expressed non-coding RNAs, encoded by Gene trap locus 2 (Gtl2) and Imprinted maternally expressed transcript (H19), co-located as imprinted gene clusters. Using deoxyoligonucleotide microarrays and real-time RT-PCR analysis we showed Dlk1 and Gtl2 to exhibit a time-course of expression during tooth development that was similar to that of Igf2 and H19. Western blot analysis of proteins encoded by Dlk1 and Igf2 suggested that the levels of these proteins reflected those of the corresponding mRNAs. Immunohistochemical studies of DLK1 in murine molars detected the protein in both epithelial and mesenchymal regions, in developing cusp mesenchyme, and in newly synthesized enamel and dentin tubules. IGF2 protein was detected primarily at prenatal stages, suggesting that it may be active before birth. Analysis of methylation of cytosine-phosphate-guanine (CpG) islands in both Dlk1 and Igf2 suggested the presence of an increasing fraction of hypermethylated bases with increasing time of development. The increased levels of hypermethylation coincided both with the diminished levels of expression of Dlk1 and Igf2 and with decreased levels of DLK1 and IGF2 proteins in the tooth germ, suggesting that their expression is regulated via methylation of CpG islands present in these genes.

Effects of in vivo transfection with anti-miR-214 on gene expression in murine molar tooth germ.

MicroRNAs (miRNAs) are an abundant class of noncoding RNAs that are believed to be important in many biological processes through regulation of gene expression. Little is known of their function in tooth morphogenesis and differentiation. MicroRNA-214 (miR-214), encoded by the polycistronic Dnm30os gene, is highly expressed during development of molar tooth germ and was selected as a target for silencing with anti-miR-214. Mandibular injection of 1-100 pmol of anti-miR-214 close to the developing first molar in newborn mice resulted in significant decrease in expression of miR-214, miR-466h, and miR-574-5p in the tooth germ. Furthermore, levels of miR-199a-3p, miR-199a-5p, miR-690, miR-720, and miR-1224 were significantly increased. Additionally, the expression of 863 genes was significantly increased and the expression of 305 genes was significantly decreased. Among the genes with increased expression was Twist-1 and Ezh2, suggested to regulate expression of miR-214. Microarray results were validated using real-time RT-PCR and Western blotting. Among genes with decreased expression were Amelx, Calb1, Enam, and Prnp; these changes also being reflected in levels of corresponding encoded proteins in the tooth germ. In the anti-miR-214-treated molars the enamel exhibited evidence of hypomineralization with remnants of organic material and reduced surface roughness after acid etching, possibly due to the transiently decreased expression of Amelx and Enam. In contrast, several genes encoding contractile proteins exhibited significantly increased expression. mRNAs involved in amelogenesis (Ambn, Amelx, Enam) were not found among targets of miRNAs that were differentially expressed following treatment with anti-miR-214. It is therefore suggested that effects of miR-214 on amelogenesis are indirect, perhaps mediated by the observed miR-214-dependent changes in levels of expression of numerous transcription factors.

Expression of prion gene and presence of prion protein during development of mouse molar tooth germ.

In order to gain insight into possible cellular functions of the prion protein (PrP) during normal development, the expression of Prnp (encoding the PrP) and the distribution of the PrP were studied in murine tooth germs. Expression of Prnp in the mouse first molar tooth germ was highly dynamic, increasing several-fold during the secretory phase of odontogenesis, exhibiting a time-course of expression similar to that of genes coding for other extracellular proteins [e.g. enamel matrix proteins (Amelx, Ambn, Enam), Aplp1, Clstn1, and Clu]. Western blot analysis suggested that the amounts of PrP and amyloid beta (A4) precursor-like protein 1 (APLP1) in the tooth germ followed time-courses similar to those of the corresponding mRNAs. Immunohistochemical studies of the distribution of PrP in murine molar and incisor tooth germs at embryonic day (E)18.5 suggested that this protein was located in the cervical loop, outer enamel epithelium, pre-ameloblasts, and dental papilla. Different degrees of immunolabelling of pre-ameloblasts on the mesial and distal aspects of a lower molar cusp may be related to different enamel configurations on the two aspects. It is concluded that the dynamic patterns of expression of Prnp, and of distribution of PrP, suggest that PrP may have functions during secretory odontogenesis, perhaps in relation to amelogenesis.

Gene expression and dental enamel structure in developing mouse incisor.

At the mouse incisor tip the initially differentiated ameloblasts produce a thin, prism-free enamel, while further apically, in the immediate adjacent segment, the enamel thickness increases and the four-layered enamel of mouse incisor is formed. Comparative gene-expression profiling was carried out on RNA isolated from these two segments of incisor tooth germs at embryonic day (E)17.5 and at postnatal days (P)0, 1, 2, and 10 using microarrays to measure messenger RNA (mRNA) and microRNA (miRNA) species present in the segments. Validation of expression data was achieved using real-time reverse transcription-polymerase chain reaction (RT-PCR) and western blotting. Bioinformatic data suggested enhanced cellular apoptosis in the incisal tip segment, which, together with diminished expression of the Amelx and Enam genes, may contribute to the production of the thin enamel seen in this tooth segment. For genes exhibiting higher levels of expression in the adjacent segment where complex enamel is being formed, bioinformatic analysis suggested significant associations with cellular functions involving the actin cytoskeleton, cellular development, morphology, and movement. This is suggested to reflect that ameloblasts with Tomes' process are being organized in transverse rows, facilitating the transverse movement that results in prism decussation in the inner enamel of the adjacent segment. Bioinformatic analysis of miRNA expression data lends support to these suggestions.

Differential gene expression profiling of the molar tooth germ in peroxisome proliferator-activated receptor-alpha (PPAR-alpha) knockout mouse and in wild-type mouse: molar tooth phenotype of PPAR-alpha knockout mouse.

Gene expression profiling of the first molar tooth germ at embryonic days (E)17.5 and 18.5, and at postnatal days (P)0, 2, and 6 from peroxisome proliferator-activated receptor-alpha (PPAR-alpha) knockout mouse and from wild-type mouse was carried out using microarrays and validated using real-time reverse transcription-polymerase chain reaction (RT-PCR) and western blotting. When comparing expression profiles at each time-point, a total of 1,235 genes showed significantly different expression, 772 of which exhibited significantly decreased expression in tooth germ from knockout mouse. With genes exhibiting significantly decreased levels of expression in tooth germ from PPAR-alpha knockout mouse, bioinformatic analysis using ingenuity pathway analysis yielded significant associations to cellular functions related to cellular growth/proliferation and to networks related to regulation of calcium homeostasis. Using scanning electron microscopy to investigate molars from adult PPAR-alpha knockout mouse, the molar size was found to be slightly reduced, the enamel structure was found to be normal, but cervical molar enamel exhibited evidence suggesting hypomineralization. Although the PPAR-alpha knockout had no significant effect on molar morphology, the results suggest that active PPAR-alpha signaling is required to achieve normal mineralization of molar enamel, most probably through regulation of calcium homeostasis and metabolism of vitamin D. Cyp27b1 was expressed in tooth germ, suggesting that tooth germ can synthesize active vitamin D. Expression of Cyp27b1 was significantly enhanced in postnatal PPAR-alpha knockout tooth germ.

Distribution and structure of the initial dental enamel formed in incisors of young wild-type and Tabby mice.

Mouse incisor enamel can be divided into four layers: an inner prism-free layer; an inner enamel with prism decussation; outer enamel with parallel prisms; and a superficial prism-free layer. We wanted to study how this complex structural organization is established in the very first enamel formed in wild-type mice and also in Tabby mice where enamel coverage varies considerably. Unworn incisors from young female wild-type and Tabby mice were ground, etched, and analyzed using scanning electron microscopy. In both wild-type and Tabby mice, establishment of the enamel structural characteristics in the initially formed enamel proceeded as follows, going from the incisal tip in an apical direction: (i) a zone with prism-free enamel, (ii) a zone with occasional prisms most often inclined incisally, and (iii) a zone where prism decussation was gradually established in the inner enamel. The distribution of enamel in Tabby mice exhibited considerable variability. The sequence of initial enamel formation in mouse incisors mimics development from a primitive (prism-free) structure to an evolved structure. It is suggested that genes controlling enamel distribution are not associated with genes controlling enamel structure. The control of ameloblast configuration, life span, organization in transverse rows, and movement is important for establishing the characteristic mature pattern of mouse incisor enamel.

Changes in gene-expression during development of the murine molar tooth germ.

In a matter of a few days the murine tooth germ develops into a complex, mineralized, structure. Murine 30K microarrays were used to examine gene expression in the mandibular first molar tooth germs isolated at 15.5dpc and at 2DPN. Microarray results were validated using real-time RT-PCR. The results suggested that only 25 genes (3 without known functions) exhibited significantly higher expression at 15.5dpc compared to 2DPN. In contrast, almost 1400 genes exhibited significantly (P<0.015) higher expression at 2DPN compared to 15.5dpc, about half of which were genes with unknown functions. More than 50 of the 783 known genes exhibited higher than 10-fold increase in expression at 2DPN, amongst these were genes coding for enamel matrix proteins which were expressed several 100-fold higher at 2DPN. GO and KEGG analysis showed highly significant associations between families of the 783 known genes and cellular functions relating to energy metabolism, protein metabolism, regulation of cell division, cell growth and apoptosis. The use of bioinformatics analysis therefore yielded a functional profile in agreement with known differences in tissue morphology and cellular composition between these two stages. Such data is therefore useful in directing attention towards genes, or cellular activities, which likely are worthy of further studies as regards their involvement in odontogenesis.

Distribution and structure of dental enamel in incisors of Tabby mice.

OBJECTIVE: In Tabby mice, the Ta (EDA) gene is mutated. The resulting syndrome is homologous to hypohidrotic ectodermal dysplasia in humans. The Tabby phenotype is characterized by developmental defects of ectodermally derived structures. The teeth show aberrations in number, size and morphology. Dental enamel is a product of specialized epithelial cells, the ameloblasts. It was the aim of the present study to investigate the dental enamel phenotype in Tabby incisors, with emphasis on its distribution and structure. DESIGN: The incisors from five female Tabby and three female wild-type mice were sectioned and ground transversely, etched for 45s with 0.1% nitric acid, sputter-coated with gold-palladium, and observed in SEM. RESULTS: All measured dimensions were more variable in Tabby mice, as was the outline of the enamel-dentin junction. Maxillary incisors were wider in Tabby mice, while mandibular incisors were wider in wild-type mice. No significant difference in enamel thickness was observed. The enamel on the mesial aspect tended to extend further lingually in Tabby incisors in both jaws. On the lateral aspect, this tendency was only significant in mandibular incisors. The enamel-dentin junction often lacked the mesial concavity. Instances of hypoplastic enamel were observed. The complex mouse enamel structure was generally well preserved in Tabby mice, only few instances of aberrant structure were observed. CONCLUSIONS: It is suggested that the reciprocal expression pattern of Ta and Edar (the Ta ligand receptor gene) in outer and inner enamel epithelium, respectively, may influence the position of the enamel-cementum junction.

SEM observations of Retzius lines and prism cross-striations in human dental enamel after different acid etching regimes.

OBJECTIVE AND DESIGN: Sectioned and ground enamel surfaces need to be etched with acid in order for the enamel structure to be exposed for study in the SEM. In the present study Retzius lines and prism cross-striations were observed in variably ground specimens after etching with nitric acid of different concentrations (0.1, 1, 2.5 and 5%) and for different time periods (15, 30, 45, 90 and 180s). The aims were to improve our interpretation of these structures and to find the best etching regime. RESULTS AND CONCLUSIONS: Retzius lines in the superficial enamel were more readily exposed than Retzius lines in deeper parts of the enamel. The former showed discontinuity defects that facilitated their identification, while the latter lacked such defects. Few deep Retzius lines were exposed and could first be identified with certainty after etching for 30s in 1% acid. The structural and compositional basis for the visibility of deep Retzius lines may be subtle and difficult to identify. Scratches from the sectioning/grinding procedures may disturb their identification, but in general the grinding procedure was of minor importance for interpretation of enamel structure. Prism cross-striations were more readily exposed with the three stronger acid concentrations than with the weakest. They appeared as alternate light and dark bands, with a lower crystal concentration in the dark bands. No clear association with the prism varicosities/undulations was found. Etching with 1% nitric acid for 30s seems to be a good compromise for the study of Retzius lines and prism cross-striations.

A comparison of resins for embedding teeth, with special emphasis on adaptation to enamel surface as evaluated by scanning electron microscopy.

OBJECTIVE: Eight commercially available resins, Epoxy Embedding Medium ("Epon"), Spurr, Epofix, Epo-Kwick, Epo-Thin, Technovit 9100 New, Unicryl, and Vestopal W, all intended for embedding specimens for microscopy, were tested and compared for binding and adaptation to the enamel surface of human third molar crowns that had been embedded in them. DESIGN: After sectioning, grinding and acid etching, the specimens were observed in the SEM. The presence or absence of a gap between enamel and resin was recorded together with the width of the gap, when present. RESULTS: A pilot study, where gaps were observed before and after etching, indicated that the etching did not widen the gap appreciably. The resins could be divided into three groups according to gap width: "Epon" and Spurr performed best with a mean gap width of only 0.6microm. Vestopal W, Epofix, Epo-Kwick, Epo-Thin, and Technovit 9100 New constituted a group of medium performance with mean gap widths of 2.3, 3.4, 3.8, 3.9 and 3.9microm, respectively. Less satisfactory performance was observed for Unicryl, with a mean gap width of 7.4microm. CONCLUSIONS: It is concluded that "Epon" and Spurr should be chosen for high quality embedding of undecalcified teeth, while Epo-Kwick is an acceptable alternative if a faster procedure is needed.