Scanning electron microscopy of early microbial colonization of human enamel and root surfaces in vivo

Abstract – This study describes the pattern of microbial colonization in six dental students, who carried test pieces of enamel and root surface in intraoral acrylic appliances for 4, 8, 12, 24, and 48 h, respectively. Oral hygiene was discontinued during the experimental periods. The results showed that oral microorganisms colonized enamel and root surfaces individually. The pattern of colonization mimicked the surface structure of the tooth; on enamel surfaces the first bacteria appeared in pits and surface irregularities followed by proliferation along the perikymata, while on root surfaces bacterial colonization was characterized by a haphazard distribution. Within the initial 24-h period root surfaces were more heavily colonized than were enamel surfaces. This pattern was characteristic irrespective of differences in the intra- and interindividual rates of bacterial coverage. It is concluded that early microbial colonization in vivo is determined by the surface structure of the tooth. Consequently, natural tooth surfaces should not be replaced by artificial surfaces in structural studies of microbial deposits in the oral cavity.     

Transmission electron microscopy of early microbial colonization of human enamel and root surfaces in vivo

This study describes the early microbial colonization of teeth by the use of light- and transmission electron microscopy. Six dental students carried a total of 60 test pieces of unerupted enamel and root surface in intraoral acrylic appliances for 4, 8, 12, 24 and 48 h, during which periods oral hygiene was abandoned. Pronounced variations were recorded in structure and thickness of the pellicle across the individual surfaces of both dental tissues. Bacterial single-cell colonization increased the electron density of the adjoining pellicle. Micro-colonies of bacteria were observed in relation to enamel surface irregularities such as perikymata, while the distribution on root surfaces appeared incidental. Root surfaces were generally colonized by thicker deposits than homologous enamel surfaces although the structural composition of the microbiota was similar. Gram-positive bacteria with thick cell walls appeared in coccoid or rod-shaped configurations depending on the age of the bacterial deposit. These bacteria were further characterized by selective invasion between collagen fibers. After 48 h the complexity of the microbiota was increased by the establishment of new bacterial species in the superficial layer. It is concluded that the pattern and composition of the early microbiota on teeth is more complex and variable than hitherto assumed.     

Scanning electron microscopy of early microbial colonization of human enamel and root surfaces in vivo

This study describes the pattern of microbial colonization in six dental students, who carried test pieces of enamel and root surface in intraoral acrylic appliances for 4, 8, 12, 24, and 48 h, respectively. Oral hygiene was discontinued during the experimental periods. The results showed that oral microorganisms colonized enamel and root surfaces individually. The pattern of colonization mimicked the surface structure of the tooth; on enamel surfaces the first bacteria appeared in pits and surface irregularities followed by proliferation along the perikymata, while on root surfaces bacterial colonization was characterized by a haphazard distribution. Within the initial 24-h period root surfaces were more heavily colonized than were enamel surfaces. This pattern was characteristic irrespective of differences in the intra- and interindividual rates of bacterial coverage. It is concluded that early microbial colonization in vivo is determined by the surface structure of the tooth. Consequently, natural tooth surfaces should not be replaced by artificial surfaces in structural studies of microbial deposits in the oral cavity.     

Scanning electron microscopic observations of early plaque formation in vivo on enamel specimens treated with delmopinol

Abstract The aim of this double blind, cross-over, scanning electron microscopic (SEM) study was to compare the effect of topical application of 0.5% delmopinol HCl (aqueous solution) with placebo on early supragingival plaque formation on enamel specimens attached to a maxillary premolar. 3 subjects underwent 2 × (placebo and delmopinol treatment, respectively) 7 treatment periods (1/2, 1, 2, 8, 24 hours; 3, 7 days) of undisturbed dental plaque accumulation, during which 2 ml of placebo and delmopinol HCl 0.5%, respectively, were applied topically to all teeth 2 × daily. At the end of each period, the specimen was retrieved and processed for SEM. For the specimens obtained after 24 h or less, the number of visible micro-organisms was estimated. For the 3- and 7-day specimens, the area covered by plaque was calculated planimetrically. Large inter-individual differences were observed in the number of bacteria attaching during the first 24 h, with apparently no major differences between delmopinol and placebo. 1 subject showed a bacterial colonization predominated by curved rods throughout the first 8 h. Compared to placebo, the area of the specimens covered by plaque was reduced after 3 and 7 days of delmopinol use, with coccoid cells as the predominant morphotype. Plaque maturity, characterized by the appearance of corncob formations and high numbers of different morphotypes, was not observed on the delmopinol specimens, in contrast to the placebo specimens. The study shows that topical application of 0.5% delmopinol HCl delays and interferes with dental plaque maturation.     

Microbial invasion and subsurface colonization of rat enamel in early fissure caries observed by scanning electron microscopy

Abstract – In order to find out whether microbes can be detected in the incipient rat caries lesions underneath the visibly intact enamel surface, the maxilla of OM-rats, fed a cariogenic diet, were fixed and partially bisected sagittally with a thin saw. The individual teeth were finally split with a knife to get a crack surface from the fissure region. Selected tooth halves showing slight carious changes at the dentin-enamel junction were prepared for SEM and examined. In many preparations, colonies of organisms were found firmly attached to the crack surface. Single, predominantly coccus-like organisms and a few rods could be seen mostly located in the interprismatic furrows at varying depth throughout the enamel. Somc lacuna-like hollows, with or without the occupant organisms, could be seen carved in the prism structure. The enamel side of the DEJ in the lesion area could be completely covered by the microbes. In samples representing arrested caries, amorphous precipitates were found at the interprismatic substance of the DEJ region.     

Topographic distribution of subgingival plaque along root surfaces of human periodontally diseased teeth

Abstract The purpose of this study was to characterize the topographical distribution and organization of subgingival plaque in periodontally diseased teeth. 26 extracted teeth were fixed and processed for undecaicified histological evaluation. The sections were cut perpendicular to the long axis of the tooth and analyzed by phase-contrast microscopy. The coronal portion of the analyzed roots showed a dense accumulation of filamentous forms, fusiform rods, coccoid forms and loosely aggregated spirochetes. The middle and apical portions showed a non-uniform distribution of the microflora, with microorganisms representing all the known morphotypes. Furthermore, plaque was detected below undisturbed periodontal fibers, indicating that plaque not only forms apically, but also in a lateral direction, penetrating and colonizing below areas where periodontal fibers are inserted into the root surface.     

Exaggerated abrasion/erosion of human dental enamel surfaces: a case report

An atypical, rapidly proceeding abrasion/erosion of the labial enamel surfaces of the maxillary and mandibular incisors and canines in a 27-yr-old man is reported. Ultrastructural examination of a replica of the teeth showed a practically structureless enamel surface both at the initial examination and after 12 months. However, at the end of the period, minor areas of dentin tubules became visible, indicating that a substantial loss of the tooth substance had taken place. The patient's occupation involved daily environmental contact with proteolytic enzymes. In vitro study of enamel exposed to one of the actual proteolytic enzymes showed dissolution of enamel substance, and it cannot be excluded that enzymatic decomposition of the organic enamel matrix is a contributing cause of the observed exaggerated loss of tooth substance.     

Microbiology of the early colonization of human enamel and root surfaces in vivo

This study describes the predominant early microflora on human teeth on the basis of microbiologic identification of 1742 fresh isolates. The isolates were obtained from four dental students who carried test pieces of enamel and root surface in the oral cavity for 4, 8, 12, and 24 h. During the experimental periods oral hygiene was discontinued. Under equal conditions root surfaces were more heavily colonized than were enamel surfaces. However, the composition of the microbiota was the same. Within the first 24 h the microflora was dominated by streptococci and Gram-positive pleomorphic rods. S. sanguis contributed only 6-18% of the early colonizers whereas S. mitis and S. oralis varied between 24-42% and 1-27% (mean values), respectively. The relative proportion of S. oralis increased significantly within the observation period while the proportion of S. salivarius and arginine-positive S. mitis showed a declining tendency. Actinomyces species adsorbed to the tooth surfaces within the first 4 h but did not increase their relative proportions until after 8-12 h, possibly due to a long doubling time. In one individual, encapsulated bacteria resembling Stomatococcus mucilaginosus were observed among the early colonizers. The time-dependent shifts in the bacterial populations within 24 h corroborate parallel ultrastructural findings.     

Scanning electron microscopy of fluorideinduced disturbances on the enamel surface of rat molars

Abstract — Rats were given a single high dose of fluoride at the age of 5 days and killed after 24 h, 10 or 15 days. The maxillary Erst molars were prepared for scanning electron microscopic examination. It was concluded that a single dose of fluoride, preferentially affecting ameloblasts with a high secretory activity, leads to the formation of subameloblastic cysts and enamel hypoplasias covered with granular deposits.     

Effect of plasma on composition of human enamel and cementum pellicle

Abstract – Slabs of human enamel and cementum were incubated with plasma alone or with various mixtures of plasma and saliva. Proteins and glycoproteins that adsorbed to the surface of the slabs in 0 to 60 min were labeled by lactoperoxidase-catalyzed ¹²⁵I-iodination and by mild periodate oxidation followed by NaB³H₄ reduction. The labeled components were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and visualized by autoradiography or fluorography. From plasma alone, a 58 and a 66 kDa protein (probably albumin) were adsorbed to the enamel surface in relatively equal amounts, but no ¹²⁵I-labeled components were detected on the cementum surface in the absence of saliva. Adding 10% saliva to the incubation mixture promoted the adsorption of the 58 and 66 kDa components to cementum. In addition, another set of proteins, including components of 44, 47, 29, and 25 kDa, was adsorbed to both cementum and enamel in the presence of saliva. These six proteins were the major ¹²⁵I-labeled species in all of the pellicles formed from mixtures of plasma and saliva. The electrophoretic mobility of the major 120 and 140 kDa ³H-labeled sialoglycoproteins adsorbed to both cementum and enamel was similar to that of the low-molecular-weight mucin of submandibular/sublingual saliva.     

Circumferential continuity of perikymata in human dental enamel investigated by scanning electron microscopy

Abstract – Theoretically the perikymata may represent one of two possible configurations: closed circles or continuous spirals. In the present study one randomly selected perikyma groove in one randomly selected tooth (the mandibular first premolar of a 12-yr-old girl) was tracked around the circumference of the crown in the scanning electron microscope. It was found that the perikymata geometrically represented closed circles. The possibility of methodologic error was excluded.     

Organic structures of developmental origin in human surface enamel

Abstract – The aim of this study was to examine the distribution of organic material in mature enamel surfaces immediately prior to eruption. Thirty-six samples of buccal or lingual enamel from unerupted third molars were prepared for transmission electron microscopy by a method involving demineralization of the enamel after embedding in Epon. The results showed that at time of eruption human surface enamel is a highly porous structure containing large amounts of developmental protein which appear as a variety of triangular, funnel-shaped, or invaginated configurations extending into the enamel. The implications of this finding may be of importance to the understanding of early caries lesion formation in the enamel since organic structures may modify the diffusion of ions in and out of the tissue.     

A scanning electron microscopic and microradiographic study of pits in fluorosed human enamel

Abstract— The aim of the present study has been to correlate the surface appearance of pitted, fluorosed enamel in the scanning electron microscope (SEM) with the microradiographic features of the underlying tissue. Intact enamel surfaces of severely fluorosed teeth appeared similar to that of normal enamel. In the unabraded surfaces discrete pits were sharply demarcated from the surrounding intact enamel surface leaving steep walls of parallel running rods. The microradiographic appearance of sections made through pits indicated that focal loss of surface enamel occurred corresponding to the inner highly hypomineralized part of the fluorotic subsurface lesion. The abrupt wall formation and the finding that the striae of Retzius never changed direction along the margins of the lesions indicated that pits in fluorosed enamel may be secondarily produced defects rather than true hypoplasias. Further evidence of the posteruptive origin of the defects was derived from the observation that enamel lamellae occasionally formed part of the lateral border. The relatively higher degree of radiopacity observed in the tissue surrounding the pit indicates a posteruptively acquired deposition of minerals in the exposed porous enamel.     

Light and transmission-electron microscopy study of enamel organ cells in germectomized human third mandibular molars

Abstract— Using nine serially sectioned germectomized mandibular third molars it was possible to examine light microscopical (LM) and transmission-electron microscopical (TEM) features of maturing human enamel organ cells. The degree of enamel mineralization was estimated by quantitative imbibition studies in polarized light. It was possible to distinguish between three progressive stages of enamel mineralization. The most advanced stage was characterized by external enamel porosity. In the least advanced stages the enamel porosity appeared more extensive beneath a less porous surface layer. Ruffle- and smooth-ended ameloblasts were identified corresponding to the maturing enamel. Smooth-ended ameloblasts were the most frequently observed. However, no preferences for one of the two cell types could be observed in relation to the different stages of enamel mineralization. The maturing human enamel organ cells broadly revealed the same characteristics with respect to morphology features, intracellular organization, and junctional complexes as described in the maturation zone of the rat incisor enamel organ. Our findings therefore add to the view that the basic pattern of amelogenesis is identical in human and rat incisor enamel.     

Uncoated specimen of human enamel obeerved in the scanning electron microscope

Abstract – A1-mm-thick section of human premolar was acid etched and observed uncoated in the SEM at accelerating voltages of 3, 5, 10 and 15 k v . Prisms and interprismatic substance were easily distinguishable. Low-voltage operation (3 and 5 k v ) gave the best results. Specimen charging was detectable at 5 k v and caused reduced image quality at 10 and 15 k v . Application of silver paste did not reduce charging appreciably. Prolonged observation at high magnification (x 10 000) resulted in contamination of the specimen with consequent charging and reduced resolution. Dental enamel seems to be a material which is well suited when uncoated for obervation in the SEM. This may be due both to the high content of the relatively heavy atom calcium, giving good secondary electron emission, and possibly to a certain degree of conductivity caused by diffusible icons.     

The ultrastructure of fluorosed human dental enamel

abstract — More than 80 specimens of enamel from 20 ground sections of eight teeth showing severe dental fluorosis have been examined in the transmission electron microscope. The study has shown that the prisms have normal size and form, but are surrounded by large periprismatic gaps. The individual crystals commonly appear in cross sections as regular, flattened hexagons with an average width of 91 nm and average thickness of 45 nm. Particularly in the tail regions and at prism peripheries, extensive intercrystalline spaces were found. The crystal lattice demonstrated a spacing of 0.82 nm. Demineralization of thin sections with PTA showed a preferential dissolution of the crystal centers. Based on the ultra-structural observations it is proposed that the hypomineralized regions of fluorosed enamel are produced by an arrest of enamel maturation, possibly due to toxic damage to the ameloblasts in the absorptive or maturative phase of their life cycle.     

In vivo early human dental plaque formation on different supporting substances. A scanning electron microscopic and bacteriological study.

Different studies have shown that various substances may have an influence on early human dental plaque formation. The purpose of the present study was to compare on tooth substances and supporting prosthetic materials the amount of plaque deposition by SEM and the quantity of selected bacteria using anaerobic culturing techniques. 5 bridges, replacing a missing molar or premolar, were incorporated in 3 patients. In the midbuccal area of each pontic, a semi-precision attachment was placed allowing the insertion of the following test facings: enamel, dentine, non gamma 2-amalgam, alloys of 85% and 55% gold, silver-palladium, chrome-cobalt, chrome-cobalt-titanium, and ceramic. For each material, 2 facings were fabricated. After 4 and 24 hours in situ, bacteriological samples were taken and processed for further identification. After a 2nd period of 4 and 24 hours in situ, the same facings were carefully removed and prepared for SEM-examination. All 4-hour specimens exhibited various areas covered by plaque, the amount of which varied with the different supporting substances. The very smooth surfaces (e.g., gold) harbored sparse deposits, while the rougher (e.g., amalgam) were covered by more plaque. After 24 hours of plaque development, an increase in the number of micro-organisms was noted for all the specimens. After 4 and 24 hours of plaque accumulation, no specific trends suggesting a preferential colonization on the different substances were observed. This study has shown that the amount of early deposits on different substances seems to be related to the degree of their surface roughness, while plaque formation was qualitatively similar.     

人牙釉质酸蚀过程中三维形貌的原子力显微镜观察

目的 比较不同酸蚀时间处理后牙釉质表面的三维形貌并分析粗糙度的改变,探讨酸蚀对牙釉质微观形貌的影响.方法 将15颗新鲜前磨牙包埋、切割、打磨后得到光滑牙釉质切片,按单纯随机法分为5组,用37％磷酸凝胶分别酸蚀0(对照组)、5、10、20和30s,原子力显微镜观察釉质表面形貌,分析轮廓改变,计算图像的算术平均粗糙度(Ra)、均方根粗糙度(Rq)、十点高度平均差(Rz)值及表面积、容积并对5组的结果数据进行单因素方差分析,用Turkey's法进行组间两两比较,以P＜0.05为差异有统计学意义.结果 镜下形貌观察显示,0～20 s内釉质的釉柱结构逐渐明显,釉质表面由较光滑逐渐变为典型鱼鳞状;20 s后表面形态变化不明显.表面轮廓观察显示,随着酸蚀时间由0s增加至30 s,釉柱间隙首先被溶解,酸蚀深度逐渐增加到2.8μm,30 s后釉柱顶部宽度减小至1.8μm,且高度略降低.从0s到30 s,Ra值由(19.69±3.42) nm增加到(359.51±75.79)nm,Rq值由(22.02±5.57) nm增加到(431.02±83.09) nm,Rz值由(0.24±0.08) μm增加到(2.38±0.26) μm;组间比较显示,Ra和Rq值的20s组与30 s组和Rz值的10、20和30 s组差异无统计学意义(P＞0.05),其余各组间差异均有统计学意义(P＜0.05).同时釉质表面积由(406.77±3.88) μm2 (0 s)增大到(546.69±84.02) μm2 (30 s),容积由(65.73±14.46) μm3 (0 s)增大到(474.63±52.50)μm3(30 s).结论 随着酸蚀时间的增加,釉质表面酸蚀深度不断增大,表面粗糙度增加,表面积及容积增大,酸蚀会造成釉质微观形貌的明显改变.