Histological Evaluation of Allium sativum Oil as a New Medicament for Pulp Treatment of Permanent Teeth.

OBJECTIVE: The objective of this study was to evaluate the histo pathology effects of two medicaments Allium sativum oil and formocresol on the remaining pulp tissue of the permanent teething children. MATERIALS AND METHODS: A total of 18 premolars were included in this study. Two sound premolars were extracted and subjected to histological examination to show the normal pulp tissue. Pulpo tomy procedure was performed in the rest of the remaining 16 premolars; half of them using Allium sativum oil and the rest of the tested premolars were medicated using formocresol and all were sealed with suitable restoration. Then, premolars extracted at variable intervals (48 hours, 2 weeks, 1 month, 2 months), stained using hemotoxylin and eosin etain (H&E) and prepared for histopathology examination. RESULTS: Histological evaluation seemed far more promising for Allium sativum oil than formocresol. Histological evaluation revealed that teeth treated with Allium sativa oil showed infammatory changes that had been resolved in the end of the study. On the contrary, the severe chronic infammation of pulp tissue accompanied with formocresol eventually produced pulp necrosis with or without fibrosis. In addition, pulp calcification was evidenced in certain cases. CONCLUSION: Allium sativum oil is a biocompatible material that is compatible with vital human pulp tissue. It offers a good healing potential, leaving the remaining pulp tissue healthy and functioning.

Nigella sativa oil as a pulp medicament for pulpotomized teeth: a histopathological evaluation.

PURPOSE: Concerns about the safety of formocresol (FC) as a pulpotomy agent in Pediatric Dentistry have lead to the search of new capping medicaments. Indigenous plant medicines such as Nigella Sativa (NS) have been the focus of many researches. Therefore the purpose of this study was to investigate histo-pathologically the pulp response to NS oil and FC in dogs. METHOD: Forty teeth in 4 male dogs of undefined breed aging 12-14 months were used in this study. Coronal access cavities were performed on the upper and lower premolars so that both medicaments were tested in the same animal in alternate sides of the mouth. Four weeks after treatment the animals were sacrificed, paraffin sections were prepared for histological, histochemical and immuno-histochemical staining. RESULTS: Specimens in the NS group showed mild to moderate vasodilatation. Few specimens showed scattered inflammatory cell infiltration and the odontoblastic layer was continuous. While the FC group showed moderate to severe vasodilatation with high inflammatory cell infiltrate and degenerative changes. CONCLUSIONS: NS possesses an anti-inflammatory effect and the pulp maintains its vitality after its application, which could qualify its use as a pulp medicament for pulpotomized teeth in clinical practice.

Effects of low-level laser therapy and orthodontic tooth movement on dental pulps in rats.

OBJECTIVES: To describe the microscopic pulpal reactions resulting from orthodontically induced tooth movement associated with low-level laser therapy (LLLT) in rats. MATERIALS AND METHODS: Forty-five young male Wistar rats were randomly assigned to three groups. In group I (n = 20), the maxillary right first molars were submitted to orthodontic movement with placement of a coil spring. In group II (n = 20), the teeth were submitted to orthodontic movement plus LLLT at 4 seconds per point (buccal, palatal, and mesial) with a GaAlAs diode laser source (830 nm, 100 mW, 18 J/cm(2)). Group III (n = 5) served as a control (no orthodontic movement or LLLT). Groups I and II were divided into four subgroups according to the time elapsed between the start of tooth movement and sacrifice (12 hours, 24 hours, 3 days, and 7 days). RESULTS: Up until the 3-day period, the specimens in group I presented a thicker odontoblastic layer, no cell-free zone of Weil, pulp core with differentiated mesenchymal and defense cells, and a high concentration of blood vessels. In group II, at the 12- and 24-hour time points, the odontoblastic layer was disorganized and the cell-free zone of Weil was absent, presenting undifferentiated cells, intensive vascularization with congested capillaries, and scarce defense cells in the cell-rich zone. In groups I and II, pulpal responses to the stimuli were more intense in the area underneath the region of application of the force or force/laser. CONCLUSIONS: The orthodontic-induced tooth movement and LLLT association showed reversible hyperemia as a tissue response to the stimulus. LLLT leads to a faster repair of the pulpal tissue due to orthodontic movement.

Ca-binding domains in the odontoblast layer of rat molars and incisors under normal and pathological conditions.

We recently reported the presence of high concentrations of a Ca-binding matrix in the circumpulpal dentin of rat incisors which had been prevented from mineralization by a systemic administration of 1-hydroxyethylidene-1,1-bisphosphonate (HEBP), a type of bisphosphonates, thus suggesting the role of the putative Ca-binding matrix in the appositional mineralization of circumpulpal dentin (TAKANO et al., 1998, 2000; OHMA et al., 2000). In this study, we examined the distribution of Ca-binding domains in the pulp tissue of normal rat teeth and its changes under the influence of HEBP, in order to identify and clarify the role of the Ca-binding matrix in the physiological process of dentin mineralization. Observation of the normal rat tooth pulp showed occasional, tiny extracellular deposits of Ca-enriched material in the odontoblast layer, associated primarily with pericapillary regions. Such deposits were immunopositive for dentin sialoprotein (DSP), displayed high levels of X-ray peaks for calcium and phosphorus, and showed a drastic increase in amount by daily injections of HEBP. A brief vascular perfusion of high Ca-containing solution in normal animals caused the extensive deposition of Ca-P complexes along the basolateral membranes of odontoblasts but not in the other regions of the pulp tissue. These data suggest the existence of DSP-enriched extracellular Ca-binding domains in the odontoblast layer and also indicate a novel Ca-binding property of the basolateral membranes of odontoblasts. Since DSP is primarily synthesized as dentin sialophosphoprotein (DSPP) and later cleaved into dentin phosphophoryn (DPP) and DSP in odontoblasts, and since DSP has no notable affinity for Ca, the sites of DSP-immunopositive Ca-P deposits in the odontoblast layer may also contain DPP, a highly phosphorylated acidic protein having a strong binding property for calcium. Characteristic Ca-binding properties seen in the odontoblast layer appear to be related to the regulation of the appositional mineralization of circumpulpal dentin.

The anti-inflammatory effects of human recombinant copper-zinc superoxide dismutase on pulp inflammation.

Inflammation in the dental pulp is accompanied by release of a wide variety of highly oxidative molecules known as reactive oxygen species (ROS). ROS concentrations are controlled in vivo by an antioxidant enzyme scavenger system that may be overwhelmed by the increases in ROS production seen during inflammation. Supplementation of the antioxidant defense system, therefore, may limit the severity of the inflammatory response to injury due to this component. To test this hypothesis, this study examined the effects of superoxide radical scavenging on pulpal inflammation induced in rat molars by standardized cavity preparation. The extent of pulp inflammation was compared histomorphometrically between animals treated with exogenous administration of a human recombinant antioxidant enzyme, copper-zinc superoxide dismutase, conjugated to polyethylene glycol (hr-CuZn-SOD), versus saline-vehicle controls. There was a statistically significant reduction in area of inflammation involvement in those animals treated with hrCuZn-SOD, compared with controls. Although hrCuZn-SOD administration did not completely eliminate inflammation in all animals treated, there was a statistically significant lessening of the severity of the inflammatory response, as well as a greater degree of reparative dentin observed in the hrCuZn-SOD-treated animals.

Human cementum tumor cells have different features from human osteoblastic cells in vitro.

Cells obtained from human cementoblastoma and alveolar bone were isolated and cultured. Initial and late stages of mineralization were assessed by using atomic force microscopy, scanning electron microscopy and X-ray microanalysis. In cultures of cementoblastoma-derived cells the initial stages of mineralization showed well-defined spherical-shaped structures, while the osteoblastic cells showed plaque-like deposits. These morphological patterns of mineral deposition could serve as nucleation centers for hydroxyapatite crystals. Late stages of mineralization at 28 and 35 d maintained those morphological differences established in initial cultures. The material deposited by cementoblastoma and osteoblastic cells, analyzed by EDX spectra, revealed similar Ca/P ratios for both cell types. These values were similar to those reported for hydroxyapatite in enamel and bone. Alkaline phosphatase specific activity (AlP), of osteoblastic cells at 3, 7 and 11 d, showed an increase of 27.9, 50.9 and 37.0% (p < 0.001), respectively. However, at 15 and 19 d there was an increase of AlP activity of cementoblastoma cells by 39.4 and 34.5% over osteoblastic cells (p < 0.001). Immunostaining of cementoblastoma and osteoblastic cells using a specific mAb against a cementum-derived attachment protein revealed strong immunostaining of cementoblastoma cells which was localized to the cell membrane and fibril-like structures (96.2 +/- 1.3). A few osteoblastic cells also stained weakly with the anti-CAP mAb (6.4 +/- 0.6). Sections of decalcified paraffin embedded cementoblastoma specimens, when immunostained with anti-CAP mAb, showed strong immunostaining of the cells surrounding the regular and irregularly-shaped calcified masses of the tumor. Putative cementocytes also stained positively. Immunostaining with a polyclonal antibody against osteopontin strongly stained the osteoblastic cells (89.0 +/- 3.6). Cementoblastoma cells showed weaker staining (54.2 +/- 2.4). The results suggest that cementoblastoma cells could be a major source of specific cementum proteins. These cells could provide the opportunity to elucidate the regulation of the cementogenesis process.

Effects of essential fatty acid deficiency on rat molar pulp cells.

In rats fed an essential fatty acid deficient (EFAD) diet, either during pregnancy (DN) or for 4 wk postnatally (ND), the cell density in the central part of the pulp increased about two- and threefold, respectively, of that in rats who had received a conventional diet containing sunflower oil. Cells were especially numerous around capillaries. The cell density was also increased twofold in the subodontoblastic layer in the outer part of the pulp, cells being smaller in ND compared with DN. In contrast, the odontoblasts were reduced in height, and the Höhl cells formed a thin layer in EFAD rats. This emphasizes some aspects of pulp specificity which reacted differently from odontoblasts. We suggest that the function of killer cells which normally destroy cells at the periphery of the pulp may be impaired by the diet, leading to cell accumulation.

The occurrence of interglobular dentin in incisors of hypophosphatemic mice fed a high-calcium and high-phosphate diet.

The incisor dentin of hypophosphatemic (Hyp) mice was examined histopathologically to determine whether the multiple occurrences of interglobular dentin would be influenced by the serum phosphate level. Both normal and Hyp mice (12 weeks of age) were divided into two groups. The mice in one group were given a control diet (1.42% Ca, 1.16% P) and the other a high-calcium and high-phosphate diet (2.00% Ca, 3.00% P) for 30 days. Blood was collected from the mice every fifth day for measurement of the calcium and phosphate concentrations in serum. Both ground and decalcified cross-sections were prepared from incisors from the mandible and maxilla for microscopic examination. The levels of serum Ca and P were almost constant in normal mice, regardless of diet. On the other hand, serum P levels in Hyp mice fed the control diet were significantly lower than those in normal mice. The ten days' feeding of the high-Ca/-P diet significantly elevated the serum P level in Hyp mice, and it reached a level similar to that of the normal mice. However, histopathological examination showed no significant changes in incisor dentin of Hyp mice fed the high-Ca/-P diet, and interglobular dentin still occurred. These results suggest that the multiple formations of interglobular dentin, which is the most outstanding feature of X-linked hypophosphatemic vitamin-D-resistant rickets, are not influenced in Hyp mice by the short-time normalization of the serum phosphate level.

The role of ascorbic acid on the structural integrity of developing tooth germs.

Tooth germs grown in ascorbate deficient medium for up to 20 days underwent progressive and widespread changes. Proliferation and differentiation of preameloblasts and preodontoblasts progressed normally. Newly differentiated odontoblasts, however, became vacuolated when they began secreting: this suggested a metabolic disturbance. Failure to maintain differentiated odontoblasts, ameloblasts and pulpal cells resulted in aberrant dentin matrix, cessation of dentin production, and finally overall structural collapse with loss of normal morphology. Biochemical studies then were undertaken to define the lesion involved. The relative rate of collagen synthesis in ascorbate deficient cultures was comparable to that of ascorbate supplemented cultures, but the collagen was found to be underhydroxylated. In this state it would be unstable at 37 degrees and subject to preferential degradation. This correlates with the observation that a major fraction of the hydroxyproline in the scorbutic cultures was found in the medium as small molecular weight peptides. The overall effect of ascorbate deficiency was to deprive the tooth germ of the normal quality and quantity of collagen resulting in the characteristic histological and structural abnormalities observed. Flattening and deterioration due to structural failure most likely resulted from abnormal extracellular matrix synthesis in the supportive pulp and dentin due to the aberrant collagen.