Requirement of proper occlusal force for morphological maturation of neural components of periodontal Ruffini endings of the rat incisor

The present study examined the effect of reduced occlusal force on morphological maturation of periodontal Ruffini endings, primary mechanoreceptors in the periodontal ligament, of the rat incisor. The reduction of occlusal force was induced by grinding the cutting edges of unilateral incisors of the rat from postnatal day 14 (PN14d), when periodontal Ruffini endings are immature. Under normal development, the axon terminals of Ruffini endings gradually ramified with the passage of time, and showed ruffled outlines having numerous dot-like structures around PN28d. When the mechanical stimulation was reduced, appearance of dot-like structures at the axon terminals delayed. Quantitative analysis elucidated that the percentages of immunoreactive areas for protein gene product 9.5, a marker protein of neural elements, at ground side were significantly smaller than those at non-ground side 14 days following the initial grinding. The distribution and morphology of terminal Schwann cells was not apparently affected. The present results indicate that the proper mechanical stimulation to the ligament contributes to the morphological maturation of the periodontal Ruffini endings.     

Reinnervation in the canine periodontal ligament of replanted teeth using an antibody to protein gene product 9.5: an immunohistochemical study

Abstract— The re-innervation process in the periodontal ligament of replanted canine teeth was examined by immunohistochemistry for protein gene product 9.5 (PGP 9.5), a general marker for neurons, and by electron microscopy. Within 1 week of replantation, the periodontal fibers had regenerated, filling the narrow spaces between the alveolar bone and the root surface around the cervical and apical regions. Near the root apex, however, no PGP 9.5-im-munopositive nerve fibers were found in the regenerated periodontal ligament except for those in the alveolar half of the ligament. At 2 weeks after replantation, many nerve fibers positive for PGP 9.5 had ascended the periodontal ligament from the thick nerve bundles located near the root apex. Fine nerve endings showing complicated ramification were also present in the apical region. By 3 or 4 weeks after replantation, the vascular network was regenerated and principal periodontal fibers were re-established throughout the entire length of the periodontal ligament. The extensively ramified PGP 9.5-immunopositive structures had increased in thickness and density and showed characteristic treelike profiles by 3 weeks. Electron microscopy confirmed that most of these structures were Ruffini-like endings, and demonstrated that such nerve terminals were almost regenerated by 4 weeks post-replantation. These results indicated that, in the periodontal ligament of replanted canine teeth, the regeneration of the nerve fibers including mechanosensory receptors first showed signs of regeneration by 2 weeks following tooth replantation and proceeded rapidly thereafter. Regeneration of the periodontal ligament including fiber architecture as well as vascular and neural elements was almost complete by 4 weeks after replantation.     

Re-innervation in the canine periodontal ligament of replanted teeth using an antibody to protein gene product 9.5: an immunohistochemical study

The re-innervation process in the periodontal ligament of replanted canine teeth was examined by immunohistochemistry for protein gene product 9.5 (PGP 9.5), a general marker for neurons, and by electron microscopy. Within 1 week of replantation, the periodontal fibers had regenerated, filling the narrow spaces between the alveolar bone and the root surface around the cervical and apical regions. Near the root apex, however, no PGP 9.5-immunopositive nerve fibers were found in the regenerated periodontal ligament except for those in the alveolar half of the ligament. At 2 weeks after replantation, many nerve fibers positive for PGP 9.5 had ascended the periodontal ligament from the thick nerve bundles located near the root apex. Fine nerve endings showing complicated ramification were also present in the apical region. By 3 or 4 weeks after replantation, the vascular network was regenerated and principal periodontal fibers were re-established throughout the entire length of the periodontal ligament. The extensively ramified PGP 9.5-immunopositive structures had increased in thickness and density and showed characteristic tree-like profiles by 3 weeks. Electron microscopy confirmed that most of these structures were Ruffini-like endings, and demonstrated that such nerve terminals were almost regenerated by 4 weeks post-replantation. These results indicated that, in the periodontal ligament of replanted canine teeth, the regeneration of the nerve fibers including mechanosensory receptors first showed signs of regeneration by 2 weeks following tooth replantation and proceeded rapidly thereafter. Regeneration of the periodontal ligament including fiber architecture as well as vascular and neural elements was almost complete by 4 weeks after replantation.     

Growth-associated protein-43 immunohistochemical and ultrastructural changes in jaw muscle spindles of the rat following loss of occlusion

The effects of complete loss of occlusion on the structural and functional status of these muscle spindles were investigated by immunohistochemistry either for protein gene product 9.5 (PGP 9.5) or growth-associated protein-43 (GAP-43) by light and electron microscopy. All the upper molars of 4-week-old Wistar rats were extracted and the erupted portions of the upper and lower incisors of the same animals were cut-off at the level of the interdental papilla every other day. In a control group, immunoreactivity for GAP-43 was positive in the developing annulospiral endings of 2-week-old rats, but was not detected in any of the muscle spindles after 3 weeks of age. At 4 weeks of age, the PGP 9.5 immunostained spindles had well-differentiated annulospiral endings. Ultrastructurally, these afferent endings showed lenticular or circular profiles in cross-sections, and were differentially indented into the intrafusal-fibres. The inner surfaces of the terminals formed rather smooth myoneural junctions, while the outer surfaces were covered only by basal lamina continuous with that of the underlying intrafusal muscle fibres. After the experimental elimination of occlusal contact, GAP-43 immunoreactivity reappeared in some nerve endings of muscle spindles by 3 days, and persisted for at least 28 days. During this period, the afferent-terminals exhibited various fine structural abnormalities such as irregular outlines and invaginated neuromuscular interfaces. Some sensory-terminal (ST) profiles were completely engulfed by intrafusal-fibres. However, GAP-43 expression and ultrastructural alterations became undetectable within a week of the end of incisal cutting and the recovery of incisal-contact. These data indicate that remodelling of nerve terminals in muscle spindles, as assessed by GAP-43 expression and ultrastructural changes, occurs soon after a loss of occlusion, and ceases if incisal-contact is restored. It is concluded that possible changes in jaw muscle function, as well as a sudden loss of proprioceptive sensory input from the periodontal mechanoreceptors of molars and incisors, induce the structural reorganisation of nerve terminations in jaw muscle spindles that is associated with the appearance and disappearance of GAP-43 immunoreactivity.     

Evidence for the existence of intraepithelial nerve endings in the junctional epithelium of rat molars: an immunohistochemical study using protein gene product 9.5 (PGP 9.5) antibody

Innervation of the junctional epithelium was investigated in rat molars by means of immunohistochemistry for protein gene product 9.5 (PGP 9.5) at light and electron microscopic levels. In comparison with our previous study on same tissues using neurofilament protein (NFP)-antibody, the PGP 9.5-immuno-staining further disclosed numerous nerve fibers in the gingiva of rat molars and revealed the existence of a well-developed plexus of PGP 9.5-positive nerve fibers. The interproximal portion also contained numerous nerve fibers.
Observation of horizontal sections revealed a denser innervation toward the inner junctional gingival epithelium than the outer marginal epithelium. The nerve fibers, beaded in appearance and extending from the nerve bundles in the lamina propria, penetrated into the junctional epithelial layer and were distributed throughout the junctional epithelium, with some nerves being located near the epithelial surface. Non-neuronal cells showing PGP 9.5-immunoreactivity were absent in the junctional epithelium. In immunoelectron microscopy, the axoplasm of nerves in the gingiva was filled with electron-dense reaction products of PGP 9.5, except for the cell organellae. The nerve fibers were devoid of Schwann cell investment and terminated among the epithelial cells in the junctional epithelium, frequently beneath the epithelial surface. The intraepithelial nerve endings contained various kinds of vesicles including large-cored ones, supporting the presence of peptidergic innervation shown by previous studies. These findings confirmed the usefulness of PGP 9.5-immunohisto-chemistry for the identification of delicated nerve fibers in dental tissue, and suggested the dense network of nerve fibers that may serve as sensory receptor and other functions in the junctional epithelium.     

大鼠磨牙及牙周组织中神经纤维分布:神经丝蛋白免疫组织化学研究

目的：观察大鼠磨牙及牙周组织神经纤维分布特点。方法：大鼠磨牙及其牙槽骨脱钙后切征并行神经丝蛋白（ＮＦＰ）免疫,组织化学染色。结果：大鼠磨牙、牙周膜及牙龈中神经纤维丰富,神经丝蛋白阳性神经纤维在髓角处形成牙本质细胞下丛,其中一些神经纤维进入牙本质。在牙周膜,神经丝蛋白阳性神经纤维在牙根下段密集分布 。因牙周膜内存在两型神经来梢：游离神经来梢及Ｒｕｆｆｉｎｉ终末。前者在整个牙周膜内皆有分布。后者常成组     

Altered expression level of calbindin D28k in the periodontal ligament of rat molar in response to changes in occlusal force

The present immunohistochemical study was designed to investigate the alteration in the expression level of calbindin D28k in the periodontal ligament of the rat molar in response to changes in occlusal force to clarify the physiological role(s) of this protein in the ligament. In normal periodontal ligament of the lower first molar, immunoreactivity for calbindin D28k was found in the spindle-shaped cells, presumably fibroblasts, at the alveolar portion of the ligament at the distal side of the mesial root and mesial side of the distal root. Following the overload of occlusal force to the upper first molar by bite-raising, the number and immunoreactivity of the positive cells in the periodontal ligament of the lower first molar increased gradually. A more significant increase was detected at 7 d following the bite-raising compared to the normal animals. When occlusal force was removed by the extraction of the upper first molar, the expression level of calbindin D28k in the periodontal ligament of the lower first molar rapidly decreased, however a subsequent gradual increase was recognized. Statistical analysis of the spatial immunoreactivity of calbindin D28k in the periodontal ligament was performed and showed statistically significant differences. The present results suggest that calbindin D28k may play important roles in the homeostasis and cytoprotection of the periodontal fibroblasts against occlusal force.     

Morphological changes in periodontal mechanoreceptors of mouse maxillary incisors after the experimental induction of anterior crossbite: a light and electron microscopic observation using immunohistochemistry for PGP 9.5

Ruffini nerve endings (mechanoreceptors) in the periodontal ligament (PDL) of mouse incisors were examined to elucidate whether experimentally-induced crossbites cause any changes or abnormalities in their morphology and distribution. Anterior guiding planes were attached to the mandibular incisors of 3-week-old C3H/HeSlc mice. At 3 days and 1, 2, 4, 6, and 8 weeks post-attachment of the appliance, the mice were sacrificed by perfusion fixation. Frozen sagittal cryostat sections of the decalcified maxillary incisors were processed for immunohistochemistry of protein gene product 9.5, followed by histochemical determination of tartrate-resistant acid phosphatase activity to reveal sites of alveolar bone resorption. Despite the absence of bone resorption within the lingual PDL of control mice, distinct resorption sites were seen in the respective regions of the experimental animals. Unlike the controls, many Ruffini endings showing vague and swollen contours, with unusually long and pedunculated micro-projections were observed in the affected lingual PDL of the incisors in the experimental animals with short-term anterior crossbite induction. Club-shaped nerve terminations with few, if any, micro-projections were observed in the lingual PDL of experimental animals with long-term induction, as well as in aged control mouse incisors. Differences in the distribution of Ruffini endings were also observed. These results indicate that changing the direction of the force applied to the PDL results in rapid and prolonged changes in the morphology of Ruffini-like mechanoreceptors.     

Development and terminal differentiation of pulp and periodontal nerve elements in subcutaneous transplants of molar tooth germs and incisors of the rat

Ectopic tooth transplants are known to receive rich innervation of local neurons, but the precise location and structural features of neurites in the pulp and periodontal ligament (PDL) of such transplants are unclear. In this experiment, the molar tooth germs of rat embryos and incisors of young rats were subcutaneously transplanted into the dorsal regions of rats and processed, at various time intervals, for immunohistochemical demonstration of neural elements. Teeth with periodontal tissue elements developed in most of the molar transplants in 6 or 8 wk and received rich innervation, including some autonomic fibres, in the pulp. Nerve elements were also confirmed to be present in the PDL of these transplants, including specialized nerve ending-like structures reminiscent of the periodontal Ruffini endings. Mechanoreceptor-like structures were also induced in the regenerated PDL of similarly transplanted incisors, although the success rate was low. We conclude that rich and highly ordered innervation of the pulp, and occasional development of mechanoreceptors in the regenerated PDL of ectopic dental transplants, imply a high probability of successful induction of teeth with both nociceptive and mechanical sensations in the ectopic tooth and/or tooth germ transplant systems, although differentiation of mechanoreceptor-like nerve endings occured in only a few rare cases.     

Immunohistochemical localization of SNARE core proteins in intrapulpal and intradentinal nerve fibers of rat molar teeth

ObjectiveThe present study was designed to elucidate whether three soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor (SNARE) core proteins, syntaxin-1, synaptosomal-associated protein of 25 kDa (SNAP-25), and vesicle-associated membrane protein-2 (VAMP-2), are present in the dental pulp of the rat molar at both the light and electron microscopic levels.DesignImmunohistochemistry for protein gene product 9.5 (PGP 9.5), a pan-neuronal marker, syntaxin-1, SNAP-25, and VAMP-2 was performed on decalcified rat molars for light and electron microscopic analyses. Double-immunolabeling of PGP 9.5 and the SNARE core proteins, as well as combinations of the SNARE core proteins, was also carried out.ResultsPGP 9.5-immunoreactive nerve fibers ran toward the coronal region, ramified at the subodontoblast layer, and formed the subodontoblastic nerve plexus. Most nerve fibers penetrated the predentin and dentin along the dentinal tubules. Most, if not all, nerve fibers displayed immunoreactivity for syntaxin-1, SNAP-25, and VAMP-2. Immunoelectron microscopic analyses confirmed the presence of immunoreactivity for the SNARE core proteins within the intradental axonal elements.ConclusionsThe present findings suggest that, since SNARE core proteins participate in the docking and exocytosis of synaptic vesicles in the central nervous system, they may contribute to vesicle exocytosis from the dental nerve fibers even though there are no apparent synapses.     

Effect of experimentally induced occlusal trauma on substance p expression in human dental pulp and periodontal ligament

Introduction

The purpose of this study was to quantify the effect of occlusal trauma experimentally induced with occlusal interferences on substance P (SP) expression in healthy human dental pulp and periodontal ligament.

Methods

Twenty-eight human dental pulp and periodontal ligament samples were obtained from healthy premolars in which extraction was indicated for orthodontic reasons. Before extraction, occlusal trauma was induced with experimental occlusal interferences in half of these premolars by placing a resin block over their occlusal surface and submitting patients to chew gum for 30 minutes. The remaining healthy premolars were extracted without occlusal trauma and served as a control group. All dental pulp and periodontal ligament samples were processed, and SP was measured by radioimmunoassay.

Results

There was 45% and 120% greater SP expression in dental pulp and periodontal ligament, respectively, of teeth with experimentally induced occlusal trauma. Paired t test showed statistically significant differences for both human dental pulp and periodontal ligament (P = .02 and P < .001, respectively) of teeth submitted to occlusal trauma when compared with control group values.

Conclusions

SP expression in human dental pulp and periodontal ligament increases when teeth are submitted to occlusal trauma experimentally induced with occlusal interferences.     

Serotonin-immunoreactive Epithelial Cells in the Main Excretory Ducts of Rat Submandibular Glands

Although several morphological studies of main excretory duct of the submandibular gland have been performed, few reports present immunohistochemical data. Some epithelial cells or basal cells contain dense granules of an array shape by electron microscopic observation. However, the details of granulated cells have not been clarified immunohistochemically. In this study, both protein gene product 9.5 (PGP 9.5) and chromogranin or serotonin immunoreactive cells in the main excretory duct (MED) of the rat submandibular gland were observed. Some nerve endings were also observed among the epithelium of the MED. Therefore, the modifying mechanism of primary saliva in the MED may be regulated by both serotonergic nerve and endocrine cells with serotonin.     

Nerve-epithelium association in the periodontal ligament of guinea pig teeth

Several lines of evidence have suggested that periodontal nerves have other roles besides sensory function. Exploring the distribution pattern of nerves in relation to other structures within the periodontal ligament of various species should be important to understand their roles within the ligament. This study investigated whether any association exists between the nerves and the epithelial cells in the periodontal ligament of continuously erupting guinea pig molars, which show distinct enamel epithelium layers among the cementum pearls. Ten guinea pigs were fixed by vascular perfusion and jaw sections were processed for immunohistochemistry of protein gene product 9.5 (PGP 9.5), growth-associated protein-43 (GAP-43) and glia-specific S-100 protein, and for enzyme histocytochemistry of cholinesterase. Nerves that were immunopositive for the above neuronal markers were located predominantly in the alveolus-related part of the periodontal ligament. Some nerves, immunoreactive for PGP 9.5 and GAP-43, were also found in the tooth-related part (TRP) of the periodontal ligament close to the tooth surface. PGP 9.5-positive nerves in the TRP appeared very thin and terminated by making loops or plexus-like structures in close apposition to the epithelium layers, overlying the enamel surface in between cementum pearls. Such an intimate association between nerves and the enamel epithelium was not found in the labial periodontal tissue of incisors or the apical growing end of the molar, where periodontal fibre attachment was indistinct. The association between nerves and epithelium in the periodontal ligament of guinea pig molar is site specific and is only seen in the presence of cementum, suggesting that this association is related to the attachment function of the ligament.     

Morphology of neural endings in the human periodontal ligament: An electron microscopic study

The ultrastructure of sensory nerve endings in the human periodontal ligament from 43 extracted teeth was studied using serial sections. Three types of nerve endings were found: free nerve endings (FNE), Ruffini-like endings and lamellated corpuscles. Free nerve endings stem from unmyelinated or from myelinated nerve fibers. The endings contain neurotubuli, neurofilaments and vesicles. Ruffini-like receptors were mostly found in the apical part of the periodontal ligament. In these Ruffini-like receptors a particularly abundant concentration of mitochondria appears. In some cases desmosome-like junctions are present between neurite and ensheathing cell. Lamellated corpuscles were also found in the periodontal ligament. The lamellae are extremely endocytotic and are in close contact with each other.     

The absence of proprioceptive nerve endings in the human periodontal ligament: the role of periodontal mechanoreceptors in the reflex control of mastication.

A review of the literature was conducted to determine the presence or absence of proprioceptive nerve endings in the human periodontal ligament. A histologic review of the periodontal ligament innervation concluded that nerve endings found were those mediating pain, pressure, or touch and that there is no histologic evidence of any "classic" proprioceptive nerve ending in the periodontal ligament. A summary is given concerning the precise role of nerve endings in the periodontal membrane, their afferent pathways, and the role of masticatory muscle proprioception, jaw reflexes, and the temporomandibular joint in the coordinated control of mastication and mandibular proprioception.     

Microanatomical changes and biomolecular expression at the PDL‐entheses during experimental tooth movement

The novel aspect of this study was to contextualize the co‐localization of biomolecular expression in widened and narrowed periodontal ligament (PDL)‐space within a mechanically activated periodontal complex. The PDL is unique as it is the only ligament with both innervation and vascularization. Maxillary molars in 6‐week‐old male C57BL/6 mice (N = 5) were experimentally translated for 2 weeks using an elastic spacer. Contralateral teeth were used as controls. Mechanical testing of the periodontal complex of a mouse in situ and imaging using X‐ray micro‐computed tomography (micro‐XCT) illustrated deformations within blood vessels (BV) of the PDL. PDL‐bone and PDL‐cementum entheses at the widened and narrowed PDL‐spaces following experimental tooth movement (ETM) illustrated osterix (OSX), bone sialoprotein (BSP), cluster of differentiation 146 (CD146), and protein gene product 9.5 (PGP9.5), indicating active remodeling at these sites. PGP9.5 positive nerve bundles (NBs) were co‐localized with multinucleated cells (MCs), Howship's resorption lacunae, and CD146 positive BVs. Association between nerves and MC was complemented by visualizing the proximity of osmium tetroxide stained NBs with the ultrastructure of MCs by performing scanning transmission electron microscopy. Spatial association of NB with BV, and NB with MC, provided insights into the plausible co‐activation of NBs to initiate osteoclastic activity. Resorption of mineral occurred as an attempt to restore PDL‐space of the load‐bearing complex, specifically at the PDL‐entheses. Mapping of anatomy‐specific structural elements and their association with regenerative molecules by correlating light and electron micrographs provided insights into the use of these extracellular matrix molecules as plausible targets for pharmacological interventions related to tooth movement. Within the realm of tissue regeneration, modulation of load can reverse naturally occurring mineral formation to experimentally induced resorption, and naturally occurring mineral resorption to experimentally induced formation at the enthesial sites to permit tooth translation.     

Innervation of the periodontal ligament in the alligatorid Caiman crocodilius

The mode of development and structure of crocodilian teeth and periodontium parallels that of mammals, but the teeth are continuously replaced throughout the lifetime of those animals. In this report, the innervation and fibres of the crocodilian periodontal ligament were examined using histology, immunohistochemistry for S-100 protein and transmission electron microscopy. Crocodilian periodontal ligaments had the following characteristics: (1) horizontal fibres, which connect the alveolar bone to the root cementum and (2) longitudinal fibres, which ran parallel to the tooth axis, with nerves and blood vessels in the middle layer of the ligament. The apex of root and tooth germs were both embedded in thick circular fibres. S-100 protein was detected in neural elements including terminal portions which were densely distributed in the periodontal ligament and dental follicle. The S-100 positive neural elements formed a periodontal plexus. We found two types of nerve endings; free endings and simple encapsulated corpuscles as described in mammals. The presence of such nerve endings in caiman suggests that these teeth, in addition to having a biting function, may also act as highly sensitive sensory organs.     

大鼠磨牙正常牙周膜感觉神经分布

选用雄性ＳＤ大鼠５只，将左右上颌第一磨牙冠状方向作冰冻切片，厚度４０μｍ，采用免疫化学技术，观察牙周膜中含ＣＧＲＰ的神经纤维分布特点。结果表明：牙周膜神经纤维分布不均匀，以根尖１／３神经分布最多，其次是牙龈邻近的颈１／３区域。牙周膜中含ＣＧＲＰ的神经纤维形态各异：多数为单根或多根纤维沿血管走行，少数单根纤维走行无规律，常以游离端形式终止。纤维丰富区多呈“分枝状”、“簇”、“网”分布。大部分神经纤维靠近牙槽骨侧。     

Investigation of periodontal ligament reaction upon excessive occlusal load--osteopontin induction among periodontal ligament cells

OBJECTIVE: The purpose of the present study was to investigate the reaction of the periodontal ligament to excessive occlusal loading by observing the histological changes and osteopontin induction. The possibility of ligand for receptor activator of nuclear factor kappaB (RANKL) participation in osteopontin induction was also discussed. BACKGROUND: The precise mechanism of periodontal ligament breakdown by excessive occlusal loading remains unclear. We established an experimental model for excessive occlusal loading in vivo. Osteopontin is known to be produced upon mechanical loading and is considered to induce the migration of osteoclasts to the resorption site. RANKL is one of the essential factors for osteoclast maturation and induces the constitutive induction of intracellular osteopontin in vitro. METHODS: The occlusal surface of the upper left first molars of rats was raised by steel wire bonding in order to induce occlusal trauma. The destruction of the periodontal ligament was observed and the production of osteopontin and RANKL by periodontal ligament cells was detected via immunohistochemistry. RESULTS: Our model produced wide-ranging destruction of the periodontal ligament. From day 3 to day 7, prominent compression of the periodontal ligament and osteoclast migration were observed at the apical interradicular septum. Osteopontin was detected in some osteoclasts, surrounding fibroblasts, and osteoblasts adjacent to the compression area. RANKL was observed from day 1 to day 7 around the osteoblasts and osteoclasts. CONCLUSIONS: Our model was useful for the detailed investigation of periodontal ligament breakdown during excessive occlusal loading. Although intracellular osteopontin was produced in osteoclasts with intermittent occlusal loading, the role of this protein in the cells was not clear. No correlation between RANKL distribution and osteopontin production in osteoclasts could be found.     

Merkel-like cells in Malassez epithelium in the periodontal ligament of cats: an immunohistochemical,confocal-laser scanning and immuno electron-microscopic investigation

The cellular heterogeneity of Malassez epithelium (ME) residing in the periodontal ligament has recently been reported, and the presence and coexistence of the neuropeptides calcitonin gene-related peptide (CGRP), substance P (SP) and vasoactive intestinal peptide (VIP) in single cells in ME has been shown (1). However, the identity of these neuroendocrine cells has so far not been verified. This study was undertaken in order to elucidate the identity of the neuroendocrine cells in ME by means of transmission electron microscopy, confocal scanning microscopy and immunohistochemistry using antibodies to protein gene product (PGP) 9.5 and cytokeratin 20 (CK). Gingival tissue was included in the study as a positive control for identification of Merkel-like cells in oral epithelium. CK 20 immunopositive cells were present in both Malassez epithelium and in basal cell layers of gingival epithelium showing a distribution consistent with PGP 9.5 labelled cells in both epithelia. The results from PGP 9.5 immuno electron microscopy clearly evidenced the presence of single, intensely labelled cells and some nerve fibres invested between the Malassez epithelial cells. The conformity of the immunopositive cells in Malassez and gingival epithelium verified by double immunolabelling with PGP 9.5 and CK 20, indicates that the labelled neuroendocrine cells are identical in ME and in gingival epithelium. This demonstrates that Malassez epithelium not only exhibits neuroendocrine cells, but additionally that the neuroendocrine cells represent Merkel-like cells.