Epithelial Cell Rests Of Malassez Research Articles

Role of the epithelial cell rests of Malassez in the development, maintenance and regeneration of periodontal ligament tissues

Periodontitis is a highly prevalent inflammatory disease that results in damage to the tooth-supporting tissues, potentially leading to tooth loss. Periodontal tissue regeneration is a complex process that involves the collaboration of two hard tissues (cementum and alveolar bone) and two soft tissues (gingiva and periodontal ligament). To date, no periodontal-regenerative procedures provide predictable clinical outcomes. To understand the rational basis of regenerative procedures, a better understanding of the events associated with the formation of periodontal components will help to establish reliable strategies for clinical practice. An important aspect of this is the role of the Hertwig's epithelial root sheath in periodontal development and that of its descendants, the epithelial cell rests of Malassez, in the maintenance of the periodontium. An important structure during tooth root development, the Hertwig's epithelial root sheath is not only a barrier between the dental follicle and dental papilla cells but is also involved in determining the shape, size and number of roots and in the development of dentin and cementum, and may act as a source of mesenchymal progenitor cells for cementoblasts. In adulthood, the epithelial cell rests of Malassez are the only odontogenic epithelial population in the periodontal ligament. Although there is no general agreement on the functions of the epithelial cell rests of Malassez, accumulating evidence suggests that the putative roles of the epithelial cell rests of Malassez in adult periodontal ligament include maintaining periodontal ligament homeostasis to prevent ankylosis and maintain periodontal ligament space, to prevent root resorption, to serve as a target during periodontal ligament innervation and to contribute to cementum repair. Recently, ovine epithelial cell rests of Malassez cells have been shown to harbor clonogenic epithelial stem-cell populations that demonstrate similar properties to mesenchymal stromal/stem cells, both functionally and phenotypically. Therefore, the epithelial cell rests of Malassez, rather than being 'cell rests', as indicated by their name, are an important source of stem cells that might play a pivotal role in periodontal regeneration.

The characteristics of epithelial cell rests of Malassez during tooth eruption of development mice

It has been demonstrated that Hertwig's epithelial root sheath (HERS) has an important role in root development, closely related to development of cementum epithelial rests of Malassez (ERM) as the residuum after HERS fragment, and is the exclusive epithelial structure in the mature periodontal ligament. Some studies reported that ERM may play a role in maintaining a stable environment of periodontal, and likely to be involved in regeneration of periodontal tissue, especially of cementum. However, the function of the ERM is not well understood. In this study, we observed the morphology and biological characteristics of ERM of the maxillary 1st molar with surrounding periodontal tissues of BALB/c mice during the period of tooth cusp erupted out of the gingiva to occlusion stability. Immunohistochemistry revealed ERM predominately located at the cervical and root furcation regions of the periodontal ligament. The number of ERM cells at the cervical and root furcation regions of the post-built occlusion stage decreased compared to pre-built occlusion stage and occlusion building stage. Transmission electron microscopy analysis showed that epithelial cell nuclei with typical features of apoptosis were observed at the post-built occlusion stage, and consistent with positive bodies labeled by TUNEL(terminal deoxy-transferase (TdT)-mediated dUTP-biotin nick end labeling), while proliferating cell nuclear antigen positive bodies mainly located at occlusion built stage. It suggests that ERM may regulate in alveolar bone remodeling in association with the periodontal ligament during tooth erupting to occlusion stability and may play important roles potentially in regeneration and homeostasis of the periodontal tissues.

The effects of epithelial rests of Malassez cells on periodontal ligament fibroblasts against centrifugal forces in vitro

ObjectivesThe purpose of this study was to investigate the effects of epithelial rests of Malassez (ERM) cells on PDL fibroblasts in vitro against centrifugal forces using a co-culture system. Materials and methodsPDL fibroblasts and ERM cells derived from porcine were used. PDL fibroblasts were seeded in 6-well dishes. ERM cells plated in a chamber with a 0.4μm pore membrane were placed into the wells for 5days, and then were centrifuged as a mechanical stress. Vascular endothelial growth factor (VEGF), heat shock protein (HSP) 27, osteocalcin (OCN), bone sialoprotein (BSP), receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) mRNA levels were analyzed, and RANKL expression was analyzed at the protein level. ResultsVEGF mRNA expression by PDL fibroblasts co-cultured with ERM cells with or without force was higher than those cultured without ERM cells. HSP27 mRNA expression by PDL fibroblasts cultured without ERM cells with force was higher than those co-cultured with ERM cells. OCN and BSP mRNA expression by PDL fibroblasts co-cultured with ERM cells was significantly lower than without ERM cells with or without force. The expressions of RANKL by PDL fibroblasts co-cultured with ERM cells with force were higher than those without ERM cells. OPG mRNA expression by PDL fibroblasts co-cultured with ERM cells without force were lower than those without ERM cells. ConclusionThese results demonstrate that ERM cells play an important role in preventing PDL cells from being susceptible to dento-alveolar ankylosis during compression of the PDL.

Epithelial Cell Rests of Malassez Contain Unique Stem Cell Populations Capable of Undergoing Epithelial–Mesenchymal Transition

The epithelial cell rests of Malassez (ERM) are odontogenic epithelial cells located within the periodontal ligament matrix. While their function is unknown, they may support tissue homeostasis and maintain periodontal ligament space or even contribute to periodontal regeneration. We investigated the notion that ERM contain a subpopulation of stem cells that could undergo epithelial-mesenchymal transition and differentiate into mesenchymal stem-like cells with multilineage potential. For this purpose, ERM collected from ovine incisors were subjected to different inductive conditions in vitro, previously developed for the characterization of bone marrow mesenchymal stromal/stem cells (BMSC). We found that ex vivo-expanded ERM expressed both epithelial (cytokeratin-8, E-cadherin, and epithelial membrane protein-1) and BMSC markers (CD44, CD29, and heat shock protein-90β). Integrin α6/CD49f could be used for the enrichment of clonogenic cell clusters [colony-forming units-epithelial cells (CFU-Epi)]. Integrin α6/CD49f-positive-selected epithelial cells demonstrated over 50- and 7-fold greater CFU-Epi than integrin α(6)/CD49f-negative cells and unfractionated cells, respectively. Importantly, ERM demonstrated stem cell-like properties in their differentiation capacity to form bone, fat, cartilage, and neural cells in vitro. When transplanted into immunocompromised mice, ERM generated bone, cementum-like and Sharpey's fiber-like structures. Additionally, gene expression studies showed that osteogenic induction of ERM triggered an epithelial-mesenchymal transition. In conclusion, ERM are unusual cells that display the morphological and phenotypic characteristics of ectoderm-derived epithelial cells; however, they also have the capacity to differentiate into a mesenchymal phenotype and thus represent a unique stem cell population within the periodontal ligament.

Responses of BrdU label-retaining dental pulp cells to allogenic tooth transplantation into mouse maxilla

Recently, we demonstrated that a pulse of BrdU given to prenatal animals reveals the existence of slow-cycling long-term label-retaining cells (LRCs), putative adult stem or progenitor cells, which reside in the dental pulp. This study aims to clarify responses of LRCs to allogenic tooth transplantation into mouse maxilla using prenatal BrdU-labeling, in situ hybridization for osteopontin and periostin, and immunohistochemistry for BrdU, nestin, and osteopontin. The upper-right first molars were allografted in the original socket between BrdU-labeled and non-labeled mice or between GFP transgenic and wild-type mice. Tooth transplantation caused degeneration of the odontoblast layer, resulting in the disappearance of nestin-positive reactions in the dental pulp. On postoperative days 5-7, tertiary dentin formation commenced next to the preexisting dentin where nestin-positive odontoblast-like cells were arranged in the successful cases. In BrdU-labeled transplanted teeth, dense LRCs were maintained in the center of the dental pulp beneath the odontoblast-like cells including LRCs, whereas LRCs disappeared in the area surrounding the bone-like tissue. In contrast, LRCs were not recognized in the pulp chamber of non-labeled transplants through the experimental period. Tooth transplantation using GFP mice demonstrated that the donor cells constituted the dental pulp of the transplant except for endothelial cells and some migrated cells, and the periodontal tissue was replaced by host-derived cells except for epithelial cell rests of Malassez. These results suggest that the maintenance of BrdU label-retaining dental pulp cells play a role in the regeneration of odontoblast-like cells in the process of pulpal healing following tooth transplantation.

Capturing the Regenerative Potential of Periodontal Ligament Fibroblasts

The cell population within the periodontal ligament (PDL) tissue is remarkably heterogeneous1. Fibroblasts, a mixed population of cells, are the main cellular component of the PDL and the cell type most often studied for periodontal regeneration. Osteoblasts and osteoclasts are found on the bone side, while fibroblasts, macrophages, undifferentiated adult/mesenchymal stem cells, neural elements, and endothelial cells are found throughout the PDL. Epithelial rests of Malassez cells and cementoblasts are focused near the root surface. PDL tissue also includes loose connective tissue between dense fiber bundles that contain branches of the periodontal blood vessels and nerves2. The complexity of the PDL tissue, with its various cell types and cell progenitor components, explains the challenges involved in therapies to restore tissue following periodontal disease. Cementoblasts, osteoblasts, and endothelial cells must migrate, differentiate, and coordinately interact with a variety of soluble mediators to regenerate the periodontium3. Stem cells located in the PDL tissue are key contributors to this process4. Stem cells in the PDL are important not only for formation and maintenance of the tissue but also for repair, remodeling, and regeneration of adjacent alveolar bone and cementum5. Our laboratory has shown that progenitor cells isolated from PDL tissue by selection with cell surface markers STRO-1+ and CD146+ are capable of differentiating into chondrogenic, osteogenic, and adipogenic phenotypes under appropriate culture conditions

The effects of epithelial rests of Malassez cells on periodontal ligament fibroblasts: A co-culture investigation for epithelialmesenchymal interactions

=Background: The major function of epithelial rests of Malassez (ERM) cells is to maintain the homeostasis of the periodontal ligament (PDL). The purpose of this study was to characterize the effects of ERM cells on PDL fibroblasts in vitro using a co-culture system. Methods: PDL fibroblasts and ERM cells derived from porcine tissues were used. PDL fibroblasts were seeded in 6-welldishes. ERM cells plated in a chamber with a 0.4 μm pore membrane, were placed into the wells for 5 days. Osteocalcin (OCN), bone sialoprotein (BSP), osteoprotegerin (OPG) and receptor activator of NF-k B ligand (RANKL) mRNA levels in the PDL fibroblasts were then analyzed using real time RT-PCR. Alkaline phosphatase (ALP) activity was also measured. PDL fibroblasts cultured without ERM cells were used as a control. Results: OCN, BSP and OPG mRNA levels in PDL fibroblasts co-cultured with ERM cells were lower than the levels in the control group. Meanwhile, the RANKL mRNA level in PDL fibroblasts co-cultured with ERM cells was significantly higher than that of the controls (P<0.01). ALP activity of PDL fibroblasts co-cultured with ERM cells was significantly lower than in the controls (P<0.01). Conclusion: This study shows that ERM cells affect the functions of PDL fibroblasts, which decrease hard tissue formation and increase bone resorption. Therefore, ERM cells prevent dento-alveolar ankylosis of the PDL.

Epithelial Cell Rests of Malassez Modulate Cell Proliferation, Differentiation and Apoptosis via Gap Junctional Communication under Mechanical Stretching in vitro

Epithelial cell rests of Malassez (ERM) are involved in the maintenance and homeostasis of the periodontal ligament. The objective of this study was to investigate the effect of mechanical stretching on cell growth, cell death and differentiation in the ERM. Cultured porcine ERM were stretched for 24 hr in cycles of 18% elongation for 1 sec followed by 1 sec relaxation. The numbers of cells and TUNEL-positive cells were then counted. The expression of mRNAs encoding gap junction protein α1 (Gja1), ameloblastin, bone morphogenetic protein 2 (BMP2), bone morphogenetic protein 4 (BMP4) and noggin were evaluated using quantitative real-time PCR. The number of cells in the stretching group was approximately 1.3-fold higher than that in the non-stretching controls at 24 hr (p<0.01). Apoptotic cells ranged from 1.9-2.5% in the stretching group at 24 hr, but were only 0.6% in the control group (p<0.01). The expression of Gja1, ameloblastin and noggin mRNAs in the stretching group was decreased at 24 hr compared with in the non-stretching group (p<0.01), whereas the expression of BMP2 and BMP4 mRNAs in the stretching group was significantly higher than that in the control group (p<0.01). Incorporation of 18 α-glycyrrhetinic acid (18GA, a gap junction inhibitor) promoted proliferation and apoptosis and confirmed both the increase of BMP2 and BMP4 and the decline of Gja1, ameloblastin and noggin in ERM. Thus, the ERM modulate cell proliferation and apoptosis, and inhibit differentiation by reducing expression of Gja1 under mechanical stretching.

Disruption of periodontal integrity induces expression of apin by epithelial cell rests of Malassez

It has been suggested that epithelial cell rests of Malassez (ERM) may express enamel matrix proteins and play an important role in periodontal regeneration. Two novel proteins, apin (APIN) and amelotin (AMTN), produced by maturation-stage ameloblasts and junctional epithelium, have recently been identified. The objective of this study was to evaluate whether the ERM express APIN and AMTN under normal conditions and after periodontal challenge. Gingivectomy and orthodontic tooth movement were carried out on the left side of the maxillae of rats. The control group included the untreated contralateral side of these animals and the maxillae of normal, untreated rats. Animals were sacrificed by intracardiac perfusion on days 3 and 5 after the experimental procedures and maxillary molars were decalcified and processed for paraffin embedding. Immunohistochemistry was used to evaluate the expression of various ameloblast products, including APIN, AMTN, ameloblastin (AMBN) and amelogenin (AMEL). At 3 and 5 days after periodontal challenge, ERM were more evident in the periodontal ligament along the root surface and in the root furcations. Immunodetection of APIN, but not of the other three proteins, was observed in the ERM following the disruption of periodontal integrity. No immunolabeling for APIN, AMTN, AMBN and AMEL was detected in the ERM under normal conditions. The expression of APIN at an early time-point following disruption of periodontal integrity suggests that this protein may be part of the cascade of events leading to the activation of ERM during periodontal healing and regeneration.

Disease-Associated Prion Protein in the Dental Tissue of Mice Infected with Scrapie

Transmissible spongiform encephalopathies (TSEs) induce fatal neurodegenerative diseases in man and animals. The present study demonstrates immunohistochemically the presence of disease-associated prion protein (PrP(Sc)) in the epithelial cell rests of Malassez (ERM) of mice experimentally infected with ME7 scrapie by the intracerebral route. Mouse bioassay of scrapie-infected dental tissue revealed prolonged incubation periods, suggesting that there are relatively low amounts of infectious agent in dental tissue compared with the brain. These findings indicate that PrP(Sc) may spread from the brain to the ERM along the cranial nerves via the trigeminal ganglion that innervates the dental tissues. Dental tissue might therefore be a potential source of PrP(Sc) for horizontal transmission of TSEs.

Induction of MMP-2 at the interface between epithelial cells and fibroblasts from human periodontal ligament

MMP-2 can degrade type IV collagen and MMP-14 can activate pro MMP-2. The present study was undertaken to examine the expression of MMP-2 and MMP-14 with respect to interaction between the cells of the epithelial rests of Malassez and fibroblasts from human periodontal ligament. Explants of human periodontal ligament tissues produced outgrowths containing both putative epithelial rests of Malassez cells and human periodontal ligament fibroblasts after incubation in a modified serum-free medium. The distribution and expression of MMP-2 and MMP-14 were analysed using immunohistochemistry, in situ hybridization and RT-PCR analysis. The conditioned media and cell extracts were collected for western blot analysis for MMP-2. Putative epithelial rests of Malassez cells at the interface between the cells of the epithelial rests of Malassez and fibroblasts expressed MMP-2 and MMP-14 strongly. However, in situ hybridization analysis revealed that human periodontal ligament fibroblasts expressed MMP-2 mRNA while putative epithelial rests of Malassez cells expressed MMP-14 mRNA at the interface. The RT-PCR analysis showed that the expression of MMP-2 mRNA was significantly higher when putative epithelial rests of Malassez cells and human periodontal ligament fibroblasts were cultured together than when cultured alone. Western blot analysis showed that the active form of MMP-2 was detected at higher levels in the conditioned medium of the co-cultured cells. These findings indicate that putative epithelial rests of Malassez cells stimulate the production of MMP-2 in human periodontal ligament fibroblasts. Up-regulated proMMP-2 bound by MMP-14 expressed in epithelial rests of Malassez cells can degrade matrix molecules, such as type IV collagen, in the basal membrane between putative epithelial rests of Malassez cells and human periodontal ligament fibroblasts.

Quiescent epithelial cell rests of Malassez can differentiate into ameloblast‐like cells

Epithelial cell rests of Malassez (ERM) are quiescent epithelial remnants of Hertwig's epithelial root sheath (HERS) that are involved in the formation of tooth roots. After completion of crown formation, HERS are converted from cervical loop cells, which have the potential to generate enamel for tooth crown formation. Cervical loop cells have the potential to differentiate into ameloblasts. Generally, no new ameloblasts can be generated from HERS, however this study demonstrated that subcultured ERM can differentiate into ameloblast-like cells and generate enamel-like tissues in combination with dental pulp cells at the crown formation stage. Porcine ERM were obtained from periodontal ligament tissue by explant culture and were subcultured with non-serum medium. Thereafter, subcultured ERM were expanded on 3T3-J2 feeder cell layers until the tenth passage. The in vitro mRNA expression pattern of the subcultured ERM after four passages was found to be different from that of enamel organ epithelial cells and oral gingival epithelial cells after the fourth passage using the same expansion technique. When subcultured ERM were combined with subcultured dental pulp cells, ERM expressed cytokeratin14 and amelogenin proteins in vitro. In addition, subcultured ERM combined with primary dental pulp cells seeded onto scaffolds showed enamel-like tissues at 8 weeks post-transplantation. Moreover, positive staining for amelogenin was observed in the enamel-like tissues, indicating the presence of well-developed ameloblasts in the implants. These results suggest that ERM can differentiate into ameloblast-like cells.

In Vitro Characterization of the Cytokine Profile of the Epithelial Cell Rests of Malassez

The epithelial cell rests of Malassez (ERM) are an integral part of the periodontal ligament and are considered to play an important role in dental pathology. Surprisingly, this cell type is poorly described and is often disregarded in the context of periodontal research. The aim of this study was to establish primary cell cultures of human ERM, characterize the cytokine profile, and compare it to other periodontal cell entities. ERM-derived epithelial cells were isolated from the periodontal ligament of three subjects. A cytokine antibody array, including 120 cytokines in two membranes, was used to determine the cytokine profile of conditioned medium from the ERM-derived epithelial cells. The results were compared to those of gingival epithelial cells and periodontal ligament fibroblasts. ERM-derived epithelial cells expressed 29 of 120 cytokines in significant amounts, including cytokines, chemokines, growth factors, and related proteins, such as interleukin (IL)-1, -6, -8, and -10; granulocyte macrophage-colony stimulating factor; monocyte chemoattractant protein (MCP)-1, -2, and -3; amphiregulin; glial-derived neurotrophic factor; vascular endothelial growth factor; and insulin-like growth factor binding protein-2. The cytokine profile of ERM cells was similar to that of gingival epithelial cells but strikingly different from the profile of periodontal ligament fibroblasts. The results indicated that, via paracrine secretion of a variety of soluble factors, the ERM cells actively take part in the homeostasis of the periodontium. Therefore, future research on the pathophysiology of periodontal tissue should include this often overlooked cell type.

Observations on continuously growing roots of the sloth and the K14‐Eda transgenic mice indicate that epithelial stem cells can give rise to both the ameloblast and root epithelium cell lineage creating distinct tooth patterns

Root development is traditionally associated with the formation of Hertwig's epithelial root sheath (HERS), whose fragments give rise to the epithelial cell rests of Malassez (ERM). The HERS is formed by depletion of the core of stellate reticulum cells, the putative stem cells, in the cervical loop, leaving only a double layer of the basal epithelium with limited growth capacity. The continuously growing incisor of the rodent is subdivided into a crown analog half on the labial side, with a cervical loop containing a large core of stellate reticulum, and its progeny gives rise to enamel producing. The lingual side is known as the root analog and gives rise to ERM. We show that the lingual cervical loop contains a small core of stellate reticulum cells and suggest that it acts as a functional stem cell niche. Similarly we show that continuously growing roots represented by the sloth molar and K14-Eda transgenic incisor maintain a cervical loop with a small core of stellate reticulum cells around the entire circumference of the tooth and do not form a HERS, and still give rise to ERM. We propose that HERS is not a necessary structure to initiate root formation. Moreover, we conclude that crown vs. root formation, i.e. the production of enamel vs. cementum, and the differentiation of the epithelial cells into ameloblasts vs. ERM, can be regulated independently from the regulation of stem cell maintenance. This developmental flexibility may underlie the developmental and evolutionary diversity in tooth patterning.

Profiling of differentially expressed genes in porcine epithelial cells derived from periodontal ligament and gingiva by DNA microarray

Objective It is unknown which genes are differentially expressed in cultured epithelial cells derived from the epithelial rests of Malassez (ERM) in periodontal ligament and oral gingival epithelium (OGE). This study analysed the different gene expression of OGE and ERM cells using a DNA microarray technique. Design Epithelial cells from ERM and OGE were isolated from porcine periodontal ligament and oral gingival epithelium. Each RNA sample extracted from the cells was reverse transcribed into cDNA and labelled with either cytidine 5-dUTP (Cy5) or cytidine 3-dUTP (Cy3). These labelled cDNA probes were then mixed and simultaneously hybridised to the Pig 13K microarray plate bearing 13,295 different genes (Operon, AL). Cellular enzyme-linked immunosorbent assay (CELISA) was performed to confirm the expression at protein level. Results There were nine genes common to the triplicate microarrays in ERM cells and one in OGE cells. Four of the nine genes including tissue factor (TF), FAT cadherin (FAT) and two unknown genes were expressed at levels more than threefold higher in ERM cells than in OGE cells. The protein levels of both TF and FAT in ERM cells were significantly higher than those in OGE. Conclusion TF and FAT may act as markers to distinguish ERM cells from OGE cells in vitro.

Epithelial Cell Rests of Malassez and OX6‐Immunopositive Cells in the Periodontal Ligament of Rat Molars: A Light and Transmission Electron Microscope Study

It has been shown that human and cat epithelial cell rests of Malassez (ERM) consist of heterogeneous cell populations. Immunohistochemical and immunoelectron microscopic analyses have verified the presence of neuroendocrine and Merkel-like cells in both of these epithelia. During experimental orthodontic tooth movement, immunocompetent cells have also been found in the vicinity of ERM in rat periodontal ligament (PDL), but have not been characterized in normal rat PDL. The aim of this study was to investigate the presence and distribution of MHC class II antigen presenting cells by using OX6 antibody in ERM of rat molars by light and transmission electron microscopy. Immunohistochemical and immunoelectron microscopic observations of rat maxillary molars confirmed the presence of OX6-positive cells in contact with ERM. Some immunopositive cytoplasmic processes containing vesicles interdigitated with cells of the Malassez epithelial clusters. Based on these findings it can be concluded that immunocompetent cells are localized close to Malassez epithelial clusters in normal rat PDL. Furthermore, the ultrastructural evidences indicate a possible interaction between the epithelial and immunocompent cells and suggest morphological and functional properties for ERM.

Mutual induction of noncollagenous bone proteins at the interface between epithelial cells and fibroblasts from human periodontal ligament

Epithelial-mesenchymal interactions are responsible for cell differentiation during periodontal regeneration. The present study was undertaken to examine the expression of alkaline phosphatase and noncollagenous bone proteins, such as osteopontin, osteocalcin and bone sialoprotein, with respect to interaction between the cells of the epithelial rests of Malassez and fibroblasts from human periodontal ligament. Explants of human periodontal ligament tissues produced outgrowths containing both putative epithelial rests of Malassez cells and human periodontal ligament fibroblasts in a modified serum-free medium. Putative epithelial rests of Malassez cells cultured alone, and human periodontal ligament fibroblasts cultured alone, were used as controls. The expression levels of amelogenin were analyzed by in situ hybridization. The expression and distribution of alkaline phosphatase and noncollagenous bone proteins in both cell populations at the interface between putative epithelial rests of Malassez cells and human periodontal ligament fibroblasts were analyzed by immunohistochemistry, in situ hybridization and reverse transcription-polymerase chain reaction. Amelogenin mRNA was detected at high levels only in putative epithelial rests of Malassez cells at the interface. Alkaline phosphatase and bone sialoprotein mRNAs were detected significantly at the interface between putative epithelial rests of Malassez cells and human periodontal ligament fibroblast cells. In particular, bone sialoprotein and its mRNA were expressed significantly in human periodontal ligament fibroblasts at the interface between putative epithelial rests of Malassez cells and human periodontal ligament fibroblast cells. The expressions of osteopontin and its mRNA were not different between putative epithelial rests of Malassez cells and human periodontal ligament fibroblasts at the interface. Osteocalcin and its mRNA were expressed strongly in putative epithelial rests of Malassez cells at the interface between putative epithelial rests of Malassez cells and human periodontal ligament fibroblasts. These findings indicate that the epithelial-mesenchymal interaction modulates the expression of alkaline phosphatase, osteocalcin and bone sialoprotein in putative epithelial rests of Malassez cells and human periodontal ligament fibroblasts, suggesting that epithelial-mesenchymal interactions play a role in the maintenance of periodontal ligament.

Immunohistochemical localization of epithelial rests of Malassez in human periodontal membrane

The aim of the present study was to describe the localization and extension of epithelial rests of Malassez (ERM) in the periodontal membrane (PDM) in normal human third molars. The material consisted of 24 normally developed human third molars surgically removed from patients with an age range from 15 to 27 years (six females and six males). The root lengths were developed from close to half-length to complete apex closure. The extracted teeth were fixed in 10 per cent neutral-buffered formalin, decalcified in ethylene diamine tetra acetic acid EDTA, paraffin embedded and cut sagittaly in 5 microm serial sections. Immunohistochemistry was performed using polyclonal rabbit anti-bovine cytokeratin (wide-spectrum screening, WSS) and the EnVision+ dual link system. The results were based on the visual comparison of WSS in the tissue sections using a light microscope. It was demonstrated that the ERM cells were distributed in the PDM in a network-shaped manner along the root surface and in the furcation region. The distribution of ERM was more prominent in teeth with incomplete root formation. The ectodermal tissue layer might influence not only the morphology of the tooth but also tooth eruption. The reaction of this epithelial layer in connection with ankylosis and orthodontic tooth movement may be of future interest.

In vitro differentiation of epithelial cells cultured from human periodontal ligament

Alkaline phosphatase and noncollagenous bone proteins are produced prior to cementum formation. While it has been suggested that epithelial rests of Malassez are involved in cementum formation, little is known about the relationship between epithelial rests of Malassez and cementum formation. The purpose of the present study was to determine whether the epithelial rests of Malassez cells cultured from human periodontal ligament can produce alkaline phosphatase and noncollagenous bone proteins, such as osteopontin, osteocalcin and bone sialoprotein. An outgrowth of putative epithelial rests of Malassez cells was produced from periodontal ligament explant, and second passage cultures were used in the experiments. Human gingival epithelial cells and periodontal ligament fibroblasts were used as controls. The expression levels of amelogenin were analyzed by immunostaining and in situ hybridization. Furthermore, the expression levels of alkaline phosphatase and noncollagenous bone proteins were assessed by immunostaining and reverse transcription-polymerase chain reaction. Amelogenin, alkaline phosphatase and osteopontin proteins and their corresponding mRNAs were detected at high levels in putative epithelial rests of Malassez cells. Osteocalcin and bone sialoprotein were not expressed in putative epithelial rests of Malassez cells. Alkaline phosphatase and noncollagenous bone proteins were seen in periodontal ligament fibroblasts, but not in gingival epithelial cells. Our results suggest that putative epithelial rests of Malassez cells cultured alone do not transform into maturing cells to form the cementum, but may play a potential role in the mineralization process.

Proliferation of Epithelial Cell Rests, Formation of Apical Cysts, and Regression of Apical Cysts after Periapical Wound Healing

There is continuing controversy regarding the potential for inflammatory apical cysts to heal after nonsurgical endodontic therapy. Molecular cell biology may provide answers to a series of related questions. How are the epithelial cell rests of Malassez stimulated to proliferate? How are the apical cysts formed? How does the lining epithelium of apical cysts regress after endodontic therapy? Epithelial cell rests are induced to divide and proliferate by inflammatory mediators, proinflammatory cytokines, and growth factors released from host cells during periradicular inflammation. Quiescent epithelial cell rests can behave like restricted-potential stem cells if stimulated to proliferate. Formation of apical cysts is most likely caused by the merging of proliferating epithelial strands from all directions to form a three-dimensional ball mass. After endodontic therapy, epithelial cells in epithelial strands of periapical granulomas and the lining epithelium of apical cysts may stop proliferating because of a reduction in inflammatory mediators, proinflammatory cytokines, and growth factors. Epithelial cells will also regress because of activation of apoptosis or programmed cell death through deprivation of survival factors or by receiving death signals during periapical wound healing.