Rat Molar Periodontal Ligament Research Articles

Function of chemokine (CXC motif) ligand 12 in periodontal ligament fibroblasts.

The periodontal ligament (PDL) is one of the connective tissues located between the tooth and bone. It is characterized by rapid turnover. Periodontal ligament fibroblasts (PDLFs) play major roles in the rapid turnover of the PDL. Microarray analysis of human PDLFs (HPDLFs) and human dermal fibroblasts (HDFs) demonstrated markedly high expression of chemokine (CXC motif) ligand 12 (CXCL12) in the HPDLFs. CXCL12 plays an important role in the migration of mesenchymal stem cells (MSCs). The function of CXCL12 in the periodontal ligament was investigated in HPDLFs. Expression of CXCL12 in HPDLFs and HDFs was examined by RT-PCR, qRT-PCR and ELISA. Chemotactic ability of CXCL12 was evaluated in both PDLFs and HDFs by migration assay of MSCs. CXCL12 was also immunohistochemically examined in the PDL in vivo. Expression of CXCL12 in the HPDLFs was much higher than that in HDFs in vitro. Migration assay demonstrated that the number of migrated MSCs by HPDLFs was significantly higher than that by HDFs. In addition, the migrated MSCs also expressed CXCL12 and several genes that are familiar to fibroblasts. CXCL12 was immunohistochemically localized in the fibroblasts in the PDL of rat molars. The results suggest that PDLFs synthesize and secrete CXCL12 protein and that CXCL12 induces migration of MSCs in the PDL in order to maintain rapid turnover of the PDL.

Histomorphometric evaluation of periodontal compression and tension sides during orthodontic tooth movement in rats

OBJECTIVE: The purpose of this study was to evaluate the thickness of the periodontal ligament of rat molars during orthodontic tooth movement (OTM). METHODS: Thirty Wistar rats were divided into three groups of 10 animals each: GI, GII and GIII and the mice were euthanized at 7, 14 and 21 days, respectively. Experimental subjects were compared to their respective controls by the Mann-Whitney test. Comparison of values between compression and tension sides were performed during the same and different time periods through Analysis of Variance (ANOVA), Kruskal-Wallis test and, subsequently, Tukey's test. RESULTS: Groups GI and GII showed decreased PDL size in the apical regions of the mesiobuccal root and in the cervical region of the distobuccal root. There was also an increased PDL in the cervical regions of the mesiobuccal root, apical region of the distobuccal root and middle region of both roots. CONCLUSION: The reduction and increase in PDL size were seen in the same root, which characterizes tooth inclination. The apical, middle and cervical regions were compared with one another in each time period and at three times: 7, 14 and 21 days. They were also compared in each region, confirming a tipping movement in GI and GII and a gradual decreased intensity between GI to GII, reaching normal dimension in GIII.

Development of the oxytalan fiber system in the rat molar periodontal ligament evaluated by light- and electron-microscopic analyses

In the elastic fiber system of the periodontal ligaments only oxytalan fibers can be identified, whereas all three types of fibers, oxytalan, elaunin and elastic fibers, are present in the gingiva. However, little information is available concerning their organization in the developing periodontal ligament. In the present study, growth and distribution of the oxytalan fiber system were examined in the developing periodontal ligament of rat molars using the specific staining for oxytalan, elastic and collagen fibers, and electron-microscopic analyses. Oxytalan staining clearly confirmed the earliest oxytalan fibers in a bell-staged tooth germ at embryonic day 18, which were tiny violet-colored fibers in the dental follicle. Their cross images were made up of dot-like microfibrils of 10-15nm in diameter close to fibroblasts in the dental follicle of the rat molars aged 1 day. These microfibrils appeared to be linked to one another through delicate filaments in 3-nm-diameter. At the beginning of root formation, the cross figures of oxytalan fibers were found as dot-like structures around the root sheath as well as in areas very close to blood vessels. As development proceeded, longer oxytalan fibers were produced in the apico-occlusal direction along with blood vessels. In addition, the immunoreactive products to anti-amyloid β protein on the surface of blood vessels suggest that this molecule might be involved in the adhesion of oxytalan fibers to vascular basement membranes. Thus, the oxytalan fiber system might regulate periodontal ligament function through tensional variations registered on the walls of the vascular structures.

Immunoexpression of aquaporin-1 in the rat periodontal ligament during experimental tooth movement

This study examined the immunoexpression pattern of aquaporin-1 (AQP1), first identified as a water channel protein, in the periodontal ligament of rat molars during experimental tooth movement to clarify its role in periodontal responses in an overloaded model by the insertion of a piece of elastic band. In the control group without any treatment, the cementoblasts and osteogenic cells as well as the vascular endothelial cells showed AQP1 immunoreaction. In the experimental group, hyalinized tissue and intensely AQP1 positive amorphous structures which were identified as degenerated endothelial cells by immunoelectron microscopy, occurred at the compression side on Days 1 and 3. AQP1 immunoreaction came to be stronger in the intact endothelial cells around the hyalinized tissue. The hyalinized tissue had almost disappeared by Day 5 when many macrophages reactive to acid phosphatase activity appeared. The periodontal width on Day 7 became almost the same as that in the control group. These findings indicate that the hyalinized tissue and damaged AQP1 positive endothelial cells are phagocytized by macrophages which have temporally migrated, and suggest that the surviving endothelial cells with intense AQP1 reaction are involved in periodontal regeneration by capillary sprouting.

Immunolocalization of lubricin in the rat periodontal ligament during experimental tooth movement

Lubricin is a protein which contributes to the boundary lubrication, facilitating low friction levels at the interfacing surfaces of joints. In tendons and ligaments it facilitates the relative movement of collagen bundles. Its expression is affected by mechanical signals and cytokines. During application of orthodontic forces to teeth, there is a transduction of mechanical forces to the cells of the periodontal ligament (PDL), which triggers several biological reactions causing the synthesis of prostaglandins, cytokines and growth factors. The aim of the present study was to examine the immunolocalization of lubricin and to evaluate if it is time-dependently and differentially detected within the PDL following the application of orthodontic forces to create areas of compression and tension. This was achieved by placing elastic bands between the maxillary first and second molars of 16 male Sprague-Dawley rats (each weighing 120–200g) for 12 and 24h. The molar-bearing segments were dissected and processed for histological and immunohistochemical examination. Binding of a monoclonal antibody was used to evaluate lubricin localization using an indirect streptavidin/biotin immunperoxidase technique. Lubricin, was constitutively expressed in the PDL of rat molars. After the experimental force was applied to the tooth, lubricin was down-regulated, on both sides (compression and tension) of the PDL, in a time-dependent fashion, although to a different extent, being at any time more expressed on the tension side. Furthermore, in every sample, almost all PDL cells in the adjacent tooth cementum and alveolar bone, were more heavily immunolabeled by lubricin antibody, contrary to those located in the central portion of the PDL. Lubricin expression therefore seems related to PDL remodeling and tooth displacement following the application of an orthodontic force, and it appears that lubricin may play an important role during tooth movement.

Characterization and induction of cementoblast cell proliferation by bioactive glass nanoparticles

Cementum is a mineralized tissue that lines the surface of the tooth root enabling attachment of the periodontal ligament to the root and surrounding alveolar bone. Studies examining the mechanisms involved in the formation of root cementum have been hindered by an inability to isolate and culture the cells required for cementum production (cementoblasts). This study isolated and characterized cementoblast cells derived from rat molar periodontal ligament. It was observed that the isolated cells expressed F-Spondin, a cementoblast marker, while F-Spondin expression was not observed in the cells of other tissues such as gingival fibroblasts and osteoblasts. As expected, the isolated cementoblast cells also expressed osteocalcin (OC), bone sialoprotein (BSP), alkaline phosphatase (ALP), and type I collagen, demonstrating the presence of mineralized tissues genes in cementoblast cells. These cells showed high ALP activity and calcified nodule formation in vitro. Since cementogenesis could be a critical event for regeneration of periodontal tissues, this study investigated whether bioactive glass particles could affect the proliferation of cementoblasts since they are known to enhance osteoblast proliferation. It was found that the ionic products from bioactive glass nanoparticles increased cementoblast viability, mitochondrial activity, and induced cell proliferation. Together, these results show the characterization of cementoblast cells from rat molar periodontal ligament. Additionally, it was shown that bioactive glass nanoparticles induced cementoblast to proliferate, indicating that they could be a potential material for use in cement regeneration through tissue engineering.

Expressions of RANKL/RANK and M-CSF/c-fms in root resorption lacunae in rat molar by heavy orthodontic force

The differentiation and functions of osteoclasts are regulated by receptor activator of nuclear factor-κB (RANK)/receptor activator of nuclear factor-κB ligand (RANKL) system that stimulates osteoclasts formation. Macrophage colony-stimulating factor (M-CSF) is also essential for osteoclastogenesis. A recent immunocytochemical study reported that RANKL/RANK and M-CSF/c-fms were localized in the periodontal ligament of rat molars during experimental orthodontic tooth movement. The present study focused on the expressions of RANKL/RANK and M-CSF/c-fms in root resorption area during experimental tooth movement in rats. Forty 6-week-old male Wistar rats were subjected to an orthodontic force of 10 or 50 g with a closed coil spring (wire size: 0.005 inch, diameter: 1/12 inch) ligated to the maxillary first molar cleat by a 0.008 inch stainless steel ligature wire to induce a mesial tipping movement of the upper first molars. Experimental tooth movement was undertaken for 10 days. Each sample was sliced into 6 μm continuous sections in a horizontal direction and prepared for haematoxylin and eosin (H and E) and immunohistochemistry staining for tartrate-resistant acid phosphatase (TRAP), RANK, RANKL M-CSF, and c-fms in root resorption area. Statistical analysis was carried out using a Mann-Whitney U-test with a significance level of P<0.01. On days 7 and 10, immunoreactivity for RANKL/RANK and M-CSF/c-fms was detected in odontoclasts with an orthodontic force of 50 g, but not 10 g. Therefore, RANKL/RANK and M-CSF/c-fms systems may be involved in the process of root resorption by heavy orthodontic force.

Alteration of bFGF expression with growth and age in rat molar periodontal ligament

To test the hypothesis that immunohistochemical changes in expression of basic fibroblast growth factor in the periodontal ligament do not change with age. Thirty male Wistar-ST rats were divided into growing groups (5, 9, and 15 weeks of age) and aging groups (6, 12, and 18 months of age). Serial sagittal sections (5 microm thick) were cut parallel at the distobuccal roots of the maxillary first molar. Paraffin-embedded tissue sections were stained with hematoxylin and eosin and rabbit polyclonal antibodies to basic fibroblast growth factor. The number and the area of basic fibroblast growth factor-immunoreactive cells in the periodontal ligament of the maxillary first molar decreased with age. The number of basic fibroblast growth factor-immunoreactive cells was much greater in the root furcation area, which experiences the greatest effect of occlusal force. Regardless of age, the production of basic fibroblast growth factor in the periodontal ligament may occur subject to functional demand. The hypothesis is rejected. The expression of basic fibroblast growth factor in the periodontal ligament decreased with age.

Changes in the Distribution of Nerve Fibers Immunoreactive to Calcitonin Gene-Related Peptide According to Growth and Aging in Rat Molar Periodontal Ligament

To analyze the age-dependent changes in nerve fibers immunoreactive to calcitonin gene-related peptide (CGRP-ir) in the periodontal ligaments of rats. Thirty male Wistar-ST rats were divided into growing groups (5, 9, and 15 weeks of age) and aging groups (6, 12, and 24 months of age) (n = 5 in each group). Eight serial sagittal sections, 5 microm thick, were cut parallel to the distobuccal root of the maxillary right first molar. These tissues were stained with a rabbit monoclonal antibody against CGRP. The observation area was divided into three parts (mesial, apical, and distal) and observed using a light microscope. CGRP-ir nerve fibers were primarily distributed in the apical periodontal ligament in the growing group, with significantly more fibers than in the aging group. CGRP-ir nerve fibers in the periodontal ligament are dense during the growth period and decrease gradually with aging, indicating that CGRP may affect periodontal tissue with growth and aging.

Immunohistochemical Characterization of Elastic System Fibers in Rat Molar Periodontal Ligament

Among elastic system fibers, oxytalan fibers are known as a ubiquitous component of the periodontal ligament, but the localization and role of elastin-containing fibers, i.e., elastic and elaunin fibers, has yet to be clarified. In this study, we immunohistochemically investigated the localization of elastin and fibrillin, major proteins of elastin-containing fibers in the periodontal ligament of rat lower first molars. At the light microscope level, distribution of elastin-positive fibers was not uniform but often concentrated in the vicinity of blood vessels in the apical region of the ligament. In contrast, fibrillin-positive fibers were more widely distributed throughout the ligament, and the pattern of their distribution was comparable to the reported distribution of oxytalan fibers. At the ultrastructural level, assemblies or bundles of abundant fibrillin-containing microfibrils were intermingled with a small amount of elastin. This observation indicated that elastin-positive fibers observed under the light microscope were elaunin fibers. No mature elastic fibers, however, were found in the ligament. These results show that the major components of elastic system fibers in the periodontal ligament of the rat mandibular first molar were oxytalan and elaunin fibers, suggesting that the elastic system fibers play a role in the mechanical protection of the vascular system.

Characteristics of periodontal ligament subpopulations obtained by sequential enzymatic digestion of rat molar periodontal ligament

Periodontal ligament (PDL) consists of different cell populations in various differentiation stages. In the present study, we isolated cell populations from rat molar PDL by sequential enzymatic digestion and characterized growth potential and mineralization activity of the PDL subpopulations (PDL-SP) to throw light on the mechanism of PDL remodeling and, in its turn, periodontal tissue regeneration. PDL attached to extracted rat molars was digested 2 mg/ml collagenase and 0.25% trypsin at 37°C for 30 min. Then four consecutive digestions were performed for 20 min each in a fresh digestive solution. The solutions were centrifuged to collect released cells and 5 PDL subpopulations (30M-, 50M-, 70M-, 90M-and 110M-PDL-SP) were obtained. Light microscopic observation showed that about a half of PDL in width attached on the root surface of extracted teeth and 30M-PDL-SP was considered to contain cells mainly from middle portion of PDL. Scanning electron microscopic examination indicated that 110M-PDL-SP was enriched by root lining cementoblastic cells. 30M-PDL-SP showed a high level of proliferative activity. Although the growth potential of a subpopulation decreased in PDL-SP toward the root surface, 110M-PDL-SP had a high proliferative activity equivalent to that of 30M-PDL-SP. Analyses of alkaline phosphatase (ALP) and mineralization activities showed that higher activities in PDL-SP toward the surface of roots and that 110M-PDL-SP had the highest activity of ALP and the largest number of mineralization nodules. The present study shows as supposed by previous studies on cell kinetics in PDL that subpopulations with larger growth potential were generally located in the middle portion of PDL and those with higher mineralization activities toward the surface of the roots. It is suggested, however, that a possible pathway of PDL cell turnover may exist within the PDL-SP on the root surface in addition to the generally recognized pathway from the middle area of PDL to root surface.

Effects of age on the stress–strain and stress–relaxation properties of the rat molar periodontal ligament

Objective: We examined the stress–strain and stress–relaxation properties of the periodontal ligament (PDL) in the rat molar at 2, 6, 12, and 24 months of age to elucidate age-related changes in the tooth support function of the PDL. Design: From the dissected left and right mandibles in each rat, a pair of transverse sections (ca. 0.45 mm in thickness) of the first molar was cut at the middle part of the mesial root. We then obtained a load–deformation curve for the PDL, using one of the paired sections. The other section was loaded to as much as 50% of the maximum load as determined from the contralateral section, and keeping the deformation constant for 10 min, a load–relaxation curve was obtained and analysed. Results: The maximum shear stress and tangent modulus decreased between 2 and 24 months of age. As the maximum shear strain increased with age ( P<0.001), the failure strain energy density did not change between 2 and 24 months of age. The stress–relaxation during the 10 min period decreased from 2 to 24 months of age ( P<0.01). The relaxation process of the PDL in each age was well described by a sum of three exponential decay functions. The age-related decrease in the relaxation was found to be mainly due to the increase in the relaxation time for the long-term relaxation component. Conclusion: These results indicate that the maximum shear stress and stiffness of the rat molar PDL decrease between 2 and 24 months of age; but its toughness remains unchanged due to an increase in the extensibility. Our findings further indicate that the fluid flow and movements of macromolecules within the stretched PDL fibres during the stress–relaxation decrease with advancing age.

Phosphatidylinositol-dependent bond between alkaline phosphatase and collagen fibers in the periodontal ligament of rat molars.

Alkaline phosphatase (ALP) is anchored to the outer leaflet of the lipid bilayer via phosphatidylinositol (PI) and ALP activity has been localized in the plasma membrane of numerous tissues. In the periodontal ligament ALP activity is found in the collagen fibers in addition to the plasma membrane of the osteoblasts and fibroblasts. In this study, we examined the distribution of ALP activity in the periodontal ligament of rat molars and also examined whether the bond between ALP and collagen fibers is dependent on PI by using phosphatidylinositol-specific phospholipase C (PI-PLC). ALP activity was distributed in the periodontal ligament. The activity mirrored the distribution of collagen fibers in the periodontal ligament. Cytochemical analysis also demonstrated that ALP activity was located not only in the plasma membrane of fibroblasts, but also in the collagen fiber bundles and fibrils in the periodontal ligament. After treatment with PI-PLC, the loss of ALP activity in the periodontal ligament was observed histochemically, and the loss of ALP activity in the fibroblasts as well as in the collagen fiber bundles and fibrils was observed cytochemically. These results strongly indicate that the bond between ALP and the collagen fibers is also dependent on PI.

Development of oxytalan fibers in the rat molar periodontal ligament.

Although oxytalan fibers are known to be a ubiquitous component of the periodontal ligament, little information has been available concerning their organization in the developing periodontal ligament. In the present study, growth and distribution of oxytalan fibers were examined in the developing periodontal ligament of rat molars aged 11, 14, 19, 21 and 28 days. A quantitative analysis of the fibers was made and the spatial relationship between the fibers and blood vessels was studied by means of a three-dimensional reconstruction of serial sections. At the beginning of root formation, oxytalan fibers appeared at first as dot-like structures around the root sheath as well as in areas very close to blood vessels. These structures were resolved in the electron microscope to be made up of 12-nm-wide microfibrils in the vicinity of the surface of the cells of the root sheath. In the process of development, these dot-like structures elongated into entities with helical appearances. As the development further proceeded, longer oxytalan fibers were produced in the apico-occlusal direction along with blood vessels. Quantitative analysis showed that an increase in oxytalan fibers coincided with an increase in the density of the vascular network in the developing periodontal ligament. Based on the results of the present study, the role of oxytalan fibers in the developing periodontal ligament may be in the maintenance of the integrity of the vascular system as previously suggested.

Distribution pattern of versican, link protein and hyaluronic acid in the rat periodontal ligament during experimental tooth movement.

The ability of the periodontal ligament (PDL) to rapidly remodel is the basis of orthodontic tooth movement. During the tooth movement, matrix proteoglycans (PGs) may play important roles in spatial, mechanical and biological aspects for the maintenance and repair of the PDL. The aim of this study was to characterize the distribution of a large hyaluronic acid (HA)-binding proteoglycan, versican, link protein (LP) and HA in the rat molar PDL during experimental tooth movement by histochemical and immunohistochemical methods. Experimental tooth movement was performed according to Waldo's method. Histologically, regressive changes, such as decrease of fibroblasts and collagen fibers and exudative change of edema were observed in the compressive side and progressive changes, such as proliferation of fibroblasts and collagen fibers, in the strain side one day after treatment. By 3 days after tooth movement, regressive or progressive changes were not observed in either side. Using monoclonal antibodies specific to versican core protein or LP, the positive immunoreactivity for both molecules was constantly observed throughout the PDL. After the experimental force was applied to the tooth, however, the immunostainings of versican and LP became significantly intense only in the compressive side but decreased in the strain side. The intensity in the compressive side was strongest one day after the force was applied and gradually diminished thereafter. HA of both sides did not change during experimental tooth movement. Since HA is present in the PDL, large amounts of versican and LP expressed in the compressive side may create large hydrated aggregates via their association with HA that dissipates the compressive force applied to this tissue.

Endothelin-1 expression in the microvasculature of normal and 3-hour continuously loaded rat molar periodontal ligament.

Endothelin-1 (ET-1) distribution is described in the microvascular bed (MVB) of normal rat molar periodontal ligament (PDL). Five male Sprague-Dawley rats, aged 90 days, were anaesthetized and an external pressure of 100 +/- 20 g, maintained for 3 hours, was transmitted occlusally to randomly allocated left or right molars. The controls were the contralateral molars. Rats were perfused for 5 minutes with 5 per cent paraformaldehyde, and the mandibles post-fixed and stored in 30 per cent sucrose. Sagittal, undemineralized, mandibular jaw sections, approximately 150 microns thick, were immunolabelled with ET-1 and alpha-smooth muscle actin (alpha-SMA) primary antibodies, and IgG/CY5 and IgG/CY3 secondary antibodies, respectively. Serial images were captured with a Bio-Rad MRC-1000 UV confocal laser scanning microscope (CLSM). In the control PDL, the ET-1 immunolabelling occurred sporadically in all categories of PDL blood vessels. ET-1 showed a punctate distribution within endothelial cells, producing longitudinal and circumferential pan-endothelial labelling. Immunoreactivity to the ET-1 antibody occurred at or adjacent to vessel branching sites, and affected blood vessels with and lacking alpha-SMA immunolabelling. Treatment effects on ET-1 immunofluorescence were analysed for vascular endothelium, socket bone surface cells, cementum surface cells and PDL background in the cervical, inter-radicular and apical regions. Significant (P < 0.05) region by treatment interactions occurred for the endothelium and bone. Cementum showed a significant (P < 0.05) region effect and a significant (P < 0.05) treatment effect. However, the region by treatment interaction was not significant. Background immunofluorescence showed significant (P < 0.05) region by treatment effects for the endothelium and bone. ET-1 activity is the default state for normal PDL vascular endothelium.

Developmental changes of the vasculature in the periodontal ligament of rat molars: a scanning electron microscopic study of microcorrosion casts.

Vascularization of the periodontal ligament was examined in developing upper first molars of rats from 5 to 30 d after birth with light and scanning electron microscopy. Formation of the vascular network in the periodontal ligament (PDL) started with the beginning of root formation. The PDL vessels derived from the basal region of the tooth germ ran parallel to the long axis of the root and connected with the vascular network of the enamel organ at the cervical end. The boundary of these 2 networks was initially indistinct but became clearer with the progress of root formation. The PDL vessels further elongated longitudinally and connected with each other by lateral branches to form a coarse mesh. Other vessels derived from the alveolar bone via Volkman's canals also contributed to the vascular construction of the PDL. The vessels from the alveolar bone provided branches to the existing mesh of the PDL. Consequently, the vascular network of the PDL consisted of vessels from 2 sources: 1 derived from the basal region of the tooth germ, and the other from the alveolar bone. The density of the vascular network reduced with the progress of root formation, especially at the middle part of the root, but the mesh at the apical region maintained a basket-like structure.

In Vitro Effects of Collagenase on Biomechanical Properties and Morphological Features of the Rat Molar Periodontal Ligament.

Biomechanical properties and morphological features of the rat molar periodontal ligament were examined after treatment with various concentrations of bacterial collagenase. Treatment with collagenase caused dose-dependent decreases in the maximum shear stress, maximum shear strain, tangent modulus, and failure strain energy density of the periodontal ligament. Light microscopic analysis after mechanical testing revealed that in controls, the periodontal ligament ruptured in the middle region or near the bone surface, while in the collagenase-treated specimens, it ruptured at the region adjacent to the cementum surface. Scanning electron microscopy revealed considerable degradation of the periodontal collagen fibres and fibrils by the enzyme. Treatment with collagenase markedly reduced the phase retardation values of collagens and the areas occupied by birefringent collagen fibres within the ligament. We suggest that collagenase treatment causes a decrease in the amount and density of collagen fibres and a disorganization of the fibres that reduces the strength, extensibility, stiffness, and toughness of the ligament. Our findings indicate that the periodontal collagens are load-bearing, key structural components providing structural and functional integrity to the ligament.

Biomechanical and Morphological Studies on the Periodontal Ligament of the Rat Molar after Treatment with α-amylase in vitro

Biomechanical properties and morphological features of the rat molar periodontal ligament were examined after treatment with α-amylase. Treatment with α-amylase induced dose-dependent decreases in the maximum shear stress, tangent modulus, and failure strain-energy density of the periodontal ligament; in addition, it weakened the alcian-blue staining of the periodontal ligament and exposed periodontal collagen fibrils as revealed by scanning electron microscopy. Azan staining and polarized microscopic observations of the periodontal collagen fibers were not markedly different between the control and α-amylase treated specimens. These results suggest that decreases in the strength of the periodontal ligament due to α-amylase digestion are largely due to removal of interfibrillar substances such as acid glycosaminoglycans and neutral polysaccharides from the periodontal ligament. It is also suggested that the interaction of the interfibrillar substances with collagen fibrils is involved in determining the biomechanical properties of the periodontal ligament.

Different localizations of growth-associated protein (GAP-43) in mechanoreceptors and free nerve endings of adult rat periodontal ligament, dental pulp and skin.

Distributions of growth-associated protein-43 (GAP-43) in the periodontal ligament and dental pulp of adult rats were studied by light and electron microscopy. The mature periodontal ligament and dental pulp contained numerous GAP-43-positive neural elements, comprising periodontal Ruffini endings and thin nerve fibers, but expression patterns differed among the kinds of nerves. In the periodontal ligament of rat molars, immunoelectron microscopy revealed that GAP-43 like immunoreactivity in the Ruffini ending, an essential mechanoreceptor, was confined to the Schwann sheaths around the axon terminals and was not in the axon terminals themselves, unlike free endings that revealed axonal GAP-43. However, the lamellar Schwann cells associated with the cutaneous receptors did not exhibit any GAP-43 like immunoreactivity though they were intensely reactive for low affinity nerve growth factor receptor (p75-NGFR), a marker for lamellar Schwann cells in mechanoreceptors. The characteristically uniform expression of GAP-43 in the Schwann lamellae that surround the Ruffini mechanoreceptors of rat molar ligament suggests that Schwann cells are involved in the GAP-43 mediated plasticity of these receptors. On the other hand, the pulpal nerves were filled with the reaction products in their axonal spaces, suggesting the potential for neuronal plasticity during normal function and after tooth injury.