TGF-beta3 induces ectopic mineralization in fetal mouse dental pulp during tooth germ development

Several members of the transforming growth factor (TGF)-beta superfamily are expressed in developing teeth from the initiation stage through adulthood. Of those, TGF-beta1 regulates odontoblast differentiation and dentin extracellular matrix synthesis. However, the molecular mechanism of TGF-beta3 in dental pulp cells is not clearly understood. In the present study, beads soaked with human recombinant TGF-beta3 induced ectopic mineralization in dental pulp from fetal mouse tooth germ samples, which increased in a dose-dependent manner. Further, TGF-beta3 promoted mRNA expression, and increased protein levels of osteocalcin (OCN) and type I collagen (COL I) in dental pulp cells. We also observed that the expression of dentin sialophosphoprotein and dentin matrix protein 1 was induced by TGF-beta3 in primary cultured dental pulp cells, however, not in calvaria osteoblasts, whereas OCN, osteopontin and osteonectin expression was increased after treatment with TGF-beta3 in both dental pulp cells and calvaria osteoblasts. Dentin sialoprotein was also partially detected in the vicinity of TGF-beta3 soaked beads in vivo. These results indicate for the first time that TGF-beta3 induces ectopic mineralization through upregulation of OCN and COL I expression in dental pulp cells, and may regulate the differentiation of dental pulp stem cells to odontoblasts.     

Upregulation of bone-like extracellular matrix expression in human dental pulp stem cells by mechanical strain

There are many different types of periodontal diseases. One such disease causes a defect of alveolar bone that is considered serious. Hence, researchers have examined potential treatments for this type of disease using tissue engineering techniques. Periodontal tissues are exposed to mechanical stress caused by occlusion and mastication, and both the cells and extracellular matrix in these tissues undergo architectural modifications to compensate for the applied stress. Therefore, in this study we analyzed the effect of mechanical tension on the osteogenesis of human dental pulp stem cells (DPSCs). To identify osteogenesis induced by mechanical stress in dental pulp, we examined the effects of tension on DPSCs. We evaluated the effects of mechanical stimuli on the osteogenesis of human dental pulp cells grown on silk scaffolds subjected to 10% strain using a bioreactor. The tension was applied with 0.2 Hz over the course of 5 days and was then continuously applied for 10 more days. We evaluated cell differentiation by RT-PCR, Western blotting and immunohistochemistry. Applying 10% tension to the culture resulted in increases in collagen type I, fibronectin, osteoprotegerin, and bone sialoprotein expression and decreases in a-smooth muscle actin expression. These data suggest that mechanical stimulation promotes osteogenesis in DPSCs.     

Inhibition of jaw opening reflexes by stimulation of the central gray matter and raphe nuclei in cats

The effect of stimulation of the mesencephalic central gray matter and raphe nuclei on jaw opening reflexes evoked by excitation of high-threshold (dental pulp) and low-threshold (A-alpha) fibers of the infraorbital nerve afferents was studied in cats anesthetized with chloralose and pentobarbital. The jaw opening reflex evoked by stimulation of the dental pulp was shown to be effectively suppressed by conditioning stimulation of the central gray matter and raphe nuclei. The reflex evoked by stimulation of low-threshold infraorbital nerve afferents also was depressed (but less deeply and for a shorter period than the reflex evoked by stimulation of the dental pulp) during stimulation of the raphe nuclei and caudal zone of the central gray matter, but was unchanged after stimulation of the points located in the rostral zone of the central gray matter. Application of single stimuli or bursts of five stimuli with a frequency of 100 Hz had no effect on the reflexes studied. Short-term stimulation with a burst of 10–20 stimuli with a following frequency of 200–400 Hz led to inhibition of the reflexes, which lasted 450–1000 msec. Long-term stimulation of the central gray matter and raphe nuclei for 30 sec with a frequency of 50 Hz caused inhibition of jaw opening reflexes evoked by stimulation of both high- and low-threshold afferents for 60 min. Impulses from the central gray matter and raphe nuclei thus have a mainly inhibitory action on the jaw opening reflex evoked by stimulation of high-threshold afferents, but they act less effectively on the reflex evoked by stimulation of low-thres-hold afferents. The duration of inhibition depends essentially on the parameters of stimulation.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 16, No. 3, pp. 374–387, May–June, 1984.     

Ulex europaeus agglutinin-I binding to dental primary afferent projections in the spinal trigeminal complex combined with double immunolabeling of substance P and GABA elements using peroxidase and colloidal gold

Ulex europaeus agglutinin I (UEA-I) is a plant lectin with an affinity for L-fucosyl residues in the chains of lactoseries oligosaccharides associated with medium- and smaller-diameter dorsal root ganglion neurons and their axonal processes. These enter Lissauer's tract and terminate within the superficial laminae of the spinal cord overlapping projections known to have a nociceptive function. This implies that the surface coatings of neuronal membranes may have a relationship with functional modalities. The present investigation further examined this concept by studying a neuronal projection with a nociceptive function to determine whether fucosyl-lactoseries residues were incorporated in its primary afferent terminals. Transganglionic transport of horseradish peroxidase (HRP) following injection into tooth pulp chambers was employed to demonstrate dental pulp terminals in the trigeminal spinal complex, while peroxidase and fluorescent tags were used concomitantly to stain for UEA-I. Double immunolabeling for substance P (SP) and gamma-aminobutyric acid (GABA) using peroxidase and colloidal gold allowed a comparison of the distribution of a known excitatory nociceptive transmitter with that of UEA-I binding in specific subnuclei. Synaptic interrelationships between UEA-I positive dental pulp primary afferent inputs and specific inhibitory terminals were also examined. SP immunoreactivity occurred in laminae I and outer lamina II (IIo) of subnucleus caudalis (Vc) and in the ventrolateral and lateral marginal region of the caudal half of subnucleus interpolaris (Vi), including the periobex area in which Vi is slightly overlapped on its lateral aspect by cellular elements of Vc. The adjacent interstitial nucleus (IN) also showed an intense immunoreactivity for this peptide antibody. UEA-I binding displayed a similar distribution pattern in both Vc and Vi, but extended into lamina IIi and the superficial part of Lamina III in Vc. Dental pulp terminals were found to have a comparable distribution; however, many extended into the dorsal portion of the caudal half of Vi and the ventromedial quadrant of rostral Vi. Electron-microscopic analysis showed that transganglionically labeled dental pulp terminals contained ovoid, complex membrane-bound vacuoles laden with transported HRP. The preterminal axon and synaptic membranes of those dental pulp terminals located in zones of Vc and Vi displaying an affinity for UEA-I were usually characterized by a patchy, electron-dense coating of the peroxidase tag. SP was demonstrated ultrastructurally with Protein-A colloidal gold (3-nm particles), whereas GABA immunoreactivity was revealed by the avidin-biotin-peroxidase method.(ABSTRACT TRUNCATED AT 400 WORDS)     

Human recombinant gamma-interferon stimulates proliferation and inhibits collagen and fibronectin production by human dental pulp fibroblasts

Human recombinant-gamma-interferon was tested on human dental pulp fibroblast activity in vitro. Fibroblast proliferation was estimated by a colorimetric test. Type I and type III collagens and fibronectin were quantified by radioimmunoassay in culture supernatant from confluent fibroblasts. A dose dependent stimulation of the proliferation was observed when fibroblasts were treated with recombinant-gamma-interferon. In contrast, an inhibition of the synthesis of soluble types I and III collagen and fibronectin by confluent cell cultures treated with recombinant-gamma-interferon occurred without apparent modification of the insoluble collagen level in the cell layer. Quantimetric analysis of type I collagen immunoperoxidase labelling have demonstrated that there was no intracellular storage of type I collagen in these cultured fibroblasts. These data support the view that human recombinant-gamma-interferon can affect human dental pulp fibroblast functions and thus may play an important part in the regulation of fibrosis.     

Wnt5a Promotes Inflammatory Responses via Nuclear Factor κB (NF-κB) and Mitogen-activated Protein Kinase (MAPK) Pathways in Human Dental Pulp Cells

Wnt5a has been found recently to be involved in inflammation regulation through a mechanism that remains unclear. Immunohistochemical staining of infected human dental pulp and tissue from experimental dental pulpitis in rats showed that Wnt5a levels were increased. In vitro, Wnt5a was increased 8-fold in human dental pulp cells (HDPCs) after TNF-α stimulation compared with control cells. We then investigated the role of Wnt5a in HDPCs. In the presence of TNF-α, Wnt5a further increased the production of cytokines/chemokines, whereas Wnt5a knockdown markedly reduced cytokine/chemokine production induced by TNF-α. In addition, in HDPCs, Wnt5a efficiently induced cytokine/chemokine expression and, in particular, expression of IL-8 (14.5-fold) and CCL2 (25.5-fold), as assessed by a Luminex assay. The cytokine subsets regulated by Wnt5a overlap partially with those induced by TNF-α. However, no TNF-α and IL-1β was detected after Wnt5a treatment. We then found that Wnt5a alone and the supernatants of Wnt5a-treated HDPCs significantly increased macrophage migration, which supports a role for Wnt5a in macrophage recruitment and as an inflammatory mediator in human dental pulp inflammation. Finally, Wnt5a participates in dental pulp inflammation in a MAPK-dependent (p38-, JNK-, and ERK-dependent) and NF-κB-dependent manner. Our data suggest that Wnt5a, as an inflammatory mediator that drives the integration of cytokines and chemokines, acts downstream of TNF-α.     

Effects of electrical stimulation of ventral septal area on firing rates of pyrogen-treated thermosensitive neurons in preoptic anterior hypothalamus from rabbits

Although there is considerable evidence supporting that fever evolved as a host defense response, it is important that the rise in body temperature would not be too high. Many endogenous cryogens or antipyretics that limit the rise in body temperature have been identified. Endogenous antipyretics attenuate fever by influencing the thermoregulatory neurons in the preoptic anterior hypothalamus (POAH) and in adjacent septal areas including ventral septal area (VSA). Our previous study showed that intracerebroventricular (I.C.V.) injection of interleukin-1beta (IL-1beta) affected electrophysiological activities of thermosensitive neurons in VSA regions, and electrical stimulation of POAH reversed the effect of IL-1beta. To further investigate the functional electrophysiological connection between POAH and VSA and its mechanisms in thermoregulation, the firing rates of thermosensitive neurons in POAH of forty-seven unit discharge were recorded by using extracellular microelectrode technique in New Zealand white rabbits. Our results show that the firing rates of the warm-sensitive neurons decreased significantly and those of the cold-sensitive neurons increased in POAH when the pyrogen (IL-1beta) was injected I.C.V. The effects of IL-1beta on firing rates in thermosensitive neurons of POAH were reversed by electrical stimulation of VSA. An arginine vasopressin (AVP) V1 antagonist abolished the regulatory effects of VSA on the firing rates in thermosensitive neurons of POAH evoked by IL-1beta. However, an AVP V2 antagonist had no effects. These data indicated that VSA regulates the activities of the thermosensitive neurons of POAH through AVP V1 but not AVP V2 receptor.     

Interleukin-6 expression on inflamed rat dental pulp tissue after capped with Trigona sp. propolis from south Sulawesi,Indonesia

Background: Propolis is a natural product of plant resins collected by honeybees from various plant sources. It is used as a remedy in folk medicine since ancient times because of its several biological and pharmacological properties. Recently, propolis has been used by dentist to treat various oral diseases. It was always mentioned as an anti-inflammatory agent. Cytokines are proteins that provide communication between cells and play a critical role in a wide variety of processes. It released from cells in an inflammatory process that active, mediate or potential actions of other cells or tissues. When dental pulp has inflammation, several pro-inflammatory cytokines including Interleukin-6 (IL-6) was released by innate immune cells. Objective: To analyse the expression of IL-6 on inflamed rat dental pulp tissue following application of propolis. Material and methods: Trigona sp. propolis was obtained from Luwu Regency, south Sulawesi Province, Indonesia. Flavonoid and non-flavonoid extracts were purified from propolis using thin layer chromatography. The study was applied on 80 male Sprague Dawley rats, 10–12 weeks of age, divided randomly and equally into 5 groups. Group I, as negative control group was not conducted any treatment. At group II, III, IV and V. A Class I cavity (Black Classification) were made on the occlusal surface of right maxillary first molar. The dental pulp was perforated using dental explorer and allowed in the oral environment for 1 h, after that, Ethanolic Extract Propolis (EEP) (Group II), Extract Flavonoid-Propolis (EFP) (Group III), Extract Non-Flavonoid Propolis (ENFP) (Group IV), or Calcium Hydroxide (Ca(OH)₂) (Group V) were applied on dental pulp. All cavities were then filled with Glass Ionomer Cement as permanent filling. The rats being sacrificed in 6 h, 2 days, 4 days and 7 days. Sample biopsy were obtained, IL-6 expression was detected by using immunohistochemistry method. Data was analyzed statistically using Freidman and Kruskal Wallis tests with significance level of P < 0.05. Results: All agent showed IL-6 expression in inflamed rat dental pulp tissue, and this expression was decreased with the longer of observation time periods. EEP more stronger to decreased IL-6 expression on inflamed rat dental pulp tissue than other agent. There is significant difference (P < 0.05) of IL-6 expression between group I and other groups in 6 h and 2 days but not in 4 and 7 days time periods. Conclusion: Trigona sp. propolis from south Sulawesi, Indonesia could suppressed the expression of IL-6 on inflamed rat dental pulp tissue.     

Type I collagen regulated dentin matrix protein-1 (Dmp-1) and osteocalcin (OCN) gene expression of rat dental pulp cells

In this study, we investigated the effect of type I collagen on dentin matrix protein-1 (Dmp-1) and osteocalcin (OCN) gene expression of dental pulp cells. The mRNA level of Dmp-1 gene was down-regulated; however, OCN gene expression was up-regulated by the culture of dental pulp cells with type I collagen. These findings imply that type I collagen regulates mRNA level of Dmp-1 and OCN gene that are predominantly expressed in active odontoblasts. The change of gene expression by type I collagen was suppressed by the blocking of collagen-integrin interaction. We could conclude that the effect of type I collagen was mediated via binding of collagen to integrin receptors.     

Noxious stimuli do not determine reflex cardiorespiratory effects in anesthetized rabbits

Raimondi, G., J. M. Legramante, F. Iellamo, G. Frisardi, S. Cassarino, and G. Peruzzi. Noxious stimuli do not determine reflexcardiorespiratory effects in anesthetized rabbits. J. Appl. Physiol. 81(6): 2421-2427, 1996.Themain purpose of this study is to examine whether the stimulation of anexclusively pain-sensing receptive field (dental pulp) could determinecardiorespiratory effects in animals in which the cortical integrationof the peripheral information is abolished by deep anesthesia. In 15 anesthetized (-chloralose and urethan) rabbits, low (3-Hz)- andhigh-frequency (100-Hz) electrical dental pulp stimulation wasperformed. Because this stimulation caused dynamic and static reflexcontractions of the digastric muscles leading to jaw opening[jaw-opening reflex (JOR); an indirect sign of algoceptive fiberactivation], experimentally induced direct dynamic and staticcontractions of the digastric muscle were also performed. The low- andhigh-frequency stimulation of the dental pulp determined cardiovascular[systolic arterial pressure (SAP): 21.7 ± 4.6 and 10.8 ± 4.7 mmHg, respectively] andrespiratory [pulmonary ventilation(E): 145.1 ± 44.9 and 109.3 ± 28.4 ml /min, respectively] reflexresponses similar to those observed during experimentally induceddynamic (SAP: 17.5 ± 4.2 mmHg;E: 228.0 ± 58.5 ml /min) and static (SAP: 5.8 ± 1.5 mmHg;E: 148.0 ± 75.3 ml /min) muscular contractions. The elimination ofdigastric muscular contraction (JOR) obtained by muscular paralysis didaway with the cardiovascular changes induced by dental pulpstimulation, the effectiveness of which in stimulating dental pulpreceptors has been shown by recording trigeminal-evoked potentials insix additional rabbits. The main conclusion was that, indeeply anesthetized animals, an algesic stimulus is unable to determinecardiorespiratory effects, which appear to be exclusively linked to thestimulation of ergoreceptors induced by muscular contraction.

     

Tea catechins reduce inflammatory reactions via mitogen-activated protein kinase pathways in toll-like receptor 2 ligand-stimulated dental pulp cells

AimsIn this study, we evaluated whether catechins could inhibit the expression of pro-inflammatory mediators induced by dental caries-related bacteria, Streptococci, or pathogen-associated molecular patterns (PAMPs) stimulation in human dental pulp fibroblasts (HDPF). We further determined the mechanisms of the anti-inflammatory activity of catechins.Main methodsStreptococci or PAMP-stimulated HDPF were treated with catechin, and then the expression and production of pro-inflammatory mediators were determined by RT-PCR and ELISA. Furthermore, the signal transduction pathways activated with toll-like receptor (TLR)2 ligand were assessed by Immunoblot and ELISA using blocking assay with specific inhibitors.Key findingsIncreased expressions of pro-inflammatory mediators are found in inflamed dental pulp, especially in HDPF. We recently reported that dental pulpal innate immune responses may mainly result from the predominantly-expressed TLR2 signaling. Catechins, polyphenolic compounds in green tea, exert protective and healing effects through multiple mechanisms, including antioxidative and anti-inflammatory effects. However, there are no reports concerning the effects of catechins on dental pulp. In this study, we demonstrated that the up-regulated expressions of IL-8 or PGE₂ in Streptococci or PAMP-stimulated HDPF were inhibited by catechins, (?)-epicatechin gallate (ECG) and (?)-epigallocatechin gallate (EGCG). In TLR2 ligand-stimulated HDPF, specific inhibitors of extracellular signal regulated kinase (ERK)1/2, p38, c-jun NH₂-terminal kinase (SAP/JNK), NF-κB or catechins markedly reduced the level of pro-inflammatory mediators and the phosphorylation of these signal transduction molecules was suppressed by catechins.SignificanceThese findings suggest that catechins might be useful therapeutically as an anti-inflammatory modulator of dental pulpal inflammation.     

Ulex europaeus Agglutinin-I Binding to Dental Primary Afferent Projections in the Spinal Trigeminal Complex Combined with Double Immunolabeling of Substance P and GABA Elements Using Peroxidase and Colloidal Gold

Ulex europaeus agglutinin I (UEA-I) is a plant lectin with an affinity for L-fucosyl residues in the chains of lactoseries oligosaccharides associated with medium and smaller-diameter dorsal root ganglion neurons and their axonal processes. These enter Lissauer's tract and terminate within the superficial laminae of the spinal cord overlapping projections known to have a nociceptive function. This implies that the surface coatings of neuronal membranes may have a relationship with functional modalities. The present investigation further examined this concept by studying a neuronal projection with a nociceptive function to determine whether fucosyl-lactoseries residues were incorporated in its primary afferent terminals. Transganglionic transport of horseradish peroxidase (HRP) following injection into tooth pulp chambers was employed to demonstrate dental pulp terminals in the trigeminal spinal complex, while peroxidase and fluorescent tags were used concomitantly to stain for UEA-I. Double immunolabeling for substance P (SP) and γ-aminobutyric acid (GABA) using peroxidase and colloidal gold allowed a comparison of the distribution of a known excitatory nociceptive transmitter with that of UEA-I binding in specific subnuclei. Synaptic interrelationships between UEA-I positive dental pulp primary afferent inputs and specific inhibitory terminals were also examined.

SP immunoreactivity occurred in laminae I and outer lamina II (II_o) of subnucleus caudalis (Vc) and in the ventrolateral and lateral marginal region of the caudal half of subnucleus interpolaris (Vi), including the periobex area in which Vi is slightly overlapped on its lateral aspect by cellular elements of Vc. The adjacent interstitial nucleus (IN) also showed an intense immunoreactivity for this peptide antibody. UEA-I binding displayed a similar distribution pattern in both Vc and Vi, but extended into lamina II; and the superficial part of Lamina III in Vc. Dental pulp terminals were found to have a comparable distribution; however, many extended into the dorsal portion of the caudal half of Vi and the ventromedial quadrant of rostral Vi.

Electron-microscopic analysis showed that transganglionically labeled dental pulp terminals contained ovoid, complex membrane-bound vacuoles laden with transported HRP. The preterminal axon and synaptic membranes of those dental pulp terminals located in zones of Vc and Vi displaying an affinity for UEA-I were usually characterized by a patchy, electron-dense coating of the peroxidase tag. SP was demonstrated ultrastructurally with Protein-A colloidal gold (3-nm particles), whereas GABA immunoreactivity was revealed by the avidin—biotin—peroxidase method. This combined approach labeled a variety of simple axodendritic to large complex scalloped dental terminals which contained SP and were shown to have a UEA-I affinity. In addition, many of the larger terminals formed contacts with GABA-ergic dendrites and received inputs from GABA-ergic synapses. These complexes were most concentrated in lamina II_o of Vc and the ventrolateral zone of Vi. Many terminals in laminae II_i; and III with a UEA-I-positive surface coating failed to bind with the antiserum for SP, indicating that other transmitters may colocalize with UEA-I and suggesting that absolute correlations between specific oligosaccharide plasmalemmal coatings and functional modalities should be approached with caution. Further studies employing antisera to different transmitters are currently underway to better define the relationship between transmitter localization and anatomical substrates within this circuitry. These studies should eventually provide additional clues about relationships between functional properties and oligosaccharide coatings of primary afferent projections.     

Demonstration of glutamate in primary sensory trigeminal neurons innervating dental pulp in rats]

Primary sensory trigeminal neurons supplying the dental pulp of incisors in rats were labelled by retrograde axonal transport. Using an auto-metallographic intensification procedure, 48 hrs. after injection of wheat-germ colloidal gold in the pulp, gold particles were detected in the cytoplasm of the neurons as black granulations. Glutamate was found in 45-60% of the neurons by submitting ganglion slices to an anti-glutamate immuno-serum revealed by immunocytochemistry. Among the neurons supplying the dental pulp of incisors by their peripheral process, 70% are Glu+, 30% Glu-. These observations suggest that the population of neurons supplying the dental pulp is not functionally homogeneous and that Glu- neurons use a different neurotransmitter. The coexistence of Glu+ and Glu- neurons could also indicate that glutamate expression is modulated during the life of these neurons.     

Change in the nociceptive reactions caused by dental pulp stimulation during excitation of the midbrain]

It was demonstrated in chronic experiments on cats that stimulation of certain midbrain regions decreased or fully depressed the pain reaction evoked by dental pulp stimulation. The antinociceptive effect depending on the parameters of the brain stimulation was shown in differential change of the separate motor and vegetative and emotional - behaviour components, forming a general pain reaction. A poststimulation analgesia was revealed and the dynamic of restoration of different pain manifestations after the cessation of brain stimulation was traced. Possible mechanism of the realization of the antinociceptive effect are discussed.     

Angiotensin mechanism of auriculo-acupuncture dental analgesia

In unanaesthetized rabbits auriculo-acupuncture electrostimulation with frequency of 15 Hz decreased the amplitude of somatosensory EP second component in response to the tooth pulp electrostimulation which was blocked by intravenous injection of naloxone but not by intraventricular injection of saralasin. The same effect of auriculo-acupuncture electrostimulation with frequency 100 Hz was blocked by saralasin, was increased by angiotensin II, was diminished by methysergide but wasn't changed by naloxone. It's suggested that there is angiotensinergic antinociceptive mechanism of dental pain which is activated by auriculo-acupuncture electrostimulation with frequency 100 Hz.     

The relationship between cell proliferation and differentiation and mapping of putative dental pulp stem/progenitor cells during mouse molar development by chasing BrdU-labeling

Human dental pulp contains adult stem cells. Our recent study demonstrated the localization of putative dental pulp stem/progenitor cells in the rat developing molar by chasing 5-bromo-2’-deoxyuridine (BrdU)-labeling. However, there are no available data on the localization of putative dental pulp stem/progenitor cells in the mouse molar. This study focuses on the mapping of putative dental pulp stem/progenitor cells in addition to the relationship between cell proliferation and differentiation in the developing molar using BrdU-labeling. Numerous proliferating cells appeared in the tooth germ and the most active cell proliferation in the mesenchymal cells occurred in the prenatal stages, especially on embryonic Day 15 (E15). Cell proliferation in the pulp tissue dramatically decreased in number by postnatal Day 3 (P3) when nestin-positive odontoblasts were arranged in the cusped areas and disappeared after postnatal Week 1 (P1W). Root dental papilla included numerous proliferating cells during P5 to P2W. Three to four intraperitoneal injections of BrdU were given to pregnant ICR mice and revealed slow-cycling long-term label-retaining cells (LRCs) in the mature tissues of postnatal animals. Numerous dense LRCs postnatally decreased in number and reached a plateau after P1W when they mainly resided in the center of the dental pulp, associating with blood vessels. Furthermore, numerous dense LRCs co-expressed mesenchymal stem cell markers such as STRO-1 and CD146. Thus, dense LRCs in mature pulp tissues were believed to be dental pulp stem/progenitor cells harboring in the perivascular niche surrounding the endothelium.     

Analysis of time- and concentration-mortality relationship of nitenpyram against different larval stages of Nilaparvata lugens (Hemiptera: Delphacidae)

Bioassay of nitenpyram was conducted on the planthopper Nilaparvata lugens (St?l) (Hemiptera: Delphacidae) in the laboratory by using the rice-stem dipping method. The resulting data were analyzed using the complementary log-log time-concentration-mortality modeling technique, yielding the parameters for time and concentration effects of nitenpyram against N. lugens instars. The LC50 values to instars I and II estimated for 24, 48, and 72 h after treatment were 1.03, 0.27, and 0.11 mg/liter, respectively. These values were lower than those for instars III and IV and instars V at the same times. The estimates of LT50 for instars I and II were decreased from 69.06 to 16.22 h with the increasing concentration from 0.125 to 2 mg/liter; the same estimates for both instars III and IV and instars V at the concentrations of 1-2 mg/liter were close to each other, ranging from 33.11 to 44.16 h, much higher than the values of instars I and II at the same concentration. Based on the time-concentration-mortality relationships fitted and the virulence indices (LC50 and LT50) estimated, nitenpyram is a promising pesticide for N. lugens control.     

Fluid mechanics in dentinal microtubules provides mechanistic insights into the difference between hot and cold dental pain

Dental thermal pain is a significant health problem in daily life and dentistry. There is a long-standing question regarding the phenomenon that cold stimulation evokes sharper and more shooting pain sensations than hot stimulation. This phenomenon, however, outlives the well-known hydrodynamic theory used to explain dental thermal pain mechanism. Here, we present a mathematical model based on the hypothesis that hot or cold stimulation-induced different directions of dentinal fluid flow and the corresponding odontoblast movements in dentinal microtubules contribute to different dental pain responses. We coupled a computational fluid dynamics model, describing the fluid mechanics in dentinal microtubules, with a modified Hodgkin-Huxley model, describing the discharge behavior of intradental neuron. The simulated results agreed well with existing experimental measurements. We thence demonstrated theoretically that intradental mechano-sensitive nociceptors are not "equally sensitive" to inward (into the pulp) and outward (away from the pulp) fluid flows, providing mechanistic insights into the difference between hot and cold dental pain. The model developed here could enable better diagnosis in endodontics which requires an understanding of pulpal histology, neurology and physiology, as well as their dynamic response to the thermal stimulation used in dental practices.     

Selective analgetic effects of angiotensin and bombesin during the action of dental and cutaneous nociceptive irritants

In non-anesthesized rabbits intraventricular injection of angiotensin II reduced the amplitude of somatosensory evoked potential to nociceptive tooth pulp, but not to nociceptive electrocutaneous stimulation. The same injection of bombesin induced the contrary analgetic effect. The systemic naloxone (0.1 mg/kg) injection didn't reverse the peptides analgetic effects. It's suggested that selective analgetic effects of angiotensin II and bombesin are determined by the presence of the specific different peptide mechanisms for nociception with the different pain genesis.