Biological characteristics of stem cells from deciduous tooth pulp of Beagle dog / 实用口腔医学杂志

Objective:To culture and characterize the deciduous tooth pulp stem cells(DTPSCs)of Beagle dog.Methods:DTPSCs were cultured using enzyme tissue block method from Beagle dog aged 6 weeks.The cells were purified by cloning culture,identified by immunohistochemistry and flowcytometry.The biological characteristics were studied by CCK-8 assay,osteogenetic induction,li-pogenic induction and dentinogenic induction assays.Results:Beagle stem cells from deciduous tooth pulp were obtained,the cell colony formation rate was 32%.The cells were STRO-1 and CD146 positive,CD14,CD45 and CD86 negative.After multiple induc-tion culture the cells were positive for alizarin red staining,oil red staining,ALP expression and DSSPP expression.Conclusion:The deciduous tooth pulp stem cells of Beagle dog have multilineage differentiation abilities.

Quantitative Ultrastructural Analysis of Endings Presynaptic to the Tooth Pulp Afferent Terminals in the Trigeminal Oral Nucleus

The ultrastructural parameters related to synaptic release of endings which are presynaptic to tooth pulp afferent terminals (p-endings) were analyzed to understand the underlying mechanism for presynaptic modulation of tooth pulp afferents. Tooth pulp afferents were labelled by applying wheat-germ agglutinin conjugated horseradish peroxidase to the rat right lower incisor, whereafter electron microscopic morphometric analysis with serial section and reconstruction of p-endings in the trigeminal oral nucleus was performed. The results obtained from 15 p-endings presynaptic to 11 labeled tooth pulp afferent terminals were as follows. P-endings contained pleomorphic vesicles and made symmetrical synaptic contacts with labeled terminals. The p-endings showed small synaptic release-related ultrastructural parameters: volume, 0.82 ± 0.45 µm³ (mean ± SD); surface area, 4.50 ± 1.76 µm²; mitochondrial volume, 0.15 ± 0.07 µm³; total apposed surface area, 0.69 ± 0.24 µm²; active zone area, 0.10 ± 0.04 µm²; total vesicle number, 1045 ± 668.86; and vesicle density, 1677 ± 684/µm². The volume of the p-endings showed strong positive correlation with the following parameters: surface area (r=0.97, P<0.01), mitochondrial volume (r=0.56, P<0.05), and total vesicle number (r=0.73, P<0.05). However, the volume of p-endings did not positively correlate or was very weakly correlated with the apposed surface area (r=-0.12, P=0.675) and active zone area (r=0.46, P=0.084). These results show that some synaptic release-related ultrastructural parameters of p-endings on the tooth pulp afferent terminals follow the "size principle" of Pierce and Mendell (1993) in the trigeminal nucleus oralis, but other parameters do not. Our findings may demonstrate a characteristic feature of synaptic release associated with p-endings.

Pain behavior to electroacupuncture in rabbit tooth pulp

Aim: The aim of this study was to verify the pain behavior to electroacupuncture (EACP) in rabbit tooth-pulp assay. Methods: Albino rabbits weighing 1.5-2.0 kg) were pretreated with saline or morphine (5mg/kg, e.v.) 10 min before the nociceptive test (NT). In another group, EACP (rectangular pulses, f1=2 Hz, f2=0.1 s, 3 mA) was applied in acupoints and sham points, before and during the NT. After 120 min, EACP was withdrawn and the nociceptive threshold was measured every 10 min until the initial nociceptive threshold was achieved. Results: EACP, using the Yintang, ST4 and ST5 acupoints, induced an increased in the nociceptive threshold and this effect persisted for up to 2 h, even after the removal of electric stimulation. Application of EACP at sham points did not show significant analgesic activity. The present results demonstrated that males presented a higher initial level of analgesia, but a poorer maintenance of analgesic effect after the EACP procedure, while females demonstrated a long lasting analgesic effect even after discontinuation of EACP. Conclusions: EACP presented an analgesic effect in a rabbit tooth pulp assay that was probably due to the release of endogenous opioids. The duration of this analgesic effect seems to be different for males and females.

Processing Mechanism of Sensory Information Originated from the Oral Cavity in the Trigeminal Nucleus Oralis / 대한해부학회지

To analyze the synaptic characteristics of axon terminals originated from the tooth pulp in the trigeminal nucleus oralis, labeling of tooth pulp afferents with wheat-germ agglutinin conjugated horseradish peroxidase and morphometric analysis with electron microscopic photographs were performed. The results obtained from 23 labeled endings were as follows. All of the labeled boutons contained clear and round synaptic vesicles (dia. 45~55 nm). 3 (13.64%) out of 23 labeled endings have 20~105 dense cored vesicles and do not make synaptic contacts with p-endings. But remaining 20 labeled endings (86.36%) almost do not have dense cored vesicles and 12 of them make synaptic contacts with p-endings. The mean number of synaptic contacts was 2.61+/-2.06 and the postsynaptic profiles were usually middle or distal dendrite and dendritic spine (1.74+/-1.36) rather than soma or proximal dendrite. The mean number of synaptic contacts with pendings was 0.87+/-1.01. And the frequency of the synaptic triads were 0.39+/-0.58. The vesicle density was 993.23+/-267.41/mum(2). The volume of labeled bouton was 3.54+/-2.20 mum(3) and highly correlated (P < 0.01) with surface area (11.78+/-4.92 mum(2), r = 0.95), total apposed surface area (2.90+/-1.56 mum(2), r=0.72), total active zone area (0.61+/-0.37 mum(2), r = 0.82), mitochondrial volume (0.75+/-0.53 mum(3), r = 0.94), the number of synaptic vesicles (2621.30+/-1473.61, r= 0.91) and the number of synaptic contacts (r = 0.76). These results suggest that there are two groups of tooth pulp afferent terminals according to the presence of dense cored vesicles in the trigeminal nucleus oralis. And the sensory processing mechanism of each groups may be different. And the "size principle" of Pierce & Mendell (1993) is also applicable to these terminals.

Quantitative and Ultrastructural Analysis of Afferent Terminals from Tooth Pulp in the Trigeminal Principal Sensory Nucleus / 대한해부학회지

To analyze the synaptic characteristics of axon terminals originated from tooth pulp in the trigeminal principal sensory nucleus, and find morphological evidence that p-endings on these terminals use GABA as neurotransmitter, labeling of tooth pulp afferents with wheat -germ agglutinin conjugated horseradish peroxidase, postembedding immunocytochemistry and morphometric analysis were performed. The results obtained were as follows. The labeled boutons contained clear and round synaptic vesicles, and the mean number of synaptic contacts was 4.21 +/- 2.58. The postsynaptic profiles were usually middle or distal dendrites and dendritic spines rather than soma or proximal dendrites. The mean number of synaptic contacts with p -endings was 2.62 +/- 2.01. The volume of labeled bouton was 4.39 +/- 3.08 micro meter 3 and highly correlated (P<.01) with surface area (r = 0.96), total apposed surface area (r = 0.84), total active zone area (r = 0.84), mitochondrial volume (r = 0.95), the number of synaptic vesicles (r = 0.92), the number of synaptic profiles (r = 0.76) and the number of synaptic p -endings (r = 0.67). The p -endings contained pleomorphic synaptic vesicles. The volume of p -ending was 0.91 +/- 0.47 micro meter 3 and highly correlated (P<.01) with surface area (r = 0.94), total apposed surface area (r = 0.76), total active zone area (r = 0.62), mitochondrial volume (r = 0.70) and the number of synaptic vesicles (r = 0.94). P -endings were GABA immunoreactive. These results suggest that tooth pulp afferent terminals in trigeminal principal sensory nucleus may be related to complex processing of sensory information and presynaptic p -endings using GABA as neurotransmitter, are important in this sensory processing. The "size principle" of Pierce and Mendel (1993) is also applicable to tooth pulp afferent terminals and presynaptic p -endings.