Antagonistic effect of D-myo-inositol-1,2,6-trisphosphate (PP56) on neuropeptide Y-induced vasoconstriction in the feline dental pulp.

Intra-arterial injection of neuropeptide Y (NPY) (1.3-2.0 micrograms/kg) resulted in decreases of pulpal blood flow by 37.7 +/- 5.7% (mean +/- SEM). The intra-arterial injection of D-myo-inositol-1,2,6-trisphosphate (PP56) (0.3 mg/kg) alone changed pulpal blood flow by 1.0%. The effect of NPY in the presence of PP56 resulted in significantly smaller decreases in pulpal blood flow ranging from 27.2 +/- 5.4 to 16.6 +/- 3.5% from control as compared with NPY alone. In effect, PP56 partially blocked the decreases in pulpal blood flow caused by NPY. The electrical stimulation of the sympathetic nerve alone resulted in decreases in pulpal blood flow of 41.7 +/- 6.2%. The electrical stimulation of the sympathetic nerve following the intra-arterial administration of PP56 decreased pulpal blood flow by 23.1 +/- 6.0% from control, significantly less than the sympathetic nerve stimulation alone. PP56 attenuated the decrease in pulpal blood flow caused by the sympathetic nerve stimulation by 44.4 +/- 11.0%. Similarly, the combination of PP56 and phentolamine followed by electrical stimulation of the sympathetic nerve reduced the decrease in pulpal blood flow caused by the sympathetic nerve stimulation alone by 43.0 +/- 8.6%. These results provide evidence that the non-peptide PP56 is capable of antagonizing vasoconstriction caused by NPY in the feline dental pulp. In addition, they show functional evidence that NPY as well as noradrenaline are released from the sympathetic nerve endings during its stimulation and cause vasoconstriction.

Effects of CO2 laser irradiation on pulpal blood flow.

CO2 laser may be beneficially applied to teeth in preventive and restorative dentistry, but its physiopathologic effects on the dental pulp have not been reported. To address the dynamic pulpal changes induced by CO2 laser the purpose of this study was to examine such changes in pulpal microcirculation. The enamel surfaces of 4 cat canines were irradiated by CO2 laser beam, delivered by either a handpiece or microslad with a focal spot diameter of 0.21mm and 0.33mm, respectively. Exposure powers of 2.6-20 W were used for durations of 0.05-0.2 sec, resulting in energy density range of 304-1440 J/cm2. Non-invasive measurement of pulpal blood flow (PBF) by a laser Doppler flowmeter was maintained continuously before and following CO2 laser irradiation. CO2 laser irradiation resulted in an immediate increase in PBF, which subsided after 2-3 min. Large pulps responded with a significantly higher PBF increase than small pulps. These results confirm that CO2 laser irradiation of the tooth surface has a thermal effect on the pulp, which is moderated by the insulating capability of dentin. It appears that the associated thermal damage is not extensive when laser exposure parameters are such as used herein.

Effects of selected inflammatory mediators on blood flow and vascular permeability in the dental pulp.

One of the major events involving inflammatory processes is the alteration of microcirculatory hemodynamics by inflammatory mediators released from tissue components. Using modern macrocirculatory techniques, 15 mu radioisotope labeled microspheres, 133Xe washout, laser Doppler flowmetry and double isotopes, 125 and 131I-albumin, and microcirculatory methods, intravital fluorescence microscopy with FITC labeled dextran, we have examined the effects of selected mediators, e.g. 5-hydroxytryptamine (5-HT), prostaglandin E2 (PG-E2), bradykinin (BK), substance P (SP), calcitonin gene related peptide (CGRP) and histamine on blood flow and vascular permeability in the pulp of experimental animals. Surprisingly, SP and CGRP caused weak albumin leakage in the pulp, while the opposite is true in high compliance tissues, such as muscles, suggesting that the vessels in a low compliance environment, such as the pulp, may not be as permeable in response to selected mediators. Intraarterial injection of 5-HT caused a strong vasoconstriction which was mediated by 5-HT1p receptor subtype. The pulpal 5-HT receptor subtype was identified by immunocytochemistry, receptor autoradiography and functional investigations. Intravital fluorescence microscopy observations of the rat incisor preparation showed that histamine, BK and PGE2 increased permeability, whereas isoproteranol caused partial inhibition of the BK-induced increase. In an induced pulpal inflammation model using plaque extract, blood flow increased over 40% in the moderately inflamed pulp, which demonstrated severe vasodilation and polymorpholeukocyte accumulation. In the partially necrotic pulp, blood flow decreased nearly 35%. Results of this study clearly show that there is a high structural/functional correlation in pulpal microcirculation in inflammation. As demonstrated in this presentation, the effects of inflammatory mediators on pulpal microcirculatory hemodynamics are complex.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of capsaicin on pulpal blood flow.

One of the initial events in pulpal inflammation has been characterized as neurogenic inflammation: the release of neuropeptides following excitation of sensory C-fibers by noxious stimuli which alters microcirculatory parameters, that is, vasodilation and plasma extravasation. Thus, the purpose of the study was to investigate the effect of capsaicin on pulpal blood flow (PBF) with the aim of understanding neurogenic inflammation in the dental pulp by characterizing the response of the pulpal vasculature to repeated applications of various concentrations of capsaicin. Experiments were performed on canine teeth of cats anesthetized with sodium pentobarbital. PBF was measured by the laser Doppler flowmeter following the application of capsaicin into the dentinal cavities of the canine teeth. The increases of PBF to capsaicin were 31.8 +/- 6.3% (n = 6) and 54.2 +/- 6.2% (n = 9), for 1 microM and 100 microM capsaicin, respectively. The increase of PBF peaked at about 50 sec after the capsaicin placement and the PBF returned to control level within 20 min following capsaicin removal. Four repeated applications of 1 microM capsaicin caused a small change in the peak PBF amplitude. In contrast, a second application of 100 microM capsaicin caused a significantly smaller increase of PBF than the first application (26.8 +/- 6.4% vs. 54.2 +/- 6.2%). Results of the present study show that local application of capsaicin caused the increase of PBF and that repeated capsaicin stimulations inhibited PBF responses. Smaller PBF increases in response to high concentrations of capsaicin also suggested the depletion of vasoactive substances, e.g. substance P, from the sensory nerve terminals, which appears to be dose-dependent. The PBF response to capsaicin suggests that neurogenic inflammation in the dental pulp involves capsaicin-sensitive nerve endings.

Functional alterations in pulpal microcirculation in response to various dental procedures and materials.

Using the quantitative 15 microns microsphere injection method, the effects of several restorative procedures on pulpal microcirculation in dog canine teeth were investigated. Pulpal blood flow (PBF) decreased steadily as the remaining dentin thickness (RD) became smaller with crown preparation without water spray. One hour after the preparation wit 1/3 RD (approximately 1 mm) PBF was reduced to 10% of the control, indicating that dry deep preparation has a deleterious effect on PBF. A careful preparation to an RD of 1 mm under copious water spray had a negligible effect on PBF. Dry preparation to an RD of more than 50%, on the other hand, caused a significant increase in blood flow through shunt vessels, AVAs and "U"-turn loops as determined with the 9 microns and 15 microns microspheres. These shunt vessel activities were especially prominent in the apical portion of the pulp, suggesting that the activation of the shunt vessels may be one of the compensatory mechanisms of the pulp in response to inflammation. In rat incisor teeth observed by intravital microscope, shunt vessels opened up as the incisor tips were drilled. Impression procedures after tooth preparation with water spray using the copper band with wax caused severe flow fluctuation as compared to rubber base material impression. In anesthetics, the use of epinephrine was found to be the single most important factor affecting pulp circulation. Whether it is by infiltration or mandibular block, the use of epinephrine containing anesthetic caused a severe reduction in PBF.(ABSTRACT TRUNCATED AT 250 WORDS)

In situ testing of CO2 laser on dental pulp function: effects on microcirculation.

The effect of CO2 laser irradiation on pulpal microcirculation was studied in cat canines. The enamel surfaces of 4 teeth were exposed with energy densities of 304-1440J/cm2, using either a handpiece or a microslad, with a focal spot of 0.21mm and 0.33mm respectively. Pulpal blood flow (PBF) before and following lasing was recorded through the intact tooth surface by a laser Doppler flowmeter. CO2 laser irradiation caused an increase in PBF, which was immediate and transient. The PBF increase was higher in a large pulp than in a small pulp, and it was inversely related to the focal spot size. These findings confirm that the dental pulp is thermally affected by CO2 lasing of the tooth surface, however, without extensive pulp coagulation. It is concluded that the effects of laser irradiation on the pulpal microcirculation may be studied in situ by means of the presented methodology.

Comparison of pulpal blood flow in dog canine teeth determined by the laser Doppler and the 133xenon washout methods.

Pulpal blood flow measurements by laser Doppler flowmetry were compared with those made simultaneously and on the same teeth by the 133Xe washout method. Pulpal method flow in the resting state was 82.4 ml/min per 100 g as calculated from the fast component of the 133Xe washout, whereas laser Doppler flowmetry recorded 263 mV. The close intra-arterial (i.a.) injection of the vasoconstrictor norepinephrine (10(-2)-10(-1) micrograms/kg) caused a reduction of pulpal blood flow by 20.2% with the 133Xe washout and by 9.7% with the laser Doppler flowmetry in 6 observations in 4 dogs. The laser Doppler flowmetry measurements of these norepinephrine-induced changes correlated significantly with changes measured by the 133Xe washout method as analysed by linear regression (p = 0.004).

Comparison of the effects of intra-arterial and locally applied vasoactive agents on pulpal blood flow in dog canine teeth determined by laser Doppler velocimetry.

The vasoactive agents norepinephrine, 5-hydroxytryptamine, isoproterenol and bradykinin, at concentrations which changed local arterial pressure without changing systemic arterial pressure significantly, were injected intra-arterially (i.a.) into the maxillary artery or applied directly in a deep dentinal cavity on the buccal surface of canine teeth. The probe of a laser Doppler velocimeter was placed in the cavity to monitor pulpal blood flow. Bolus i.a. injections of the vasoconstrictors norepinephrine and 5-hydroxytryptamine produced a statistically significant (p less than 0.05) reduction in pulpal blood flow, 21.1 +/- 3.7 and 30.7 +/- 15.2%, and the local arterial pressure from the lateral nasal artery increased with norepinephrine but decreased with 5-hydroxytryptamine. The i.a. injections of the vasodilators isoproterenol and bradykinin were found to decrease both local arterial pressure and pulpal blood flow, 17.7 +/- 6.0 and 22.7 +/- 4.2%, respectively (p less than 0.05). However, local application of isoproterenol and bradykinin caused a biphasic response: an increase in pulpal blood flow, 8.6 +/- 1.6 and 9.4 +/- 1.1% (p less than 0.05), followed by a decrease, 19.1 +/- 11.9 and 5.3 +/- 2.1% (p greater than 0.005). Local application of norepinephrine and 5-hydroxytryptamine caused a decrease in pulpal blood flow, 23.7 +/- 5.2% (p less than 0.05) and 9.3 +/- 5.2% (p less than 0.05), respectively. These findings were in accordance with those from other reliable methods, such as injections of radioisotope-labelled 15 microns microspheres and the 133Xe washout technique, making laser Doppler flowmetry a reliable alternative. In addition, the biphasic response to the locally applied vasodilators and the reduction of pulpal blood flow after i.a. injection of vasodilators suggest that regulation of pulpal blood flow is determined by the combined effects of the low compliance environment and the stealing of perfusion to the pulp by the neighbouring tissues.

Hemodynamic regulation of the dental pulp in a low compliance environment.

Pulpal hemodynamic regulation has been investigated in experimental animals using 133Xe washout, 15-microns radioisotope-labeled microsphere injection, and intravital microscope methods. Three distinct types of pulp blood flow reductions were observed. Type I was characterized by reduction in response to direct stimulation of the sympathetic nerve; intraarterial infusion of norepinephrine, 5-HT, or prostaglandin F2 alpha; and indirect stimulation of the sympathetic nerve. The Type II response, an initial increase in flow followed by a decrease, was observed with isoproterenol, prostaglandin E2, substance P, and bradykinin. This biphasic flow response is caused by the low compliance environment of the tooth and may play a role in pulp inflammatory processes. The Type III response follows administration of histamine and is characterized by a gradual decrease in pulpal blood flow.

Quantitative assessment of microcirculation in the rat dental pulp in response to alpha- and beta-adrenergic agonists.

Responses to intra-arterial injections of the alpha-agonist, norepinephrine, the beta-agonist, isoproterenol, and to selected blockers were investigated in vivo by intravital microscopy. Luminal diameters (D) of pulpal microvessels and red cell velocities (Vrbc) were measured simultaneously to facilitate calculation of volumetric blood flow (Q). Norepinephrine caused a decrease in D, mean red cell velocity (Vm) and Q within individual microvessels. These responses were blocked by the alpha-antagonist, phenoxybenzamine. In response to the arteriolar D increase and venular D and Q decrease with isoproterenol, a transient increase in arteriolar Q was followed by a decrease within the first 60 s. No such effects were observed when the beta-antagonist, propranolol, was administered before the isoproterenol injection. The biphasic response in Q in response to isoproterenol is most likely related to the low compliance of the pulp. In the low-compliance environment a passive compression of venules could result from an active dilation of arterioles with an attendant rise in extravascular tissue pressure.

Microcirculation of the dental pulp and its autonomic control.

Pulpal vessels are innervated with sympathetic adrenergic vasoconstrictor fibers. Electrical stimulation of the cervical sympathetic nerve causes pulpal arteriolar constriction and a reduction of arteriolar and venular flow rate (in mm3/s) in rat incisor pulps and a decrease in pulpal blood flow (in ml/min/100 g) in canine pulps of cats and dogs. The alpha-antagonist phenoxybenzamine and the alpha-1 antagonist prazosin attenuated the decrease in pulpal blood flow (PBF) caused by sympathetic stimulation. Reflex excitation of the sympathetic nerve system by hemorrhage caused pulpal vasoconstriction and a reduction of PBF. Induction of hemorrhagic hypotension in dogs subjected to cervical sympathectomy and adrenalectomy caused less pronounced pulpal vasoconstriction and flow reduction than in normal dogs. Pulpal vessels are also equipped with alpha-1, alpha-2 and beta-adrenergic receptors. Activation of alpha-1 and alpha-2 receptors by intraarterial injection of phenylephrine and clonidine caused a reduction in PBF in dogs and decreases in arteriolar and venular diameters and volumetric flow in rats. These responses were blocked by the alpha-1 and alpha-2 antagonists. Activation of beta-2 receptors by i.a. injection of isoproterenol caused a paradoxical reduction of PBF in dogs. In rats isoproterenol caused a transient increase in the flow rate followed by a reduction, and arteriolar dilation was accompanied by venular constriction. These flow responses to isoproterenol were blocked by the beta-2 antagonist propranolol. Results of microcirculatory studies in dogs and rats indicate that pulpal hemodynamics are regulated significantly by the sympathetic adrenergic system.

Biphasic pulp blood-flow response to substance P in the dog as measured with a radiolabelled, microsphere injection method.

A dose of the vasoactive substance P (SP), affecting local circulation only (0.0001-0.001 micrograms/kg body weight) when given by a close injection into the maxillary artery, caused a biphasic flow response in pulpal blood flow (PBF), as measured by a radioisotope-labelled 15 micron microsphere injection technique. Immediately after the SP injection, PBF increased some 66 per cent (p less than 0.02) and vascular hindrance (Z) decreased to 40 per cent against control values (p less than 0.02), indicating vasodilation. Thirty seconds after the SP injection, PBF decreased by 53 per cent (p less than 0.05), and pulpal Z increased 139 per cent against control values (p less than 0.02), indicating decreased blood flow. This biphasic flow response may be due to the active vasodilation of arterioles by SP, which causes a compression of venules and hence a reduction of PBF in what is a low-compliance system.

The influence of 5-hydroxytryptamine (serotonin) on blood flow in the dog pulp.

The effect of intra-arterial injection of 5-hydroxytryptamine (5-HT) on pulpal blood flow of the dog was determined using the 15-micron radioisotope-labeled microsphere injection method. Pulpal blood flow was significantly reduced following the 5-HT injection. This decrease in blood flow appeared to be due to vasoconstriction as determined by an increase in pulpal vascular hindrance. However, our findings do not preclude the possibility that blood flow was reduced as a result of passive compression of venules produced by vasodilation in a low-compliance environment.