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The effects of thermal and mechanical stimulation on blood flow in healthy and inflamed gingiva in man.

The purpose of this study was to compare the state of the microcirculation in healthy gingiva with that in untreated, inflamed gingiva in humans. Reflection photoplethysmography (RP) was used to record blood flow from 13 clinically healthy, attached gingival sites in seven adults and 11 inflamed sites in seven patients. Changes in the pulse amplitude of the RP signal were measured before and after application of thermal (cold and hot water) and mechanical (brushing) stimuli to the gingiva. Hot water and brushing, but not cold water, produced a significant increase in the pulse amplitude in healthy gingiva (p < 0.05, Friedman analysis and Student-Newman Keuls test). None of the stimuli produced a significant change in inflamed gingiva. In neither group of participants was there a significant correlation (Spearman rank correlation coefficient) between vascular responsiveness and age. The results indicated that vasomotor control may be impaired in inflamed gingiva.

Adult↗

Autonomic and sensory innervation of cat molar gland and blood vessels in the lower lip, gingiva and cheek.

Innervation of the molar gland and blood vessels in the lower lip, gingiva and cheek mucous membrane was investigated in the cat with the aid of whole mount acetylthiocholinesterase (WATChE) histochemistry and retrograde neuronal tracing methods with horseradish peroxidase (HRP) and HRP-conjugated wheat germ agglutinin (WGA-HRP). The molar gland was found to be supplied from the buccal nerve and branches of the mylohyoid nerve on the basis of microdissection of WATChE-stained mandibular preparations under a dissecting microscope. The rostral half of the lower lip-gingiva was innervated by mental branches from the inferior alveolar nerve. The caudal half of the lower lip-gingiva and cheek mucous membrane were observed to be supplied from the buccal nerve. Following injections of HRP/WGA-HRP into the molar gland, lower lip-gingiva and cheek, many retrogradely labeled ganglion neurons were observed in the ipsilateral main and accessory otic ganglia, superior cervical ganglion and mandibular division of the trigeminal ganglion. In the pterygopalatine ganglion, a small number of positive neurons were found, but in a few cases in which the injected tracer was restricted to the lower lip-gingiva and anterior half of the molar gland, labeled neurons were not detected in the main ganglion nor in its accessory microganglia. These findings indicate that the cat molar gland receives a postganglionic parasympathetic supply from the otic ganglia, postganglionic sympathetic input from the superior cervical ganglion and sensory innervation from the trigeminal ganglion by way of the buccal nerve and mylohyoid nerve. Vessels in the rostral half of the lower lip-gingiva receive the same inputs from the inferior alveolar nerve, and vessels in the caudal half receive inputs from the buccal nerve. The vessels in the cheek mucous membrane receive dual parasympathetic supplies from the otic ganglia and the pterygopalatine ganglion by way of the buccal nerve.

Acetylcholinesterase↗

Hyperaemic response to cigarette smoking in healthy gingiva.

BACKGROUND AND AIM: Cigarette smoking is currently considered as a risk factor for periodontal disease. Controversy exists as to whether the vasoconstrictive property of nicotine is one of the pathogenic mechanisms. To this end we tested the hypothesis that cigarette smoking is causing vasoconstriction in the healthy human gingiva. MATERIALS AND METHODS: Gingival blood flow was continuously measured with laser Doppler flowmetry in healthy (n=13) casual consumers of tobacco. Simultaneously, recordings were made of skin blood flow in the forehead and the thumb as well as heart rate (HR) and blood pressure (BP). In another session infraorbital nerve block anaesthesia (INB) with 1.0 ml of Carbocain without vasoconstrictive additives was used to identify nervously mediated vascular responses to cigarette smoking (n=8). RESULTS: Cigarette smoking induced a modest hyperaemic response in the gingiva that was lower than the relative increases in BP and HR, and the calculated gingival vascular conductance decreased. In the forehead, flow responses were similar to those in the gingiva, while in the thumb a powerful vasoconstriction was observed. During the later part of the 10-min recovery period, BP and HR tended to decrease while blood flow in the gingiva and forehead remained high. INB potentiated the hyperaemic response to cigarette smoking in gingiva. CONCLUSIONS: The present results help to shed some light on the understanding of the vasoactive mechanisms induced by cigarette smoking, and to support the hypothesis that cigarette smoking causes nervously mediated vasoconstriction in the healthy human gingiva. However, the degree of vasoconstriction was far less than in the thumb skin, and in our subjects was overcome by the evoked rise in arterial perfusion pressure. As a consequence, gingival blood flow increased during smoking. It is speculated that small repeated vasoconstrictive attacks due to cigarette smoking may in the long run contribute to gingival vascular dysfunction and periodontal disease.

Adult↗

A longitudinal evaluation of varying widths of attached gingiva.

32 patients with bilateral areas of inadequate attached gingiva on the facial surface of homologous contralateral teeth have been followed for 6 years. Treatment consisted of scaling, root planing, oral hygiene and maintenance at 3- to 6-month intervals or as needed to control inflammation. A free gingival graft was placed on one side (experimental), while the other side served as the unoperated control. Areas of inadequate attached gingiva on control sides, to include those with recession and no attached gingiva, did not demonstrate additional recession or further loss of attachment. Gingival inflammation and plaque were significantly reduced. On experimental sides, the dimension of keratinized and attached gingiva increased and was stable over 6 years. Areas which began with recession and no attached gingiva exhibited a reduction in recession and gain in clinical attachment following the placement of a gingival graft. Examination of patients who had discontinued participation in the study for a period of 5 years revealed a re-establishment of gingival inflammation on the control sides associated with additional recession. Similar changes were not observed in areas treated by a free graft. The findings demonstrate that it is possible to maintain periodontal health and attachment through control of gingival inflammation despite the absence of attached gingiva.

Dental Scaling↗

Mechanical stimulation by toothbrushing increases oxygen sufficiency in human gingivae.

We studied the immediate effect of mechanical stimulation with a toothbrush on oxygen sufficiency in human gingivae. Teeth were brushed with a modified Bass technique at various forces (100, 200, and 300 g) for various times (5, 10, and 30 s). Hemoglobin (Hb) oxygen saturation was measured before and intermittently for 60 min after brushing, by non-invasive tissue reflectance spectrophotometry. Hb oxygen saturation increased within a few min after brushing and then slowly returned to the initial level. Toothbrushing at 200 g for 10 s caused the longest-lived increase in Hb oxygen saturation; saturation remained about 6% above the baseline level for more than 25 min. Before brushing, Hb oxygen saturation was significantly lower in inflamed gingivae than in healthy gingivae (p < 0.05). After brushing of inflamed gingivae, the Hb oxygen saturation almost reached the level measured in healthy gingivae before brushing. These findings suggest that stimulation with a toothbrush increases oxygen sufficiency in both healthy and inflamed gingivae.

Adolescent↗

Immunohistochemical study of neuronal markers in human gingiva with phenytoin-induced overgrowth.

The immunohistochemical occurrence of several different neuronal markers has been investigated in human gingiva with phenytoin-induced overgrowth. The material was compared to gingival material taken following surgical orthodontic treatment. Gingiva obtained from the phenytoin-treated groups seemed to have a reduced number of neurofilament (NF) immunoreactive nerve fibers in the propria compared to control material. In both phenytoin as well as control gingiva sparsely distributed, thin, calcitonin gene-related peptide (CGRP) and substance P (SP) immunoreactive fibers were found in the propria. No obvious differences between the two groups could be observed for CGRP and SP. Immunoreactive cells for somatostatin (SOM) with a dendritic cell shape were found in the propria in both groups, sometimes in densely packed clusters. A tendency for increase of SOM-immunoreactive cells in the phenytoin-treated gingiva was observed. A few gamma-melanocyte stimulating hormone (gamma-MSH)-immunoreactive cells with a round appearance were found in control as well as phenytoin-affected gingiva. In one instance, however, a heavy gamma-MSH-immunoreactive cell infiltration was seen in the phenytoin sample. No immunoreactivity in either the phenytoin-treated group or in the control group was seen for proctolin or galanin. The results indicated that gingiva with phenytoin-induced overgrowth has a reduced innervation density revealed with NF immunohistochemistry.

Adolescent↗

Advanced glycation endproducts (AGEs) induce oxidant stress in the gingiva: a potential mechanism underlying accelerated periodontal disease associated with diabetes.

We hypothesized that one mechanism underlying advanced periodontal disease in diabetes may involve oxidant stress in the gingiva, induced by the effects of Advanced Glycation Endproducts (AGEs), the irreversible products of non-enzymatic glycation and oxidation of proteins and lipids which accumulate in diabetic plasma and tissue. Infusion of AGE albumin, a prototypic ligand, into mice resulted in increased generation of thiobarbituric acid reactive substances (TBARS) compared with infusion of non-glycated albumin in the gingiva, as well as in the lung, kidney and brain. Pretreatment of the animals with the antioxidants probucol or N-acetylcysteine (NAC) prevented the generation of TBARS in the gingiva. Affinity-purified antibody to AGEs demonstrated increased immunoreactivity for AGEs in the vasculature and connective tissues of the gingiva in streptozotocin-induced diabetic mice compared to non-diabetic controls. Increased immunoreactivity for AGEs was also demonstrated in the gingiva of diabetic humans compared with non-diabetic individuals via immunohistochemistry and ELISA. Consistent with these data, immunohistochemistry for heme oxygenase-1, a marker of enhanced oxidant stress, was increased in the gingival vasculature of diabetic mice and humans compared with non-diabetic controls. These data suggest that AGEs present in diabetic gingiva may be associated with a state of enhanced oxidant stress, a potential mechanism for accelerated tissue injury.

Adult↗

Identification of the Fas antigen in human gingiva.

The Fas antigen is a cell-surface glycoprotein that mediates apoptosis from the cell surface into the cytoplasm. Polyclonal antibody (Fas D) was raised against a synthetic polypeptide selected from the extracellular part of the human Fas antigen (amino acid residues 104-114) and was used to detect the Fas antigen in human gingiva. Biopsy specimens of human gingiva were prepared, and the paraffin sections were reacted with the Fas D antibody by an immunohistochemical method. The antibody localized to the prickle-cell layer and to granular layer keratinocytes of human gingiva. Proteins were also prepared from human gingiva and subjected to SDS-PAGE, followed by Western-blotting analysis with the Fas D antibody. The antibody interacted with a band corresponding to an estimated molecular weight of 35 kDa. The incidence of the immunoreactive 35-kDa protein was detected in the gingiva of 90% of the 20 individuals examined. The Fas antigen detected in human gingiva may be related to the physiological turnover of oral mucosa.

Amino Acid Sequence↗

Fordyce's granules of the incisor and molar gingiva in F344 rats.

Fordyce's granules were observed in the gingiva of the upper incisor and molar teeth in F344 rats. The data were based on 734 males and 722 females that were used as control and treated animals in 26-week, 65-week, and 2-year studies by the National Toxicology Program. The incidence of Fordyce's granules was markedly different when comparing sex, age, and site of the lesion. Fordyce's granules were very common in the midsagittal gingiva of the upper incisor in males and increased in incidence with age (34.2, 50, and 56.3% in 26-week, 65-week, and 2-year studies, respectively). The granules of the incisor gingiva were rare in females (0,0, and 2.8% in 26-week, 65-week, and 2-year studies, respectively). Fordyce's granules of the molar gingiva were very rare in both sexes and were found only in 9/734 (1.2%) males and in 3/722 (0.4%) females. Only three unilateral granules of the molar were grossly recognized as focal swelling of the gingiva or a white nodule with a huge cyst in the third upper molar. Histologically, Fordyce's granules were arranged as a collection of sebaceous glands unassociated with hair follicles. In addition, the granules of the molar gingiva were associated with cystically dilated ducts filled with sebum. Ultrastructurally, the sebaceous cells were characterized by varying numbers of cytoplasmic lipid droplets and occasional desmosome and hemidesmosome formation. Fordyce's granules previously reported in rats of other strains were also reviewed and compared with those in F344 rats in regard to incidence, location, and age.

Animals↗

Levels of immunoglobulins IgG, IgA, and IgM in the human inflamed gingiva.

Resected inflamed gingival tissue obtained from 16 periodontal patients and a pooled sample of noninflamed gingiva from five edentulous patients were assayed for immunoglobulins IgG, IgA, and IgM using low-level immunodiffusion plates. Findings based on the gingival assays include:(1) IgA and IgG are present in both inflamed and normal gingiva and although their levels are substanitally higher in the inflamed gingiva, their ratio, one to another, remains the same; (2) IgM can not be consistently demonstrated in inflamed gingiva with the assay technique employed: (3) local immune response exists in the inflamed gingiva of humans with chronic periodontal disease; and (4) there is an unknown homeostatic mechanism which regulates the globulin levels in states of health and inflammation. Further study is needed to correlate clinical impressions with histological and quantitative measure of immunoglobulin levels in the inflamed gingiva and to investigate the role of delayed hypersensitivity in the disease process. Work must also be directed toward the detection of specific antigens responsible for elicting the immune response in the gingival tissue of the periodontal patient.

Adult↗

The width of lingual mandibular attached gingiva.

One hundred and twenty individuals in good oral health were divided into six groups according to age. Measurements were made on the depth of the gingival sulcus, and the distance from the margin of the free gingiva to the mucogingival junction on the lingual mandibular teeth. The mean, extreme, standard deviation and analysis of variance were computed. A distinct pattern of the width of healthy lingual attached gingiva was found. The largest width was found lingual to the first and second molars with a mean height of 4.7 mm. This was followed by the third molar [3mm] second bicuspid [2.5 mm] first bicuspid [2 mm], and cuspid, lateral and central [1.9mm]. Lingual attached gingiva showed that there was a variation in width with each tooth and between individuals. There was no variation in width of lingual attached gingiva between sexes. The extreme measuremtns ranged from less than 1 mm to 8 mm. The data from the study gives the dentist some indication of the mean and extreme ranges of lingual attached gingiva. This data can serve as a guide in diagnoiss and treatment planning and in selecting procedures to reestablish these zones of attached gingiva.

Female↗

A stereomicroscopic and immunohistochemical study of vasculature in gingiva bleeding after probing.

This study investigated the stereomicroscopic appearance of gingival sulcular surfaces and the histologic alterations of subepithelial vasculature of gingiva bleeding after probing. Sixty-four pairs of gingival biopsies from 49 patients were obtained and the results of stereomicroscopically visible vasculature were observed in half of the specimens. The number of biopsies showing that the vasculature of bleeding gingiva was significantly greater than those of non-bleeding gingiva. The immunohistochemical marker UEA-1 was used to identify endothelial cells on the vascular walls of 34 gingival biopsies from 15 patients. Following stereomicroscopic examination, tissue preparation with routine and UEA-1 stainings was performed. Volumetric and numerical analyses of vasculature in 6 divided portions of subepithelial connective tissue were measured. The results showed that significantly greater numbers of UEA-1 staining vessels were found within the apico-superficial portion of bleeding gingiva than those of non-bleeding gingiva. However, there was no specific correlation between the UEA-1 positive vessels within gingival tissue and stereomicroscopic visibility of vasculature on the surface appearance. It is suggested that increased vascularity in gingiva may be responsible for bleeding on clinical probing.

Adult↗

Increased expression of interleukin-1 receptors on fibroblasts derived from inflamed gingiva.

We investigated the expression of interleukin-1 receptors (IL-1R) on the surfaces of cultured gingival fibroblasts derived from healthy and inflamed gingiva and the effects of IL-1 alpha and IL-1 beta and prostaglandin E2 (PGE2) on IL-1R expression. Fibroblasts were obtained from explant culture of both healthy and inflamed gingiva. IL-1R on cell surfaces was detected immunohistochemically using an anti-human IL-1R monoclonal antibody. IL-1R expression was assessed quantitatively using an enzyme linked immunosorbent assay (ELISA). Positive staining for IL-1R was more evident on cells from inflamed gingiva compared with cells from healthy gingiva. The ELISA showed a significantly increased number of IL-1R on cells from inflamed gingiva compared with cells from healthy gingiva (P < 0.05). Treatment with IL-1 but not PGE2 increased expression of IL-1R on fibroblasts. These findings suggest that gingival fibroblast responses to IL-1 may represent a mechanism for amplification of gingival inflammation.

Adult↗

[Photoplethysmograph obtained from a small area of human gingiva].

We have established a method to observe circulation in a small area of the human gingiva. Reflected light photo-plethysmographs (RLP) and transilluminated light photo-plethysmographs (TLP) were recorded from healthy gingiva in 2 young adults of twenties. A tungsten-lamp, connected to a stabilized power source, was used to illuminate the gingiva. The reflected or transilluminated light was collected using a fibre-optic bundle. A CdSe photo-conductive cell was used as the photo-detector. The ECG was recorded simultaneously. The results were as follows: 1) RLP and TLP were both synchronous with the heart beat and showed a dicrotic-notched wave form. 2) When the light-collecting fibre was 0.5 mm from the surface of the gingiva, the dicrotic notch with TLP was clearer than that with RLP. 3) When the light-collecting fibre was placed less than 0.5 mm from the gingival surface, clear dicrotic notches were seen in RLP. 4) When the surface of the gingiva was covered with white opaque paint to prevent transilluminated light, definite dicrotic notches were observed with RLP. 5) RLP was produced mainly by the pulsation of the gingival surface, and the pulsatile movement of the tooth had a little effect on RLP. 6) RLP was consisted from the light reflected from the surface of the gingiva and also from the light reflected after penetrating some distance in the gingival tissue.

Adult↗

Edema-preventing mechanisms in rat gingiva.

The long-term effect of increased local venous pressure (Pv) on interstitial fluid pressure (Pi), colloid osmotic pressure (COPi), and fractional removal rate of 125I-labeled human serum albumin (kAlb) was studied in rat gingiva. Measurements were performed on experimental animals and sham-operated controls up to 4 days after ligation of jugular veins. On the day of ligation Pv in the facial veins rose from 2.5 +/- 0.3 (SD) to 15.8 +/- 2.8 mm Hg and stayed at about this level for 2 days before a decrease to 7.4 +/- 0.9 mm Hg on day 4. In free gingiva Pi rose from an average of 3.5 +/- 0.4 to a maximum of 6.3 +/- 0.7 mm Hg, whereas in attached gingiva the corresponding increase in Pi was from 6.0 +/- 0.7 to 11.1 +/- 2.1 mm Hg. One day after the ligation COPi in wick fluid from gingiva was reduced from the control level of 10.6 +/- 1.4 to 4.5 +/- 0.9 mm Hg. COP in plasma and COPi in subcutaneous tissue on the back were unaffected. The removal rate of 125I-labeled albumin (kAlb) from the gingiva showed a nearly threefold increase after venous ligation, from 0.073 +/- 0.01 to 0.211 +/- 0.06 h-1. It is concluded that in free and attached gingiva, both a rise in Pi and a decrease in COPi will counteract the increased filtration pressure and thus prevent edema formation during venous stasis. The fall in COPi is most likely due to increased lymph flow and not dilution, as venous stasis significantly increased kAlb without any visible increase in gingival volume.

Animals↗

Histometric analysis of cell populations in gingiva with bleeding on probing by immunohistochemistry.

The present study was performed by immunohistochemical methods in order to morphometrically characterize and compare the composition of cellular infiltrates in human gingivae with bleeding and non-bleeding upon probing. A total of 64 gingival biopsies (33 bleeding and 31 non-bleeding gingivae) were obtained from 23 patients. Routine Hematoxylin & Eosin and immunohistochemical stains were performed following tissue preparations. On each section, three zones were histologically divided. For each zone, the percentage and histometric analysis of plasma cells, phagocytic cells, B-cells and T-cells was calculated to characterize the composition of cellular infiltrates in gingiva associated with bleeding on probing. Significant difference was found in the middle and apical zones of infiltrated connective tissue when compared with plasma cells expressed by kappa- or lambda-chain positive immunostaining between bleeding and non-bleeding gingivae. Significant difference was also found in the coronal zone when compared with phagocytic cells and B lymphocytes between bleeding and non-bleeding gingivae. It is suggested that clinical bleeding tendency of gingiva may be highly associated with histological alterations of underlying inflammatory cell populations and a complicated immunological mechanism appears to be involved.

B-Lymphocytes↗

Local and systemic factors in periodontal disease increase matrix-degrading enzyme activities in rat gingiva: effect of micocycline therapy.

We previously reported that both local and systemic factors relevant to the pathogenesis of periodontal disease can increase gingival collagenase activity in rats. Since the degradation of extracellular matrix is an essential feature of periodontal disease and this tissue breakdown requires multiple enzyme interactions, the current study was carried out to determine the effects of bacterial endotoxin (LPS) (a local factor) and diabetes (a systemic factor) on a panel of matrix-degrading enzymes (collagenase, gelatinase, elastase, and beta-glucuronidase) in the gingiva of rats. In addition, the effects of therapy with a semisynthetic tetracycline (minocycline) were investigated. Ten male, Sprague-Dawley rats were made diabetic by IV injection of streptozotocin. Four of the ten rats then received minocycline (10 mg/day) by oral gavage on a daily basis for 3 weeks. Nineteen nondiabetic rats served as controls and 9 of them received 10 microliters of E. coli LPS (10 mg/ml) by injection into the labial gingiva every other day during the last week of the study. The other 10 nondiabetic rats were sham injected with saline into the gingiva. At the end of the 3 week experimental period, gingival tissue and skin were dissected from each rat and extracted for enzyme analysis. Our results showed that diabetes markedly increased the four matrix-degrading enzyme activities in both gingiva and skin. In contrast, local LPS injection increased these enzyme activities in the gingiva alone. Systemic therapy with minocycline completely ameliorated these elevated enzyme levels in diabetic rats in both gingiva and skin. Minocycline added in vitro to the enzyme assay systems containing skin extract from diabetic rats also inhibited collagenase and gelatinase activities, but no inhibition was observed for elastase and beta-glucuronidase activities, indicating that the MMPs and other enzymes were inhibited by minocycline, during diabetes, by indirect and indirect mechanisms, respectively.

Alveolar Bone Loss↗

A study on the removal of the melanin pigmentation of dog gingiva by CO 2 laser irradiation.

Removal of melanin pigmentation of the gingiva with CO 2 laser irradiation was investigated. The melanin-pigmented gingiva of 5 dogs were classified according to the degree of gingival pigmentation (slight, moderate, severe) and its extent (local, extensive). CO 2 laser was irradiated on the surface of the pigmented gingiva under the following conditions; defocused beams of spot size 4 mm, irradiation time 0.2(S), output 6-8(W)). That is, the irradiation energy density was 9.6-12.8 J/cm 2. The irradiated gingiva was examined macroscopically and histopathologically. The histopathological changes were investigated by hematoxylin-eosin staining and Masson staining for the differential diagnosis of melanin pigmentation cells. These examinations were done one week and three weeks after the laser irradiation. Immediately after irradiation, the gingiva surface showed white color and bulla-like appearance. One week after irradiation, the pigmentation of the lased gingiva disappeared in all cases macroscopically. Histologically, the pigmented-laden cells could not be found and no inflammatory cell infiltration was observed. Three weeks after irradiation, no reappearance of pigmentation was observed in either macroscopical or microscopical investigations excluding severe/extensive cases. These findings suggested that it is possible to remove gingival melanin pigmentation by CO 2 laser irradiation.

Animals↗