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In the process of host defence against microbial challenge, neutrophils release granule contents with the potential side effect of damaging structural tissues. In the junctional epithelium such damage may contribute to the degeneration and renewal of the epithelial cells attached directly to the tooth (DAT cells), and subsequently to periodontal pocket formation. This study reports on lactoferrin, one of the substances released by neutrophils, and its effects on epithelial cell adhesion, growth, DNA synthesis and spreading of cell colonies at concentrations recorded in the crevicular fluid. We show that, in opposition to what has been reported on bacterial cells, lactoferrin has no effect on the DNA synthesis of attached epithelial cells in model systems attempting to simulate the DAT cells in vivo. However, both iron-saturated and unsaturated lactoferrin hampered cell adhesion, growth and spreading of cell colonies in a dose-dependent manner. These findings suggest that lactoferrin does not affect epithelial cell proliferation but it may have a role in delaying the repair of the DAT cell population during inflammation by interfering with cell adhesion.

The structural and functional barrier preventing the free advancement of microbial plaque subgingivally along the tooth surface is formed by the junctional epithelial (JE) cells directly attached to the tooth (DAT cells). The mechanism leading to degeneration of the DAT cells is not known. In the present study we examined the possible role of short chain fatty acids (SCFAs) on epithelial cells by making use of 2 epithelial cell cultures (HaCaT and ERM) and an explant culture model of human JE. The SCFAs butyrate and propionate were used in concentrations found in human plaque and gingival crevicular fluid (0.25-16.0 mM). The SCFAs had no effect on primary cell adhesion nor on the epithelial attachment apparatus (EAA). By contrast, even 0.25 mM of butyrate significantly retarded epithelial cell growth. Similar effects with propionate were first observed at concentrations higher than 1.0 mM. The retardation of epithelial cell growth was found to be due to inhibition of cell division. Furthermore, after butyrate treatment dense accumulations of intermediate filaments and cytoplasmic vacuolization were characteristically seen in cells adjacent to cells of normal appearance. This suggests that some cells of the growing epithelial cell population are more sensitive to the SCFAs than others, and agrees with previous reports on the DAT cells of periodontally-involved teeth in vivo. The results suggest that SCFAs are microbial factors that play a role in the initiation and progression of periodontal pocket formation by impairing epithelial cell function rather than having a direct effect on the EAA.

This study examined the directly attached cells (DAT cells) of the human junctional epithelium from teeth extracted for advanced periodontal disease. The monolayer nature of the DAT cells remaining on the tooth surface after extraction offers a unique opportunity to study their morphology and activity in situ. We collected DAT cells and examined them by transmission electron microscopy and by autoradiography of cells labeled with 3H-thymidine. Our morphological results suggest that degenerative changes associated with pocket formation affect individual cells rather than regions of the DAT cell population at one time. The epithelial attachment apparatus (EAA) appeared to be the most resistant structure persisting on the tooth surface during the degeneration of the individual DAT cells. The new technique developed for two-dimensional observation of the sheet of DAT cells labeled in situ on the tooth surface in culture showed that the attached cells, even in periodontal disease, exhibit proliferative ability, suggesting a regenerative role for the DAT cells.

Pregnant hamsters were treated by topical application of formaldehyde solution on day 8, 9, 10 or 11 of gestation. Fetuses recovered on day 15 were weighed, measured, and examined for teratogenic effects of formaldehyde exposure. The resorption rate was increased in the treated groups, but formaldehyde treatment did not significantly affect weight or length, nor did any malformations which could be related to treatment appear. It was concluded that fetal risk due to maternal topical exposure to formaldehyde is minimal in this model system.

Chicken eggs of 4 or 6 days of incubation were injected with 10 mug 6-aminonicotinamide (6-AN) or 6-AN plus various doses of sodium ascorbate, calcium ascorbate, or ascorbic acid; 11-day embryos were examined grossly and histologically. 6-AN-treated embryos had various degrees of micromelia and were reduced in overall size. All three ascorbates inhibited 6-AN teratogenesis but not completely. The extent of inhibition was dose related. Increased amounts of intercellular matrix and decreased necrosis of chondrocytes in the limb cartilage of protected embryos correlated with the gross findings.

Amino acids methionine and histidine, which are soluble in propylene glycol, were investigated for their purported beneficial effects on aspirin-induced gastric mucosal damage in the rat. The pathognomonic changes observed microscopically in the fundic region of the stomach of animals administered daily doses (100 mg/kg), for up to 15 days, of aspirin solutions (0.36 M) in propylene glycol incorporated with the amino acids were compared with those of animals given equivalent quantities of aspirin in an aqueous suspension combined with an aluminum hydroxide antacid. A "delayed" onset of aspirin-induced cellular damage due to the presence of amino acids, analogous to that associated with the use of antacids, was found as determined partly by differences in the staining ability of injured cells with hematoxylin and eosin.

The incidence of spontaneous avian spondylopathy was maintained between 58 and 66% in the progeny of one flock (Flock A) and less than 40% in the progeny of another (Flock B), by proper selection and breeding for 4 generations. Spondylolisthesis, the major type of spondylopathy present, was observed at the first and sixth vertebral levels. Spondylolisthesis was progressive in nature and consisted of a ventrodorsal rotation of the 1st and/or 6th thoracic vertebrae leading to spinal cord injury and paraplegia or partial crippling in some birds. There was occasionally extensive muscle degeneration and connective tissue proliferation around the distorted vertebrae and the spinous ligaments bridging the defect were stretched and appeared heavier than normal. Spondylopathy was also observed in 20-day chick embryos. Spondylolisthesis was observed in 33% of the embryos examined from Flock A-G2 and the other conditions present consisted of scoliosis and abnormal lordosis and kyphosis. Using a grading scale of +1 (mild) to +3 (severe), all cases of embryonic spondylopathy were graded as +1. The back defects present in Flock B-G2 embryos were entirely due to abnormal kyphosis.

The ability of the isolated gill epithelium of Mytilus edulis to incorporate [14C]glucosamine as a precursor in the biosynthesis and secretion of mucous glycoproteins was investigated. Localization of mucous cells in the gill filament was achieved using histochemical staining techniques. Mucus cells containing neutral and acidic mucins were found in the lateral region, whereas mucus cells containing primarily neutral or sulfated mucins were found in the abfrontal region. Autoradiographic results showed that in both regions, the mucous cells were rich in content of the incorporated radiolabel. The secreted glycoproteins containing the incorporated radiolabel were analyzed by column chromatography using Bio-Gel P-2 and P-6. Two populations of the glycoproteins differing in molecular size were isolated. Upon alkaline reductive borohydride cleavage of the O-glycosidic linkages of the high molecular weight protein, about 70% of the radiolabel and 85% of the carbohydrate content were removed from the protein. The alkaline borohydride cleavage resulted in the formation of at least six oligosaccharide chains of various lengths of sugar units. Gas chromatographic analysis of the carbohydrate composition shows that the glycoproteins contain N-acetylglucosamine, N-acetylgalactosamine, and galactose, fucose, and mannose as the neutral monosaccharides. The above results indicate that the isolated gill epithelium of M. edulis is capable of incorporating [14C]glucosamine in the synthesis of secretable mucin-type glycoproteins.

Methyl salicylate was administered topically to pregnant hamsters at 7d9h and the teratogenic results were compared with those obtained following oral treatment with the same compound. Both treatments produced the same defect in embryos recovered at day 9: failure of fusion of the neural tube, especially in the area of the developing brain. Analysis of serum salicylate levels following both treatments produced similar curves and indicated that teratogenic levels of salicylate can reach the maternal circulation after topical exposure.