Reconstruction of severely resorbed alveolar ridge crests with dental implants using a bovine bone mineral for augmentation.

This study reviews the outcome of implant placement in 61 patients after augmentation of severely atrophic alveolar bone with a bovine bone mineral, Bio-Oss. Bone augmentation was performed at 4 different sites: alveolar crest width, alveolar crest height, antral cavity, or nasal cavity. After a mean healing time of 11.9 months, 231 implants were placed in Bio-Oss bone. The time of loading of the implants varied between 12 and 113 months. Calculated from the time of implant placement and irrespective of loading time, a survival rate of 80.5% for the individual implants was estimated. In most patients (73%), Bio-Oss was mixed with autogenous bone from the chin. However, the results indicated that autogenous bone may be excluded from the Bio-Oss graft.

A 1-year clinical report of a one-piece implant abutment.

PURPOSE: Solid one-piece standard and conical abutments, which do not involve the external implant hex when seated, have been designed. The present clinical study represents a 1-year follow-up of the use of these abutments for anchorage of implant-supported fixed prostheses for oral rehabilitation of 36 completely and eight partially edentulous patients. MATERIALS AND METHODS: After 1 year of loading, the implant-supported dentures were removed, and the seating of the 214 one-piece abutments was inspected clinically and radiographically. RESULTS: After 1 year of loading, no loose abutments were observed. One complication was reported; it involved fracture of two abutment screws within the same fixed denture. Prosthesis mobility and gold screw loosening accompanied this complication. Ninety-one percent of the patients exhibited healthy gingiva, and 9% showed erythema/edema. No periimplantitis was identified. CONCLUSION: The results of this study--healthy marginal tissue, a mean bone loss of 0.3+/-0.6 mm, and no abutment retightening 1 year after loading--support the use of the one-piece abutment design in implant-supported screw-retained restorations.

Expression of the peroxisome proliferator-activated receptor (PPAR) in the mouse colonic mucosa.

The peroxisome proliferator-activated receptor (PPAR), a member of the steroid nuclear receptor superfamily, has been shown to be activated by various compounds such as fibrates, thiazolidinediones, prostaglandins, and fatty acids. Here we demonstrate expression of PPAR in mouse colonic and small intestinal mucosa by Western blot analysis and immunohistochemistry, indicating a higher expression level in the differentiated colonic epithelial cells facing the intestinal lumen. Quantification of PPAR mRNA by ribonuclease protection assay revealed relatively high expression of PPAR gamma and Nuc1 in the colon as compared to the small intestine. In contrast, PPAR alpha expression was higher in the small intestine as compared to the colon. These results demonstrate the presence of PPAR in the intestinal mucosa; however, the physiological roles of the various isoforms in the intestine remain to be established.

Increased c-fos mRNA and binding to the AP-1 recognition sequence accompanies the proliferative response to deoxycholate of HT29 cells.

To further understand the molecular mechanisms of bile acid-mediated colon tumor promotion, we have examined the possible role of AP-1 activity in this process. The AP-1 complex has been reported to play an important role in control of cell growth. Our studies show that lithocholate, deoxycholate and ursodeoxycholate exhibited marked proliferative effects on a human adenocarcinoma cell line (HT29), while cholate was without effect. The proliferative effects appeared to be confined to narrow concentration windows which differed for the different bile acids. We demonstrate that deoxycholate caused an increase in expression of c-fos mRNA and increased binding to the AP-1 site, effects which were maximum at the concentration at which the bile acid induced the maximum proliferative effect on the cells. Cholate was without effect on AP-1 binding activity. In addition, we show that the AP-1 complex induced by treatment of the cells with the bile acid contained the c-fos protein. This could suggest that prolonged deregulated expression of AP-1 activity in colonic cells by certain bile acids may contribute to tumor promotion in the colon.

Role of protein kinase C in growth stimulation of primary mouse colonic epithelial cells.

The phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (160 nM) and the secondary bile acid, deoxycholic acid (50 microM) stimulated DNA synthesis in quiescent primary epithelial cells from the normal mouse colon as measured by autoradiographic analysis of [3H]thymidine incorporation. The purpose of this present study was to investigate the involvement of protein kinase C in the proliferative response of the normal colonic cells. The protein kinase C inhibitor, bisindolyl-maleimide GF 109203X, efficiently blocked the proliferative response of the cells to the phorbol ester and caused a dose-dependent decrease in the response to deoxycholic acid. While the phorbol ester-induced proliferation was unaffected by another inhibitor, H-7, the response of the cells to deoxycholic acid was blocked. Pretreatment of the cells with the phorbol ester (160 nM) for 24 h blocked the proliferative response to deoxycholic acid. Measurement of the intracellular distribution of protein kinase C activity showed a time-dependent and significant translocation of the enzyme activity from the soluble to the particulate cell fractions after exposure to 12-O-tetradecanoylphorbol-13-acetate. While exposure to the bile acid indicated a similar time-dependent translocation of the enzyme activity, the effect was not significant. The phorbol ester induced a time-dependent accumulation of c-fos mRNA and protein was measured by solution hybridization and immunocytochemistry, respectively. No effect of deoxycholic acid on c-fos expression could be observed in the present study. The data support a role for protein kinase C in the growth stimulating effect of physiological concentrations of deoxycholic acid on normal colonic epithelial cells. However, differences in the mechanisms underlying phorbol ester- and bile acid-induced proliferation are indicated.

Activation and effects of the food-derived heterocyclic amines in extrahepatic tissues.

Humans frequently inhale as well as ingest cooked-food mutagens, among which the heterocyclic amines are the quantitatively most important. An extensive systemic distribution of these mutagens implies that most tissues in the body are exposed. Tissues containing cytochrome P450 (CYP) may be particularly susceptible to DNA damage. Accordingly, animal experiments have shown that oral exposure to heterocyclic amines leads to tumor formation at multiple sites. CYP1A2, which has only been demonstrated in the liver, seems to be the isozyme most efficient in metabolically activating the heterocyclic amines. In extrahepatic tissues, however, other CYP forms are likely to be important. Using Salmonella mutagenicity as an endpoint, we have studied the metabolic activation of 2-amino-3-methylimidazo[4,5,f]quinoline (IQ), 2-amino-3,8-dimethylimidazo[4,5,f]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) by isolated lung microsomes from rats and mice. Our studies show that CYP2A3, an isozyme that has hitherto not been investigated with regard to its capacity to activate heterocyclic amines, catalyses a major part of the IQ activating reactions in the uninduced lung. The formation of mutagens during cooking of meat is highly temperature dependent and meat extracts heated at 200 degrees C show a strong mutagenic activity in the Ames Salmonella assay. These extracts caused mutations at the HPRT locus in normal human fibroblasts as well as a pronounced decrease in survival of the cells. Furthermore, the heated meat extracts caused a decreased proliferative activity in primary cultures of normal mouse colonic epithelial cells as measured by autoradiography.

Effect of lipid extracts of fecal water from colorectal tumor patients on DNA synthesis in Swiss 3T3 cells.

BACKGROUND: Alterations in the proliferative activity of the colonic epithelium play a central role in the development of human colon cancer. It has been suggested that crypt cell hyperproliferation may partly be due to cell regeneration after exposure to cytotoxic surfactants in fecal water. METHODS: The present study determined whether lipid extracts of fecal water from colorectal tumor patients had the capacity to influence DNA synthesis in growing and quiescent Swiss 3T3 cells, at concentrations at which no cytotoxicity was evident. RESULTS: Four of five such extracts significantly increased DNA synthesis in growing cells but were without effect on quiescent cells. Comparison of the effects of the lipid extracts on the cells with that of bile acids, compounds that have received much attention as potential regulators of proliferation, indicated that the effect of the lipid extracts could not solely be attributed to bile acids. CONCLUSIONS: Lipid components of fecal water may also influence cell proliferation through mechanisms other than cytotoxicity.

Increases in 13-hydroxyoctadecadienoic acid dehydrogenase activity during differentiation of cultured cells.

Recently, oxidation products of linoleic acid such as 13-hydroxyoctadecadienoic acid (HODE) have been implicated in the regulation of cellular physiology including the proliferative response to growth factor treatment. In addition, an NAD(+)-dependent 13-HODE dehydrogenase was recently described. To evaluate the contribution of this enzyme to cellular processes we have examined the behavior of the enzyme under different conditions. In the present report, changes in the activity of 13-hydroxyoctadecadienoic acid dehydrogenase during in vitro differentiation of two different cell lines were examined. The cell line HT-29 undergoes induced differentiation via manipulation of the medium while the Caco-2 line undergoes spontaneous differentiation upon attainment of confluence. In both cell lines, longer culture times were accompanied by increases in 13-HODE dehydrogenase activity. The increase in enzyme activity continued even after cell proliferation had ceased. Cellular differentiation was verified by the observation of increases in sucrase and alkaline phosphatase activities. In addition, the activity of 13-HODE dehydrogenase was measured in growing, early confluent and late confluent cultures of undifferentiating Swiss mouse 3T3 fibroblasts. In the fibroblast line, no significant changes in 13-HODE dehydrogenase activity were observed during the course of the experiment. The specific activity of 13-HODE dehydrogenase was also significantly different between the three cell lines, consistent with the extent of differentiation. Highest levels of activity were found in Caco-2 cells (200-400 pmol/min/mg) and barely detectable levels in the fibroblasts (0.6-2 pmol/min/mg). The correlation between 13-HODE dehydrogenase and cell differentiation suggests the enzyme may have a role to play in the partitioning of cells between proliferation and differentiation pathways.

Proliferative potential and expression of cell type specific functions in primary mouse colonic epithelial cells.

Primary cultures of mouse colonic epithelial cells have been obtained that are typically epithelial by morphology and moreover express keratins and endogenous beta-galactosidase; this latter activity was also demonstrated in the epithelial lining of the mouse colonic mucosa. The proliferative response of the primary colonic epithelial cells to epidermal growth factor, insulin, and the bile acid, deoxycholic acid, has been studied. Using primary cultures maintained at suboptimal growth conditions, which yielded 96 to 100% quiescent cells, epidermal growth factor, insulin, and the bile acid, deoxycholic acid, at concentrations at which it normally occurs in the aqueous phase of human feces, stimulated proliferation as measured by autoradiography. Exposure of the cells to combinations of these factors resulted in additive increases in growth. In conclusion, cells from the normal mouse colon can now be cultured while retaining at least two normal marker functions and moreover respond to some known mitogens and the potential tumor promoter deoxycholic acid. The cells can also be subcultivated while maintaining their epithelial morphology and marker functions for at least 3 passages.

The influence of Streptococcus mutans on adhesion of Candida albicans to acrylic surfaces in vitro.

Adhesion of Candida albicans and Streptococcus mutans was studied by incubation of radiolabelled cells with acrylic test specimens in a chemically defined growth medium. Strep. mutans adhered firmly in the presence of sucrose, while C. albicans was only loosely attached to the acrylic in both glucose and sucrose media. Firm adhesion of C. albicans occurred when the yeast was incubated simultaneously with Strep. mutans, in the presence of sucrose. The adhesion of C. albicans was also stimulated by incubation with Strep. mutans culture supernatants. Adhesion was not affected by the presence of partially purified glucosyltransferase from Strep. mutans IB. Coaggregation between C. albicans and Strep. mutans upon growth in sucrose medium was observed by light and scanning electron microscopy. No coaggregation was observed in the presence of glucose.

Water-soluble glucans from Streptococcus mutans strain Ing-Britt as an energy source for bacterial growth.

Two fractions of water-soluble glucans with different molecular weight were produced by Streptococcus mutans IB. The larger glucan had a molecular weight of about 40,000. The molecular weight of the small glucan was estimated to be 4,100 by gel filtration chromatography and by biochemical methods. These glucans were tested for their ability to initiate and support growth of S. mutans IB and Streptococcus sanguis 903. S. sanguis 903 could grow with the low molecular weight glucan at a reduced rate compared with glucose. S. mutans IB could not utilize any of the glucans. No endo- or exo-glucanase activities could be detected in culture supernatants of any of the strains. In addition to maltose S. sanguis 903 could also utilize maltotriose, maltopentaose and maltoheptaose for growth while S. mutans IB could not grow with maltopentaose or maltoheptaose.

Adhesion of Streptococcus rattus and Streptococcus mutans to metal surfaces.

The adhesion of Streptococcus rattus BHT and Streptococcus mutans IB to metal specimens of amalgam, silver, tin and copper was studied using (6-3H)thymidine labeled cells. In the standard assay the metal specimens were suspended by a nylon thread in an adhesion solution containing a chemically defined bacterial growth medium (FMC), sucrose, and radiolabeled bacteria. Maximum amounts of adhering bacteria were obtained after about 100 min of incubation. Saturation of the metal specimens with bacteria was not observed. Both strains also adhered in the absence of sucrose, indicating that glucan formation was not necessary for adhesion. However, in the presence of glucose, adhesion was only 26-45% of that observed in the presence of equimolar sucrose. Sucrose-dependent stimulation of adhesion seemed to be due to increased cell-to-cell adhesion capacity. Isolated radiolabeled water-insoluble and water-soluble polysaccharides produced from sucrose by S. rattus BHT were not adsorbed to the metal surfaces.