Prediction of permeability of FD-4 through porous poly (2-hydroxyethyl methacrylate) membrane by multiple linear regression and artificial neural network.

The aim of this study was to predict the permeability through porous poly (2-hydroxyethyl methacrylate) (pHEMA) membranes of fluorescein isothiocyanate-labeled dextran molecular weight 4400 (FD-4) as a model of peptide and protein drug movement. Homogeneous standard membranes were prepared by redox polymerization. Permeability data were predicted by an artificial neural network (ANN) as a function of polymerization factors, and the accuracy was compared with that of conventional multiple linear regression (MLR). Good linearity was observed with each model, with the correlation coefficient of a leave-one-out cross-validation (Rcross) being 0.857 for the MLR model and 0.876 for the ANN model. The mean bias and mean accuracy for the ANN were somewhat smaller than those of the MLR. The ANN method provides an accurate quantitative approximation of the permeability coefficient of FD-4, as judged by conventional MLR, and could be applied to prediction of the non-linear relation between polymerization factors and the permeability of FD-4.

Correlations between salivary protease and supragingival or subgingival calculus index.

Significant positive correlations between protease activities in saliva and supragingival/subgingival calculus indices, periodontal pocket depth, or P-M-A index were confirmed. When the partial correlation was calculated, the subgingival calculus index correlated well with the sediment protease at pH 8.5, and the supragingival calculus index showed a significant correlation with the supernate protease at pH 4.5.

Correlation between the protease activities and the number of epithelial cells in human saliva.

The positive significant correlation (r = 0.788, n = 37) was shown between the protease activity and the number of epithelial cells (not of leukocytes) in saliva. There was little protease activity observed in glandular saliva. It is possible to conclude from the results that a main source of salivary protease is the epithelial cells in saliva. It was also found that the oral prophylaxis reduced the numbers of epithelial cells and leukocytes and the protease activity.

Enhanced rectal absorption of [Asu1,7]-eel calcitonin in rats using polyacrylic acid aqueous gel base.

The effects of polyacrylic acid aqueous gel on the absorption of rectally administered [Asu1,7]-eel calcitonin, a calcitonin analogue, were investigated in rats. The [Asu1,7]-eel calcitonin (1 U/kg) was given into the rectal loop in gel bases at various pH (5.5-8.5) and polyacrylic acid concentrations (0.01-1.0% w/v). The maximum hypocalcemic effect was obtained in approximately 30 min after administration of the analogue in a 0.1% w/v polyacrylic acid gel base at pH 5.5. The plasma calcium level decreased by approximately 18% from the initial level. Rectal administration in vehicles such as polyethylene glycol 1000, triglyceride fatty acid mixture base, or saline solution had little or no hypocalcemic effect at a dose of 5 U/kg. The results indicated that a polyacrylic acid aqueous gel base significantly improved the absorption of this analogue. Furthermore, rectal administration in a polyacrylic acid gel base (0.1% w/v; pH 5.5) required a dose 35 times greater than an intravenously administered dose to achieve an equivalent hypocalcemic effect.

Oral delivery of insulin using pH-responsive complexation gels.

The goal of oral insulin delivery devices is to protect the sensitive drug from proteolytic degradation in the stomach and upper portion of the small intestine. In this work, we investigate the use of pH-responsive, poly(methacrylic-g-ethylene glycol) hydrogels as oral delivery vehicles for insulin. Insulin was loaded into polymeric microspheres and administered orally to healthy and diabetic Wistar rats. In the acidic environment of the stomach, the gels were unswollen due to the formation of intermolecular polymer complexes. The insulin remained in the gel and was protected from proteolytic degradation. In the basic and neutral environments of the intestine, the complexes dissociated which resulted in rapid gel swelling and insulin release. Within 2 h of administration of the insulin-containing polymers, strong dose-dependent hypoglycemic effects were observed in both healthy and diabetic rats. These effects lasted for up to 8 h following administration.

Effects of sex hormones on production of prostaglandin E2 by human peripheral monocytes.

The effects of sex hormones on the vitro production of prostaglandin (PG) E2 by monocytes were investigated. Monocytes were obtained from heparinized peripheral blood of healthy adults and incubated for 24 hours with lipopolysaccharide (LPS) and sex hormones. After incubation, the medium was assayed for PGE2 by means of radioimmunoassay. PGE2 production by monocytes was enhanced by progesterone. Estradiol reduced PGE2 production at 0.4 ng/ml, but enhanced it at 20 ng/ml. Testosterone reduced PGE2 production. The reduced PGE2 production by monocytes treated with 0.4 ng/ml of estradiol was restored to the control level by addition of progesterone at 20 ng/ml. These results suggest that sex hormones may modulate gingival inflammation mediated by PGE2.

Effects of sex hormones on production of interleukin-1 by human peripheral monocytes.

BACKGROUND: Interleukin-1 (IL-1) is a potent mediator of inflammation and is known to induce bone resorption. We studied the effects of sex hormones on the function of human monocytes and demonstrated that prostaglandin E2 (PGE2) production was enhanced by progesterone and estradiol. As PGE2 has been shown to suppress the production of IL-1 by monocytes, it was speculated that sex hormones also modify the production of IL-1 by regulating PGE2 production. Thus, the effects of sex hormones on the production of IL-1 from human peripheral monocytes and the influence of PGE2 were investigated. METHODS: Mononuclear leukocytes were obtained from 22 healthy adults. Progesterone, 17-beta estradiol (estradiol), and testosterone were used as representative sex hormones. Monocytes were incubated at 37 degrees C in air with 5% CO2 for 24 hours in RPMI 1640 medium with sex hormones at the designated concentrations. LPS (Salmonella typhimurium) was used to stimulate the monocytes at a concentration of 10 microg/ml. The concentrations of IL-1alpha and IL-1beta in the medium were determined by enzyme-linked immunosorbent assay kits. The concentration of PGE2 was determined using a direct radio-immunoassay kit. Indomethacin was used to inhibit the synthesis of PGE2 and eliminate its effect on the production of IL-1. RESULTS: Estradiol at concentrations of 0.04 ng/ml or more significantly reduced both IL-1alpha and IL- 1beta production. Progesterone also reduced IL-1alpha and IL-1beta production significantly at concentrations of 0.1 ng/ml or more and 0.02 ng/ml or more, respectively. The reductions in IL- 1alpha and IL-1beta production by sex hormones were not affected by addition of indomethacin. CONCLUSIONS: Estradiol and progesterone inhibited the production of IL-1 from human peripheral monocytes. The inhibition was not the result of enhanced production of PGE2. Under conditions in which sex hormone levels are low, monocytes produce IL- more readily in response to stimulation by LPS than high levels of such hormones. Low concentrations of sex hormones may be considered as one of the risk factors for periodontitis.

Effects of sex hormones on chemotaxis of human peripheral polymorphonuclear leukocytes and monocytes.

The effects of sex hormones on the in vitro chemotaxis of polymorphonuclear leukocytes (PMNs) and monocytes were investigated using fMLP as the chemoattractant. PMNs, monocytes, and plasma were obtained from heparinized peripheral blood of healthy adults. Chemotaxis of PMNs or monocytes treated with sex hormones were tested using 48-well chemotaxis microchambers. The correlation between sex hormone levels in plasma and the chemotactic ability of PMNs from the same donor was also investigated. The chemotaxis of PMNs was enhanced by progesterone, while it was reduced by estradiol. Random migration of PMNs was also enhanced by progesterone and reduced by estradiol. The effect of estradiol on PMN chemotaxis was inhibited by addition of antiestrogens or progesterone. Testosterone did not have a measurable effect on PMN chemotaxis. A significant positive correlation was found between the concentration of progesterone in plasma of females and PMN chemotactic ability in vitro. For males, there was no significant relationship between plasma levels of sex hormones and PMN chemotactic ability. Estradiol and testosterone levels in plasma did not correlate with PMN chemotactic ability. Sex hormones had no effect on the chemotaxis of monocytes. These results suggest that the altered PMN chemotaxis associated with gingival inflammation may be due to the effects of sex hormones.

Immunohistochemical study of gamma delta T cells in human gingival tissues.

The distribution and the density of gamma delta T cells in human gingival tissues were examined immunohistochemically in biopsy samples obtained from 20 subjects. Few gamma delta T cells were observed in gingival tissue free from inflammatory cell infiltration, but were found, albeit in low numbers, in association with inflammatory cell infiltration, especially T cells. This relationship with T cells was confirmed statistically. The ratios of gamma delta T cells to T cells in the epithelia and in the connective tissue were calculated in the sections in which more than 500 CD3-positive cells were identified. Seven of eight such epithelial specimens showed a ratio of less than 1% and one less than 2% (mean +/- SD; 0.8% +/- 0.4). In the connective tissue, 8 of 13 such specimens showed less than 1%, three less than 2%, one 3%, and one 7% (1.4% +/- 1.9). These results suggest that basically gamma delta T cells are not resident cells in the gingival epithelium such as comprise the first defense line against exogenous irritation. They may play some role in the pathogenesis of periodontal disease collaborating with alpha beta T cells in the inflammatory response.

Analysis of oestrogen receptor mRNA by reverse transcriptase-polymerase chain reaction in human periodontal ligament cells.

Periodontal ligament (PDL) cells have osteoblast-like features and are capable of differentiating into osteogenic cells. As human osteoblasts express oestrogen receptor mRNA, it is possible that PDL cells do so also, but findings have been conflicting. To determine whether they do express oestrogen receptor mRNA, the reverse transcriptase-polymerase chain reaction was performed with two different primers. Cells were obtained from a healthy periodontal ligament of premolar extracted for orthodontic reasons. The human breast adenocarcinoma cell-line MCF7 was used as a positive control. Expression of oestrogen receptor mRNA was detected in PDL cells with one of the primers but with less intensity than in MCF7 cells. Southern hybridization confirmed these results. These findings suggest that PDL cells express oestrogen receptor mRNA at low levels.

Effect of a protease derived from the oral bacterium Bacteroides loescheii on the inhibition of calcium-phosphate precipitation by human parotid saliva.

A protease from the culture fluid of this microorganism was purified and characterized using DEAE-Sephadex and chromatofocusing and classified as a thiol protease with a mol. wt estimated as 27,500. It abolished the inhibitory activity of parotid saliva on the precipitation of calcium phosphate from supersaturated solutions. Such proteases may contribute to dental calculus formation.

The effects of oestrogen on osteocalcin production by human periodontal ligament cells.

The purpose was to investigate the effects of oestradiol on the function of periodontal ligament (PDL) cells by measuring the production of osteocalcin in vitro. Cells were obtained from the healthy periodontal ligament of teeth extracted from two males and two females for orthodontic reasons. Serum-free medium was used when testing the effects of oestradiol on PDL cells. The amount of osteocalcin in the culture medium was analysed by two-step sandwich enzyme immunoassay in the presence or absence of oestradiol. It was shown that oestradiol enhanced the production of osteocalcin by PDL cells in a time- and dose-dependent manner. PDL cells obtained from both male and female donors were affected by oestradiol. It thus appears that oestradiol is one of the factors important for PDL cells to express their function.

Effects of fluoride intake on the mineral content, acid solubility and resorption caused by experimental periodontitis of rat alveolar bone.

Adult rats were given either distilled water or drinking water containing 100 parts/10(6) of fluoride. The alveolar bone of rats given fluoride for 90 days showed an increased mineral content and decreased acid solubility compared to the bone of rats given distilled water. Experimental periodontitis was initiated in both groups after 110 days of treatment to cause alveolar bone resorption. Fourteen days later, the rats were killed and it was found that the alveolar bone resorption caused by experimental periodontitis was significantly smaller in the rats given fluoride in their drinking water than in those given distilled water. The findings suggest that fluoride intake might have a protective effect on rapidly progressing alveolar bone resorption.

Absorption of insulin from pluronic F-127 gels following subcutaneous administration in rats.

The main objective of this work was to evaluate the use of Pluronic (PF127) gels, polylactic-co-glycolic acid (PLGA) nanoparticles and their combination for parenteral delivery of peptides and proteins having short half-lives using insulin as a model drug. The in vitro insulin release profiles of various PF127 formulations were evaluated at 37 degrees C using a membraneless in vitro model. In vivo evaluation of the serum glucose and insulin levels was performed following subcutaneous administration of various insulin formulations in normal rats. The in vitro results demonstrated that the higher the concentration of PF127 in the gel, the slower the release of insulin from the matrices, independent of the vehicle used. The acute hypoglycemic peak resulting from administration of an insulin solution between 0.5 and 2.0 h after administration (peak at 1 h) is replaced after administration of insulin-PLGA nanoparticles by an almost constant hypoglycemic effect with a slower recovery of the serum glucose levels at about 2 h after administration. By loading insulin into PF127 gels, a slower and more prolonged hypoglycemic effect of insulin was obtained in inverse proportion to the polymer concentration. PF127 gel formulations containing insulin-PLGA nanoparticles had the most long-lasting hypoglycemic effects of all formulations. From the current in vitro and in vivo study, we concluded that PF127 gel formulations containing either drug or drug-nanoparticles could be useful for the preparation of a controlled delivery system for peptides and proteins having short half-lives.

Enhanced rectal absorption of insulin-loaded Pluronic F-127 gels containing unsaturated fatty acids.

The objective of this study was to prepare and to evaluate Pluronic F-127 (PF127) gel containing unsaturated fatty acids such as oleic acid (18:1), eicosapentaenoic acid (20:5) and docosahexaenoic acid (22:6) as a potential formulation for rectal delivery of insulin. The hypoglycemic effect of insulin was examined following rectal administration of the various formulations in normal rats. Rectal insulin absorption was markedly enhanced, and marked hypoglycemia was induced by all PF127 gels (insulin dose, 5 U/kg) containing different unsaturated fatty acids. PF127 gels containing unsaturated fatty acids presented low tmax mean values indicating that the absorption of insulin occurred very rapidly in the rectum. The relative hypoglycemic efficacy of PF127 gel formulations containing fatty acids such as oleic acid, eicosapentaenoic (EPA) and docosahexaenoic (DHA) were 28.4+/-8.1, 26.8+/-14.3 and 23.1+/-5.7%, respectively. The finding demonstrated that 20% PF127 gels containing unsaturated fatty acids are potential formulations for rectal delivery of insulin.

Pluronic F-127 gels incorporating highly purified unsaturated fatty acids for buccal delivery of insulin.

The present study investigated the release profiles of insulin from Pluronic F-127 (PF-127) gel containing unsaturated fatty acids such as oleic acid (18:1), eicosapentaenoic acid (20:5) or docosahexaenoic acid (22:6) and the hypoglycemic effect of insulin following the buccal administration of the gel formulations in normal rats. Insulin release from the gels decreased in the presence of unsaturated fatty acids. Remarkable and continuous hypoglycemia was induced by all PF-127 gels (insulin dose, 25 IU/kg) containing unsaturated fatty acids. PF-127 gels containing oleic acid showed the highest pharmacological availability (15.9+/-7.9%). Our finding demonstrate that 20% PF-127 gels containing unsaturated fatty acids are potential formulations for the buccal delivery of insulin.

Antibacterial effect of silver-zeolite on oral bacteria under anaerobic conditions.

OBJECTIVES: The purpose of this study was to evaluate the antibacterial effect of silver-zeolite (SZ) against oral bacteria under anaerobic conditions. METHODS: The antibacterial activity of SZ was evaluated by determining the minimum inhibitory concentrations (MICs) using two-fold serial dilutions of SZ in Brain Heart Infusion broth. Release of Ag+ into the broth was measured by an atomic absorption technique. RESULTS: SZ inhibited the growth of the bacteria tested under anaerobic conditions. The MIC of SZ ranged between 256 and 2048 micrograms/ml, which corresponded to a range of 4.8-38.4 micrograms/ml of Ag+. All strains grew in broth containing 16,384 micrograms/ml of type-A zeolite. SIGNIFICANCE: These results suggested that SZ may be a useful vehicle to provide antibacterial activity to dental materials used even under anaerobic conditions such as deep in the periodontal pocket.

Development of an oral health indicator in infants.

OBJECTIVE: To develop a simple predictive indicator for 1.5-year-old infants' oral health as an aid for maternal preventive efforts. DESIGN: The development of an indicator was based on the relationship between life style factors of the 1.5-year-old infants and the caries incidence of the same children at three years of age. Data gained at examination of the 3-year-old subjects were subjected to analysis with the caries-free information on the same subjects at 1.5 years. SETTING: A community health centre in Hiroshima Japan. SUBJECTS: 1575 children received an oral check-up at 3 years of age at a community health centre. OUTCOME MEASURES: The category scores of the six items which were extracted retrospectively from life style factors at 1.5 years affecting caries prevalence at 3 years of age were quantified and transformed into integral numbers so as to make up a total of 100. The sum of the six transformed scores was named an Infants Dental Index (IDI). RESULTS: The IDI score was associated significantly with the dmft score at 3 years of age (r = -0.18 P < 0.001). When the subjects were classified into four groups with respect to the IDI score these four groups were significantly related to the caries onset between 1.5 and 3 years of age (chi 2 = 36.74, P < 0.001 CONCLUSIONS: The IDI developed in this study appeared to be valid so that it could be applied in the field of the community dental health to identity higher risk infants and direct more effective health education to mothers.

Periodontal regeneration: myth or reality?

Recent scientific advances have enabled scientists to understand some of the basic biological events leading both to an accelerated and improved healing of many tissues. These advances are now being applied to periodontal wound healing. Previous studies have indicated that various oncogenes are sequences related to growth factors and, that when these sequences are altered, cellular phenotype and especially cellular proliferation is altered. Data are presented using various human cell lines, including a c-myc-transfected periodontal ligament (PDL) cell line, which delineate the relationship between oncogenes, mRNA oncogene transcripts and polypeptide growth factors as inducers of cell phenotypic alterations, including adhesion, migration and proliferation. The polypeptide growth factors are a unique class of molecules that regulate cell phenotype both in vitro and in vivo. These demonstrable phenotypic conversions have indicated that such factors will play an important role in hard-soft tissue repair. Additionally, new data suggest that older PDL cells are not responsive to polypeptide stimulation; both the migratory and proliferative responses are diminished. A unique growth factor has been isolated and sequenced, which when applied to older PDL cells will reverse this refractory phenotype. These recent studies are extensively reviewed with emphasis and conclusions based on growth factor-induced periodontal regeneration. These studies stress the undeniable role that these novel approaches will have in future chairside periodontal practice.

[The comparison of blood levels between peripheral vein and tooth extraction wound after the oral administration of antibiotics (author's transl)].

The oral administration of 300 mg of clindamycin was undertaken on 23 patients, of 500 mg of cefadroxil on 11 patients and of 250 mg of talampicillin on 12 patients, and then tooth extraction was performed under local anesthesia. Blood samples were taken from the extraction wound and the peripheral vein at the same time and assayed by the bioassay method. The blood levels of clindamycin and cefadroxil indicated a similar pattern between the extraction wound and the peripheral vein, but the blood level of talampicillin reached peek level rapider than clindamycin and cefadroxil. The blood levels of the extraction wound were 60 - 80% as compared with the venous blood levels with each antimicrobial agent.