Increased bFGF level in the serum of patients with phenytoin-induced gingival overgrowth.

To elucidate the involvement of bFGF (basic fibroblast growth factor) in the pathogenesis of phenytoin-induced gingival overgrowth, we measured the concentration of bFGF in the serum of 36 epileptic patients taking phenytoin and in 94 normal volunteers by enzyme-linked immunosorbent assay technique. The concentration of phenytoin in serum was determined by high-performance liquid chromatography. In 34 of 36 patients taking phenytoin in this investigation, apparent gingival overgrowth was noticed. The mean concentration of bFGF was 33.9+/-18.5 pg/ml in the overgrowth group and 10.6+/-5.2 pg/ml in the volunteer group (p<0.01). The serum phenytoin level did not correlate (r=0.22, p=0.2) with the degree of gingival overgrowth but there was a significant correlation (r=0.38, p=0.023) between the degree of gingival overgrowth and the serum bFGF level. However, no correlation was observed among age, daily phenytoin dose, total phenytoin dose, duration of phenytoin therapy, serum phenytoin level, or serum bFGF level. The results suggested that enhanced serum bFGF level was implicated in the pathogenesis of phenytoin-induced gingival overgrowth.

Fluid and protein flux across the pulpodentine complex of the dog in vivo.

Dentinal fluid volume and protein concentrations were measured from cavities prepared in the dentine of dog molars. Fluid was collected under spontaneous conditions, during the application of negative pressures, and during a recovery period. The data allowed the calculation of dentine permeability as hydraulic conductances, fluid flux, protein flux, reflection coefficients and pulpal tissue pressures. In general, the protein concentration of dentinal fluid was about one-fifth that of plasma during spontaneous conditions and it fell when fluid flux was increased by the application of external negative pressures. The collection and analysis of dentinal fluid looks promising as a non-invasive method of assessing the state of the underlying pulp.

Polyamine analysis of infected root canal contents related to clinical symptoms.

The contents of infected root canals were collected by mechanical and chemical debridement. Intracanal polyamines were identified and quantified by HPLC in all canals examined, however, no histamine was detected in any canal. The relationship between the amount of each polyamine and clinical signs of the teeth was analyzed statistically. The teeth without clinical signs tended to have limited varieties and smaller amounts of polyamines. Using chi 2, the amounts were significantly greater in teeth with spontaneous pain, swelling and putrescent odor (p less than 0.01), with exudate (p less than 0.025) and with percussion pain (p less than 0.1) than in teeth without. Amounts of putrescine (p less than 0.05) and total polyamines (p less than 0.01) were greater in teeth with spontaneous pain and percussion pain than in teeth without clinical sign, and those with root canal exudate also had greater amounts of total polyamines than those without (p less than 0.01). The amounts of cadaverine, however, from the teeth with gingival fistulae, was greater than from those without (p less than 0.05). No significant relationship was established between the amount of each polyamine and the presence of putrescent odor or gingival swelling. Intracanal polyamines, especially putrescine may leak out through apical foramen and may be implicated in pain production by eliciting acute inflammatory response in the periapical tissues.

Properties of kininase in rat dental pulp.

A kininase, capable of degrading bradykinin, was partially purified from the dental pulp of rats, and its properties were investigated. Chromatography on both Sephadex G-200 and DEAE Sephadex A-50 columns gave a single peak of kininase activity. The molecular weight of the enzyme, estimated by gel filtration, was about 67,000, and the optimum pH for the enzymatic reaction was about 7.5. The enzyme appears to contain a labile SH group(s) that is essential for its activity, because CdCl2, HgCl2 (0.1 mM each), and p-chloromercuric benzoate (0.05 mM) inhibited the enzyme completely, while dithiothreitol retarded the loss of activity during storage. Of various peptides tested, bradykinin was the substrate most sensitive for the enzyme. The enzyme released several amino acids located in the C-terminal regions of bradykinin--angiotensin I and neurotensin--but only one C-terminal amino acid from des-Arg9--bradykinin and angiotensin II. In contrast, the enzyme did not release any amino acids from substance P, of which only the two amino acids in the N-terminal region are the same as those of bradykinin, but its C-terminal is blocked by an amino group. Although the enzyme was not so highly purified as to rule out the contribution of other peptidases, these results suggest that the dental pulp of rats may contain a single enzyme that degrades bradykinin, and the enzyme may be a type of carboxypeptidase, differing from known kininases from other animal sources.

Fluorometric assay of kininase activities in rat tissues.

A simple method for the assay of bradykinin (BK)-degrading enzymes was investigated. The procedure of the method includes enzymatic degradation of BK, separation of the residual BK on a small P-cellulose column (0.6 X 3 cm), and its fluorometrical determination based on the reaction with fluorescamine. BK was separated completely from its fragments produced during enzymatic reaction by the column chromatography. The recovery rate of BK was 96 +/- 3%. Quantitative determinations could be carried out on 0.2 nmol of BK, at least in the fluorometry. This method was available for the assay of the enzymes in tissue homogenates as well as in purified preparations, and its usefulness for the study of the enzymes is presented.