Effect of prosthetic restoration on masticatory function in patients with shortened dental arches: a multicentre study.

The aim of this multicentre study was to investigate the effect of prosthetic restoration for missing posterior teeth on mastication in patients with shortened dental arches (SDAs). Partially dentate patients who had an intact teeth in anterior region and missed distal molar(s) (2-12 missing occlusal units) classified as Kennedy Class I or Class II were recruited from seven university-based dental hospitals in Japan. Of the 125 subjects who underwent baseline (pre-treatment) and follow-up/post-treatment evaluation, 53 chose no replacement of missing teeth and 72 chose treatment with removable partial dentures (n = 53) or implant-supported fixed partial dentures (n = 19). Objective masticatory performance (MP) was evaluated using a gummy jelly test. Perception of chewing ability (CA) was rated using a food intake questionnaire. In the no-treatment group, mean MP and CA scores at baseline were similar to those at follow-up evaluation (P > 0·05). In the treatment group, mean MP after treatment was significantly greater than the pre-treatment mean MP (P < 0·05). However, the mean perceived CA in the treatment groups was similar at pre- and post-treatment (P > 0·05). In a subgroup analysis of subjects in the treatment group, subjects with lower pre-treatment CA showed a significant CA increase after treatment (P = 0·004), but those with higher pre-treatment CA showed a significant decrease in CA (P = 0·001). These results suggest that prosthetic restoration for SDAs may benefit objective masticatory performance in patients needing replacement of missing posterior teeth, but the benefit in subjective chewing ability seems to be limited in subjects with perceived impairment in chewing ability before treatment.

Effect of prosthetic restoration on oral health-related quality of life in patients with shortened dental arches: a multicentre study.

The aim of this multicentre prospective study was to investigate the effect of prosthetic restoration for missing posterior teeth in patients with shortened dental arches (SDAs). SDA patients with 2-12 missing occlusal units (a pair of occluding premolars corresponds to one unit, and a pair of occluding molars corresponds to two units) were consecutively recruited from seven university-based dental hospitals in Japan. Patients chose no replacement of missing teeth or prosthetic treatment with removable partial dentures (RPDs) or implant-supported fixed partial dentures (IFPDs). Oral health-related quality of life (OHRQoL) was measured using the oral health impact profile (Japanese version - OHIP-J) at baseline and follow-up/post-treatment evaluation. Of the 169 subjects who completed baseline evaluation, 125 subjects (mean age; 63.0 years) received follow-up/post-treatment evaluation. No-treatment was chosen by 42% (53/125) of the subjects, and 58% (72/125) chose treatment with a RPD (n = 53) or an IFPD (n = 19). In the no-treatment (NT) group, the mean OHIP summary score at baseline was similar to that at follow-up evaluation (P = 0.69). In the treatment (TRT) group, the mean OHIP summary score decreased significantly after the RPD treatment (P = 0.002), and it tended to decrease, though not statistically significant (P = 0.18), after the IFPD treatment. The restoration of one occlusal unit was associated with a 1.2-point decrease in OHIP summary score (P = 0.034). These results suggest that the replacement of missing posterior teeth with RPDs or IFPDs improved OHRQoL. Prosthetic restoration for SDAs may benefit OHRQoL in patients needing replacement of missing posterior teeth.

Manometric evaluation of oral function with a hand-held balloon probe.

Tongue pressure measured with a disposable hand-held balloon probe has been used for assessing tongue function; however, no diagnostic standard for assessing other oral functions exists currently. In this study, to develop a method for multifactorial manometric evaluation of oral functions, 20 men and 20 women (21-32 years of age) were instructed to apply 7-s maximal voluntary muscular effort on a hand-held balloon probe placed against the anterior and lateral parts of the palate, buccal surface of the molars on the habitual chewing side and labial surface of the anterior teeth for measuring anterior and posterior tongue, cheek and lip pressures (LPs), respectively. Intra-session reproducibility was determined by three repeated measurements, and associations between the obtained and the conventional data on oral diadochokinesis were tested. The men exhibited higher values of all the pressure types. Further, both genders showed a positive correlation between anterior and posterior tongue pressures (PTPs) (P < 0·05), but only the women exhibited positive correlations between anterior tongue and cheek pressures, cheek and PTPs, anterior tongue and LPs, and cheek and LPs (P < 0·05). No statistically significant correlation was found between the pressures and the number of syllabic articulations, except between LP and the number of /pa/ articulations in the women (r = 0·524, P < 0·05). In conclusion, the balloon probe method enables objective manometric evaluation of oral functions and could be an effective tool for clinical epidemiological studies and evidence-based decision-making in nursing care.

Factors related to prosthetic restoration in patients with shortened dental arches: a multicentre study.

The aim of this study was to identify the factors related to prosthetic restoration in patients with shortened dental arches (SDA). SDA patients with 2-12 missing occlusal units were consecutively enrolled from seven university-based dental hospitals in Japan. Of the 145 subjects (mean age; 63·4 years), 41% chose no treatment and 59% sought to replace their edentulous spaces with removable partial dentures or implant-supported fixed partial dentures. Restoration decisions were related to tooth loss patterns. Only 3% of subjects missing just second molar(s) sought to receive prosthetic treatment, while the percentage increased to 58% in subjects who were missing first and second molars and 93% in subjects missing premolar(s). Logistic regression analyses found that young age, increased number of missing occlusal units, asymmetric arch and presence of chewing complaint were significant predictors for prosthetic restoration (P<0·05). Increased number of missing occlusal units and asymmetric arch were significant predictors for the presence of chewing complaint (P<0·05). These results suggest that perceived impairment of chewing ability owing to missing occlusal units is a critical factor for prosthetic restoration in SDA patients.

Dynamic viscoelastic properties of vinyl polysiloxane denture soft lining materials.

The aim of this study was to investigate the dynamic viscoelastic properties of seven commercially available vinyl polysiloxane denture soft lining materials. Five rectangular specimens (2 x 10 x 30 mm) were prepared from each material. The complex modulus E* (MPa) and loss tangent (tan delta) of each specimen were determined with a non-resonance forced vibration method using an automatic dynamic viscoelastometer at 1 Hz after 1 day of dry storage, and after 1, 30, 60, 90 and 180 days of wet storage at 37 degrees C. All data were analysed using one-way anova and Bonferroni/Dunn's test for multiple comparisons with a significance level of P < 0.01. All materials varied widely in terms of viscoelasticities and showed both an increase in E* and a decrease in tan delta at 1 Hz after the 1-day wet storage. After 60 days of wet storage, both E* and tan delta did not change significantly. The stiffer materials (>30% filler content) with high E* values (>2.00 MPa) showed elastic behaviour with tan delta values of around 0.03. The softer materials (6% filler content) with high tan delta values (initial value > 0.10) showed viscous behaviour and were easily affected by water absorption after the 1-day wet storage. It can be concluded that for the proper selection of vinyl polysiloxane denture soft lining materials, it is very important to evaluate the viscoelastic properties after 60 days of wet storage.

Effects of tooth loss and denture wear on tongue-tip motion in elderly dentulous and edentulous people.

The purpose of this study was to clarify quantitatively the differences in tongue-tip motion among the dentulous elderly people and also among the elderly edentulous, both with and without their dentures and, to identify the influence of tooth loss and denture wear on tongue-tip motion. Fourteen young dentulous people, 12 elderly dentulous people and 13 elderly edentulous people participated in this study. Subjects were asked to swallow a 10 mL barium sulfate solution three times. The elderly edentulous people were asked to swallow the solution while wearing dentures and with dentures removed. Functional swallowing was recorded on cine-film with a digital subtraction angiography system. Lateral cinefluorography images were obtained from seated subjects. Using a cine-projector, the movements of the tongue surface were traced as dots and lines frame by frame on a single tracing sheet within a definite period of time from the beginning of the oral phase to the end of the pharyngeal phase. With counting the number of 'trajectories' of tongue-tip motion, tongue movements were classified as 'stable' and 'hyperactive' types. The results was that significantly more 'hyperactive' type subjects were found among the elderly edentulous who were not wearing dentures (12 of 13) compared with the dentulous young (1 of 14), the elderly dentulous (1 of 13) or the elderly edentulous wearing dentures (1 of 13) (P < 0.001). The tongue-tip motion for the 'hyperactive' type was very complex and the tongue-tip anchoring against the palate was always instable.

Patterns of missing occlusal units and oral health-related quality of life in SDA patients.

The aim of this study was to explore the relationship between patterns of missing occlusal units (OUs) and oral health-related quality of life (OHRQoL) in subjects with the shortened dental arches (SDAs). Subjects with SDAs were recruited consecutively for 1 month from six university-based prosthodontic clinics. In total, 115 SDA subjects participated (mean age, 58.5 +/- 10.0 years; 71% female). The location and number of missing teeth were examined and the number of missing OUs was calculated. To evaluate OHRQoL, the Japanese version of the Oral Health Impact Profile (OHIP-J) was administered and the summary score of OHIP-J was calculated. The SDA subjects were categorized depending upon the anterior-posterior lengths of the missing or remaining OUs. Regression analyses were performed to investigate the OHIP-J differences between groups of subjects with various anterior-posterior SDA lengths. The analyses revealed that subjects who only lost the second molar contact exhibited significantly better OHRQoL than those who lost more teeth [coefficient: 11.1, 95% confidence interval (CI): 2.8-19.2, P = 0.02]. Furthermore a statistically significant group difference was observed between the groups with and without the first molar occlusal contact (coefficient: 12.8, 95% CI: 1.4 to 24.1, P = 0.03). In conclusion, although our results are of exploratory nature and need validation, patterns of missing OUs are likely to be related to the OHRQoL impairment in SDA subjects with the presence of first molar contact having a particularly important role.

Characterization of a cartilage-derived 66-kDa protein (RGD-CAP/beta ig-h3) that binds to collagen.

A 66-kDa collagen fiber-associated protein (RGD-CAP) was isolated from a fiber-rich fraction of pig cartilage by ultrafiltration and collagen-affinity chromatography. Amino acid sequencing and cDNA cloning indicated that the RGD-CAP is identical or closely related to beta ig-h3 protein which is induced in human adenocarcinoma cells by transforming growth factor-beta (TGF-beta) (Skonier, J., Neubauer, M., Madisen, L., Bennett, K., Plowman, G.D., and Purchio, A.F. (1992) DNA Cell. Biol. 11, 511-522). The RGD-CAP, as well as beta ig-h3, has the RGD sequence in the C-terminal region. The native RGD-CAP bound to type I, II, and IV collagens even in the presence of 1 M NaCl. A recombinant preparation of RGD-CAP expressed in Escherichia coli cells also bound to collagen but not to gelatin. The RGD-CAP mRNA was expressed in chondrocytes throughout all stages, although the expression level was highest during the prehypertrophic stage. In addition, TGF-beta increased the RGD-CAP mRNA level in chondrocyte cultures. Since RGD-CAP transcripts were found in most tissues, this novel collagen-binding protein may play an important role in cell-collagen interactions in various tissues including developing cartilage.

Effects of parathyroid hormone (PTH) and PTH-related peptide on expressions of matrix metalloproteinase-2, -3, and -9 in growth plate chondrocyte cultures.

The roles of PTH and PTH-related peptide (PTH-rp) in the expression of matrix metalloproteinases (MMPs) during endochondral bone formation were investigated, using various cartilages obtained from young rabbits and rabbit chondrocyte cultures. Immunohistochemical, immunoblotting, zymographical, and/or Northern blot analyses showed that MMP-2 and -9 levels were much higher in the growth plate than in permanent cartilage in vivo. In growth plate chondrocyte cultures, PTH, PTH-rp, and (Bu)2cAMP increased the amount of MMP-2 present in the culture medium, as revealed by zymograms and immunoblots, whereas the other tested growth factors or cytokines, including bone morphogenetic protein-2 and interleukin-1, did not increase the MMP-2 level. PTH also increased the MMP-2 messenger RNA level within 24 h. In addition, PTH increased MMP-3 and -9 levels in the growth plate chondrocyte cultures. However, in articular chondrocyte cultures, PTH had little effect on the levels of MMP-2, -3, and -9. In contrast to PTH, interleukin-1 induced MMP-3 and -9, but not MMP-2, in growth plate and articular chondrocytes. These findings suggest that in ossifying cartilage, PTH/PTH-rp plays a pivotal role in the induction of various MMPs, including MMP-2 (which is considered to be a constitutive enzyme), and that PTH/PTH-rp is involved in the control of cartilage-matrix degradation during endochondral bone formation.

Diverse mechanisms of osteoblast spreading on hydroxyapatite and titanium.

Hydroxyapatite (HA) is an osteoconductive implant material. We previously demonstrated that RGD peptides regulate the spreading of HOS cells on HA but not on titanium, speculating that the osteoconductivity of HA might be attributed to this RGD domain-dependent spreading of osteoblasts. To confirm this hypothesis, the molecules which regulate the spreading of HOS cells on HA and on titanium were investigated. The 50% effective dose (ED50) of RGD peptide for the spreading on HA was five fold lower comparing to titanium. Anti-alphaV integrin antibody, vitronectin, and fibronectin inhibited the spreading on HA but not on titanium. In Western blot analysis, vitronectin and fibronectin were found in components adsorbed to HA but not to titanium. Taken together, the spreading of HOS cells on HA but not on titanium requires the interaction of alphaV integrin and its ligands. The ED50 of the RGD peptides on titanium but not on HA was remarkably reduced by neuraminidase treatment, that by itself could not inhibit the spreading on both materials. This phenomenon suggests that RGD domain and sialic acid cooperatively but not independently mediate the spreading of HOS cells on titanium. Collectively, the molecules regulating the spreading on HA are apparently different from those on titanium. The spreading of osteoblasts mediated by RGD domain of vitronectin and fibronectin might contribute to the osteoconductive ability of HA.

Action of FGMgCO3Ap-collagen composite in promoting bone formation.

To improve the biological properties of materials as bone substitutes, functionally graded CO3 apatite crystals containing magnesium, FGMgCO3Ap, were synthesized to be mixed with atelocollagen and made into a composite pellet. A radio-labeled cell adhesion experiment showed that the degree of adherence of mouse MC3T3E1 osteoblast-like cells to the FGMgCO3Ap-collagen composite was better than to CO3Ap-collagen and much better than to the Ti plate. When the composites were implanted beneath the periosteum cranii of rats, the FGMgCO3Ap-collagen composite was metabolized faster than the CO3Ap-collagen composite and better formation of new bone and osteoblast arrangement at the interface between the composite and the periosteum cranii was observed. When the composites were implanted into the femur of rabbits, clear bone formation with a higher degree of bone density was observed for the FGMgCO3Ap-collagen composite. These results suggest that the Mg2+ ions taken into the apatite crystals may contribute to the acceleration of osteoblast adhesion to apatites and promote bone formation, cross-talking with osteoblasts at the molecular level.

Acid pretreatment of titanium implants.

The purpose of this study was to evaluate the effectiveness of several methods of cleaning titanium surfaces as pretreatment for surface modifications by analyzing the chemical interaction of three acids, such as Na(2)S(2)O(8),H(2)SO(4) and HCl, followed by rinsing with acetone or ultrapure water. Chemical evaluation, using X-ray photoelectron spectroscopy (XPS), and mechanical evaluation, using nanoindentation, were employed. XPS revealed that an untreated Ti surface consisted of carbon- and nitrogen-containing contaminant and titanium oxide layer on metallic titanium substrate. The method involving the combination of 10 N HCl and acetone was the most effective of all the methods investigated. Such a combination most effectively reduced values of contamination parameters C/Ti and N/Ti, as well as the intensity of the titanium oxide component in Ti 2p spectra. Chlorine was barely detected from the surface treated with HCl in any concentration. Sulfur from the residual S(2)O(8)(2-) or SO(4)(2-), however, was detected from the samples treated with either Na(2)S(2)O(8) or H(2)SO(4). The S/Ti values depended on concentration of the acidic solution. In addition, nanoindentation measurements revealed that Young's modulus of the surface treated with 0.1-10 N HCl was not significantly different from that of an untreated surface (p > 0.05). Consequently, the HCl/acetone treatment is proposed as an excellent decontamination method for the surface preparation process of Ti.

Antifungal drugs affect adherence of Candida albicans to acrylic surfaces by changing the zeta-potential of fungal cells.

The effect of sub-inhibitory concentrations of antifungal drugs on the adherence of Candida albicans to acrylic surfaces was investigated. Among five antifungals tested, azalomycin F and aculeacin A significantly enhanced the adherence. The zeta-potential of fungal cells was affected by antifungal drugs, whereas no significant change in cell surface hydrophobicity was observed. The relationship obtained between the change in the adherence and that in zeta-potential suggests that the enhanced adherence was caused by decreased electric repulsive forces.

RGD peptides regulate the specific adhesion scheme of osteoblasts to hydroxyapatite but not to titanium.

Hydroxyapatite (HA) is a bioactive dental implant material which accelerates bone formation on its surface. The mechanism of this acceleration is not clear. The elucidation of the cell adhesion might be the key to the understanding of the bioactive mechanism of HA. In this study, we analyzed the adhesion of HOS human osteoblasts onto HA and titanium to find the particular adhesion to HA. In short-term cultures in fetal bovine serum-pre-coated materials, a significantly higher number of cells adhered to HA than to titanium. In addition, serum-free conditions with phosphate-buffered saline pre-coating or bovine serum albumin pre-coating materials were tested. The results were nearly the same among all pre-coating conditions, suggesting that the quantity of cell adhesion was not affected by serum components. However, in the morphological observations by SEM, the form of adhesion was found to differ among pre-coating conditions. The osteoblasts tightly adhered and spread onto both HA and titanium with serum pre-coating, whereas the cells loosely adhered and did not spread without serum. To evaluate the Arg-Gly-Asp (RGD) sequence-specific adhesion, we used synthetic RGD peptides for a competitive inhibition test. The results showed that RGD peptides remarkably inhibited the tight adhesion and spreading of osteoblasts onto HA, whereas they did not strongly inhibit adhesion and spreading onto titanium. These results demonstrate that the regulation of cell adhesion to HA is different from that to titanium. Our study suggests that the RGD-containing serum proteins might have a major role in regulating the specific adhesion of osteoblasts to HA, and in inducing enhanced cell growth and differentiation.

Continuous administration of basic fibroblast growth factor (FGF-2) accelerates bone induction on rat calvaria--an application of a new drug delivery system.

Some studies have shown that locally applied basic fibroblast growth factor (FGF-2) enhances bone regeneration at a fracture site, while others have not been in agreement. We developed a new continuous FGF-2 delivery system designed to accelerate cytokine-induced new bone formation. A subperiosteal pocket was surgically formed in 36 eight-week-old male Wistar rats. The rats were administered 0, 1, 10, or 100 ng of FGF-2 contained in a collagen minipellet, mixed with allogeneic demineralized bone matrix in a dome-shaped Millipore filter and then placed into the pocket. New bone formation in the dome was evaluated at 2, 4, and 8 wks after placement. Soft x-ray radiographs disclosed an apparently larger radiopaque region in the 1-ng group at 4 wks compared with those in the other groups. Morphometrical analysis revealed that the new bone area in the 1-g group was significantly larger than that in the 0-g group (p<0.01). In the 100-ng FGF-2 group, new bone formation seemed suppressed. We concluded that continuous slow administration of a small amount of FGF-2 accelerates bone-derived osteogenic cytokine-induced new bone formation.

Stimulation of platelet-activating factor synthesis by neurotransmitters in salivary glands.

Platelet-activating factor (PAF), a phospholipid mediator exhibiting potent biological activities, has been shown to stimulate amylase release from the pancreas and salivary glands. The capacity of salivary glands for PAF biosynthesis in response to stimulation has also been demonstrated. To elucidate the role of PAF in salivary glands, we studied the regulation of platelet-activating factor synthesis by the autonomic nervous system in canine salivary glands. Acetylcholine and ionomycin stimulated PAF production in dispersed cells from parotid, submandibular, and sublingual glands of dogs. Norepinephrine and phenylephrine, but not isoproterenol, also stimulated PAF production in submandibular gland cells. Norepinephrine-induced PAF production was blocked by phentolamine but not by propranolol. Acetylcholine and norepinephrine increased both the PAF production and liberation of [14C]arachidonic acid from cells pre-labeled with [14C]arachidonic acid in the presence of Ca2+ in the medium. These stimulants increased [14C]arachidonic acid liberation without the accompanying production of PAF in Ca(2+)-deprived medium. No activators or inhibitors of protein kinase C produced or affected acetylcholine-induced PAF production. Lyso-PAF:acetyl-CoA acetyltransferase was activated in the cells treated with acetylcholine, norepinephrine, isoproterenol, and 8Br-cyclic AMP. Deprivation of Ca2+ in the medium markedly reduced acetylcholine-induced activation of the transferase, but little affected norepinephrine-, isoproterenol-, and 8Br-cyclic AMP-induced activation. Dithiothreitol-insensitive cholinephosphotransferase activity was also increased by acetylcholine, norepinephrine, isoproterenol, and 8Br-cyclic AMP, and the deprivation of Ca2+ in the medium further increased the activation of the enzyme activity by these agents. These results suggest that PAF synthesis in canine salivary glands is under the control of muscarinic cholinergic and alpha-adrenergic systems via Ca(2+)-dependent remodeling pathways, and that the independent activation of either phospholipase A2 or acetyltransferase is insufficient for PAF production in submandibular gland cells, i.e., the concurrent activation of these enzymes is required.

Cyclic ADP-ribose induces Ca2+ release from caffeine-insensitive Ca2+ pools in canine salivary gland cells.

Cyclic ADP-ribose (cADPR), a novel putative messenger of the ryanodine receptor, was examined regarding its ability to mobilize Ca2+ from intracellular Ca2+ stores in isolated cells of parotid and submandibular glands of the dog. cADPR induced a rapid and transient Ca2+ release in the digitonin-permeabilized cells of salivary glands. cADPR-induced Ca2+ release was inhibited by ryanodine receptor antagonists ruthenium red, ryanodine, benzocaine, and imperatoxin inhibitor but not by the inositol 1,4,5-trisphosphate (IP3)-receptor antagonist heparin. Thapsigargin, at a concentration of 3 to 30 microM, inhibited IP3-induced Ca2+ release, while higher concentrations were required to inhibit cADPR-induced Ca2+ release. Cross-potentiation was observed between cADPR and ryanodine or SrCl2, suggesting that cADPR sensitizes the Ca2+-induced Ca2+ release mechanism. Cyclic AMP plays a stimulatory role on cADPR- and IP3-induced Ca2+ release in digitonin-permeabilized cells. Calmodulin also potentiated cADPR-induced Ca2+ release, but inhibited IP3-induced Ca2+ release. Acetylcholine and ryanodine caused the rise in intracellular free Ca2+ concentration ([Ca2+]i) in intact submandibular and parotid cells. Caffeine did not produce any increase in Ca2+ release or [Ca2+]i rise in any preparation. ADP-ribosyl cyclase activity was found in the centrifuged particulate fractions of the salivary glands. These results suggest that cADPR serves as an endogenous modulator of Ca2+ release from Ca2+ pools through a caffeine-insensitive ryanodine receptor channel, which are different from IP3-sensitive pools in canine salivary gland cells. This system is positively regulated by cyclic AMP and calmodulin.

Inhibition of Na+,K(+)-ATPase by platelet-activating factor in dog submandibular glands.

Physiological stimulation of dog submandibular gland has been shown to generate platelet-activating factor (PAF). However, PAF is not released from cells in the tissue. To assess its intracellular activity, the effect of PAF on Na+,K(+)-ATPase was examined in dog submandibular gland cells. PAF inhibited Na+,K(+)-ATPase in membrane preparations, and the inhibitory effect was dependent on the protein concentration in the enzyme preparation. The inhibitory effect of a low concentration of PAF was antagonized by a PAF-receptor antagonist, BN 50,739, but at high concentrations, PAF was not antagonized. Kinetic analysis of PAF inhibition of Na+,K(+)-ATPase suggests that the inhibition of Na+,K(+)-ATPase by PAF is not due to competition by PAF at K(+)- or Na(+)-binding sites on the enzyme, but by complex inhibitory mechanisms. These results suggest that PAF may interact with specific and nonspecific site of action resulting in the inhibition of Na+,K(+)-ATPase. Ouabain increased mucin release from dog submandibular gland cells. Because Na+,K(+)-ATPase and ion exchange pathways are important in the secretory responses of acinar cells, PAF may regulate intracellularly the secretory function of acinar cells by modulating Na+,K(+)-ATPase and ionic homeostasis.

Periodontal tissue regeneration on the surface of synthetic hydroxyapatite implanted into root surface.

Several studies have demonstrated that periodontal ligament (PDL)-derived cells can form a new connective tissue attachment (NCTA) not only on denuded dentin but also on nondental inorganic materials. Although the cellular response to these materials may be different than that to denuded dentin, the process of NCTA formation on the surface of non-dental inorganic materials is still undetermined. In the present study, we compared the process of NCTA formation on the surface of an inorganic material with that on a denuded root surface by implanting a block of synthetic hydroxyapatite (HA) into a root cavity prepared on the first molar of a rat. We then observed tissue regeneration under conditions where the PDL-derived cells could populate the HA surface using an occlusive membrane. After fibrinous exudation at 3 days postoperatively, granulation tissue from the PDL filled the space between the HA block and the membrane at 1 week. Four weeks after the operation, although the granulation tissue increased in density at the interface to the HA block, no obvious deposition of cementum-like tissue was seen on the HA block. Six weeks after the operation, the deposition of cementum was observed on the surface of the HA block. The deposition of cementum became more extensive and insertion of collagen fibers into the cementum layer became more obvious 8 weeks postoperatively. Twelve weeks after the operation, the cementum covered almost the entire surface of the HA block. On the surface of the denuded dentin in control teeth, cementum deposition was seen at 4 weeks after the operation and increased in thickness from 6 weeks on.(ABSTRACT TRUNCATED AT 250 WORDS)

Isolation and serum-free culture of epithelial cells derived from epithelial rests of Malassez in human periodontal ligament.

To study biological character and function of epithelial rests of Malassez (ERM) in human periodontal ligament, we have developed a serum-free culture system of epithelial cells (ME) derived from ERM. The mitogenic effects of fibroblast growth factor (FGF)-1, FGF-2, and FGF-7/keratinocyte growth factor (KGF) on ME, human periodontal ligament-derived fibroblasts (PLF), human oral epithelial cells (OE), and human submandibular gland-derived epithelial cells (SGE) were investigated under a serum-free culture condition. FGF-1 and FGF-7/KGF stimulated the growth of both ME and SGE but FGF-2 had no effect. On the other hand, FGF-1, FGF-2, and FGF-7/KGF increased the OE proliferation. These results suggested that the divergent requirement of FGF ligands among these cells would be attributed to the different expression pattern of FGF receptor (FGFR) messenger ribonucleic acid (mRNA) isotypes. Therefore, we examined the expression of FGFR isotypes in these cells. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of ME-and SGE-derived mRNAs revealed the presence of mRNA encoding FGFR2-IIIb, a high affinity receptor for FGF-1 and FGF-7/KGF. However, no mRNAs for other FGFR isotypes were detected in both ME and SGE. On the contrary, OE expressed FGFR1-IIIc, FGFR3-IIIb, and FGFR4 mRNAs as well as FGFR2-IIb. These results indicate that FGF binding sites on ME dominantly bind to FGF-1 and FGF-7/KGF, which transduce their signals via FGFR2-IIIb. Immunohistochemical analysis, PCR-Southern, ribonuclease protection assay (RPA), and Western blotting revealed that PLF expressed FGF-7/KGF mRNA and its peptide. These observations suggest that FGF-7/KGF might mediate epithelial-mesenchymal interactions between ME and PLF to maintain normal structure and function of periodontal ligament.