Cumulative correlations of lysozyme, lactoferrin, peroxidase, S-IgA, amylase, and total protein concentrations with adherence of oral viridans streptococci to microplates coated with human saliva

Redundancy refers to the observation that many salivary proteins exhibit similar properties in vitro. It is possible that bacterial adherence to salivary pellicle occurs as a cumulative effect of multiple proteins. This study determined the joint and individual contributions of salivary amylase, S-IgA, lysozyme, salivary peroxidase, lactoferrin, and total protein concentrations to adherence by oral viridans streptococci in microplates coated with whole saliva from 123 persons. Strains used were: Streptococcus gordonii Blackburn, 10558, Streptococcus mitis 10712, 903, Streptococcus oralis 10557, 9811, and Streptococcus sanguis 10556, 13379. Rabbit antibody against 13379 was used for the detection of adherence. This antibody cross-reacted with all strains. Absorbance was standardized against saliva pooled from five donors. All saliva samples had been previously assayed for amylase, lactoferrin, lysozyme, secretory IgA, peroxidase, and total protein. Adherence scores for all strains except 13379 were significantly and positively correlated. Salivas binding high or low levels of one strain tended to bind others correspondingly. Multiple regression indicated significant contributions to 10558 adherence from total protein and lactoferrin (positive), and peroxidase and lysozyme (negative). Similar results were obtained for Blackburn and 903. Significant individual correlations were seen for 9811 and total protein (positive), 10557 and peroxidase (negative), and 13379 and lactoferrin (negative). Salivas with high adherence scores contained significantly more protein and lactoferrin, and significantly less peroxidase, than salivas with low adherence scores. These findings support the hypothesis that multiple proteins contribute to the adherence of streptococcal strains in vivo.     

Streptococcal diversity in oral biofilms with respect to salivary function

OBJECTIVE: In a previous study, we screened 149 subjects and established four groups high or low for salivary killing of oral bacteria, and for aggregation and live and dead adherence of oral bacteria (as a combined factor). Caries scores were significantly lower in both High Aggregation-Adherence groups. In this study we looked at the effects of those differences in salivary function on the quantity and diversity of oral biofilm streptococci. DESIGN: Subjects from those four groups were recalled for collection of overnight oral biofilm from buccal upper central incisors, lingual lower central incisors, buccal upper and lower first molars, and lingual upper and lower first molars. At each site, groups were compared for total biofilm (by DNA concentration), total streptococci (by quantitative PCR), and streptococcal diversity (by Streptococcus-specific denaturing gradient gel electrophoresis). RESULTS: Total biofilm DNA and total streptococci were correlated. Both were highest on buccal molar surfaces and lowest on lingual lower central incisors, and both were significantly lower in the High Aggregation-Adherence groups (particularly at the buccal molar site). Fifty distinct bands were observed in denaturing gradient gels. There was great diversity within and between sites. Three major bands were present in almost every person at every site. Densities for two of those bands were significantly lower in both High Aggregation-Adherence groups. Other less-prevalent bands also showed the same pattern. CONCLUSION: These findings are consistent with our caries results in suggesting that differences in salivary function can influence the quantity and composition of streptococci in oral biofilms.     

Saliva mediated adherence, aggregation and prevalence in dental plaque of Streptococcus mutans, streptococcus sanguis and actinomyces spp. in young and elderly humans

Salivary components in the pellicle mediate bacterial adherence to the tooth. Such components may also aggregate bacteria in saliva and prevent them becoming established in dental plaque. In the present study, the adherence and aggregation of Streptococcus mutans strain Ingbritt, S. sanguis strain 10556 and Actinomyces viscosus strain 19246 mediated by parotid and whole saliva from groups of young and elderly people were examined. Significant differences were found between test strains, salivary secretions and age groups. S. sanguis 10556 and A. viscosus 19246 generally adhered more strongly than S. mutans Ingbritt, which adhered better to pellicles from parotid saliva than from whole saliva. Strain 19246 bound in higher numbers to parotid saliva pellicles from elderly compared to young individuals. Strain 10556 adhered better to whole saliva than parotid saliva pellicles, and the difference was significant among the young individuals, indicating reduced adherence ability in elderly whole saliva. The streptococci were aggregated by parotid and whole saliva, and S. sanguis aggregation was less with whole saliva from the elderly than from the young participants. Besides a correlation between whole saliva aggregation of S. mutans and proportions of bacteria in plaque, no correlations were found for the individual binding properties of saliva and prevalence of bacteria in vivo. However, the level of saliva-mediated adherence in vitro was in the following order: S. mutans < Actinomyces S. sanguis, which corresponded to their isolation frequency in plaque. These findings emphasize the importance of initial adherence to salivary receptors in bacterial colonization on teeth. Further studies are needed to reveal if individual patterns in the in vitro binding characteristics of saliva lead to variation of colonization in vivo.     

Influence of mouse prolactin‐inducible protein in saliva on the aggregation of oral bacteria

Introduction: Mouse prolactin‐inducible protein (mPIP) is secreted in mouse saliva and has been found to bind oral bacteria, showing the highest affinity for streptococci. Comparisons between the oral flora of mPIP knockout mice and their wild‐type controls showed differences in the genera colonizing the two groups of mice. These findings suggested a role for mPIP in the colonization of the mouse oral cavity, possibly modulating the oral flora. In this in vitro study, we focused on the contribution of this protein to aggregation of oral bacteria, a process thought to promote the clearance of bacteria from the oral cavity, and one that could influence the composition of the oral bacterial community. Methods: The aggregation of selected human and mouse oral streptococci was measured spectrophotometrically. The aggregation of oral bacteria by saliva from mPIP knockout mice, which lack mPIP, was compared with that of saliva from wild‐type mice. Results: Both wild‐type saliva and mPIP knockout mouse saliva induced aggregation of human strain Streptococcus gordonii SK120 and mouse streptococci strains M105/6 and M106/2. Bacterial aggregation induced by the saliva of wild‐type mice was significantly higher than the aggregation induced by saliva from mPIP knockout mice for all the bacterial strains. Conclusion: In this study it was confirmed that mPIP plays a role in the aggregation of oral bacteria. The salivary components promoting aggregation of oral bacteria are considered to be part of the oral defense mechanisms so these findings provide insight into a possible function of mPIP in host defense by promoting aggregation of oral bacteria.     

Influence of mouse prolactin-inducible protein in saliva on the aggregation of oral bacteria

Introduction:  Mouse prolactin-inducible protein (mPIP) is secreted in mouse saliva and has been found to bind oral bacteria, showing the highest affinity for streptococci. Comparisons between the oral flora of mPIP knockout mice and their wild-type controls showed differences in the genera colonizing the two groups of mice. These findings suggested a role for mPIP in the colonization of the mouse oral cavity, possibly modulating the oral flora. In this in vitro study, we focused on the contribution of this protein to aggregation of oral bacteria, a process thought to promote the clearance of bacteria from the oral cavity, and one that could influence the composition of the oral bacterial community.
Methods:  The aggregation of selected human and mouse oral streptococci was measured spectrophotometrically. The aggregation of oral bacteria by saliva from mPIP knockout mice, which lack mPIP, was compared with that of saliva from wild-type mice.
Results:  Both wild-type saliva and mPIP knockout mouse saliva induced aggregation of human strain Streptococcus gordonii SK120 and mouse streptococci strains M105/6 and M106/2. Bacterial aggregation induced by the saliva of wild-type mice was significantly higher than the aggregation induced by saliva from mPIP knockout mice for all the bacterial strains.
Conclusion:  In this study it was confirmed that mPIP plays a role in the aggregation of oral bacteria. The salivary components promoting aggregation of oral bacteria are considered to be part of the oral defense mechanisms so these findings provide insight into a possible function of mPIP in host defense by promoting aggregation of oral bacteria.     

Saliva-induced aggregation of micro-organisms from skin, tooth surfaces, oral mucosa, and rectum

A total of 57 bacterial strains were isolated from tooth surfaces, oral mucosa, skin of the upper lip, and rectum of 3 persons. Identification of the strains indicated that each type of surface had a characteristic microflora. Aggregation of bacterial suspensions, induced by salivary agglutinins, was measured spectrophotometrically as the decrease in optical density (OD) by time. The aggregation curves for the oral strains followed a sigmoid pattern. The aggregation rates varied between individuals and strains. Most fecal strains showed an aggregation pattern which differed from that of the oral strains and was characterized by only a small, initial decrease in OD. The few strains, mainly Propionibacterium strains, collected from the skin of the upper lip did not aggregate. It appears from the data that several mechanisms are involved in the retention of bacteria to surfaces.     

Identification of a Streptococcus sanguis receptor for salivary agglutinins

The objective of this study was to characterize a fraction from oral streptococci containing receptor activity for salivary agglutinin molecules. Several species and strains of streptococci were disrupted in a Ribi press. The supernatant was nuclease-treated and subjected to differential centrifugation. Receptor activity in the fractions was measured by the inhibition of saliva-mediated bacterial aggregation. In addition, bacterial strains were tested for their ability to aggregate and to deplete saliva of agglutinin activity. Three patterns of activity were observed: Streptococcus sanguis M5 depleted saliva of agglutinin activity and aggregated well; Streptococcus sanguis CC5A depleted saliva of agglutinin but did not aggregate well; and Streptococcus faecalis S-161 neither depleted saliva of agglutinin nor did it aggregate. The 105,000 g supernatant fractions derived from Ribi-disrupted Streptococcus sanguis M5 and CC5A, but not from Streptococcus faecalis, showed dose-dependent inhibition of saliva-mediated aggregation. This inhibitory activity was non-dialyzable, had the same heat and trypsin sensitivity as that seen with intact bacteria, and was not due to enzymatic digestion of the salivary agglutinin. Iso-electric focusing revealed a single active region with a pI of 5.5 which was clearly separated from the bulk of the bacterial proteins.     

Altered bacterial aggregation and adherence associated with changes in rat parotid-gland salivary proteins induced in vivo by beta-adrenergic stimulation

Reduced adherence and aggregation were associated with protein alterations in parotid saliva after chronic treatment with the beta-adrenergic agonist isoproterenol. In contrast, saliva from animals treated with the beta-antagonist, propranolol, did not cause such changes; the protein composition of this saliva was similar to that of controls. SDS-polyacrylamide gel electrophoresis of protein in saliva samples before and after they were mixed with 10 mg of spheroidal hydroxyapatite beads (HA), as well as protein adsorbed and recovered from the HA, showed that an acidic, proline-rich protein with a molecular weight of approx. 40,000 was the predominant protein adsorbed. This protein was significantly diminished in saliva from isoproterenol-treated rats. Proteins with molecular weights between 44,000 and 48,000 and unique to the saliva from isoproterenol-treated animals were also adsorbed to HA. Thus alterations in proline-rich proteins of parotid saliva may influence the adherence and aggregation of oral bacteria, two processes considered important for in-vivo colonization of oral surfaces.     

In vitro attachment of Streptococcus sanguis to dental crown and bridge cements

The ability of a common dental plaque bacterium, Streptococcus sanguis, to adhere to dental crown and bridge cements in vitro was investigated. Cylindrical blocks of five different commercial brands of cement, with and without acquired pellicles, were indubated with buffer suspensions of S. sanguis for 1 h. Attached bacteria were counted under the microscope. S. sanguis had particularly high affinity for uncoated resin cement. In contrast, the carboxylate cement tested was a poor substrate for the adherence of this bacterium. The other cement types (zinc phosphate, zinc oxide and silico-phosphate) had intermediary qualities as adhering surfaces. The presence of an acquired pellicle, obtained by pretreatment with saliva, influenced the initial adherence of bacteria to cement in vitro. On the resin cement a salivary pellicle strongly suppressed the bacterial adhesion. For the zinc phosphate, zinc oxide and silico-phosphate cements a pellicle slightly enhanced the attachment of S. sanguis. On the carboxylate cement only few organisms attached also after pretreatment with saliva.     

Comparison of fluorescence microscopy and culture assays to quantitate adhesion of Porphyromonas gingivalis to mono- and multi-layered pocket epithelium cultures.

BACKGROUND: The present study compared 2 different methods (direct versus indirect evaluation) for the quantification of the adhesion of Porphyromonas gingivalis strains to in vitro cultured mono-layers of pocket epithelium. METHODS: The indirect culture viability assay (calculation of colony forming units) was compared to a direct microscopic evaluation using a novel fluorescent stain. The fluorescent kit was found to stain both bacteria and epithelial cells and enabled a differentiation between dead and living cells. RESULTS: Comparing the visual to the culture data, a high and significant correlation was found (Pearson's correlation = 0.75; P <0.001). The adhesion capacity was in general higher for dead epithelial cells than for living cells (P <0.01). Although comparable numbers of bacteria of 2 P. gingivalis strains (Pg 4 and Pg 5) were applied, Pg 4 showed a significantly lower adhesion capacity. This intra-strain variability was observed by the culture assay (2.3 x 10(6) versus 7.8 x 10(6)+/-2.7 x 10(6); P <0.01) and by the direct microscopy (P <0.01) for both live and dead epithelial cells. A second goal was to see whether there was a difference in the amount of bacterial adherence to mono- and multi-layers of in vitro cultured epithelium. No significant differences were found for the 5 examined P. gingivalis strains. However, interstrain differences in adhesion capacity were evident for both tissues. CONCLUSIONS: This study highlights the reproducibility of a direct microscopic evaluation of bacterial adhesion to in vitro cultured epithelial cells, and suggests both intrastrain (P. gingivalis) and inter-cell (live versus dead) variation in adhesion capacity. Studies are needed to determine the extent to which P. gingivalis strain variation is reflected in variation of other strains in humans.     

Interaction of salivary fibronectin with oral streptococci

Immunoreactive Fibronectin (Fn) has been demonstrated in stimulated human parotid saliva by western blot analysis and also found to be a component of the artificial tooth pellicles derived from hydroxyapatite (HA) beads coated with parotid saliva. Saliva depleted of gelatin-binding components showed a significantly lower degree of reactivity with anti-Fn antibodies than did the control saliva when tested by and enzyme-linked immunosorbent assay (ELISA). Depletion of gelatin-binding components from saliva was also found to affect the degree of saliva-mediated aggregation of four of the seven oral streptococci tested [Streptococcus mutans strains GS-5 and OMZ 176, S. sobrinus, and S. rattus]. Similarly, the adherence of the same four micro-organisms to the artificial tooth pellicles (derived form saliva which had previously been depleted of gelatin-binding component) was significantly inhibited (37-53%) when compared with the control saliva-coated HA beads. Pre-treatment of streptococci with 100 micrograms of soluble Fn also caused a 34-57% inhibition of adherence of the same oral streptococci to saliva-treated HA beads. Quantitation of Fn in human parotid saliva showed that the amounts of immunoreactive Fn varied form 2 to 6 micrograms/mL of parotid saliva. Furthermore, the Fn from parotid saliva was found to be adsorbed onto the bacterial surfaces, as demonstrated by immunofluorescence and ELISA. The presence of Fn in parotid saliva and its ability to bind to HA beads (artificial pellicles), in conjunction with the ability of soluble Fn to inhibit the adherence of streptococcal strains to the artificial tooth pellicles, suggest that the microbial ecology of the oral cavity may, in part, be influenced by the interactions mediated by salivary fibronectin.     

The surface free energy of oral streptococci after being coated with saliva and its relation to adhesion in the mouth

Contact angle measurements on layers of bacteria were used to determine the bacterial surface free energy (gamma b) of a variety of oral streptococcal strains, both without and after being coated with human whole saliva. At least four isolates of each species, either freshly isolated or laboratory strains, were used. The species Streptococcus mutans, S. sanguis, and S. salivarius were homogeneous, having high surface free energies, and were not affected by saliva treatment (gamma b = 106 +/- 12 and 107 +/- 10 erg X cm-2 in the absence and presence of saliva coating, respectively; n = 20). S. mitis had a very low surface free energy (46 +/- 15; n = 5), which was significantly increased after salivary adsorption (71 +/- 14 erg X cm-2; p less than 0.002). The species S. milleri contained strains with both high and low gamma b. Calculation of the interfacial free energy of adhesion (delta F adh) for bacteria from a saliva suspension to solid surfaces with various arbitrary surface free energies (gamma s) showed that, theoretically, most strains will encounter thermodynamically favorable conditions for adhesion to surfaces with a gamma s above 62 erg X cm-2. However, S. mitis strains not coated with saliva would only be able to adhere to surfaces with gamma s lower than this value. Saliva-coating reverses the calculated relationship with gamma s for these strains. The results indicate that an enamel surface with a low gamma s value would be thermodynamically unfavorable for adhesion of most oral streptococci.     

The deposition of oral bacteria at the solid/liquid and solid/liguid/air interfaces.

A technique for quantifying the deposition of microorganisms at the solid/liquid and solid/liquid/air interfaces enabled the deposition characteristics of representative strains of five oral species on glass from saline and saliva to be compared using single and mixed bacterial suspensions. The deposited bacteria were quantified microscopically and by viable counts. All the strains attached to the glass surfaces. Streptococcus sanguis NCTC 7868, Actinomyces viscosus WVU 371, and Streptococcus mutans NCTC 10449 generally attached or deposited more readily from saliva than Streptococcus salivarius NCTC 7366 or Lactobacillus casei L 8822 due perhaps to polymers on their cell surfaces forming bridges to the saliva-covered glass surfaces. The findings show that the quantity of deposited material cannot be correlated directly with its content of viable bacteria.     

Inhibition of Streptococcus mutans adherence by means of surface hydrophilization

Derivatives of polyalkylene oxide (PAO) were examined for their ability to inhibit adherence of 3H-labeled cells of Streptococcus mutans to hydroxyapatite (HA) and to plastic surfaces treated with buffer or parotid saliva from two individuals. Pellicles formed on HA with saliva from the two subjects were distinct in their binding capacity. One saliva promoted and the other saliva reduced adherence, as compared with the buffer control (BHA). Three of the PAO compounds effectively hindered binding of bacteria to BHA. However, on saliva-treated HA (SHA) the inhibition was not as effective. One compound, a phosphated polypropylene glycol, was potent in inhibiting adherence both to BHA and to the plastic surfaces treated with either buffer or saliva. However, treatment of HA with this compound followed by saliva incubation only gave a limited reduction in the number of bacteria binding. Evidently, salivary constituents are capable of interacting also with the PAO-treated surface. When 14C-labeled hydrophilizing agent was used, it was shown that the PAO was not replaced by salivary molecules. Instead, the components of the saliva that mediate the binding of bacteria seemed capable of adhering directly to the PAO layer.     

Presence of Streptococcus mutans and lactobacilli in saliva and on enamel, glass ionomer cement, and composite resin surfaces

The quantity of S. mutans, total streptococci, and lactobacilli on sound enamel surfaces and 1-yr-old glass ionomer cement and composite resin fillings with the cervical margins placed subgingivally was compared intra-individually. The amount of bacteria was compared to their number in saliva. The evaluation was done in a cross sectional study, where the patients continued to use their customary oral hygiene procedures and during a 14-day period of experimental plaque formation. The number of lactobacilli and S. mutans recovered from the test surfaces indicated that the critical salivary concentrations necessary for the isolation of S. mutans and lactobacilli from glass ionomer cement and composite resin surfaces are the same as for the enamel surfaces. The fluoride levels in plaque adjacent to glass ionomer cement will not become high enough to inhibit the accumulation of the investigated bacteria.     

血链菌死菌在生物膜再形成中的作用观察

目的　探讨血链菌死菌在生物膜更新形成中的作用。方法　利用激光共聚焦扫描显微镜和死菌 /活菌荧光染色技术相结合 ,对人工口腔内覆盖死菌 (实验组 )和未覆盖死菌 (对照组 )盖玻片表面血链菌生物膜的形成进行动态观察。结果　血链菌生物膜形成 3 0min时 ,实验组生物膜厚度和对照组相比 ,无显著性差异 ;但其细菌密度明显大于对照组 (P <0 .0 5 ) ,至 60min ,12 0min ,2 4 0min时 ,实验组生物膜厚度和对照组相比 ,有显著性差异 (P <0 .0 5 ) ,但其细菌密度几乎一致。结论　在生物膜的形成初期 ,死菌具有一定的促进作用。     

Presence of Streptococcus mutans and lactobacilli in saliva and on enamel, glass ionomer cement, and composite resin surfaces

Abstract— The quantity of S. mutans , total streptococci, and lactobacilli on sound enamel surfaces and 1-yr-old glass ionomer cement and composite resin fillings with the cervical margins placed subgingivally was compared intraindividually. The amount of bacteria was compared to their number in saliva. The evaluation was done in a cross sectional study, where the patients continued to use their customary oral hygiene procedures and during a 14-day period of experimental plaque formation. The number of lactobacilli and S. mutans recovered from the test surfaces indicated that the critical salivary concentrations necessary for the isolation of S. mutans and lactobacilli from glass ionomer cement and composite resin surfaces are the same as for the enamel surfaces. The fluoride levels in plaque adjacent to glass ionomer cement will not become high enough to inhibit the accumulation of the investigated bacteria.     

Human salivary function in relation to the prevalence of Tannerella forsythensis and other periodontal pathogens in early supragingival biofilm

OBJECTIVE: Previously, we screened 149 subjects and established four groups high or low for salivary killing of oral bacteria, and for aggregation and live and dead adherence of oral bacteria (as a combined factor). Caries scores were significantly lower in both High Aggregation-Adherence groups. Subsequently, we found that supragingival total biofilm DNA, total streptococci and two major streptococcal rRNA variants also were significantly lower in the High Aggregation-Adherence groups. In this study, we looked at the effects of those differences in salivary function on three periodontal pathogens. DESIGN: Quantitative PCR was used to determine levels of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythensis (formerly Bacteroides forsythus) in stored DNA extracts of overnight supragingival biofilm collected from buccal upper central incisors (UC), lingual lower central incisors (LC) and buccal upper and lower first molars (BM) and lingual upper and lower first molars (LM) of subjects in the four groups. RESULTS: A. actinomycetemcomitans and P. gingivalis were almost completely absent from these samples. T. forsythensis was found in 11 of 35 persons at the buccal molar site. Only two of those subjects were in the High Aggregation-Adherence groups, and that difference was statistically significant. The mean quantity of T. forsythensis also was significantly lower in the High Aggregation-Adherence groups. CONCLUSIONS: The difference between the Low and High Aggregation-Adherence groups might reflect direct interactions of salivary proteins with T. forsythensis. Alternatively, the higher levels of total biofilm and total streptococci seen in the Low Aggregation-Adherence groups might create a favourable environment for early secondary colonization of T. forsythensis.     

The vital status of human buccal epithelial cells and the bacteria associated with them

OBJECTIVE: We have shown that buccal epithelial cells (BEC) from humans can contain a polymicrobial intracellular flora. Members of that flora can induce proinflammatory responses. However, our subjects all had healthy oral mucosa. This might reflect tolerance of bacterial invasion by live BEC. Alternatively, inflammation might not occur if invaded cells were mostly dead, and thus unable to mount a response. This study addressed that issue, by determining the vital status of BEC and the bacteria associated with them. DESIGN: Initial experiments indicated that BEC were anomalously permeable to the DNA stain propidium iodide. We used that property to develop a protocol that combined the DNA stains SYTO 9 and propidium iodide (indicators of bacterial viability) with the esterase substrate calcein blue AM (an indicator of BEC viability), and Annexin V Alexa Fluor 647 conjugate (an apoptosis marker). That protocol was applied to BEC collected from 36 human subjects. RESULTS: On average, 70% of BEC displayed calcein blue staining, with no binding of Annexin V, 25% showed signs of apoptosis, and 5% did not stain with calcein blue. The mean percent of BEC with live cell-associated bacteria was 29%. Collectively, 25% of total BEC displayed calcein blue staining and live (SYTO 9 stained) bacteria. Only 1% of total BEC were negative for calcein blue and associated with live bacteria. CONCLUSIONS: Our findings suggest that live BEC are tolerant of bacterial invasion. This may be due to complex interactions between members of the polymicrobial flora and their host BEC.     

Early formation of Streptococcus sobrinus biofilm on various dental restorative materials.

OBJECTIVES: To examine the formation of dental biofilm by Streptococcus sobrinus on different types of restorative materials, using a model consisting of host and bacterial constituents. METHODS: The adsorption pattern of saliva to the restorative material was determined by means of gel electrophoresis coupled with computerized densitometry techniques. The amount of salivary proteins adsorbed onto the surfaces was measured using the Bradford method. Sucrose-dependent bacterial adhesion to the saliva-coated restorative material was tested by radioactive-labelled Streptococcus sobrinus, and viable counts of these bacteria in the biofilm was determined using bacterial culture techniques. RESULTS: Different adsorption patterns by salivary proteins to restorative materials were recorded. Durafil and acrylic dental materials demonstrated the most affinity to salivary proteins. A surface dependent adhesion profile was recorded, showing a high affinity of albumin and amylase to Acrylic and Durafil materials. Bacterial accumulation was the highest with Fuji LC and Fuji GC, which also demonstrated the highest bacterial viability. CONCLUSIONS: Our study demonstrates the specificity of biofilm formation on different brands of dental restorative materials. Formation of a variety of dental biofilms has a significant impact on the progression of dental diseases in the oral cavity.