Journal of Biomaterials and Nanobiotechnology, 2012, 3, 541-546
http://dx.doi.org/10.4236/jbnb.2012.324056 Published Online October 2012 (http://www.SciRP.org/journal/jbnb)
541
Effect of Surface Roughness and Materials Composition
on Biofilm Formation
Maryam Gharechahi, Horieh Moosavi, Maryam Forghani*
Dental Material Research Center, School of Dentistry, Mashhad University of Medical Sciences, Mashhad, Iran.
Received August 9th, 2012; revised September 13th, 2012; accepted September 28th, 2012
ABSTRACT
In the mouth, biofilm formation occurs on all soft and hard surfaces. Microbial colonization on such surfaces is always
preceded by the formation of a pellicle. The physicochemical surface properties of a pellicle are largely dependent on
the physical and chemical nature of the underlying surface. Thus, the surface structure and composition of the un-
derlying surface will influence on the initial bacterial adhesion. The aim of this review is to evaluate the influence of
the surface roughness and the restorative material composition on the adhesion process of oral bacteria. Both in vitro
and in vivo studies underline the importance of both variables in dental plaque formation. Rough surfaces will promote
plaque formation and maturation. Candida species are found on acrylic dentures, but dentures coating and soaking of
dentures in disinfectant solutions may be an effective method to prevent biofilm formation. Biofilms on gold and
amalgam are thick, but with low viability. Glass-ionomer cement collects a thin biofilm with a low viability.
Biofilms on composites cause surface deterioration, which enhances biofilm formation. Biofilms on ceramics are thin
and highly viable.
Keywords: Biofilm; Dental Plaque; Surface Roughness; Restorative Materials
1. Introduction
The oral cavity is constantly contaminated by a complex
diversity of microbial species that have a strong tendency
to colonize surfaces. The major components involved in
biofilm formation are bacterial cells, a solid surface, and
a fluid medium. Biofilm formation occurs on all hard
surfaces, e.g. the tooth surface, restorative materials and
implant components. In the formation of a biofilm to a
non-shedding surface the following stages have been
described [1-3]:
Stage 1: Conditioning layer formation
The first stage in the development of biofilm is the
adsorption of organic and inorganic molecules to the
solid surface. This conditioning layer in the oral cavity,
called pellicle, consists of numerous components includ-
ing glycoproteins, proline-rich proteins, phosphoproteins,
histidine-rich proteins, enzymes, and other molecules
that can function as receptors for bacteria.
Stage 2: Transport of bacteria to the substrate surface
The initial transport of microbes to the substrate may
occur through Brownian motion, liquid flow, or active
bacterial movement (chemotactic activity) and may in-
fluenced by many factors include pH, temperature, flow
rate of the fluid, surface energy of the substrate, bacterial
growth stage, surface hydrophobocity, etc.
Stage 3: Bacterial adhesion
The next step in biofilm formation is the adhesion of
microbial cells to the conditioning layer.
Phase 1: Initial non-specific microbial-substrate adhe-
sion. The bacterial surface structures form bridges be-
tween the bacteria and the conditioning layer [4]. Initially,
these bridges may not be strong, however with time the
bacteria-substrate bonds gains in strength.
Phase 2: Specific microbial-substrate adhesion. In this
phase polysaccharide adhesins or ligands on the bacterial
cell surface bind to receptors on the substrates [5].
Stage 4: Bacterial colonization and biofilm maturation
In this stage, the monolayer of microbes attracts sec-
ondary colonizers forming microcolony [6]. The firmly
attached microorganisms start growing, newly formed
cells remain attached, and biofilms can develop.
The physicochemical surface properties of a pellicle
are largely dependent on the physical and chemical na-
ture of the underlying hard surface [7-14]. Thus, the
characteristics of the underlying hard surface will influ-
ence on the initial bacterial adhesion.
*Corresponding author.

Effect of Surface Roughness and Materials Composition on Biofilm Formation
542
2. Influence of Surface Roughness (SR) on
Biofilm Formation
Scanning electron microscopy revealed that initial colo-
nization of the enamel surfaces starts from surface ir-
regularities such as perikymata, cracks, grooves, or abra-
sion defects, and subsequently spreads out from these
areas [15-18]. Initial adhesion preferably starts at loca-
tions where bacteria are sheltered against shear forces.
The change from reversible to irreversible attachment
can be established more easily in these sites. At surface
irregularities, attached bacteria can survive longer be-
cause they are protected against natural removal forces
and oral hygiene measures [19]. Moreover, roughening
of the surface increases the area available for bacterial
adhesion.
3. Studies on Surface Roughness
Waerhaug observed in dogs and monkeys that roughen-
ing of the subgingival enamel resulted in increased depo-
sition of dental plaque [20]. Kawai et al. found a positive
correlation between surface roughness and the amount of
plaque accumulation [21]. Sorensen that reviewed the
sequence of the initiation, formation, development, and
maturation of dental plaque, concluded that the factors
mediate plaque accumulation are 1) surface roughness; 2)
marginal fit; and 3) contour [22]. Einwag et al. examined
the influence of the surface roughness of dental filling
materials on plaque accumulation and found that S. mu-
tans adhered more frequently to rough cements than to
filling materials that take a high polish. However, the
adhesion of S. sanguis to composite materials with com-
parable roughness was only negligible different [23].
Shabzendedar et al. found that topical Acidulate Phos-
phate Fluoride (APF) gel application can accelerate the
defect of glass ionomer surface, which is susceptible to
more erosion, so gingival margins become rough. This
situation causes bacterial aggregation and gingivitis [24].
Carlén et al. stated that the unpolished glass ionomer
surfaces are rougher and bind more bacteria than unpol-
ished composite resin. Polishing of composite resin led to
an increase in bound bacteria that can be explained by a
change in surface roughness and/or electrostatic interact-
tions between the substrate and salivary components.
Polishing the glass ionomer, on the other hand, produce
little effect on surface roughness and bacterial binding
[25]. Mei et al. evaluated the streptococcal adhesion
forces with composite resins with different surface
roughness. They confirmed that Streptococcal adhesion
forces to composite increase with increasing roughness
of its surfaces [26]. Ikeda et al. also mentioned that the
surface roughness and composition of a resin composite
influenced biofilm adherence [27]. Morgan and Wilson
that investigated the effects of surface roughness and
type of denture acrylic on the early development of a
Streptococcus biofilm found that the number of bacteria
adhering to acrylic increased linearly with mean surface
roughness [28]. However, some observations were some-
what confused. Yamauchi et al. stated that the influence
of surface roughness was strain dependent. Some strains
(S. oralis, P. intermedia, and P. gingivalis C-101) were
found in higher amounts on rough sites, whereas some
strains (S. sanguis, S. mutans, S. mitis and P. gingivalis
ATCC 33277) were found in higher amounts on smooth
surfaces [29]. Azevedo et al. evaluated the effect of con-
ventional and whitening dentifrices on the weight loss,
surface roughness, and early in situ biofilm formation on
the surface of dental ceramics. They found that brushing
with both dentifrices can roughen ceramic surfaces; how-
ever the increase in roughness was not significantly con-
tributed to increased biofilm formation [30]. Park et al.
that investigated the effect of surface roughness of resin
composite on biofilm formation suggested that surface
topography (size and depth of depressions) may play a
more important role than surface roughness in biofilm
formation [31].
4. Biofilms on Dental Materials
Elevated proportions of Candida in biofilms formed on
dentures can cause stomatitis and Streptococcus mutans
accumulation on restorative materials is associated with
secondary caries. Microbial adhesion on biomaterial sur-
faces depends on the surface structure and composition
of biomaterials, and on the physicochemical properties of
the microbial cell surface, its surface charge and hydro-
phobicity [32,33].
4.1. Biofilms on Acrylic Resin
Adhesion of Candida to mucosa associated with the use
of acrylic dentures is one of the main clinical problems,
which can lead to stomatitis [34]. Also bacterial adhesion
to acrylic surfaces of dentures was seen [35]. Yeasts are
known to adhere quite strongly to denture base materials
as a result of the microporosity on the denture surface [36].
Candida adheres directly or via a layer of denture plaque
to denture base (polymethylmethacrylate—PMMA). With-
out this adherence, micro-organisms would be removed
from the oral cavity when saliva or food is being swal-
lowed [37-39]. Although Candida albicans has been
found to be the predominant oral yeast isolated from
dentures, Candida dubliniensis, Candida parapsilosis,
Candida krusei, and Candida tropicalis have also been
isolated [40]. Arai et al. investigated the effect of coating
denture base acrylic resin with titanium dioxide in order
to prevent microbial adhesion and mentioned that this
treatment method inhibited biofilm formation [41].
Soaking dentures in disinfectant solutions has been also

Effect of Surface Roughness and Materials Composition on Biofilm Formation 543
shown to be an effective method to prevent biofilm for-
mation. da Silva et al. suggested that sodium hypochlo-
rite solutions can killed Candida. albicans biofilms and
also removed them from the acrylic resin materials [42].
4.2. Biofilms on Metallic Biomaterials
In conducting materials, like gold and amalgam, elec-
tron-transfer plays a role in bacterial adhesion [43]. This
is attributed this to attraction between the negatively
charged bacteria and their positive image charges in the
conducting material, which cannot develop in a noncom-
ducting material or in the presence of a nonconductive
protein layer on the stainless steel surface [44]. Auschill
et al. found that five-day-old oral biofilms on gold and
amalgam surfaces were thick and fully covering the sub-
stratum surfaces [45]. Leonhardt placed pieces of three
restorative materials intra-orally for 24 and 72 hr and
showed that amalgam attracted about half the number of
viable bacteria than titanium oxide [46]. They said that
the low viability of biofilms on amalgam surfaces is due
to the release of toxic compounds from the alloy. How-
ever, it is possible that bacteria develop resistance against
mercury. In vitro, more bacteria resistant to mercury
were found in oral biofilms grown on amalgam than on
enamel. The levels of these mercury-resistant bacteria
remained elevated for a period of 48 hr, but after 72 hr,
the proportions returned to baseline levels. According to
study that performed by Ready, of the 42 mercury resis-
tant bacterial strains isolated, 98% were streptococci,
with Streptococcus mitis predominating. They docu-
mented that resistance to mercury was concurrent with
resistance to several antibiotics, most notably tetracycline
[47]. Auschill et al. reported that oral biofilms have low
viability (less than 2%) on gold but this cannot be due to
the release of toxic compounds, because gold is com-
pletely inert. They demonstrated that possibly, full cov-
erage by a relatively thick biofilm hampers the supply of
nutrients to the biofilm, leading to low viability [45].
4.3. Biofilms on Glass-Ionomer Cements
Glass-ionomer cements potentially reduce microleakage
by adhering to tooth structure and enhance fluoride re-
lease with a potential impact on oral biofilm formation.
Fluoride can act as a buffer to neutralize acids produced
by bacteria [48] and suppresses the growth of caries-
related oral bacteria [49]. Glass-ionomer cement indeed
collects a thin biofilm with a low viability (2% to 3%),
possibly as a result of fluoride release [45]. However, an
in vitro study also showed that glass-ionomer cements
containing fluoride did not reduce the amount of bacte-
rial growth and biofilm formation on the surfaces bathed
in saliva [50]. This suggests that either fluoride is not a
dominant factor in controlling biofilm formation, or that
its concentration is too low to be effective, depending on
the ratio between cement area and fluid volume in which
the experiments were carried out. In the oral cavity, the
large volume of saliva present, which is subject to wash-
out, makes the build-up of an effective fluoride concen-
tration difficult [51].
4.4. Biofilms on Resin Composites
Surface deterioration of resin composites has been dem-
onstrated by increased roughness, effects on filler parti-
cle exposure, and sometimes by a decreased microhard-
ness of the materials upon exposure to biofilms in vitro
[52]. Clearly, the in vivo presence of biofilm is just one
of the factors that may stimulate surface degradation,
other factors being acidic fluid intake, temperature fluc-
tuations, or simply the presence of an aqueous environ-
ment. Hansel suggested that especially the release of
ethyleneglycol dimethylacrylate and triethyleneglycol
dimethacrylate from composite resins may enhance the
growth of cariogenic bacteria, like mutans streptococci
and lactobacilli, organisms found mostly along the mar-
gins of composite fillings [53]. Schmalz reported that
components of dentin-bonding agents, such as hydro-
xyethyl methacrylate or triethyleneglycol dimethacry-
late, stimulated the growth of cariogenic organisms like S.
sobrinus and Lactobacillus acidophilus [54]. Effects of
monomer release became smaller when the light-curing
time of the composites was increased [55]. Methods to
inhibit biofilm growth on dental material have been
sought for several decades. It is demonstrated that zinc
oxide nanoparticles blended into resin composites dis-
play antimicrobial activity and reduce growth of bacterial
biofilms [56]. chlorhexidine gluconate (CHX) has been
incorporated into some dental materials in order to en-
hance the antibacterial activity [57,58]. Cheng et al. de-
veloped a nanocomposite containing amorphous calcium
phosphate or calcium fluoride nanoparticles and CHX
particles, and reported that the novel nanocomposite
could be reduced biofilm formation [59].
4.5. Biofilms on Ceramics
Hahn et al. found that inlays of two types of ceramic
surfaces collected less plaque with reduced viability over
a three-day period of no oral hygiene than did the natural
tooth surface [60]. Auschill showed that biofilms on ce-
ramic biomaterials formed in vivo during 5 days were
relatively thin (1 - 6 μm), but highly viable (from 34% to
86%). According to their study, gold and amalgam at-
tracting 11- to 17-μm-thick biofilms. They suggested that
thick biofilms are less viable than thin ones, due to a
hampered supply of nutrients to a thick biofilm [45]. The
effect of surface glazing and polishing of ceramics on
early dental biofilm formation was evaluated and found

Effect of Surface Roughness and Materials Composition on Biofilm Formation
544
that glazed surfaces tended to accumulate more biofilm
compared to polished surfaces [61]. Bremer et al. men-
tioned that Biofilm formation on various types of dental
ceramics differed significantly; and found that zirconia
exhibited low plaque accumulation [62].
5. Conclusion
The general conclusion can be drawn from the studies:
Rougher surfaces (crowns, dentures, and restorations)
accumulate and retain more plaque. The structure and
composition of biomaterials have also an important effect
on microbial colonization.
6. Acknowledgements
Authors would like thanks to Dental Material Research
Center of Mashhad Dental School for supporting and
bringing the opportunity for writing the paper.
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