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Background. Mucositis and peri-implantitis are pathologies that may be encountered during dental implant rehabilitation. Therapeutic strategies for their resolution range from non-surgical to surgical treatments and aimed at eliminating the biofilm from the implant’s surface, through mechanical, chemical or photodynamic agents.Aim. The aim was to evaluate the effect of the electric field generated by the Ximplant machine on the bacterial load and on the biofilm grown on dental implants.Materials and Methods. Ten dental implants were brought into contact with a donor’s saliva, then five implants were treated with the electric field and four were not treated.Bacterial biofilm was then measured by resazurin assay, both on treated and untreated implants.Results. The study showed the preliminary success of the electrofield in reducing the microbial population and destroying the clinical biofilm, compared with a sterile implant as control.
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Paola  Di Giacomo
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Annali di stomatologia
biofilm removal
Use of the electrical device on dental implant’s bacterial biofilm: a preliminary in vitro study
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Paola  Di Giacomo
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Department of Oral and Maxillo-Facial Sciences, Sapienza University of Rome, Italy
1971-1441
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biofilm removal
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Use of the electrical device on dental implant’s bacterial biofilm: a preliminary in vitro study
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Use of the electrical device on dental
implant’s bacterial biofilm:
a preliminary in vitro study

Giovanni Falisi1                                                                                Authors
Gianluca Botticelli1                                                                            Giovanni Falisi - Department of Life,
Eduardo Basanes Rivera2                                                                         Health and Environmental Sciences,
Antonio Scarano3                                                                                University of L’Aquila, L’ Aquila, Italy
Roberto Gatto1                                                                                  Gianluca Botticelli - Department of Life,
Sofia Rastelli1                                                                                 Health and Environmental Sciences,
Carlo Di Paolo4                                                                                 University of L’Aquila, L’ Aquila, Italy
Paola Di Giacomo4                                                                               Eduardo Basanes Rivera - Department of
                                                                                                Dentistry, University ANAHUAC of North
1
  Department of Life, Health and Environmental Sciences, University of L’Aquila, L’ Aquila,     Mexico
Italy                                                                                           Antonio Scarano - Department of
2
  Department of Dentistry, University ANAHUAC of North Mexico                                   Innovative Technology in Medicine and
3
  Department of Innovative Technology in Medicine and Dentistry, University of Chieti-          Dentistry, University of Chieti-Pescara,
                                                                                                Chieti, Italy
Pescara, Chieti, Italy
4
  Department of Oral and Maxillo-Facial Sciences, Sapienza University of Rome, Italy.           Roberto Gatto - Department of Life,
                                                                                                Health and Environmental Sciences,
                                                                                                University of L’Aquila, L’ Aquila, Italy
Corresponding author: Gianluca Botticelli                                                       Sofia Rastelli - Department of Life, Health
                                                                                                and Environmental Sciences, University
                                                                                                of L’Aquila, L’ Aquila, Italy
Abstract
                                                                                                Carlo Di Paolo - Department of Oral
Background. Mucositis and peri-implantitis are pathologies that may be encoun-                  and Maxillo-Facial Sciences, Sapienza
tered during dental implant rehabilitation. Therapeutic strategies for their resolution         University of Rome, Italy.
range from non-surgical to surgical treatments and aimed at eliminating the biofilm             Paola Di Giacomo - Department of Oral
from the implant’s surface, through mechanical, chemical or photodynamic agents.                and Maxillo-Facial Sciences, Sapienza
Aim. The aim was to evaluate the effect of the electric field generated by the Xim-             University of Rome, Italy.
plant machine on the bacterial load and on the biofilm grown on dental implants.
Materials and Methods. Ten dental implants were brought into contact with a do-
nor’s saliva, then five implants were treated with the electric field and four were
not treated.
Bacterial biofilm was then measured by resazurin assay, both on treated and un-                 License
treated implants.                                                                               This work is licensed under a Creative
Results. The study showed the preliminary success of the electrofield in reduc-                 Commons Attribution-NonCommercial-
ing the microbial population and destroying the clinical biofilm, compared with a               NoDerivatives 4.0 International License.
sterile implant as control.
                                                                                                Authors contributing to Annali di
                                                                                                Stomatologia agree to publish
Key words: mucositis, periimplantitis, biofilm removal.                                         their articles under the Creative
                                                                                                Commons Attribution-NonCommercial-
                                                                                                NoDerivatives 4.0 International License,
Introduction                                                                                    which allows third parties to copy and
The treatment of choice for the resolution of both partial and total edentulism of patients     redistribute the material providing
                                                                                                appropriate credit and a link to the
consists in the use of dental implants (1-5). In recent decades, their use has exponen-
                                                                                                license but does not allow to use the
tially evolved. Mucositis and peri-implantitis are pathologies that may be encountered          material for commercial purposes and to
during dental implant rehabilitation. (6-10).                                                   use the material if it has been remixed,
The first one is characterized by all the signs of inflammation without radiographic signs      transformed or built upon.
of bone loss, while the second one is also characterized by purulent exudate and radio-
graphic signs of peri-implant bone loss.                                                        How to Cite
                                                                                                G Falisi, G Botticelli, E Basanes Rivera,
When we talk about peri-implant bone loss we must differentiate it from biological fac-
                                                                                                A Scarano, R Gatto, S Rastelli,
tors, such as physiological remodeling or mechanical stress. The inflammatory-bacterial         C Di Paolo, P Di Giacomo.
etiology determines resorption between the interface of the bone and implant and its            Use of the electrical device on dental
consequent loss (11-16).                                                                        implant’s bacterial biofilm: a preliminary
In the literature, therapeutic strategies for the resolution of these two pathological condi-   in vitro study.
tions range from non-surgical to surgical treatment and aimed at eliminating the superfi-       Annali Di Stomatologia, 15(1), 43-47.
                                                                                                https://doi.org/10.59987/ads/2024.1.43-47
cial biofilm, through mechanical and/or chemical agents (7).
As demonstrated in many scientific papers, these techniques present critical issues
linked to the partial elimination of bacteria and surface contaminants.
In recent years the metallurgical industry has conducted studies on the electrochemical



10.59987/ads/2024.1.43-47                                                                                                      43
     Use of the electrical device on dental implant’s bacterial biofilm: a preliminary in vitro study

     cleaning of metal surfaces to eliminate and decontami-        sinki. The informed consent was obtained from all indi-
     nate biofilm and prevent its formation.                       vidual participants included in the study.
     The electric current would act on the electrochemical         The salivary collection was performed in the morning and
     bonds of the polysaccharide particles of the biofilm          the subject was asked not to practice the oral hygiene
     layer, determining a reduction in hydrogen bonding,           routine before the collection 5 ml of saliva was collected.
     also breaking the bonds that determine adhesion to            Contamination of the implants was performed using a
     surfaces. (17)                                                bacterial culture in the logarithmic phase of growth, pre-
     This new technique would allow the elimination and            pared by growing ten colonies of saliva in 5 ml of Brain
     decontamination of the metal implant surfaces while           Hearth Infusion (BHI) broth (Oxoid ThermoFisher Sci-
     keeping them intact, which does not happen with me-           entific, US) supplemented with 5% defibrinated sheep
     chanical procedures that alter their shape (10).              blood in an anaerobic environment for 96 hours at 37°C.
     The aim of the work was to evaluate the effect of the         The bacterial suspension was adjusted to the OD of 0.5
     electric field on the reduction and decontamination of        McFarland scale and subsequently diluted 1:1000. The
     bacterial plaque on the implant surface.                      formation of bacterial biofilm on the implants occurred
                                                                   by incubating the devices in a sterile vial (Eppendorf
     MATERIALS AND METHODS                                         Safe-Lock Tubes, Eppendorf Italy) with 900 μL of bacte-
                                                                   rial suspension prepared as described above. The sam-
     Bacterial contamination of dental implants                    ples were incubated in an upright position for 48 hours
     Ten dental implants in grade 4 titanium (sandblasted and      at 37°C. After incubation, the implants were washed
     etched with double acid attack) with a length of 13 and       three times in a sterile 0.9% NaCl solution to remove the
     a diameter of 4,2 (SEVENTIN-ONE by company Maco               planktonic form of non-adherent bacteria.
     dental care Salerno Buccino) were used.
     A healthy volunteer who presented with active peri-im-        Electrical treatment of implants
     plant pathology, with signs of inflammation, suppuration
                                                                   After bacterial contamination, five implants were trans-
     and with radiographic signs of peri-implant bone loss
                                                                   ferred to the treatment chamber with the addition of 100
     was recruited to donate his saliva.
                                                                   μl of 0.9% NaCl solution and treated using the “Peri-
     The present study has been conducted in accordance
                                                                   implantitis Protocol” of the X-IMPLANT instrument. (Fig-
     the principles and guidelines of the Declaration of Hel-
                                                                   ure 1, 2)
                                                                   This protocol consisted of four cycles of electrical cur-
                                                                   rent (alternating electrical current at 625 kHz, 260 Vpp,
                                                                   15 W and 180 mA) performed on the implant according
                                                                   to the programmed times of the machine, the electrode
                                                                   was positioned in 4 tangential positions peripherally
                                                                   at 90 ° from the previous position. Once the treatment
                                                                   phase with the Ximplant instrument was completed, the
                                                                   implants were further washed with 0.9% NaCl solution.
                                                                   Four implants were not treated. One implant was sterile
                                                                   and incubated with 900 μl of BHI was used as a nega-
                                                                   tive control.
                                                                   Both treated and untreated implants were added to a re-
                                                                   agent, resazurin (Labbox italia srl) and incubated for vi-
                                                                   sual evaluation at 2 hours, one day, two days and the final
                                                                   third day of the experimental procedures. (19) Table 1.

                                                                   Statistical analysis
                                                                   Descriptive statistics was performed. Chi-squared test
                                                                   was used to assess significant differences between the
     Figure 1. Treatment chamber                                   two groups (treated vs not treated) in each reference
                                                                   time with p<0.05. (Figure 1 and Table 2)

                                                                   Results
                                                                   All treated implants did not show any color change, as
                                                                   the control sterile implant.
                                                                   All not-treated implants showed a color change already
                                                                   after two hours.

                                                                   Discussion
                                                                   It is now proven that the presence of bacteria leads to
                                                                   the formation of biofilm on all surfaces, both biological
                                                                   and non-biological, as demonstrated by scientific studies
                                                                   on biofilm, the first to be formed is made up of beneficial
                                                                   bacteria called commensals. (12-15)
                                                                   However, the reduced host response and environmental
     Figure 2. Device                                              modifications caused by clinical alterations can lead to a



44                                                                                                    10.59987/ads/2024.1.43-47
G. Falisi et al.

Table 1. Row data. Negative = no color change; Positive= color change
  Implants #                After 2 hrs            After 1 day           After 2 days             After 3 days
   C1                       Negative               Negative              Negative                 Negative
   2                        Negative               Negative              Negative                 Negative
   3                        Negative               Negative              Negative                 Negative
   4                        Negative               Negative              Negative                 Negative
   5                        Negative               Negative              Negative                 Negative
   6                        Negative               Negative              Negative                 Negative
   7                        Positive               Positive              Positive                 Positive
   8                        Positive               Positive              Positive                 Positive
   9                        Positive               Positive              Positive                 Positive
   10                       Positive               Positive              Positive                 Positive


Table 2. Chi-square analysis. dF= degree of freedom.
  Time                            χ2 value                       dF                            P value
   T1 =after 2 hrs                 9000                          1                             0.003
   T2 = after 1 day                9000                          1                             0.003
   T3= after 2 days                9000                          1                             0.003
   T4= after 3 days                9000                          1                             0.003



shift in the commensal microbial flora towards the devel-        such as 12% chlorhexidine after appropriate mechanical
opment of pathogenic species, an event called dysbiosis.         debridement was used, it did not improve the scores of
Dysbiosis causes an increase in the production of in-            gingival bleeding after probing (BOP) compared to control
flammatory mediators, which induce the production of             groups where mechanical debridement alone was used.
toxic products in the host cell which in turn lead to the        Even the potential beneficial effects (reduction of BOP
destruction of the tissues around the implant.                   and deep bleeding) hypothesized using systemic anti-
In the literature, various surgical and non-surgical strate-     biotics (azithromycin) failed three/six months after treat-
gies have been introduced for the elimination of patho-          ment, just as the use of probiotics had no benefit com-
logical biofilm from surfaces. (20-24)                           pared to mechanical therapy. (28, 29)
Both are based on periodontal treatments and prevention          On the other hand, the use as an alternative to mechani-
because it is considered essential to give appropriate hy-       cal therapy such as, for example, the use of ultrasound
giene instructions to the patient to reduce the bacterial        instruments, glycine sandblasting sprays or YAG lasers
load keeping the peri-implant tissues healthy (25-27).           has a good result in clinical terms with reduction of BOP,
In some scientific works where the use of oral antiseptics       compared to mechanical debridement alone.




Contingency tables. PP= peri-implantitis protocol yes/no.



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     Use of the electrical device on dental implant’s bacterial biofilm: a preliminary in vitro study

     The use of both photonic and laser techniques, however,              7.    Gosau, M.; Hahnel, S.; Schwarz, F.; Gerlach, T.; Reichert,
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