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Table of Contents
ORIGINAL ARTICLE
Year : 2019  |  Volume : 7  |  Issue : 4  |  Page : 88-91

Comparative evaluation of biofilm formation among three differently treated surfaces on titanium samples along with gentamicin coating


1 Department of Prosthodontics, Noorul Islam College of Dental Sciences, Thiruvananthapuram, India
2 Department of Prosthodontics, Sri Sankara Dental College, Varkala, India
3 Department of Periodontics, PMS College of Dental Science & Research, Thiruvananthapuram, India
4 Department of Periodontics and Oral Implantology, PMS College of Dental Science & Research, Thiruvananthapuram, Kerala, India

Date of Submission25-Nov-2019
Date of Acceptance27-Nov-2019
Date of Web Publication24-Dec-2019

Correspondence Address:
Dr. R Arun
Department of Prosthodontics, Noorul Islam College of Dental Sciences, Neyyattinkara, Thiruvananthapuram, Kerala.
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/INJO.INJO_39_19

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  Abstract 

Background: For many years, osseointegrated oral implants have been successfully used as an excellent method for replacement of missing teeth. Biofilm formation on oral implants can cause inflammation of peri-implant tissues, which can affect the long-term success of osseointegrated implants. Aims and Objectives: This study aimed to evaluate biofilm formation on gentamicin-treated implants and to conduct a comparative evaluation process of biofilm formation among three differently treated surfaces on titanium samples along with gentamicin coating. Materials and Methods: Samples were blasted and later loaded with gentamicin drug by vacuum drying and evaluation of the strains was carried out for biofilm. Bacterial adhesion was evaluated at different time intervals of 0, 1, 4, 24, and 48h. Results: Bacterial adhesion was sequentially increasing in polished samples. Initial bacterial adhesion was higher on surface-modified samples as compared to polished samples in the first hour. Bacterial adhesion was retarded in gentamicin-coated hydroxyapatite (HA)-blasted samples up to 24h. Bacterial adhesion was considerably less on TiO2-blasted samples up to 48h. Conclusion: Implant surface modified with TiO2 and gentamicin showed delayed biofilm formation even up to 48h. Surface modification with HA has gained considerable osteoconductive surface, which is a boon for the production of future implants with less expense; however, further studies need to be carried out to prove its efficacy and effectiveness.

Keywords: Biofilm, dental implants, gentamicin, titanium alloy samples


How to cite this article:
Arun R, Rajan NS, George NE, Chandrathara T, Krishnan HR, Gayathri S. Comparative evaluation of biofilm formation among three differently treated surfaces on titanium samples along with gentamicin coating. Int J Oral Care Res 2019;7:88-91

How to cite this URL:
Arun R, Rajan NS, George NE, Chandrathara T, Krishnan HR, Gayathri S. Comparative evaluation of biofilm formation among three differently treated surfaces on titanium samples along with gentamicin coating. Int J Oral Care Res [serial online] 2019 [cited 2020 Jan 19];7:88-91. Available from: http://www.ijocr.org/text.asp?2019/7/4/88/273973




  Introduction Top


Pure titanium and titanium alloys are commonly used as implant materials in dentistry because of their favorable combination of mechanical strength, chemical stability, and biocompatibility.[1],[2],[3] The soft-tissue surrounding healthy osseointegrated dental implants share anatomic and functional features with the gingiva around teeth. Antibiotic-loaded implant coatings are used for the prevention of implant-associated infections. These coatings can provide an immediate response to the threat of implant contamination; however, these coatings do not necessitate use of an additional carrier for the antibacterial agent. Antibiotics can be loaded on to the surface of implants using the following two ways: the first is passive method and the other is active method. The passive coating technique aims to reduce bacterial adhesion by altering the physiochemical properties of the substrate, so that bacteria–substrate interactions are not favorable.[4],[5],[6],[7] On the contrary, active coatings are designed for temporary release of high fluxes of antibacterial agents immediately after the implantation procedure. High local doses of antibiotics against specific pathogens associated with implant infections can thus be administered without reaching systemic toxicity levels with enhanced efficacy and less probability for bacterial resistance. Gentamicin is an aminoglycoside antibiotic used to treat many types of bacterial infections.[8],[9] It is active against a wide range of bacterial infections, particularly gram-negative bacteria including the Pseudomonas and Proteus, and gram-positive bacteria including Streptococcus and Staphylococcus. Gentamicin is one of the few heat stable antibiotics that remain active even after autoclaving, which makes it particularly useful in the preparation of microbiological growth media.[10] Hence, this study is a novel approach to evaluate the effect of biofilm formation on surface-modified implants with and without coating of gentamicin.

Aims and objectives. The study aimed to evaluate biofilm formation on gentamicin-treated implants and to conduct a comparative evaluation process of biofilm formation among three differently gentamicin-treated surfaces on titanium samples.


  Materials and Methods Top


Commercially available Ti-6Al-4V (ASTMF11O8, Manher Metal Supply Corporation, Mumbai, India) was machined to 2mm thickness and 2 × 1.5cm length and breadth rectangular samples. These disks were mechanically polished by silicon carbide papers of grit sizes 240 and 600 in the grinder and polisher. Hydroxyapatite (HA) powder was prepared in-house by a wet precipitation technique using Ca(NO3)2–4H2O (calcium nitrate) and NH4H2PO4 (ammonium dihydrogen phosphate). The microfine powder was compacted at the pressure of 200MPa in a cold isostatic press. HA powder was loaded in the jar of the blasting machine of particle sizes 65, 125, and 250 μm. On each particle size, the target samples were away from the gun distance 2, 4, and 6cm. Each sample was blasted for 2, 4, and 6min. Five samples of each particle size, distance, and time were blasted. The same procedure was carried out for TiO2 as well. The samples were vacuumed for 15min at 200mbar. Titanium samples (five samples of each group) were transferred in to Eppendorf Tubes containing 1-mL Streptococcus sanguinis cultures. Bacterial concentration was about 109 UFC/mL. They were incubated for 0, 1, 4, 24, and 48h at 37°C. The viable count was plotted against roughness and plain samples.


  Results Top


The data were analyzed by Statistical Package for the Social Sciences software, version 16.0. Analysis of variance (ANOVA) was used to compare the statistically significant differences between the groups. Post hoc test followed by Dunnett’s t test was used to obtain the significant difference at 95% confidence interval. A value of P < 0.05 was considered statistically significant. The gentamicin antibiotic used is shown in [Figure 1]. [Table 1] shows the number of viable organisms in plain polished titanium + gentamicin. [Table 2] shows the number of viable organisms in HA blasted + gentamicin. [Table 3] shows the number of viable organisms in TiO2 + gentamicin.
Figure 1: Gentamicin antibiotic

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Table 1: Number of viable organisms in plain polished titanium + gentamicin

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Table 2: Number of viable organisms in HA blasted + gentamicin

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Table 3: Number of viable organisms in TiO2 + gentamicin

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  Discussion Top


This study suggests that the formation of biofilm is seen in all samples. However, it is observed that biofilm was delayed in surface-modified and gentamicin-loaded samples. Gentamicin loaded on TiO2 surface showed low concentrations of biofilm formation among all the other groups. The success rates obtained with dental implants depend on the volume and quality of the bone. It is often difficult to obtain implant anchorage when the density of bone is less.[8],[9],[10] Streptococcus sanguinis strain was used to evaluate the biofilm formation as Streptococcus was the predominant initial colonizing microbes.[11],[12] Biofilm evaluation of surface-modified implants with and without gentamicin-loaded samples was evaluated in this study. The scanning electron microscope (SEM) analysis and energy dispersive X-ray analysis (EDAX) report of samples showed surface roughness and sufficiently adhered elements, calcium and phosphorous, on HA-blasted samples. The SEM results showed that gentamicin loaded on TiO2 surface showed low concentrations of biofilm formation among all the other groups. It is noticed that within 1h biofilm formation was on plain polished surface. However, biofilm formation was delayed more than 1h on plain polished gentamicin-loaded samples. The biofilm formation was delayed on TiO2-blasted surface even up to 48h, whereas in HA-treated implants it was delayed only up to 4h.


  Conclusion Top


It can be concluded that implant surface-modified gentamicin showed delayed biofilm formation even up to 48h. These implants can retard the plaque formation and thus prevents peri-implantitis in the primary healing stage. This in turn can prevent failure of implants. This is ideal in situations where the patient is having poor bone quality and poor oral hygiene, and in patients having debilitating disease. Surface modification with HA has gained considerable osteoconductive surface, which is a boon for the production of future implants with less expense; however, further studies need to be carried out to prove its efficacy and effectiveness.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Brett PM, Harle J, Salih V, Mihoc R, Olsen I, Jones FH, et al. Roughness response genes in osteoblasts. Bone 2004;35: 124-33.  Back to cited text no. 1
    
2.
Ivanoff CJ, Hallgren C, Widmark G, Sennerby L, Wennerberg A. Histologic evaluation of the bone integration of TiO(2) blasted and turned titanium microimplants in humans. Clin Oral Implant Res 2001;12:128-34.  Back to cited text no. 2
    
3.
Astrand P, Engquist B, Dahlgren S, Engquist E, Feldmann H, Gröndahl K. Astra Tech and Brånemark system implants: a prospective 5-year comparative study. Results after one year. Clin Implant Dent Relat Res 1999;1:17-26.  Back to cited text no. 3
    
4.
Dige I, Nyengaard JR, Kilian M, Nyvad B. Application of stereological principles for quantification of bacteria in intact dental biofilms. Oral Microbiol Immunol 2009;24:69-75.  Back to cited text no. 4
    
5.
Lee KH, Maiden MF, Tanner AC, Weber HP. Microbiota of successful osseointegrated dental implants. J Periodontol 1999;70:131-8.  Back to cited text no. 5
    
6.
Auschill TM, Hellwig E, Sculean A, Hein N, Arweiler NB. Impact of the intraoral location on the rate of biofilm growth. Clin Oral Investig 2004;8:97-101.  Back to cited text no. 6
    
7.
de Groot K, Wolke JG, Jansen JA. Calcium phosphate coatings for medical implants. Proc Inst Mech Eng H 1998;212:137-47.  Back to cited text no. 7
    
8.
Slack R, Tindall A, Shetty AA, James KD, Rand C. 15-year follow-up results of hydroxyapatite ceramic-coated femoral stem. J Orthop Surg 2006;14:151-4  Back to cited text no. 8
    
9.
Slack R, Tindall A, Shetty AA, James KD, Rand C. 15-year follow-up results of the hydroxyapatite ceramic-coated femoral stem. J Orthop Surg (Hong Kong) 2006;14:151-4.  Back to cited text no. 9
    
10.
Davies JE. Mechanisms of endosseous integration. Int J Prosthodont 1998;11:391-401.  Back to cited text no. 10
    
11.
Hojo K, Nagaoka S, Ohshima T, Maeda N. Bacterial interactions in dental biofilm development. J Dent Res 2009;88:982-90.  Back to cited text no. 11
    
12.
Wood SR, Kirkham J, Shore RC, Brookes SJ, Robinson C. Changes in the structure and density of oral plaque biofilms with increasing plaque age. FEMS Microbiol Ecol 2002;39:239-44.  Back to cited text no. 12
    


    Figures

  [Figure 1]
 
 
    Tables

  [Table 1], [Table 2], [Table 3]



 

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