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Table of Contents
ORIGINAL ARTICLE
Year : 2019  |  Volume : 6  |  Issue : 4  |  Page : 73-75

A comparative evaluation in different types of treatments on titanium alloy samples with or without gentamicin and its effect on biofilm: An in vitro study


1 Reader, Department of Prosthodontcs, Noorul Islam College Of Dental Science, Neyyatinkara, Kerala, India
2 Reader, Department of Prosthodontics, Sri Sankara Dental College, Akathumuri, Varkala, Kerala, India
3 Senior Lecturer, Department of Prosthodontics, Sri Sankara Dental College, Akathumuri, Varkala, Kerala, India
4 Assistant Professor, Department of Periodontics, PMS College of Dental Science And Research, Vattapara, Thiruvananthapuram, Kerala, India
5 Senior Lecturer, Department of Periodontics And Oral Implantology, PMS College of Dental Science And Research, Vattapara, Thiruvananthapuram, Kerala, India

Date of Submission25-Nov-2019
Date of Acceptance27-Nov-2019
Date of Web Publication03-Jan-2020

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


DOI: 10.4103/INPC.INPC_55_19

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  Abstract 


Background: Use of osseointegrated oral implants has been an excellent method for the 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: The aims and objectives of this study lie in the comparative evaluation of biofilm formation among five differently treated surfaces on titanium samples and to evaluate the difference in the delay of biofilm formation among various surfaces.
Methodology: Six sets of polished titanium samples were blasted with sintered hydroxyapatite (HA) and titanium dioxide (TiO2). Another set of samples were blasted and later loaded with gentamicin drug by vacuum drying. The control group comprised plain polished and gentamicin drug-loaded samples. And, evaluation of the strains was done for biofilm. Bacterial adhesion was evaluated on time intervals of 0 h, 1 h, 4 h, 24 h, and 48 h.
Results: Bacterial adhesion was sequentially increasing in the polished samples. Initial bacterial adhesion was more on surface-modified samples when compared to polished samples in the 1st hour. Bacterial adhesion was retarded in gentamicin-coated HA-blasted samples up to 24 h. Bacterial adhesion was considerably less on TiO2-blasted samples up to 48 h.
Conclusion: Implant surface modified with TiO2 and gentamicin showed delayed biofilm formation even up to 48 h. 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 are to be carried out to prove its efficacy.

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


How to cite this article:
Arun R, Rajan NS, George NE, Chandrathara T K, Krishnan R H, Gayathri S. A comparative evaluation in different types of treatments on titanium alloy samples with or without gentamicin and its effect on biofilm: An in vitro study. Int J Prev Clin Dent Res 2019;6:73-5

How to cite this URL:
Arun R, Rajan NS, George NE, Chandrathara T K, Krishnan R H, Gayathri S. A comparative evaluation in different types of treatments on titanium alloy samples with or without gentamicin and its effect on biofilm: An in vitro study. Int J Prev Clin Dent Res [serial online] 2019 [cited 2020 Apr 10];6:73-5. Available from: http://www.ijpcdr.org/text.asp?2019/6/4/73/274676




  Introduction Top


Pure titanium and titanium alloys are commonly employed as implant materials in dentistry due to their favorable combination of mechanical strength, chemical stability, and biocompatibility.[1],[2],[3] The soft tissue surrounding healthy osseointegrated dental implants shares anatomic and functional features with the gingiva around teeth. Antibiotic-loaded implant coatings are employed for the prevention of implant-associated infections. They can provide an immediate response to the threat of implant contamination but do not necessitate the use of an additional carrier for the antibacterial agent. Antibiotics can be loaded on to the surface of implants by two ways – one by passive method and another by 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 other hand, active coatings are designed for temporary release of high fluxes of antibacterial agents immediately following 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] It is active against a wide range of bacterial infections mostly Gram-negative bacteria such as the Pseudomonas and Proteus and Gram-positive bacteria such as 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.[9] Hence, this study is a novel approach to evaluate the influence of biofilm formation on surface-modified implants with and without coating of gentamicin.

Aim

The aim was to evaluate biofilm formation on various surface-treated implants.

Objectives

  1. The objective lies in the comparative evaluation of biofilm formation among five differently treated surfaces on titanium samples
  2. To evaluate the difference in the delay of biofilm formation among various surfaces.



  Methodology Top


Commercially available Ti6Al4V (ASTMF11O8, MANHER Metal Supply Corporation, Mumbai, Maharashtra, India) was machined to 2-mm thickness and 2 cm × 1.5 cm length and breadth rectangular samples. These discs were mechanically polished by silicon carbide papers of grit size 240 and 600 in the grinder and polisher. Hydroxyapatite (HA) powder was prepared in house by a wet precipitation technique using CA (No. 3) 2–4 H2O (calcium nitrate) and NH4H2 PO4 (ammonium dihydrogen phosphate). The micro fine powder was compacted at 200 Mpa in a cold isostatic press. HA powder was loaded in the jar of the blasting machine of particle size (1) 65 μ, (2) 125 μ, and (3) 250 μm. On each particle size, the target samples were away from the gun distance of 2 cm, 4 cm, and 6 cm. Each sample was blasted for 2 min, 4 min, and 6 min. Five samples of each particle size, distance, and time were blasted. The same procedure was carried out for titanium dioxide (TiO2) also. The samples were vacuumed for 15 min at 200 mbar. Titanium samples (five samples of each group) were transferred into Eppendorf tubes containing 1 ml Streptococcus sanguis cultures. Bacterial concentration was about 109 ufc/ml. They were incubated for 0 h, 1 h, 4 h, 24 h, and 48 h at 37°C. The viable count was plotted against roughness and plain samples.


  Results Top


The data were analyzed by SPSS software 16.0 version. ANOVA was applied for comparing between the groups. Post hoc test followed by Dunnett t-test was used to find the significant difference at 95% confidence interval. P < 0.05 between the groups was considered statistically significant. The multiple comparisons of number of viable organisms of different groups are shown in [Graph 1] and the multiple comparisons of the effect of time on biofilm formation in different groups are shown in [Graph 2].




  Discussion Top


In the present study, TiO2 was also selected for modifying the surface of samples as the critical evaluation of various literatures has specified the role of this material in increasing the anchorage of implants. 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.[10] Comparative clinical studies have shown higher marginal bone levels for TiO2 grit-blasted implants, thereby increasing their survival rate.[11] Blasting with TiO2 always shows increased adhesion of the particles onto titanium surface as they are similar metals. This enhances the biomechanical fixation of implants. The antimicrobial property which is an added advantage of TiO2 was also considered in the selection of the material for this study.[12] A S. sanguis strain was used to evaluate the biofilm formation because Streptococcus was the predominant initial colonizing microbe.[13],[14] Biofilm evaluation of surface-modified implants with and without gentamicin was carried out in this study. Scanning electron microscope (SEM) analysis and Energy Dispersive X-Ray Analysis (EDAX) report of samples showed surface roughness and sufficiently adhered elements such as calcium and phosphorous on HA-blasted samples. The SEM results showed surface modifications on TiO2-blasted samples. It is seen that formation of biofilm is seen in all samples. However, it is observed that biofilm formation was delayed in surface-modified and gentamicin-loaded samples. Gentamicin loaded on TiO2 surface showed low concentrations of biofilm formation among all the other five groups. It is noticed within 1 h biofilm formation was on plain polished surface. However, biofilm formation was delayed more than 1 h on plain polished gentamicin-loaded samples. In contrast, the biofilm formation was delayed on TiO2-blasted surface even up to 48 h. In contrast, in HA-treated implants, it was delayed only up to 4 h.


  Conclusion Top


It can be concluded that implant surface modified with TiO2 and gentamicin showed delayed biofilm formation even up to 48 h. These implants can retard the plaque formation, thus preventing peri-implantitis in the primary healing stage. This, in turn, can prevent the failure of implants. This is ideal in situations where the patient is having poor bone quality and poor oral hygiene and in patients suffering from debilitating diseases. 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 are to be carried out to prove its efficacy.

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 Implants Res 2001;12:128-34.  Back to cited text no. 2
    
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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
    
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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
    
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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
    
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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 the hydroxyapatite ceramic-coated femoral stem. J Orthop Surg (Hong Kong) 2006;14:151-4.  Back to cited text no. 8
    
9.
Davies JE. Mechanisms of endosseous integration. Int J Prosthodont 1998;11:391-401.  Back to cited text no. 9
    
10.
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. 10
    
11.
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. 11
    
12.
Hauser-Gerspach I, Kulik EM, Weiger R, Decker EM, Von Ohle C, Meyer J. Adhesion of Streptococcus sanguinis to dental implant and restorative materials in vitro. Dent Mater J 2007;26:361-6.  Back to cited text no. 12
    
13.
Lindh T, Gunne J, Tillberg A. A meta-analysis of implants in partial edentulism. Clin Oral Impl Res 1998;106:721-64.  Back to cited text no. 13
    
14.
Goldstein GR, Preston JD. How to evaluate an article about therapy. J Prosthet Dent 2000;83:599-603.  Back to cited text no. 14
    




 

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