Orthodontic Appliances, RemovableNCT04139538
Microbiome and Immunogenity of Saliva During Orthodontic Therapy
A study testing orthodontic treatment in people with Orthodontic Appliances, Removable and Orthodontic Appliances, Fixed, run by Medical University of Vienna at 1 site in Austria.
Trial summary
There are two essential ways of treatment in the field of orthodontics. The fixed appliance treatment with self-ligating or conventional brackets and treatment with aligners. Fixed orthodontic treatment has been the optional therapy for decades. Read more about this study
- Who is it for?
- Ages 18 years and olderHealthy volunteers can take part
- Conditions the study is for
- Orthodontic Appliances, Removable, Orthodontic Appliances, Fixed, Oral Microbiome
- What is being tested?
- orthodontic treatment
- Type of treatment
- Surgery or procedure
- Could I receive a placebo?
- No, everyone receives an active treatment
- How many people
- 40 participants
- Where
- 1 siteAustria
- Study dates
- 1 Nov 2019to 1 Oct 2024
- Phase
- Interventional
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 years and older
- All sexes
- Healthy volunteers accepted
- Adult, Older adult
Who may be able to join
- Class I skeletal relationship
- Normo-divergent Frankfort Mandibular-Plane Angle
- Minimal mandibular crowding in range from 1 to 3 according to Little's Index (Little 1975)
Who cannot join
- Smoking habit
- Presence of extensive dental restorations in proximity to the gingival margin
- Presence of fixed bridges/crowns or partial dentures
- Previous periodontal nonsurgical treatment (such as full mouth disinfection, quadrant-by-quadrant therapy, full mouth debridement) within the past year
- Medications such as antibiotics, steroids, or nonsteroidal anti-inflammatory drugs within the past 6 months.
- Plaque Index > 20% before start of treatment Diabetes Pregnancy
Trial locations and contacts
1 site in 1 country.
About this study
Official title: Changes of Oral Microbiome and Immunogenity of Saliva During Orthodontic Therapy With Aligners and Multibracket Therapy - a Pilot Study
There are two essential ways of treatment in the field of orthodontics. The fixed appliance treatment with self-ligating or conventional brackets and treatment with aligners. Fixed orthodontic treatment has been the optional therapy for decades. Due to an increasing demand of patients for a more aesthetic treatment, the number of aligner treatments multiplied over the last years. Within the last years, a crucial role of oral microbiome in the maintenance of oral health was largely appreciated. However, to date there is no data on the effect of different orthodontic treatments on the oral microbiome. The aim of the research project is the prospective comparison of the oral microbiome in saliva of adult patients treated with Invisalign® aligners or metal brackets. We plan to include a sample of twenty healthy individuals. Half of the participants will receive aligner treatment with Invisalign® and half metal brackets. Saliva will be collected before, 3 and 6 months after the beginning of the orthodontic therapy via a stimulated a saliva collection. The 16S rDNA genes will be amplified and sequenced with Illumina MiSeq® System. Probing depth, bleeding on probing and Plaque Index will be evaluated over time and correlated with changes of the oral microbiome.
Read the detailed description
Adult patients (18 years and older) from the Clinical Division of Orthodontics at the University Clinic of Dentistry Vienna will be invited to participate in this pilot microbiologic study according to defined inclusion/exclusion criteria after giving their consent. This study is a pilot project and therefore no sample size calculation will be made. Twenty patients will be treated with Invisalign® aligners and twenty patients will receive multibracket therapy. A previous microbiological study showed that lingual orthodontic therapy has a short-term effect on oral microflora. Another microbial study on colonization on dental implants also reported bacterial shift upon implant therapy.
All patients will get instructions in oral hygiene on the day when clinical records are gathered. On the day when the treatment plan will be presented to the patient, the Plaque Index will be evaluated by a trained orthodontist. For the start of the treatment a Plaque Index of < 20% is required. On the day before the orthodontic treatment is started, T0 sample collection will be conducted and periodontal indices will be assessed. A clinical examination including periodontal probing depth, bleeding on probing, and Plaque Index (PI) will be performed before (T0), after 3 months (T1) and after 6 months of the beginning of either aligner or bracket therapy. Microbiological sampling will be performed by stimulated saliva collection according to our protocol. Saliva samples will be divided in aliquots in stored in cryo-tubes at -80°C. Molecular detection of microorganisms by sequencing of 16S rDNA genes, sequencing analysis and taxonomic assignment will be performed at the Department of Laboratory Medicine at the Medical University of Vienna.
By sequencing of 16s rDNA genes with sequencing analysis and taxonomic assignment of bacteria in saliva we expect to reveal possible shifts in microbial phyla/genera between various time points. Metagenomic studies are commonly performed by analysing the prokaryotic 16S ribosomal RNA gene (16S rRNA), which is approximately 1,500 base pairs long and contains nine variable regions interspersed between conserved regions. Variable regions of 16S rRNA are frequently used in phylogenetic classifications such as genus or species in diverse microbial populations.
DNA Extraction:
Due to better quantitative and qualitative results in comparison to commercial DNA extraction kits, DNA will be isolated with a modified CTAB SDS chloroform-based method. Mechanical analysis will be carried out using 1mm silica spheres under addition of proteinase K (Qiagen, Venlo, Netherlands) as well as the detergents SDS (sodium dodecyl sulfate) and CTAB (cetyltrimethylammonium bromide). After extraction, the DNA quantity will be determined with Qubit® 2.0 via the dsDNA HS kit (Life Technologies, Carlsbad, California) and NanoDrop 2000c spectrophotometer (Thermo Scientific, Waltham, Massachusetts). Additionally, the absorption ratios A260/280 and A260/230 will be used to estimate the purity of the DNA.
Metagenome Analysis:
The overall composition of the bacterial organisms in the samples will be evaluated by analysing microbiome. Therefore, we will use high throughput sequencing with the MiSeq® platform (Illumina, San Diego, California). We will analyse the variable V3 and V4 regions of the 16S rRNA gene. For the phylogenetic classification, the bacterial 16S rRNA will be amplified using primers 27F (5'-CCT ACG GGN GGC WGC AG-3') and 1391R (5'-GACTACHVGGGTATCTAATCC-3').
Bioinformatic analysis:
The quality of the Next Generation Sequencing run will be assessed using the software FASTQC 0.11.4. The removal of low-quality bases will be carried out with the Trimmomatic-0.35 software . For sequence analysis and clustering similar sequences into clusters (OTUs) the UPARSE pipeline will be utilized. For comparison and statistical analysis of the microbiome data we will use RStudio (Team 2016) as platform with R as language (R Development Core Team 2010) and different R-packages.
- Conditions
- Orthodontic Appliances, Removable, Orthodontic Appliances, Fixed, Oral Microbiome, Cell Culture, Periodontal Index
- Being tested
- orthodontic treatment
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
orthodontic treatmentProcedure or surgery
Malocclusion will be treated with orthodontic appliances (fixed or removable)
| Study group | Role | What they receive |
|---|---|---|
| Invisalign treatment | Experimental | Aligner patients will be treated with Invisalign® aligners to correct malocclusion Receives: Procedure: orthodontic treatment |
| selfligating bracket treatment | Active comparator | Multibracket patients will be treated with self ligating orthodontic brackets Receives: Procedure: orthodontic treatment |
What is being measured
The primary outcome is the main question the study is designed to answer.
changes of oral microbiome
bacteria of the oral microbiome including cariogenic bacteria via dna sequencing
Measured: up to 6 months
changes of periodontal parameters
plaque index
Measured: up to 6 months
changes of periodontal parameters
probing depth
Measured: up to 6 months
changes of periodontal parameters
bleeding on probing
Measured: up to 6 months
Secondary outcomes (2)
immunogenity of saliva on oral cells in vitro
changes in viability
Measured: up to 2 weeks
immunogenity of saliva on oral cells in vitro
gene expression changes via PCR
Measured: up to 2 weeks
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted25 Oct 2019
- Study start1 Nov 2019
- Primary completion1 Sep 2022
- Study completion1 Oct 2024 (estimated)
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (7)Papers the sponsor has linked to this study.
- Other trials for Orthodontic Appliances, RemovableStudies of the same condition, open and under way.
- More trials run by Medical University of ViennaThe sponsor's other registered studies.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Interventional
- Phase
- Phase N/A
- Primary purpose
- Diagnosis
- Allocation
- Non-randomised
- Intervention model
- Parallel groups
- Masking
- Single blind
- Who is masked
- Investigator
- Enrolment
- 40 participants (estimated)
Sponsor and investigators
- Lead sponsor
- Medical University of Vienna
- Sponsor type
- University, hospital or other
- Responsible party
- Principal investigator, Michael Nemec, Medical University of Vienna
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT04139538
- Sponsor's protocol ID
- 1288/2019
- Status last verified
- Mar 2023
- First submitted
- 22 Sep 2019
- Last updated
- 23 Mar 2023
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- No
Publications
Klindworth A, Pruesse E, Schweer T, Peplies J, Quast C, Horn M, Glockner FO. Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies. Nucleic Acids Res. 2013 Jan 7;41(1):e1. doi: 10.1093/nar/gks808. Epub 2012 Aug 28.
PubMed 22933715Edgar RC. UPARSE: highly accurate OTU sequences from microbial amplicon reads. Nat Methods. 2013 Oct;10(10):996-8. doi: 10.1038/nmeth.2604. Epub 2013 Aug 18.
PubMed 23955772Levrini L, Mangano A, Montanari P, Margherini S, Caprioglio A, Abbate GM. Periodontal health status in patients treated with the Invisalign((R)) system and fixed orthodontic appliances: A 3 months clinical and microbiological evaluation. Eur J Dent. 2015 Jul-Sep;9(3):404-410. doi: 10.4103/1305-7456.163218.
PubMed 26430371Demling A, Demling C, Schwestka-Polly R, Stiesch M, Heuer W. Short-term influence of lingual orthodontic therapy on microbial parameters and periodontal status. A preliminary study. Angle Orthod. 2010 May;80(3):480-4. doi: 10.2319/061109-330.1.
PubMed 20050740de Freitas AR, Silva TSO, Ribeiro RF, de Albuquerque Junior RF, Pedrazzi V, do Nascimento C. Oral bacterial colonization on dental implants restored with titanium or zirconia abutments: 6-month follow-up. Clin Oral Investig. 2018 Jul;22(6):2335-2343. doi: 10.1007/s00784-018-2334-0. Epub 2018 Jan 18.
PubMed 29349504Haririan H, Andrukhov O, Bertl K, Lettner S, Kierstein S, Moritz A, Rausch-Fan X. Microbial analysis of subgingival plaque samples compared to that of whole saliva in patients with periodontitis. J Periodontol. 2014 Jun;85(6):819-28. doi: 10.1902/jop.2013.130306. Epub 2013 Oct 21.
PubMed 24144271Ren Y, Vissink A. Cytokines in crevicular fluid and orthodontic tooth movement. Eur J Oral Sci. 2008 Apr;116(2):89-97. doi: 10.1111/j.1600-0722.2007.00511.x.
PubMed 18353001
Study at a glance
From the registry record, last updated by the sponsor on 23 Mar 2023. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Changes of Oral Microbiome and Immunogenity of Saliva During Orthodontic Therapy With Aligners and Multibracket Therapy - a Pilot Study
- ClinicalTrials.gov ID
- NCT04139538
- Acronym / sponsor ID
- 1288/2019
- Conditions
- Orthodontic Appliances, Removable, Orthodontic Appliances, Fixed, Oral Microbiome, Cell Culture, Periodontal Index
- Study type
- Interventional
- Recruitment status
- Unknown status
- Enrolment
- 40 participants (estimated)
- Lead sponsor
- Medical University of ViennaUniversity, hospital or other
- Study start
- 1 Nov 2019 (actual)
- Primary completion
- 1 Sep 2022 (actual)
- Study completion
- 1 Oct 2024 (estimated)
- First posted
- 25 Oct 2019 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
There are two essential ways of treatment in the field of orthodontics. The fixed appliance treatment with self-ligating or conventional brackets and treatment with aligners. Fixed orthodontic treatment has been the optional therapy for decades. Due to an increasing demand of patients for a more aesthetic treatment, the number of aligner treatments multiplied over the last years. Within the last years, a crucial role of oral microbiome in the maintenance of oral health was largely appreciated. However, to date there is no data on the effect of different orthodontic treatments on the oral microbiome. The aim of the research project is the prospective comparison of the oral microbiome in saliva of adult patients treated with Invisalign® aligners or metal brackets. We plan to include a sample of twenty healthy individuals. Half of the participants will receive aligner treatment with Invisalign® and half metal brackets. Saliva will be collected before, 3 and 6 months after the beginning of the orthodontic therapy via a stimulated a saliva collection. The 16S rDNA genes will be amplified and sequenced with Illumina MiSeq® System. Probing depth, bleeding on probing and Plaque Index will be evaluated over time and correlated with changes of the oral microbiome.
Detailed description
Adult patients (18 years and older) from the Clinical Division of Orthodontics at the University Clinic of Dentistry Vienna will be invited to participate in this pilot microbiologic study according to defined inclusion/exclusion criteria after giving their consent. This study is a pilot project and therefore no sample size calculation will be made. Twenty patients will be treated with Invisalign® aligners and twenty patients will receive multibracket therapy. A previous microbiological study showed that lingual orthodontic therapy has a short-term effect on oral microflora. Another microbial study on colonization on dental implants also reported bacterial shift upon implant therapy.
All patients will get instructions in oral hygiene on the day when clinical records are gathered. On the day when the treatment plan will be presented to the patient, the Plaque Index will be evaluated by a trained orthodontist. For the start of the treatment a Plaque Index of < 20% is required. On the day before the orthodontic treatment is started, T0 sample collection will be conducted and periodontal indices will be assessed. A clinical examination including periodontal probing depth, bleeding on probing, and Plaque Index (PI) will be performed before (T0), after 3 months (T1) and after 6 months of the beginning of either aligner or bracket therapy. Microbiological sampling will be performed by stimulated saliva collection according to our protocol. Saliva samples will be divided in aliquots in stored in cryo-tubes at -80°C. Molecular detection of microorganisms by sequencing of 16S rDNA genes, sequencing analysis and taxonomic assignment will be performed at the Department of Laboratory Medicine at the Medical University of Vienna.
By sequencing of 16s rDNA genes with sequencing analysis and taxonomic assignment of bacteria in saliva we expect to reveal possible shifts in microbial phyla/genera between various time points. Metagenomic studies are commonly performed by analysing the prokaryotic 16S ribosomal RNA gene (16S rRNA), which is approximately 1,500 base pairs long and contains nine variable regions interspersed between conserved regions. Variable regions of 16S rRNA are frequently used in phylogenetic classifications such as genus or species in diverse microbial populations.
DNA Extraction:
Due to better quantitative and qualitative results in comparison to commercial DNA extraction kits, DNA will be isolated with a modified CTAB SDS chloroform-based method. Mechanical analysis will be carried out using 1mm silica spheres under addition of proteinase K (Qiagen, Venlo, Netherlands) as well as the detergents SDS (sodium dodecyl sulfate) and CTAB (cetyltrimethylammonium bromide). After extraction, the DNA quantity will be determined with Qubit® 2.0 via the dsDNA HS kit (Life Technologies, Carlsbad, California) and NanoDrop 2000c spectrophotometer (Thermo Scientific, Waltham, Massachusetts). Additionally, the absorption ratios A260/280 and A260/230 will be used to estimate the purity of the DNA.
Metagenome Analysis:
The overall composition of the bacterial organisms in the samples will be evaluated by analysing microbiome. Therefore, we will use high throughput sequencing with the MiSeq® platform (Illumina, San Diego, California). We will analyse the variable V3 and V4 regions of the 16S rRNA gene. For the phylogenetic classification, the bacterial 16S rRNA will be amplified using primers 27F (5'-CCT ACG GGN GGC WGC AG-3') and 1391R (5'-GACTACHVGGGTATCTAATCC-3').
Bioinformatic analysis:
The quality of the Next Generation Sequencing run will be assessed using the software FASTQC 0.11.4. The removal of low-quality bases will be carried out with the Trimmomatic-0.35 software . For sequence analysis and clustering similar sequences into clusters (OTUs) the UPARSE pipeline will be utilized. For comparison and statistical analysis of the microbiome data we will use RStudio (Team 2016) as platform with R as language (R Development Core Team 2010) and different R-packages.
Study design
- Primary purpose
- Diagnosis
- Allocation
- Non-randomised
- Intervention model
- Parallel groups
- Masking
- Single blind (investigator)
- Number of arms
- 2
| Arm | Type | Receives |
|---|---|---|
| Invisalign treatment | Experimental | Procedure: orthodontic treatmentAligner patients will be treated with Invisalign® aligners to correct malocclusion |
| selfligating bracket treatment | Active comparator | Procedure: orthodontic treatmentMultibracket patients will be treated with self ligating orthodontic brackets |
Who may qualify
- Ages
- 18 years and older
- Sex
- All
- Healthy volunteers
- Accepted
- Age groups
- Adult, Older adult
Key inclusion criteria (3 listed)
- Class I skeletal relationship
- Normo-divergent Frankfort Mandibular-Plane Angle
- Minimal mandibular crowding in range from 1 to 3 according to Little's Index (Little 1975)
Key exclusion criteria (6 listed)
- Smoking habit
- Presence of extensive dental restorations in proximity to the gingival margin
- Presence of fixed bridges/crowns or partial dentures
- Previous periodontal nonsurgical treatment (such as full mouth disinfection, quadrant-by-quadrant therapy, full mouth debridement) within the past year
- Medications such as antibiotics, steroids, or nonsteroidal anti-inflammatory drugs within the past 6 months.
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| orthodontic treatmentMalocclusion will be treated with orthodontic appliances (fixed or removable) | Procedure or surgery | Invisalign treatment, selfligating bracket treatment |
Primary outcome
changes of oral microbiome
Time frame: up to 6 months
bacteria of the oral microbiome including cariogenic bacteria via dna sequencing
changes of periodontal parameters
Time frame: up to 6 months
plaque index
changes of periodontal parameters
Time frame: up to 6 months
probing depth
changes of periodontal parameters
Time frame: up to 6 months
bleeding on probing
2 secondary outcomes
immunogenity of saliva on oral cells in vitro
Time frame: up to 2 weeks
immunogenity of saliva on oral cells in vitro
Time frame: up to 2 weeks
Results and safety
Results are not available in the source record.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 23 Mar 2023.
- Background: PubMed 22933715Klindworth A, Pruesse E, Schweer T, Peplies J, Quast C, Horn M, Glockner FO. Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies. Nucleic Acids Res. 2013 Jan 7;41(1):e1. doi: 10.1093/nar/gks808. Epub 2012 Aug 28.
- Background: PubMed 23955772Edgar RC. UPARSE: highly accurate OTU sequences from microbial amplicon reads. Nat Methods. 2013 Oct;10(10):996-8. doi: 10.1038/nmeth.2604. Epub 2013 Aug 18.
- Background: PubMed 26430371Levrini L, Mangano A, Montanari P, Margherini S, Caprioglio A, Abbate GM. Periodontal health status in patients treated with the Invisalign((R)) system and fixed orthodontic appliances: A 3 months clinical and microbiological evaluation. Eur J Dent. 2015 Jul-Sep;9(3):404-410. doi: 10.4103/1305-7456.163218.
- Background: PubMed 20050740Demling A, Demling C, Schwestka-Polly R, Stiesch M, Heuer W. Short-term influence of lingual orthodontic therapy on microbial parameters and periodontal status. A preliminary study. Angle Orthod. 2010 May;80(3):480-4. doi: 10.2319/061109-330.1.
- Background: PubMed 29349504de Freitas AR, Silva TSO, Ribeiro RF, de Albuquerque Junior RF, Pedrazzi V, do Nascimento C. Oral bacterial colonization on dental implants restored with titanium or zirconia abutments: 6-month follow-up. Clin Oral Investig. 2018 Jul;22(6):2335-2343. doi: 10.1007/s00784-018-2334-0. Epub 2018 Jan 18.
- Background: PubMed 24144271Haririan H, Andrukhov O, Bertl K, Lettner S, Kierstein S, Moritz A, Rausch-Fan X. Microbial analysis of subgingival plaque samples compared to that of whole saliva in patients with periodontitis. J Periodontol. 2014 Jun;85(6):819-28. doi: 10.1902/jop.2013.130306. Epub 2013 Oct 21.
- Background: PubMed 18353001Ren Y, Vissink A. Cytokines in crevicular fluid and orthodontic tooth movement. Eur J Oral Sci. 2008 Apr;116(2):89-97. doi: 10.1111/j.1600-0722.2007.00511.x.
- Individual participant dataSharing planned: No
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Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
The record lists no contact. Use the official record to reach the sponsor.
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
A study testing orthodontic treatment in people with Orthodontic Appliances, Removable and Orthodontic Appliances, Fixed, run by Medical University of Vienna at 1 site in Austria.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 3 inclusion and 6 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: status not confirmed, as of the sponsor's last update on 23 Mar 2023. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Austria. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
Results are not available in the source record.
How can a doctor refer a patient or contact the research team?
The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
changes of oral microbiome (time frame: up to 6 months). The study lists 4 primary outcomes in all. It is what the study was designed to measure, not a finding.
Where can I read the original study information?
On the ClinicalTrials.gov record for NCT04139538, which also lists 7 linked publications.
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 23 Mar 2023.