Clinical Trial Details
— Status: Recruiting
Administrative data
| NCT number |
NCT02315222 |
| Other study ID # |
Regenium |
| Secondary ID |
|
| Status |
Recruiting |
| Phase |
Phase 2/Phase 3
|
| First received |
December 9, 2014 |
| Last updated |
December 10, 2014 |
| Start date |
November 2014 |
Study information
| Verified date |
December 2014 |
| Source |
University of Milan |
| Contact |
Giulio Rasperini |
| Phone |
335 8130194 |
| Email |
giulio.rasperini[@]unimi.it |
| Is FDA regulated |
No |
| Health authority |
Italy: Ethics Committee |
| Study type |
Interventional
|
Clinical Trial Summary
Patient with periodontal disease undergoing full mouth disinfection will be given dietary
supplements or placebo and reevaluated after 3 months.
Description:
Introduction The beneficial effect of vitamins and dietary supplements on oral wound healing
has been recently reported in clinical trials (for review see Van der Valden, JCP 2011). In
periodontal tissue, the imbalanced ratio between virulence of pathogens bacteria and
proportionate host response induce a destructive process of periodontal ligament, cementum
and alveolar bone. The importance of micronutrients including vitamins, calcium and
antioxidant for periodontal health was reported in several clinical studies and the
integration between traditional (non surgical and surgical interventions) and
micronutritional approaches were proposed for treatment of periodontitis (Van der Velden et
al. 2011).
Clinical trials reported that daily supplement of fruit, vegetable and berry juice powder,
vitamin D, Calcium seems to improve periodontal health in patients with chronic
periodontitis that were treated with non-surgical or surgical periodontal therapy or were in
maintenance program (Chapple et al 2012, Garcia et al 2011, Miley et al 2009, Bashutski et
al. 2011). However, the beneficial effects of micronutrients on periodontal health and
healing need to be more deeply investigated. Furthermore, a dietary supplement specifically
formulated for periodontal health needs to be tested.
Regenium is a dietary supplement with Q-TER (coenzyme q10 terclatrate), DHA Omega 3,
Boswellia serrata, Vitamins and mineral salts that was formulated to improve the collagen
formation and the maintenance of the health of oral mucosa. Q-TER plays a key role in the
ATP synthesis and energetic metabolism, and has a strong antioxidant function. A recent
study also reported the function of Q-10 as inhibitor of osteoclast differentiation (Moon et
al. 2012). DHA Omega 3 (Resolvine) has an important antioxidant function, affecting the
cellular aging and moderates the anti-inflammatory mechanisms. Boswellia serrata showed
beneficial effects on wound healing and contraction (Mallik et al. 2010). This micronutrient
reduces the synthesis of leukotrienes in intact neutrophils by inhibiting 5-lipoxygenase,
the key enzyme involved in the biosynthesis of leukotrienes and in the inflammatory process.
The aim of the present study is to evaluate if patients affected by severe periodontal
disease and treated with periodontal non-surgical therapy benefit from dietary supplements
(Regenium). It will be assessed the effect of Regenium on clinical parameters of periodontal
wound healing and on systemic inflammation related to periodontal disease.
1. Experimental procedure
1.1 Study population A total of 60 patients afflicted with severe periodontal disease
and that need for non-surgical periodontal therapy will be enrolled.
Inclusion criteria
- severe periodontal disease: at least 2 sites with Probing Pocket Depth (PPD)>7mm,
Bleeding On Probing (BOP)> 25%
- Age between 18 and 65 years
- Signed informed consent
Exclusion criteria:
- Systemic diseases that may affect periodontal status
- Metabolic disorders
- Nutritional conditions that may alter the formation and maturation of connective
tissue
- Pregnancy or lactation
1.2 Subject protection Informed consent will be obtained from all subjects to be
entered in the study. In obtaining the informed consent and in the conduct of the study
the principles outlined in the Declaration of Helsinki on experimentation involving
human subjects should be adhered to. At each visit, the clinician will evaluate
patients for any untoward effects. In case a patient requires any treatment during the
course of the study, the necessary treatment will be provided at the discretion of the
clinician and according to the current standard of care.
1.3 Treatment After including patient in the study, each patient will be randomly
assigned to one of the experimental groups (test, control). At T0 clinical measurements
will be harvested, saliva samples will be collected and serum analysis will be
performed. Furthermore each patient will receive oral hygiene instructions and regular
professional supragingival debridement. Therapy with Regenium or placebo will be given
to the patient and will start at T0.
Four weeks after starting the therapy (T1), clinical measurements and saliva samples
will be harvested and serum analysis will be performed. After measurements the
one-stage full-mouth disinfection will be performed as following described. Scaling and
root planing of all pockets will be performed within 24 h in combination with an
extensive application of chlorhexidine (0.20%) to all intra-oral niches such
(periodontal pockets, tongue dorsum, tonsils) and in combination with systemic
antibiotic treatment (amoxicillin cp. 1gr 2/die + 250 mg metronidazole 3/die).
To resume patients will intake Ragenium or placebo for 3 months (from 1 months before
and until 2 months after non surgical periodontal therapy), each patient will intake
one pill of Regenium Tablet in the morning (with breakfast) and one capsule of Regenium
Capsule in the evening (with dinner). Patients assigned to the control group (n=30)
will intake placebo tablets and capsules as prescribed in the test group. Compliance
will be evaluated by pill counting at the end of the study.
Three months after therapy with Regenium or placebo (T2), clinical measurements and
saliva samples will be harvested and serum analysis will be performed.
Treatment assignment To enter a patient into the study each investigator will fill in a
screening and an entry form and fax them to the Central Registrar located at the
Clinical Research support Unit of Periodontology, Dental Clinic, Dep. of Biological,
Surgical and Dental Sciences, University of Milan, Fondazione IRCCS Ca' Granda
Polyclinic Milan, Italy. Following receipt of these forms, the Registrar will proceed
to verify satisfaction of all entry criteria and will enter the subject into the study.
Patient codes will be assigned consecutively and will be made of a total of 2 digits
that will encode the subject number (from 01 to 60). These will be assigned at the
first visit, verifying the inclusion criteria and entering the patient into the study.
After having been entered patients into the study, experimental patients will be
randomly assigned by Central Registrar to one of the two treatment regimens according
to pre-defined randomization tables.
A balanced random permuted block approach will be used to prepare the randomization
tables in order to avoid unequal balance between 2 treatments. In order to reduce the
chance of unfavorable splits between test and control patients in terms of key
prognostic factors, the randomization process will take into account the following
variable: smoke. Treatment assignment will be noted in the registration and treatment
assignment form that will be kept by the central registrar. To conceal assignment from
the investigator until the time, at the end of the non-surgical periodontal therapy the
central registrar will instruct the investigator to assign a sealed envelope containing
the treatment for each patient, every envelop is endowed of a Treatment's Code,
composed by a letter (that corresponds to the clinical center) and two numbers.
When a patient enters in the study the investigator assigned him a sequential Patient's
Code, than the clinician sends Screening Form and Entry Form to the study registrar.
The central registrar will match the randomization envelope with the subject number
based on the randomization tables and he send the Registration Form, with the
correspondence between Patient's Code and Treatment's Code. Following receipt of the
registration form from the Central Registrar, the investigator will prescribe the
treatment (Regenium or placebo) to the patient and will book all the subsequent
appointments according to the protocol.
2. Measurements Clinical measurements To evaluate the inflammatory status, plaque control
and severity of periodontal disease, clinical measurements will be taken at T0
(immediately before starting with dietary supplements or placebo), at T1 (4 weeks after
starting supplementation) and at the last day of supplementation (T2= 3 months after
T0).
The following clinical parameters will be taken:
- Full Mouth Plaque Score (FMPS), Full Mouth Bleeding Score (FMBS)
- Periodontal Pocket Depth (PPD), Clinical Attachment Level (CAL), tooth mobility.
All clinical measurements will be taken by a blind calibrated examiner.
Radiographic evaluations An X-Ray status with a parallel technique will be taken at the
baseline during the diagnostic process of periodontal disease.
Serum analysis
Blood test will be performed at T0, T1 and T2 in all patients to evaluate the systemic
inflammation and to assess changes on this condition after periodontal treatment and dietary
supplements intake. The following blood markers of systemic inflammation will be evaluated
(Graziani et al. 2010):
- High sensitivity C reactive protein (hsCRP)
- D-dimers and renal function (cystatin C)
Whole Saliva Collection Unstimulated whole saliva will be collected from each subject at the
beginning of the appointment, as previously described by Ramseier et al. Subjects' will
vigorously rinsed their mouth with tap water for 20 seconds in order to remove gross debris.
The subject then expectorates the water. Following a waiting period of 2 minutes, subjects
will tip their head over the graduated test tube and passively expectorated whole saliva
into the plastic funnel placed inside the plastic tube (Mandel et al. 1976). Each tube will
be labeled with the subject's initials, harvest date and sample name. The collection will be
completed once 2mL of whole saliva is collected, or a maximum of 15 minutes of sampling time
is reached. The sample will then immediately placed on ice, supplemented with a proteinase
inhibitor combination of 1% aprotinin (1mg/ml) and 0.5% phenylmethylsulphonyl fluoride
(PMSF) (200mM in MeOH) (Sigma Chemical Company, St-Louis, MO), and aliquotted prior to
storage at -20°C until analysis.
Salivary Biomarkers Analysis Inflammatory biomarkers expression will be quantified using a
custom human cytokine protein array|| for the evaluation of matrix metalloproteinase-8 and
-9 (MMPs-8 and -9).
Upon receipt, the Quantibody® Array kits will be stored at -20°C. Prior to each assay, whole
saliva samples will be thawed at room temperature and microcentrifuged for 5 minutes to
obtain a cell-free supernatant for analysis.
Each slide containing cytokine standards used for making known serial dilutions, with sample
diluent serving as the negative control, and experimental samples will then be incubated
overnight at 4°C followed by washing unbound materials. The detection antibody will then
bound to the antigens within each well. Cy3 equivalent dye-conjugated streptavidin will be
pipetted in each well, which binds to the detection antibody associated with immune
complexes. The slides will be incubated and covered with aluminum foil to avoid light
exposure and the fluorescence from each well was detected using a laser scanner.¶ The
resultant signals of the samples will be compared to the standard curve for each of the
cytokines in order to determine the concentrations of each cytokine within the samples. Data
will be extracted and analyzed using microarray analysis software.#
Compliance and its evaluation Compliance is a serious problem for clinical trials, to
improve this fundamental aspect of the study, the clinicians will adopt some useful
technique.
Every patient will be instructed to download on his smartphone an application that will
remind him to take Regenium in the morning and in the evening; secondly the clinician will
call the patients two weeks after T0 and two and five weeks after T1, with the pretext to
remind the next appointment and to have information about their oral health, but with the
real intention to remember and to verify the regular assumption of Regenium. Lastly, at T2,
patients will have to bring back the blister, also the empty ones, for the pill counting.
It is important not to say the patients about the pill count, in order to avoid the risk of
bias created by patient who throw away the remaining pill the day before the control. To
have the blister back, clinicians will say the patients to return blister for wasting
reasons, for both package and pills.
References
J.D. Bashutski et al. The impact of vitamin D on periodontal surgery outcomes. Journal of
Dental Research, vol 90, pp. 1007-12, 2011.
M. R. Chapple, et al. Adjunctive Daily Supplementation with Encapsulated Fruit, Vegetable
and Berry Juice Powder Concentrates and Clinical Periodontal Outcomes: A Double-blind RCT.
Journal of Clinical Periodontology, vol. 39, no. 1, pp. 62-72, 2012.
M. N. Garcia, C. F. Hildebolt, D. D. Miley, D. A. Dixon, R. A. Couture, C. L. Spearie, E. M.
Langenwalter, W. D. Shannon, E. Deych, C. Mueller, and R. Civitelli, "One-year Effects of
Vitamin D and Calcium Supplementation on Chronic Periodontitis,"Journal of Periodontology,
vol. 82, no. 1, pp. 25-32, 2011.
F. Graziani, et al. Systemic Inflammation Following Non-surgical and Surgical Periodontal
Therapy Journal of Clinical Periodontology, vol. 37, no. 9, pp. 848-54, 2010.
A. Mallik et al. Evaluation of Boswellia Serrata oleo-gum resin for wound healing activity.
Der Pharmacia Lettre, , vol. 2, no. 2, pp. 457-463, 2010.
I.D Mandel, and S. Wotman, The salivary secretions in health and disease. Oral Sciences
Review, vol 8, pp. 25-47, 1976.
D. D. Miley, M. N. Garcia, C. F. Hildebolt, W. D. Shannon, R. A. Couture, C. L. Anderson
Spearie, D. A. Dixon, E. M. Langenwalter, C. Mueller, and R. Civitelli, "Cross-sectional
Study of Vitamin D and Calcium Supplementation Effects on Chronic Periodontitis,"Journal of
Periodontology, vol. 80, no. 9, pp. 1433-9, 2009.
H. Moon et al., Antioxidants, like coenzyme Q10, selenite, and curcumin, inhibited
osteoclast differentiation by suppressing reactive oxygen species generation. Biochemical
and Biophysical Research Communications vol. 418, pp. 247-253, 2012.
C.A Ramseier et al., Identification of pathogen and host-response markers correlated with
periodontal disease. Journal of Periodontology vol 80, no. 3, p. 436-46, 2009.
U. Van der Velden, D. Kuzmanova, and I. L. C. Chapple, "Micronutritional Approaches to
Periodontal Therapy,"Journal of Clinical Periodontology, vol. 38, pp. 142-158, 2011.
