Ignite Creation Date:
2024-05-06 @ 3:06 PM
Last Modification Date:
2024-10-26 @ 1:43 PM
Study NCT ID:
NCT04520438
Status:
COMPLETED
Last Update Posted:
2020-08-26
First Post:
2020-08-12
Brief Title:
L-PRF Plus Non Surgical Periodontal Treatment
Sponsor:
Universidad de los Andes Chile
Organization:
Universidad de los Andes Chile
Study Overview
Official Title:
Effect of Leucocyte and Platelet Rich Fibrin as an Adjuvant in Non-Surgical Periodontal Therapy A Split-Mouth Randomized Controlled Clinical Trial
Status:
COMPLETED
Status Verified Date:
2020-08
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
Abstract
Background Leucocyte and Platelet-Rich Fibrin L-PRF has shown to promote regenerative processes even reporting antibacterial effect The aim of this split-mouth clinical trial was to evaluate the effect of L-PRF as an adjuvant to scaling and root planing SRP
Methods 13 patients with chronic periodontitis and at least 1 bilateral periodontal pocket 6 mm were recruited The sites were randomly treated with SRP L-PRF test group or SRP alone control group The following parameters were evaluated at baseline and 6 weeks 3 and 6 months after treatment Probing pocket depth PPD clinical attachment level CAL bleeding on probing BOP gingival recession GR and root sensitivity RS Additionally the concentrations of Porphyromona gingivalis Pg Aggregatibacter actinomycetemcomitans Aa Prevotella intermedia Pi and Fusobacterium nucleatum Fn in the gingival crevicular fluid GCF were evaluated at baseline 6 weeks and 3 months after treatment
Background Leucocyte and Platelet-Rich Fibrin L-PRF has shown to promote regenerative processes even reporting antibacterial effect The aim of this split-mouth clinical trial was to evaluate the effect of L-PRF as an adjuvant to scaling and root planing SRP
Methods 13 patients with chronic periodontitis and at least 1 bilateral periodontal pocket 6 mm were recruited The sites were randomly treated with SRP L-PRF test group or SRP alone control group The following parameters were evaluated at baseline and 6 weeks 3 and 6 months after treatment Probing pocket depth PPD clinical attachment level CAL bleeding on probing BOP gingival recession GR and root sensitivity RS Additionally the concentrations of Porphyromona gingivalis Pg Aggregatibacter actinomycetemcomitans Aa Prevotella intermedia Pi and Fusobacterium nucleatum Fn in the gingival crevicular fluid GCF were evaluated at baseline 6 weeks and 3 months after treatment
Detailed Description:
Materials and Methods Study design and Patient Selection A split-mouth randomized controlled clinical trial enrolling 13 patients 8 males and 5 females mean age 523 94 years was set-up The patients were informed about the benefits and risks of the study and each participant signed informed consent
Non-surgical periodontal treatment A single periodontist LC performed all periodontal treatments NSPT consisted of two sessions of scaling and root planing SRP within 48 hours under local anesthesia using ultrasonic and hand instrumentation Additionally the sites associated with PPD 6 mm in the quadrant assigned to the test group were irrigated with L-PRF exudate and filled with longitudinal pieces of L-PRF membrane The patients received instructions about soft diet a carefully and soft mouth rinse with chlorhexidine 012 and no toothbrushing during 7 days in order to avoid the L-PRF removal After this period the following instructions were given modified Bass brushing technique dental floss and interdental brushes and chlorhexidine 012 mouth rinse for an extra 7 days Strict hygiene control by the clinician was also performed in each appointment
L-PRF membrane preparation Two samples of venous blood were collected in 2 vacutainer tubes red cup of 10 ml without anticoagulant They were centrifuged at 408 g for 12 minutes using a table centrifuge IntraSpinTM Intralock Florida USA according to the protocol described by Temmerman et al18 The L-PRF clots were removed from the tube separated from the red cells and placed in the Xpression box IntraSpinTM Intralock Florida USA to gently compress them in membranes The membranes were chopped in longitudinal pieces to fill the periodontal pockets 6mm The exudate released during the compression called L-PRF exudate was aspirated to rinse the treated pockets Figure 1
Clinical Measurements All clinical measurements were performed by a single trained and calibrated examiner NJ who was masked for the treatment assigned using a basic examination instrument kit and a University of North Carolina no 15 color-coded periodontal probe The following clinical parameters were measured at baseline and 6 weeks 3 months and 6 months after treatment probing pocket depth PPD clinical attachment level CAL gingival recession GR bleeding on probing BOP and OLeary plaque index PI19 Additionally the root sensitivity RS was evaluated using a 100-mm visual analogue scale VAS 0 indicating no pain 50 indicating average and 100 indicating an unbearable pain at 24 hrs 6 weeks 3 months and 6 months after treatment The change in PPD CAL GR BOP PI and RS difference between baseline measures and at 1 3 and 6 months were calculated
GCF collection GCF samples were collected from the deepest pocket from each quadrant before recording the clinical measurement baseline and 6 weeks and 3 months after treatment in order to evaluate the concentration of Porphyromona gingivalis Pg Aggregatibacter actinomycetemcomitans Aa Prevotella intermedia Pi and Fusobacterium nucleatum Fn The sample area was isolated with cotton rolls and contamination with saliva was avoided using an appropriate suction and air spray Then an endodontic 35 paper-point was inserted until a slight resistance was felt and it was held for 30 seconds The paper-points were immediately inserted in an Eppendorf tube and stored at -80ºC for future analysis
Microbiological processing After defrosting the samples 1ml of phosphate-buffered saline PBS was added and homogenized Then 400 μl of each sample was centrifuged at 13200 rpm for 3 minutes The obtained pellet was dispersed in 200 μl InstaGene DNA was extracted with InstaGene matrix Bio-Rad Life Science Research Hercules CA USA according to the instructions of the manufacturer Five microliters of the purified DNA were used for the quantification of Porphyromonas gingivalis Aggregatibacter actinomycetemcomitans Fusobacterium nucleatum and Prevotella intermedia A quantitative polymerase chain reaction qPCR assay was performed with a CFX96 Real-Time System Biorad Hercules CA USA using the Taqman 5 nuclease assay PCR method for detection and quantification of bacterial DNA Taqman reaction contained 125 μl Mastermix Eurogentec Seraing Belgium 45 μl sterile distilled water 1 μl of both species-specific primers and probe Table 1 and 5 μl template DNA Assay conditions for all primerprobe set consisted of an initial 2 min at 50C followed by a denaturation step for 10 min at 95C followed by 45 cycles of 95C for 15 sec and 60C for 60 sec Quantification was based on a plasmid standard curve The delta for bacterial concentration at 6 weeks 3 and 6 months difference between baseline measures and at 6 weeks 3 and 6 months were calculated
Randomization allocation concealment and calibration A coin toss method was used to assign the patients quadrants into two treatment groups SRP L-PRF or control SRP alone
Before beginning the study two recordings of the following parameters were done PPD GR and CAL in five patients with a 24-hour interval between first and second records in order to validate the intra-examiner accuracy All teeth excluding third molars were measured by periodontal probing at 6 sites mesiobuccal mediobuccal distobuccal mesiolingualpalatal mediolingualpalatal and distolingualpalatal Calibration was accepted if the measurements at baseline and after 24 hours were within 1 mm at the 95 level correlation coefficients between duplicate measurements r 095
Statistical analysis To detect a mean difference in PPD of 12 mm with a standard deviation of 10 at an α level of 005 and a β level of 010 power 09 a sample size of 10 patients was required Despite the results but to anticipate potential drop outs during the study a sample size of n 16 patients was considered
Bacterial counts were log-transformed A linear mixed model was applied with the variable patient as a random factor and time and treatment as two crossed fixed factors Smoking and PI were considered as covariables Contrasts were set up to calculated the change from baseline for the different time points between the treatments and for each treatment apart A correction for simultaneous hypothesis testing according to Sidak was applied A normal quantile plot and residual dot plot of the residual values were created showing that the assumptions underlying the statistical model could be made
Non-surgical periodontal treatment A single periodontist LC performed all periodontal treatments NSPT consisted of two sessions of scaling and root planing SRP within 48 hours under local anesthesia using ultrasonic and hand instrumentation Additionally the sites associated with PPD 6 mm in the quadrant assigned to the test group were irrigated with L-PRF exudate and filled with longitudinal pieces of L-PRF membrane The patients received instructions about soft diet a carefully and soft mouth rinse with chlorhexidine 012 and no toothbrushing during 7 days in order to avoid the L-PRF removal After this period the following instructions were given modified Bass brushing technique dental floss and interdental brushes and chlorhexidine 012 mouth rinse for an extra 7 days Strict hygiene control by the clinician was also performed in each appointment
L-PRF membrane preparation Two samples of venous blood were collected in 2 vacutainer tubes red cup of 10 ml without anticoagulant They were centrifuged at 408 g for 12 minutes using a table centrifuge IntraSpinTM Intralock Florida USA according to the protocol described by Temmerman et al18 The L-PRF clots were removed from the tube separated from the red cells and placed in the Xpression box IntraSpinTM Intralock Florida USA to gently compress them in membranes The membranes were chopped in longitudinal pieces to fill the periodontal pockets 6mm The exudate released during the compression called L-PRF exudate was aspirated to rinse the treated pockets Figure 1
Clinical Measurements All clinical measurements were performed by a single trained and calibrated examiner NJ who was masked for the treatment assigned using a basic examination instrument kit and a University of North Carolina no 15 color-coded periodontal probe The following clinical parameters were measured at baseline and 6 weeks 3 months and 6 months after treatment probing pocket depth PPD clinical attachment level CAL gingival recession GR bleeding on probing BOP and OLeary plaque index PI19 Additionally the root sensitivity RS was evaluated using a 100-mm visual analogue scale VAS 0 indicating no pain 50 indicating average and 100 indicating an unbearable pain at 24 hrs 6 weeks 3 months and 6 months after treatment The change in PPD CAL GR BOP PI and RS difference between baseline measures and at 1 3 and 6 months were calculated
GCF collection GCF samples were collected from the deepest pocket from each quadrant before recording the clinical measurement baseline and 6 weeks and 3 months after treatment in order to evaluate the concentration of Porphyromona gingivalis Pg Aggregatibacter actinomycetemcomitans Aa Prevotella intermedia Pi and Fusobacterium nucleatum Fn The sample area was isolated with cotton rolls and contamination with saliva was avoided using an appropriate suction and air spray Then an endodontic 35 paper-point was inserted until a slight resistance was felt and it was held for 30 seconds The paper-points were immediately inserted in an Eppendorf tube and stored at -80ºC for future analysis
Microbiological processing After defrosting the samples 1ml of phosphate-buffered saline PBS was added and homogenized Then 400 μl of each sample was centrifuged at 13200 rpm for 3 minutes The obtained pellet was dispersed in 200 μl InstaGene DNA was extracted with InstaGene matrix Bio-Rad Life Science Research Hercules CA USA according to the instructions of the manufacturer Five microliters of the purified DNA were used for the quantification of Porphyromonas gingivalis Aggregatibacter actinomycetemcomitans Fusobacterium nucleatum and Prevotella intermedia A quantitative polymerase chain reaction qPCR assay was performed with a CFX96 Real-Time System Biorad Hercules CA USA using the Taqman 5 nuclease assay PCR method for detection and quantification of bacterial DNA Taqman reaction contained 125 μl Mastermix Eurogentec Seraing Belgium 45 μl sterile distilled water 1 μl of both species-specific primers and probe Table 1 and 5 μl template DNA Assay conditions for all primerprobe set consisted of an initial 2 min at 50C followed by a denaturation step for 10 min at 95C followed by 45 cycles of 95C for 15 sec and 60C for 60 sec Quantification was based on a plasmid standard curve The delta for bacterial concentration at 6 weeks 3 and 6 months difference between baseline measures and at 6 weeks 3 and 6 months were calculated
Randomization allocation concealment and calibration A coin toss method was used to assign the patients quadrants into two treatment groups SRP L-PRF or control SRP alone
Before beginning the study two recordings of the following parameters were done PPD GR and CAL in five patients with a 24-hour interval between first and second records in order to validate the intra-examiner accuracy All teeth excluding third molars were measured by periodontal probing at 6 sites mesiobuccal mediobuccal distobuccal mesiolingualpalatal mediolingualpalatal and distolingualpalatal Calibration was accepted if the measurements at baseline and after 24 hours were within 1 mm at the 95 level correlation coefficients between duplicate measurements r 095
Statistical analysis To detect a mean difference in PPD of 12 mm with a standard deviation of 10 at an α level of 005 and a β level of 010 power 09 a sample size of 10 patients was required Despite the results but to anticipate potential drop outs during the study a sample size of n 16 patients was considered
Bacterial counts were log-transformed A linear mixed model was applied with the variable patient as a random factor and time and treatment as two crossed fixed factors Smoking and PI were considered as covariables Contrasts were set up to calculated the change from baseline for the different time points between the treatments and for each treatment apart A correction for simultaneous hypothesis testing according to Sidak was applied A normal quantile plot and residual dot plot of the residual values were created showing that the assumptions underlying the statistical model could be made
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None