|Detection of Bacterial DNA and Collagen Metabolism in Acutely Ruptured Achilles Tendons|
|Clinicaltrials.gov identifier||recruitment status||First Posted||Last update posted|
|NCT03931486||Recruiting||April 30, 2019||July 14, 2020|
The etiology and pathogenesis of acute Achilles tendon ruptures are complex and not fully understood. It is well known that they are associated with pre-existing pathological alterations, similar to the changes observed in tendinopathy. The present study investigates if bacteria and collagen metabolism play a role in the etiology of acute Achilles tendon rupture. During surgery, 20 patients will have taken two biopsies from the ruptured part of the tendon and two biopsies from the healthy tissue of the same tendon 2-4 cm proximal to the rupture, as a control.
|Condition or Disease:||Achilles Tendon Rupture|
1. To investigate the collagen turnover in Achilles tendons prior to a rupture.
2. To investigate the relative collagen turnover in Achilles tendons the days immediately
3. To investigate the acute collagen protein synthesis rate in acutely ruptured Achilles
4. To investigate if bacterial DNA is present in samples from the degenerative tissue in
Tendon ruptures are severe injuries that potentially lead to reduced function, reduced
The etiology and pathogenesis of spontaneous tendon ruptures are complex and not fully
Many predisposing factors have been proposed to contribute to the pathological alterations of
Heinemeier et al. found that the bulk of the collagen matrix of healthy human Achilles tendon
Recently, Rolf et al. demonstrated the presence of bacterial DNA in 25% of samples from
In this cross-sectional study, we aim to investigate if bacteria potentially are contributing
Design of the study
The study is conducted as a cross-sectional study. Patients with acute rupture of the
The procedure for the patient:
1. At the day of inclusion, the patients will have taken a blood sample and ingest 150ml of
2. At the day of the surgery, the patient will meet 3 hours before the beginning of the
3. During the surgery, the patients will have taken:
1. Two biopsies from the stump end of the ruptured tendon or ligament.
2. Two control biopsies 2-4 cm proximal to the stump end of the ruptured Achilles
Procedure for biopsies
All biopsies are taken by trained orthopedic surgeons. Every biopsy is taken during surgery
The 8 containers will hold:
1. Biopsy from the stump end of the ruptured tendon for 16S rDNA analysis.
2. Biopsy from the stump end of the ruptured tendon for 14C bomb pulse method
3. Biopsy from the stump end of the ruptured tendon for isotope analysis (deuterium oxide
4. Biopsy from the stump end of the ruptured tendon for mRNA analysis.
5. Biopsy from the ruptured tendon 2-4 cm proximal to the stump end for 16S rDNA analysis.
6. Biopsy from the ruptured tendon 2-4 cm proximal to the stump end for 14C bomb pulse.
7. Biopsy from the ruptured tendon 2-4 cm proximal to the stump end for isotope analysis
8. Biopsy from the ruptured tendon 2-4 cm proximal to the stump end for mRNA analysis
Biopsies from ruptured tendons
The biopsies are taken during open surgery. To prevent contamination of the biopsies the
1. Surgical rub and draping according to normal guidelines.
2. Skin incision and dissection according to normal guidelines.
3. As the ruptured tendon is identified, a new pack containing a sterile scalpel and a
4. 2 biopsies, 10mm long, 1mm broad and 1mm deep, are taken from the degenerated tendon
5. The first biopsy is placed in a container (for bacterial detection). On a sterile board,
6. A second new pack containing a sterile scalpel and a forceps is opened and 2 biopsies 10
7. The first biopsy is placed in a container (for bacterial detection) On a sterile board,
PCR and sequencing - Both a negative and a positive control is included in the test. Numbers
Blood culture - During surgery, blood a sample is send for culture to investigate if bacteria
Bacterial cultures - Bacterial culture tests are conducted on all patients. The investigated
- Patients skin area over the surgical site.
- The blade of the used scalpel.
The swaps are collected in separate glass tubes and send to Department of Microbiology,
Blinding of the analysis
For each included patient, the 2 containers with biopsies for bacterial detection will get a
Data registration during inclusion
- Patient ID
- Social security number
- Phone number
- Date of examination
- Date and time of insult
- Corticosteroid injection (number and time)
- History of infection
|Primary Outcome Measures:||
1. 14C concentration in acutely ruptured Achilles tendons [ Time Frame: The outcome measure will be assesed at time of surgery ]
14C concentration in biopsies from the ruptured part of the tendons and in the control biopsies proximal to the rupture are compared to the known historical values of atmospheric 14C to estimate the rate of collagen turnover. The values for turnover in the biopsies will be compared to the known rates of turnover in healthy tendons.
2. Heavy water (D2O) enrichment in acutely ruptured Achilles tendons [ Time Frame: The outcome measure will be assesed at time of surgery ]
2H isotope enrichment in the biopsies from the ruptured part of the tendons and in the control biopsies proximal to the rupture.
3. Fractional synthesis rate of protein at time of surgery in acutely ruptured Achilles tendons. [ Time Frame: The outcome measure will be assesed at time of surgery ]
The enrichment of the 15N marked proline tracer measured in the biopsies from the ruptured part of the tendons and in the control biopsies and compared to the enrichment of 15N marked proline in the blood. Based on that, the fractional synthesis rate of protein at the time of surgery is calculated.
4. Presence of bacteria in acutely ruptured Achilles tendons. [ Time Frame: The outcome measure will be assesed at time of surgery ]
Presence of bacterial DNA in the biopsies from the ruptured part of the tendons and in the control biopsies proximal to the rupture detected by 16s rDNA PCR
|Biospecimen Retention:||Samples With DNA|
|Biopsies from acutely ruptured Achilles tendonds will be taken, Two biopsies from the ruptured part of the tendon and two biopsies from the healthy tissue of the same tendon proximal to the rupture, as a control.|
|Ages Eligible for Study:||18 to 70 Years (Adult, Older Adult)|
|Sexes Eligible for Study:||All|
|Accepts Healthy Volunteers:||No|
- Age 18-70 years
- Appointment in the Outpatients Department within 4 days after injury.
- Total Acute Achilles tendon rupture
- The patient must be able to speak and understand Danish.
- The patient must be able to give informed consent.
- Rupture of the Achilles tendon either at the insertion on the calcaneus or at musculotendinous junction of the triceps surae.
- Previous rupture of the same Achilles tendon
- Undergone any surgery in the same region as the affected Achilles tendon.
- In medical treatment of diabetes.
- Present infection in the affected region.
- Contra-indication for surgery: severe arthrosclerosis with no palpable pulse in the foot, broken skin in the Achilles region of the injured leg.
- Inability to lie in prone position on the operating table.
- Rheumatoid arthritis or any other inflammatory disorder of the joints.
- Terminal illness or severe medical illness. ASA score higher than or equal to 3.
|Contacts and Locations|
Contact: Allan Cramer +4560174294 firstname.lastname@example.org
|Denmark||Department of Orthopedic Surgery, Copenhagen University Hospital, Hvidovre||Hvidovre|
|Sponsors and Collaborators|
|Copenhagen University Hospital, Hvidovre|
|Study Chair :||Kristoffer W Barfod, MD||Copenhagen University Hospital, Hvidovre|
|Responsible Party :||Copenhagen University Hospital, Hvidovre|
|ClinicalTrials.gov Identifier :||NCT03931486|
|Other Study ID Numbers :||H-18010363|
|First Posted :||April 30, 2019|
|Last Update Posted :||July 14, 2020|
|Last Verified :||July 2020|
Data (IPD) Sharing
|Plan to Share IPD:||Undecided|
|Studies a U.S. FDA-regulated Drug Product:||No|
|Studies a U.S. FDA-regulated Device Product:||No|
|Additional relevant MeSH terms :||