Protocol for an Analytical Study With Microbiological, Phenotypic, Genotypic and Multiomics Techniques

Study Overview
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Brief Summary
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The study aims to identify metabolites present in intervertebral discs colonized by C. acnes from patients with low back pain and degenerative disc disease, correlating them with their clinical, radiological and demographic profiles.

Official Title
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Cutibacterium Acnes Colonization on Intervertebral Discs of Patients With Low Back Pain: Protocol for an Analytical Study With Microbiological, Phenotypic, Genotypic and Multiomics Techniques

Conditions
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Back Pain, Low, Degenerative Disc Disease

Intervention / Treatment
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* Procedure: Lumbar Spinal Fusion Surgery
* Biological: Bacterial culture
* Genetic: Phenotypic and genotypic characterization
* Genetic: Multiomics analysis


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: low back pain lasting longer than 3 months; MRI findings of degenerative lumbar discopathy (DDL) performed less than 6 months after inclusion in the study; indication for open surgical treatment, with isolated microdiscectomy or associated with lumbar arthrodesis; failure of conservative treatment for at least 6 weeks and/or progressive neurological deficit Exclusion Criteria: History of open lumbar spine surgery at any stage of life; presence of chemotherapy or pulse therapy with corticoids; patients with immune deficiency; patients undergoing previous intradiscal therapies (nucleotomy or discography); patients undergoing previous endoscopic surgery; patients with a history of previous spinal infection treated with antibiotics in the 6 months prior to inclusion in the study; use of antibiotics in the 2 months prior to the surgical procedure

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 65 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Factorial Assignment
Masking: Quadruple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: surgical treatment<br>Lumbar Spinal Fusion Surgery | Procedure: Lumbar Spinal Fusion Surgery<br>* During surgery will be collected five disc fragments and five fragments of muscle-ligament tissue adjacent to the collected disc.<br>|
| Experimental: Bacterial culture<br>Cutibacterium acnes culture in two different atmosphere: Carbon dioxide (CO2) from 5% up to 10% CO2 10%, H2 10% and N2 80% | Biological: Bacterial culture<br>* Samples will be seeded on blood agar and anaerobic blood agar plates. After sowing in solid media, the swab will be inoculated in thioglycolate medium, where it will remain for 14 days<br>|
| Experimental: Phenotypic and genotypic characterization<br>Mass spectrometry and (Polimerase Chain Reaction) PCR | Genetic: Phenotypic and genotypic characterization<br>* The phenotypic identification of the species will be carried out through mass spectrometry, with the MALDI-TOF equipment. PCR analyzes performed in two phases, the first to confirm the presence of bacteria (target and non-target) using primers capable of amplifying eubacteria ribosomal deoxyribonucleic acid (rDNA) 16s and the second directed to confirm C. acnes through specific rDNA 16s primers.<br>|
| Experimental: Multiomics analysis<br>Genome sequencing Proteomics Metabolomics | Genetic: Multiomics analysis<br>* Whole Genome Sequencing, proteomics and metabolomics analysis<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Incidence of C. acnes on intervertebral disc | Incidence of C. acnes on intervertebral disc in patients with chronic low back pain and DDL. | Up to 5 months |