Investigating Composite Biomarkers for Pain Catastrophizing

Study Overview
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Brief Summary
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Pain is a complex, multidimensional, and subjective experience; and although, investigators use a single word pain, to describe our perception, multiple mechanisms contribute to the generation and maintenance of pain. To help diagnosing and improving pain management, there is a need for developing tools. These tools may include measurements of substances, or biomarkers, in the blood; e.g. small molecules called microRNA and proteins. In these experiments, the investigators would like to investigate how the psychological response to stress and pain alters the impulses in the brain and the content of microRNA and proteins in the blood. The future aim is to identify patients in high risk of developing and maintaining chronic pain and to be able to treat chronic pain efficiently.

Official Title
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Investigating Composite Biomarkers for Pain Catastrophizing

Conditions
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Pain, Acute

Intervention / Treatment
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* Drug: hypertonic saline
* Drug: isotonic saline


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Healthy men and women in the age 18-80 years Speak and understand English Exclusion Criteria: Acute and chronic pain Pregnancy or breastfeeding Drug addiction defined as the use of cannabis, opioids or other drugs Present or previous history of neurological, dermatological, immunological, musculoskeletal, cardiac disorder or mental illnesses that may affect the results (e.g. Neuropathy, muscular pain in the upper extremities, etc.) Focal and generalized seizure Surgery or any other therapy for epilepsy Present or previous AEDs (anti-epileptic drugs) administration Present or previous use of epileptic devices (<1 year prior the enrolment) Lack of ability to cooperate Current use of medications that may affect the trial, such as antipsychotics and pain killers as well as systemic or topical steroids and anti-inflammatory drugs. Skin diseases Consumption of alcohol or painkillers 24 hours before the study days and between these Participation in other trials within 1 week of study entry (4 weeks in the case of pharmaceutical trials)

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 80 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

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: Parallel Assignment
Masking: Single


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: hypertonic saline<br> | Drug: hypertonic saline<br>* The site of injections will be determined by palpation of the contracted First Dorsal Interosseus (FDI) muscle. The skin will be cleaned with alcohol before injection. A bolus injection of hypertonic saline (7% NaCl) will be administered to the FDI muscle using a 1 mL syringe with a disposable needle (27G), and 30 the volume of the bolus will be 0.2 mL .<br>|
| Placebo Comparator: isotonic saline<br> | Drug: isotonic saline<br>* The site of injections will be determined by palpation of the contracted First Dorsal Interosseus (FDI) muscle. The skin will be cleaned with alcohol before injection. A bolus injection of 0.2 mL isotonic saline (9 mg/mL) will be administered to the FDI muscle as control.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Oscillations of the main electroencephalogram (EEG) frequency | Electroencephalographic (EEG) recording will be performed placing a 64-electrodes cap over the scalp according to the 10-20 international system. Oscillations of the main EEG frequency bands (delta, alpha, beta, gamma) from the frontal and parietal lobes will be detected and correlated to the individual pain rate as well as the dimensions of the psychological questionnaires. | 10 minutes of recording before isotonic/hypertonic injection | 
| Oscillations of the main electroencephalogram (EEG) frequency | Electroencephalographic (EEG) recording will be performed placing a 64-electrodes cap over the scalp according to the 10-20 international system. Oscillations of the main EEG frequency bands (delta, alpha, beta, gamma) from the frontal and parietal lobes will be detected and correlated to the individual pain rate as well as the dimensions of the psychological questionnaires. | 10 minutes of recording after isotonic/hypertonic injection | 
| Perturbation of the electroencephalogram (EEG) rhythms | Electroencephalographic (EEG) recording will be performed placing a 64-electrodes cap over the scalp according to the 10-20 international system. Perturbation of the above-mentioned EEG rhythms will be measured in response to the hypertonic/isotonic saline injection and compared to a resting state baseline recording. | 10 minutes of recording before isotonic/hypertonic injection | 
| Perturbation of the electroencephalogram (EEG) rhythms | Electroencephalographic (EEG) recording will be performed placing a 64-electrodes cap over the scalp according to the 10-20 international system. Perturbation of the above-mentioned EEG rhythms will be measured in response to the hypertonic/isotonic saline injection and compared to a resting state baseline recording. | 10 minutes of recording after isotonic/hypertonic injection | 
| Collection of blood samples | Whole blood samples (5 mL per each time point, a total of 20 mL per subject) will be collected into SST™ II Advance Serum Separation Tubes containing anticoagulant EDTA. | time 0 (baseline) | 
| Collection of blood samples | Whole blood samples (5 mL per each time point, a total of 20 mL per subject) will be collected into SST™ II Advance Serum Separation Tubes containing anticoagulant EDTA. | Time 1 (3 hours) | 
| Collection of blood samples | Whole blood samples (5 mL per each time point, a total of 20 mL per subject) will be collected into SST™ II Advance Serum Separation Tubes containing anticoagulant EDTA. | time 2 (24 hours) | 
| Collection of blood samples | Whole blood samples (5 mL per each time point, a total of 20 mL per subject) will be collected into SST™ II Advance Serum Separation Tubes containing anticoagulant EDTA. | time 3 (48 hours) | 
| Collection of blood samples | Whole blood samples (5 mL per each time point, a total of 20 mL per subject) will be collected into SST™ II Advance Serum Separation Tubes containing anticoagulant EDTA. | time 4 (72 hours) | 
| MicroRNAs (miRNAs) expression analysis | From the blood samples previously collected, miRNA will be isolated for a subsequent miRNA library preparation. Subsequently, extracted miRNA will be reversely transcribed, and cDNA will be used for total miRNAs sequencing. | time 0 (baseline) | 
| MicroRNAs (miRNAs) expression analysis | From the blood samples previously collected, miRNA will be isolated for a subsequent miRNA library preparation. Subsequently, extracted miRNA will be reversely transcribed, and cDNA will be used for total miRNAs sequencing. | time 1 (3 hours) | 
| MicroRNAs (miRNAs) expression analysis | From the blood samples previously collected, miRNA will be isolated for a subsequent miRNA library preparation. Subsequently, extracted miRNA will be reversely transcribed, and cDNA will be used for total miRNAs sequencing. | time 2 (24 hours) | 
| MicroRNAs (miRNAs) expression analysis | From the blood samples previously collected, miRNA will be isolated for a subsequent miRNA library preparation. Subsequently, extracted miRNA will be reversely transcribed, and cDNA will be used for total miRNAs sequencing. | time 3 (48 hours) | 
| MicroRNAs (miRNAs) expression analysis | From the blood samples previously collected, miRNA will be isolated for a subsequent miRNA library preparation. Subsequently, extracted miRNA will be reversely transcribed, and cDNA will be used for total miRNAs sequencing. | time 4 (72 hours) | 
| Proteome analysis | From the blood samples previously collected, the protein concentration will be determined. Protein sequencing will be done using mass spectrometers. | time 0 (baseline) | 
| Proteome analysis | From the blood samples previously collected, the protein concentration will be determined. Protein sequencing will be done using mass spectrometers. | time 1(3 hours) | 
| Proteome analysis | From the blood samples previously collected, the protein concentration will be determined. Protein sequencing will be done using mass spectrometers. | time 2 (24 hours) | 
| Proteome analysis | From the blood samples previously collected, the protein concentration will be determined. Protein sequencing will be done using mass spectrometers. | time 3 (48 hours) | 
| Proteome analysis | From the blood samples previously collected, the protein concentration will be determined. Protein sequencing will be done using mass spectrometers. | time 4 (72 hours) | 
| Metabolome analysis | From the blood samples previously collected. Metabolites will be investigated and identified by a 4D feature finding using Metaboscape. | time 0 (baseline) | 
| Metabolome analysis | From the blood samples previously collected. Metabolites will be investigated and identified by a 4D feature finding using Metaboscape. | time 1(3 hours) | 
| Metabolome analysis | From the blood samples previously collected. Metabolites will be investigated and identified by a 4D feature finding using Metaboscape. | time 2 (24 hours) | 
| Metabolome analysis | From the blood samples previously collected. Metabolites will be investigated and identified by a 4D feature finding using Metaboscape. | time 3 (48 hours) | 
| Metabolome analysis | From the blood samples previously collected. Metabolites will be investigated and identified by a 4D feature finding using Metaboscape. | time 4 (72 hours) | 
| Plasma cortisol levels measurement | From the blood samples previously collected, plasma levels of cortisol will be evaluated through enzyme-linked immunosorbent assay (ELISA). | time 0 (baseline) | 
| Plasma cortisol levels measurements | From the blood samples previously collected, plasma levels of cortisol will be evaluated through enzyme-linked immunosorbent assay (ELISA). | time 1(3 hours) | 
| Plasma cortisol levels measurements | From the blood samples previously collected, plasma levels of cortisol will be evaluated through enzyme-linked immunosorbent assay (ELISA). | time 2 (24 hours) | 
| Plasma cortisol levels measurements | From the blood samples previously collected, plasma levels of cortisol will be evaluated through enzyme-linked immunosorbent assay (ELISA). | time 3 (48 hours) | 
| Plasma cortisol levels measurements | From the blood samples previously collected, plasma levels of cortisol will be evaluated through enzyme-linked immunosorbent assay (ELISA). | time 4 (72 hours) | 
| Plasma Interleukin-6 (IL-6) levels measurements | From the blood samples previously collected, plasma levels of IL-6 will be evaluated through enzyme-linked immunosorbent assay (ELISA). | time 0 (baseline) | 
| Plasma Interleukin-6 levels measurements | From the blood samples previously collected, plasma levels of IL-6 will be evaluated through enzyme-linked immunosorbent assay (ELISA). | time 1(3 hours) | 
| Plasma Interleukin-6 levels measurements | From the blood samples previously collected, plasma levels of IL-6 will be evaluated through enzyme-linked immunosorbent assay (ELISA). | time 2 (24 hours) | 
| Plasma Interleukin-6 levels measurements | From the blood samples previously collected, plasma levels of IL-6 will be evaluated through enzyme-linked immunosorbent assay (ELISA). | time 3 (48 hours) | 
| Plasma Interleukin-6 levels measurements | From the blood samples previously collected, plasma levels of IL-6 will be evaluated through enzyme-linked immunosorbent assay (ELISA). | time 4 (72 hours) | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Measuring Pain using VAS | The subject will rate the pain intensity continuously for 20 minutes and VAS will start from zero (0), representing no pain, and will end at one hundred (100) representing worst pain imaginable. | 20 minutes |