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Authors
The Journal of Visualized Experiments (JoVE) is a peer reviewed, PubMed-indexed video journal. Our mission is to increase the productivity of scientific research.
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Department of Anesthesiology, Stanford University School of Medicine
This article has been retracted at the request of the authors due to duplication of text in J Pain (Carvalho B, Clark DJ, Angst MS (2008) Local and Systemic Release of Cytokines, Nerve Growth Factor, Prostaglandin E2, and Substance P in Incisional Wounds and Serum Following Cesarean Delivery. J Pain 9: 650-657.) due to an honest mistake of the authors.
Important: There has been an erratum issued for this article. Read more…
Carvalho, B., Clark, D. J., Yeomans, D., Angst, M. S. Collecting and Measuring Nociceptive and Inflammatory Mediators in Surgical Wounds . J. Vis. Exp. (20), e962, doi:10.3791/962 (2008).
De doelstellingen van deze studie zijn om de haalbaarheid van het verzamelen en het meten van inflammatoire en nociceptieve biochemische bemiddelaars van de operatiewond site te testen, om de relatie tussen de wond en serum niveaus te evalueren, en om eventuele associaties tussen mediator release, pijn en pijnstillende consumptie na keizersnede te bepalen levering. Twintig gezonde vrouwen die een electieve keizersnede met een spinale anesthesie werden ingeschreven. Wondvocht en serum mediatoren, pijnscores en analgetica verbruik werden gemeten op 1, 6, 24, en 48 uur na de keizersnede. In wondvocht, werden 19 van de 20 mediatoren betrouwbaar gedetecteerd waaronder IL-1β, IL-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL -17, TNFa, INFγ, G-CSF, GM-CSF, MCP-1 en MIP-1β, zenuw groeifactor (NGF), prostaglandine E2 (PG-E2) en substantie P. Wound PG-E2 en diverse cytokines piekte begin van de , terwijl NGF toonde een meer vertraagde afgifte. Er waren geen correlaties tussen de concentratie versus tijd profiel van de wond en serum cytokines. Deze studie toont de haalbaarheid van het verzamelen en meten van nociceptieve en inflammatoire mediatoren in chirurgische wonden op specifieke tijdstippen. Het gebrek aan significante correlaties tussen de wond en de serumspiegels benadrukt het belang van het bepalen van site-specifieke versie indien gelokaliseerd pathologieën moeten worden bestudeerd.
Nociceptieve en inflammatoire biochemische mediator collectie
Assay analyse
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De On-Q ® PainBuster ® Hulp van de Pijn-systeem moet worden ingevoegd over de hele incisie in subcutane laag vlak voor sluiting wond. Dit vergemakkelijkt aspiratie via de drieweg-kraan op de aangegeven tijd intervallen. De On-Q ® systeem levert continu fysiologische zoutoplossing subcutaan in de wond met een snelheid van 2 ml / h. Dit voorkomt dat de katheter stolling en verbetert de betrouwbaarheid van het systeem om exsudaat monsters te produceren. Als aspiratie van exsudaat is moeilijk (ongeveer 5% van de gevallen), overweeg dan het veranderen van het onderwerp positie (bijvoorbeeld, zitten de patiënt omhoog of liggend ze plat), zachtjes boven de wond, met behulp van een 0,5-1ml fysiologisch zout flush of intrekking van de katheter 1-2 cm.
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Dr Carvalho Het werk wordt ondersteund door een gebouw Interdisciplinair vacatures in de gezondheid van vrouwen onderzoek subsidie van het Office of Research op de gezondheid van vrouwen en de National Institute of Child Health and Human Development van de National Institutes of Health (5K12 HD043452). Dr Angst levert (de On-Q ® PainBuster ® Post-Op Hulp van de Pijn System) en de financiering ontvangen voor de uitvoering van de biochemische testen van I-Flow (Lake Forest, CA).
| Name | Company | Catalog Number | Comments |
| On-Q® PainBuster® Post-Op Pain Relief System | I-Flow, Lake Forest, CA | ||
| Complete proteinase inhibitor | Roche Group | ||
| 17-multiplex bead immunoassay Bio-PlexTM plate | Bio-Rad | ||
| Bio-Plex amine coupling kit | Bio-Rad | ||
| The NGF antibody DY256 | R&D Systems | ||
| Prostaglandin E2 and substance P ELISA Kits | Assay Designs |
1. Elshal MF, McCoy JP. Multiplex bead array assays: performance evaluation and comparison of sensitivity to ELISA. Methods. 38, 317-323 (2006).
2. Heijmans-Antonissen C, Wesseldijk F, Munnikes RJ, Huygen FJ, van der Meijden P, Hop WC, Hooijkaas H, Zijlstra FJ. Multiplex bead array assay for detection of 25 soluble cytokines in blister fluid of patients with complex regional pain syndrome type 1. Mediators Inflamm. 2006, 28398, 1-8 (2006).
3. Buvanendran A, Kroin JS, Berger RA, Hallab NJ, Saha C, Negrescu C, Moric M, Caicedo MS, Tuman KJ. Upregulation of prostaglandin E2 and interleukins in the central nervous system and peripheral tissue during and after surgery in humans. Anesthesiology. 104, 403-410 (2006).
4. Holzheimer RG, Steinmetz W. Local and systemic concentrations of pro- and anti-inflammatory cytokines in human wounds. Eur J Med Res. 5, 347-355 (2000).
5. Carvalho, B., Clark, D. J. & Angst, M. S. Local and Systemic Release of Cytokines, Nerve Growth Factor, Prostaglandin E2, and Substance P in Incisional Wounds and Serum Following Cesarean Delivery. The Journal of Pain : official journal of the American Pain Society 9 (7), 650-657 (2008).
Formal Correction: Erratum: Collecting and Measuring Nociceptive and Inflammatory Mediators in Surgical Wounds
Posted by JoVE Editors on 03/07/2012.
Citeable Link.
A correction was made to: Collecting and Measuring Nociceptive and Inflammatory Mediators in Surgical Wounds. A key reference was excluded.
A fifth reference:
5. Carvalho, B., Clark, D. J. & Angst, M. S. Local and Systemic Release of Cytokines, Nerve Growth Factor, Prostaglandin E2, and Substance P in Incisional Wounds and Serum Following Cesarean Delivery. The Journal of Pain : official journal of the American Pain Society 9 (7), 650-657 (2008).
was added. The abstract was updated to :
We describe a methodology by which we are able to collect and measure inflammatory and nociceptive biochemical mediators at the surgical wound site. Collecting site-specific biochemical markers allows us to evaluate the relationship between surgical wound and serum levels; determine any associations between mediator release, pain and analgesic consumption; and evaluate the effect of systemic and peripheral drug administration on surgical wound biochemistry.
This methodology has been applied to healthy women undergoing elective cesarean delivery with spinal anesthesia. Wound exudate and serum mediators, in conjunction with pain scores and analgesics consumption were measured at 1, 6, 24, and 48 hours post-cesarean delivery. Biochemical mediators that were detected included IL-1β, IL-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, TNFα, INFγ, G-CSF, GM-CSF, MCP-1 and MIP-1β, nerve growth factor (NGF), prostaglandin E2 (PG-E2) and substance P. We found no correlations between wound and serum cytokines concentrations or time-release profiles (J Pain. 2008 Jul 9(7):650-7). This article describes and demonstrates the feasibility of collecting and assaying nociceptive and inflammatory mediators in surgical wounds at specific time points. The lack of significant correlations between serum and wound levels shows the importance of determining site-specific release if surgical wounds and localized pathologies are to be studied.
from
The objectives of this study were to test the feasibility of collecting and measuring inflammatory and nociceptive biochemical mediators at the surgical site; to evaluate the relationship between wound and serum levels; and to determine any associations between mediator release, pain and analgesic consumption post-cesarean delivery. Twenty healthy women undergoing elective cesarean delivery with spinal anesthesia were enrolled. Wound exudate and serum mediators, pain scores and analgesics consumption were measured at 1, 6, 24, and 48 hours post-cesarean. In wound exudate, 19 out of 20 mediators were reliably detected including IL-1β, IL-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, TNFα, INFγ, G-CSF, GM-CSF, MCP-1 and MIP-1β, nerve growth factor (NGF), prostaglandin E2 (PG-E2) and substance P. Wound PG-E2 and various cytokines peaked early, whereas NGF showed a more delayed release. There were no correlations between the concentration versus time profile of wound and serum cytokines. This study demonstrates the feasibility of collecting and measuring nociceptive and inflammatory mediators in surgical wounds at specific time points. The lack of significant correlations between wound and serum levels emphasizes the importance of determining site-specific release if localized pathologies are to be studied.
The technique appears robust, even if intuitive.
However, as shown in the video paper, the data are collected in the presence of superfusion with local anaesthetic solution (in this case bupivacaine) which is very likely to modify the response and the local concentrations of mediators both as a direct pharmacological action of the local anaesthetic as well as through the fluid diluent effect. Moreover, adsoprtion of the various substances by the medical plastics would need evaluation. Hence a lot of control work needs to be done so that the numerical results obtained can be interpreted correctly. Perhaps the data are semiquantitative at best.
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ReplyPosted by: Laurence E Mather, Emeitus Professor of AnaesthesiaMarch 4, 2009, 6:37 PM