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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.
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This article has been retracted at the request of the authors due to duplication of text in J Pain (Carvalho B, Clark DJ, Angst MS (2008) Local and Systemic Release of Cytokines, Nerve Growth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).
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.
Nociceptive and inflammatory biochemical mediator collection
Assay analysis
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The On-Q® PainBuster® Pain Relief System should be inserted across the entire incision in subcutaneous layer just prior to wound closure. This facilitates aspiration via the three-way stopcock at the specified time intervals. The On-Q® system continuously delivers normal saline subcutaneously into the wound at a rate of 2 ml/h. This prevents the catheter clotting and improves the reliability of the system to produce exudate samples. If aspiration of exudate is difficult (approximately 5% of cases), consider changing the subject's position (e.g., sitting the patient up or lying them flat), pushing gently above the wound, using a 0.5-1ml normal saline flush or withdrawing the catheter 1-2 cm.
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The authors have nothing to disclose.
Dr. Carvalho’s work is supported by a Building Interdisciplinary Careers in Women's Health research grant from the Office of Research on Women's Health and National Institute of Child Health and Human Development of the National Institutes of Health (5K12 HD043452). Dr Angst received supplies (the On-Q® PainBuster® Post-Op Pain Relief System) and funding for the conducting the biochemical assays from 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