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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This article has been retracted at the request of the authors due to duplication of text in J Pain (Carvalho B, Clark DJ, Angst MS (2008) Local and Systemic Release of Cytokines, Nerve Growth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).
Målen för denna studie var att testa möjligheterna att samla in och mäta inflammatoriska och nociceptiv biokemiska mediatorer i operationsområdet, för att utvärdera förhållandet mellan såret och serum, och för att fastställa eventuella samband mellan medlaren release, smärta och smärtstillande konsumtion efter kejsarsnitt leverans. Tjugo friska kvinnor som genomgår elektiv kejsarsnitt med spinal anestesi inkluderades. Sårvätska och serum medlare, poäng smärta och analgetika konsumtion mättes vid 1, 6, 24 och 48 timmar efter kejsarsnitt. I sårvätska, var 19 av 20 medlare upptäckas tillförlitligt inklusive IL-1β, IL-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL -17, TNF, INFγ, G-CSF, GM-CSF, MCP-1 och MIP-1β, nerv tillväxtfaktor (NGF), prostaglandin E2 (PG-E2) och substans P. Wound PG-E2 och olika cytokiner nådde tidigt , medan NGF visade en mer fördröjd frisättning. Det fanns inga samband mellan koncentrationen mot tiden profil sår och serum cytokiner. Denna studie visar på möjligheterna att samla in och mäta nociceptiv och inflammatoriska mediatorer i kirurgiska sår vid specifika tidpunkter. Avsaknaden av signifikanta korrelationer mellan såret och serum betonar vikten av att fastställa platsspecifika utsläppet om lokaliserade sjukdomstillstånd ska studeras.
Nociceptiv och inflammatoriska biokemiska medlare samling
Analys analys
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On-Q ® PainBuster ® Smärta system bör införas över hela snittet i subkutana vävnaden strax före tillslutning av sår. Detta underlättar strävan via trevägskran vid bestämda tidpunkter. On-Q ®-system levererar kontinuerligt koksaltlösning subkutant i såret med en hastighet av 2 ml / h. Detta förhindrar att katetern koagulering och förbättrar tillförlitligheten i systemet för att producera exsudat prover. Om aspiration av sårvätska är svårt (ca 5% av fallen), överväga att ändra motivets position (t.ex., sitter patienten upp eller ligger dem platt), pressa försiktigt över såret med en 0,5-1ml koksaltlösning spola eller dra tillbaka katetern 1-2 cm.
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Dr Carvalho arbete stöds av en byggnad Tvärvetenskap Karriär i kvinnors hälsa forskningsanslag från Office of Forskning om kvinnors hälsa och National Institute of Child Health och Human Utveckling av National Institutes of Health (5K12 HD043452). Dr Angst fick förnödenheter (On-Q ® PainBuster ® Post-Op Smärtlindring System) och finansiering för att genomföra de biokemiska analyser från I-Flow (Lake Forest, Kalifornien).
| 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.
1
ReplyPosted by: Laurence E Mather, Emeitus Professor of AnaesthesiaMarch 4, 2009, 6:37 PM