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Bead Based Multiplex Assay for Analysis of Tear Cytokine Profiles
Published on: October 13, 2017
Simultaneous detection of eight cytokines in human dermal wounds with a multiplex bead-based immunoassay for wound
Masataka Takamiya1, Sachiko Fujita, Kiyoshi Saigusa
1Department of Legal Medicine, Iwate Medical University School of Medicine, 19-1 Uchimaru, Morioka, Iwate 020-8505, Japan. mtakamiy@iwate-med.ac.jp
Insights
Multiplex cytokine analysis in human dermal wounds aids in estimating wound age. Specific cytokines like IL-6, IL-8, IFN-gamma, and TNF-alpha show strong expression patterns correlating with healing phases.
Area of Science:
- Forensic pathology
- Dermal wound healing
- Immunology
Background:
- Accurate wound age estimation is crucial in forensic pathology.
- Understanding the dynamic changes in cytokines and cellular infiltration during wound healing is essential.
Purpose of the Study:
- To quantify specific cytokines (IL-2, IL-4, IL-6, IL-8, IL-10, GM-CSF, IFN-gamma, TNF-alpha) in human dermal wounds.
- To analyze dermal cell proliferation and inflammatory cell infiltration for wound age estimation.
- To investigate the correlation between cytokine expression profiles and wound age.
Main Methods:
- Multiplex cytokine analysis of human dermal wound samples.
- Histological analysis of dermal cell proliferation and inflammatory cell infiltration (neutrophils, macrophages, T and B lymphocytes).
- Correlation analysis between cytokine levels, cellular changes, and time post-injury.
Main Results:
- Neutrophils and macrophages infiltrated within 2 hours, peaking at 33-49 hours; T and B lymphocytes infiltrated from 71 hours.
- Fibroblast proliferation and epidermal thickening were observed from 246 hours and 71 hours, respectively.
- Distinct patterns of cytokine expression were noted: IL-10, GM-CSF, IFN-gamma, and TNF-alpha increased early; IL-6 in the middle phase; IL-2, IL-4, and IL-8 from middle to late phases. IL-6, IL-8, IFN-gamma, and TNF-alpha showed strong expression.
Conclusions:
- Multiplex cytokine analysis of wound sites is a viable method for estimating wound age.
- Cytokine profiles provide valuable insights into the temporal dynamics of dermal wound healing.
- The findings are applicable to forensic pathology and other medical fields requiring wound age assessment.
Abstract:
We performed quantification of IL 2, IL 4, IL 6, IL 8, IL 10, GM-CSF, IFN gamma, and TNF alpha in human dermal wounds for wound age estimation. The proliferation of dermal cells and infiltration of inflammatory cells were also analyzed. Neutrophils and macrophages were detected from 2 h post-injury, and strong infiltrations were seen at 33-49 h. T and B lymphocytes also infiltrated simultaneously from 71 h. Strong proliferation of fibroblasts were shown from 246 h, and thickening of the epidermis from 71 h. IL 10, GM-CSF, IFNgamma, and TNF alpha increased from the early phase of dermal wound healing, IL 6 exclusively in the middle phase, IL 2, IL 4, and IL 8 from the middle phase to the late phase. Among the cytokines analyzed in the present study, IL 6, IL 8, IFNgamma, and TNF alpha were strongly expressed. Results of the present study suggest that multiplex cytokine analysis at the wound site can be useful for wound age estimation. In addition, multiplex data obtained from the same sample with a single method would demonstrate more accurate interactions of cytokines during dermal wound healing. Although the present study was oriented to practical forensic pathology, the data obtained would be informative for various fields of medicine.
