Journal of Regenerative Medicine & Tissue Engineering

Journal of Regenerative Medicine & Tissue Engineering

ISSN 2050-1218
Original Research

Application of mitogen-activated protein kinase inhibitor SP 600125 for wound healing control

Irina A Shurygina1,2*, Michael G Shurygin1,2†, Galina B Granina1† and Nikolay V Zelenin1†

*Correspondence: Irina A Shurygina irinashurygina@gmail.com

These authors contributed equally to this work.

1. Scientific Center of Reconstructive and Restorative Surgery SB RAMS, Irkutsk, Russia.

Author Affiliations

2. Irkutsk Scientific Center SB RAS, Irkutsk, Russia.

Abstract

Background: The problem of wound healing remains critical due to lack of methods for direct control of regeneration processes.

Main purpose of the study: To investigate the ability to control wound healing process by inhibiting JNK mitogenactivated protein kinase.

Methods: Skin-muscle wounds were made to Wistar rats 9 months old, 220-250 g weight. Two groups were examined, each one consisted of 30 animals: control group (inoculation of placebo plate); JNK group (inoculation of the inhibitor of JNK MAPK cascade SP 600125, the dosage per one animal was 2,6 µg/kg delivered in the form of original medicated film). The animals were taken out of the experiment in the periods from 2 hours up to 30 days. The segments of the skin-muscle wound were subjected to histological, immunohistochemical (staining for Col1A1, actin, CD34) and immunofluorescent (staining for CD34, CD45, ММР9) investigations. We measured the ratio of section area occupied by collagen fibers to the total area of tissue in micrographs our proposed method. The analysis of the contingence tables and Mann–Whitney U test were used during the investigation.

Results: The study revealed that application of JNK MAPK inhibitor alters the wound healing process considerably, stimulates early generation of connective tissue, increases its production rate and accelerates maturation of connective tissue in the wound area. Application of JNK MAPK inhibitor impacts considerably marker expression of various fibroblasts, causing higher expression of their marrow precursors in the focus, which is indicative of more active attraction of progenitor cells into the connective tissue generation zone. Early application of JNK MAPK inhibitor increases quantity of myofibroblasts in the wound and increases expression of procollagen in fibroblasts located in the zone of connective tissue formation. The risk of keloid scarring remains small because there is an early completion of the scar formation and in this area there no cells that are capable for prolonged intensive collagen synthesis. The results enable to consider local inhibition of JNK MAPK cascade associated with a wound process to be a prospective approach to acceleration of surgical wound healing.

Conclusions: It has thus been established that the application of MAPK inhibitor considerably affects marker expression of various fibroblasts, which increases the expression of marrow precursors in the focus, being indicative of a more active attraction of progenitor cells into connective tissue formation zone. The developed original method of local application of JNK MAPK inhibitor SP 600125 to surgical wound, which enables to modify the surgical wound healing process with a single application, is recommended for pre-clinical and clinical trials for further wide clinical application.

Keywords: Connective tissue, JNK mitogen-activated protein kinases, wounds and injuries

ISSN 2050-1218
Volume 2
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