Safety and efficacy of dermal fibroblast conditioned medium (DFCM) fortified collagen hydrogel as acellular 3D skin patch
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Skin substitutes are one of the main treatments for skin loss, and a skin substitute that is readily available would be the best treatment option. However, most cell-based skin substitutes require long production times, and therefore, patients endure long waiting times. The proteins secreted from the cells and tissues play vital roles in promoting wound healing. Thus, we aimed to develop an acellular three-dimensional (3D) skin patch with dermal fibroblast conditioned medium (DFCM) and collagen hydrogel for immediate treatment of skin loss. Fibroblasts from human skin samples were cultured using serum-free keratinocyte-specific media (KM1 or KM2) and serum-free fibroblast-specific medium (FM) to obtain DFCM-KM1, DFCM-KM2, and DFCM-FM, respectively. The acellular 3D skin patch was soft, semi-solid, and translucent. Collagen mixed with DFCM-KM1 and DFCM-KM2 showed higher protein release compared to collagen plus DFCM-FM. In vitro and in vivo testing revealed that DFCM and collagen hydrogel did not induce an immune response. The implantation of the 3D skin patch with or without DFCM on the dorsum of BALB/c mice demonstrated a significantly faster healing rate compared to the no-treatment group 7 days after implantation, and all groups had complete re-epithelialization at day 17. Histological analysis confirmed the structure and integrity of the regenerated skin, with positive expression of cytokeratin 14 and type I collagen in the epidermal and dermal layer, respectively. These findings highlight the possibility of using fibroblast secretory factors together with collagen hydrogel in an acellular 3D skin patch that can be used allogeneically for immediate treatment of full-thickness skin loss.
KeywordsFibroblasts Dermal fibroblast conditioned medium Acellular 3D skin patch Tissue engineering
We are thankful to our colleague Dr. Shinsmon Jose, who provided expertise that greatly assisted part of the research. Some parts of this work were performed at the UKM Bioserasi Laboratory.
Compliance with ethical standards
This study was funded by the Science Fund (02–01-02-SF0964), UKM fundamental fund (FF-2015-204) and the Tissue Engineering Centre, UKM Medical Centre.
Ethics approval and consent to participate
All procedures performed in studies involving human participants were in accordance with the ethical standards of the responsible committee on human experimentation (UKMREC), with approval code UKM FPR.4/244/FF-2015-204, and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all patients included in the study. All institutional and national guidelines for the care and use of laboratory animals were followed with approval code PP/TEC/2015/SHIPLU/20-MAY/675-MAY-2015-DEC-2016.
Conflict of interest
The authors declare that they have no conflicts of interest.
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