Self-assembling multilayer MSC-sheet promotes wound healing increasing M2 macrophage polarization

Area of Science:

• Regenerative Medicine
• Biomaterials Science
• Wound Healing Research

Background:

• Mesenchymal stem cell (MSC) therapy shows potential for chronic wound treatment.
• Limited MSC retention and cell density at the wound site hinder therapeutic efficacy.
• Novel delivery platforms are needed to improve MSC-based wound healing.

Purpose of the Study:

• To evaluate the therapeutic potential of a self-assembling multilayer MSC-sheet in a murine wound model.
• To compare the efficacy of MSC-sheets versus MSC-suspensions in promoting wound healing and modulating the immune microenvironment.
• To investigate the impact of MSC-sheets on macrophage infiltration and polarization.

Main Methods:

• Preparation and characterization of multilayer MSC-sheets from human adipose tissue.
• In vivo assessment in a murine full-thickness skin defect model using MSC-sheets, MSC-suspensions, and control.
• Evaluation of wound healing parameters (area, neoepithelialization, granulation tissue, angiogenesis) and cell retention using histological and molecular techniques.
• Analysis of macrophage infiltration and polarization (M1/M2) via immunohistochemistry and RT-PCR.

Main Results:

• MSC-sheets exhibited a multilayer structure with extracellular matrix (ECM).
• Both MSC-sheet and MSC-suspension accelerated wound closure and improved angiogenesis compared to control.
• MSC-sheets demonstrated superior MSC retention and enhanced M2 macrophage polarization compared to MSC-suspensions.
• RNA sequencing revealed MSC-sheet associated pathways for wound healing, anti-inflammation, and angiogenesis.

Conclusions:

• The self-assembling multilayer MSC-sheet is a promising scaffold-free platform for chronic wound therapy.
• MSC-sheets improve therapeutic outcomes by enhancing cell retention and modulating macrophage polarization towards an M2 phenotype.
• This technology holds potential for scalable and clinically applicable translation in regenerative medicine.