Effect of Mesenchymal Stem Cell Conditioned Media on Fibroblasts: Implications for Skin Regeneration and Wound Healing

Abstract

Introduction

Stem cell-based therapies show great promise in regenerative medicine, with bone marrow-derived mesenchymal

stem cells (BM-MSC) being particularly noteworthy for their high multilineage differentiation and paracrine

signaling. MSC, derived from sources like cord blood, adipose tissue and bone marrow, are multipotent cells

capable of differentiating into various tissues as well as known for their regenerative and immunomodulatory

properties. Conditioned media from MSC (MSC-CM), rich in bioactive factors, offers a practical therapeutic

alternative, reducing immune reactions and ensuring consistent effects. MSC-CM has shown promise in wound

healing, skin rejuvenation, and other skin applications. In this study, we investigated the effects of BM-MSC-CM

on human dermal fibroblasts (HDFs), focusing on their proliferation, migration, and the promotion of collagen

and elastin synthesis. The research underscores the potential of BM-MSC-CM in developing safer, more effective

regenerative therapies for skin applications including skin regeneration, wound healing and anti-aging.

Methods

Flow cytometry evaluated BM-MSC markers using antibodies for CD73, CD105, CD90 and CD34, CD45. BM-

MSC-conditioned media (CM) was prepared by centrifuging and filtering media collected after 48 hours.

Superficial and deep tissue dermal fibroblasts were treated with 5%, 10%, and 20% BM-MSC-CM. Proliferation

was assessed with Alamar Blue. Migration was measured using a scratch wound healing assay in confluent

fibroblasts in 6-well plates, with healing observed and analyzed using ImageJ from day 0 to 3. Collagen content

was measured using Condrex Sirius Total Collagen Kit, and elastin content was measured using Fastin Kit.

Results

Flow cytometry results showed BM-MSC demonstrate the expected phenotype for human MSC (highly positive

expression of CD105, CD73, CD90, and negative expression of CD34 and CD45). The Alamar Blue assay

demonstrated that BM-MSC-CM significantly improved proliferation of fibroblasts compared to the control

conditioned media. The scratch wound healing assay revealed that BM-MSC-CM at 5%, 10%, and 20% enhanced

migration of fibroblasts (resulted in closure of the gap) with respect to control.

Conclusions

This study demonstrated the remarkable potential of BM-MSC-CM for enhancing the functional properties of

HDFs, which are crucial for skin regeneration and wound healing. However, further studies are needed to

evaluate/identify the key bioactive factors in BM-MSC-CM as well as their effects on skin tissue repair and wound

healing in vivo.