Journal Article

Estrogen Enhances Endothelial Differentiation and Angiogenic Function of Adipose-Derived Stromal Cells to Improve Therapeutic Outcomes in Critical Limb Ischemia

Hsiao, Chang-Chun, Chiang, Hsin-Ju, Leu, Steve

2026

Overview

Background: Aging, especially after menopause, reduces the quantity and function of adult stem cells. Estrogen deficiency impairs proliferation, differentiation, and regenerative capacity. This study evaluated whether estrogen enhances endothelial differentiation of adipose-derived stromal cells (ADSCs) and improves therapeutic efficacy in critical limb ischemia (CLI). Methods: Male-derived ADSCs were assessed in vitro for endothelial differentiation using flow cytometry, biochemical assays, and angiogenesis analyses. Therapeutic effects were tested in a rat CLI model using endothelial-differentiated ADSCs (ED-ADSCs) with or without 17β-estradiol (E2). An ovariectomized (OVX) model examined estrogen deficiency and supplementation in vivo. Results: E2 promoted endothelial differentiation, increasing ERα/ERβ expression and activating PI3K/Akt/eNOS and MAPK signaling. This led to elevated VEGF expression, enhanced tube formation, and increased CD34+, KDR+, and CD31+ cell populations. In vivo, E2-pretreated ED-ADSCs significantly improved blood flow recovery. Estrogen deficiency reduced endothelial progenitor populations, which were restored by E2 supplementation. Conclusions: Estrogen modulates endothelial-associated characteristics and angiogenesis-related responses of ADSCs via ER-associated signaling, and may contribute to improved functional outcomes in ischemic conditions. E2 preconditioning may represent a potential strategy for stem cell-based therapy in estrogen-deficient settings.

• Publisher: MDPI AG,Multidisciplinary Digital Publishing Institute (MDPI)
• Subject: 1-Phosphatidylinositol 3-kinase / 17β-Estradiol / Abdomen / Adipose Tissue - cytology / adipose-derived stromal cells / AKT protein / Angiogenesis / Angiogenesis - drug effects / Animals / Blood flow / Body fat / Bone marrow / CD34 antigen / Cell differentiation / Cell Differentiation - drug effects / Cell proliferation / Cells / critical limb ischemia / Endothelial Cells - cytology / Endothelial Cells - drug effects / Endothelial Cells - metabolism / endothelial differentiation / Endothelium / Estradiol / Estradiol - pharmacology / estrogen / Estrogen receptors / Estrogens / Estrogens - pharmacology / Extremities - blood supply / Extremities - pathology / Female / Flow cytometry / Health aspects / Hindlimb - blood supply / Hindlimb - pathology / Hormone replacement therapy / Ischemia / Ischemia - drug therapy / Ischemia - pathology / Ischemia - therapy / Laboratory animals / Male / MAP kinase / Medical prognosis / Menopause / Neovascularization, Physiologic - drug effects / Oophorectomy / Ovariectomy / Ovaries / Phenols / Postmenopausal women / Rats / Rats, Sprague-Dawley / Signal Transduction - drug effects / Skin / Stem cells / Stromal cells / Stromal Cells - cytology / Stromal Cells - drug effects / Stromal Cells - metabolism / Sutures / Transplantation / Vascular endothelial growth factor / Womens health