Comparative Study of Adipose-Derived Stem Cells from Localized Scleroderma Patients and Healthy Donors in Treating Skin Fibrosis.

Abstract

Localized scleroderma (LoS) is an autoimmune disease characterized by fibrosis of the skin and atrophy of the subcutaneous fat tissue. Use of adipose-derived mesenchymal stem cells (ASCs) is a promising treatment approach for LoS. However, ASCs from scleroderma patients (LoS ASCs) have been shown to exhibit altered characteristics compared with ASCs from healthy donors (healthy ASCs). This study aimed to compare the abilities of LoS ASCs and healthy ASCs in treating skin fibrosis. The paracrine ability of ASCs was tested with cytokine array. Bleomycin-challenged mouse models received subcutaneous injection of LoS ASCs and healthy ASCs. Pathologic staining and Western blotting of collagenase type I and α-smooth muscle actin was performed. Fibroblasts derived from LoS lesions (LoS FB) were co-cultured with ASCs, and subjected to RNA sequencing to further explore the similarities and differences in the treatment mechanism. In vivo comparison revealed that healthy ASCs had a stronger proliferation ability and secreted higher levels of growth factors and cytokines, including vascular endothelial growth factor A, platelet-derived growth factor fibroblasts, and interleukin-10. Pathologic staining of the skin in mouse models treated with ASCs demonstrated that healthy ASCs were more effective in reducing dermal thickness and collagen deposition, and increasing microvessel density and the proportion of M2 macrophages. Co-culture with both healthy ASCs and LoS ASCs reduced the proliferation and migration abilities of LoS FB, and the protein expression of α-smooth muscle actin and collagenase type I. RNA sequencing and validation revealed potential difference in the canonical Wnt pathway. Healthy ASCs exhibited stronger proliferation, paracrine, antifibrosis, proangiogenesis, and immunomodulation abilities in treating skin fibrosis in scleroderma mouse models. Allogenic ASCs obtained from healthy donors are more efficient in treating skin fibrosis, and could serve as a potential alternative for patients who are not suitable candidates for liposuction surgery in the future.