Abstract
BACKGROUND: As life expectancy increases, so does the incidence of age-related disorders, including vascular diseases and chronic wounds. These conditions are often associated with impaired angiogenesis and tissue repair, posing significant clinical challenges due to prolonged healing times. Recent studies have highlighted the tissue regenerative potential of the human Mesenchymal Stem Cell-Conditioned Medium (MSC-CM). Building on this, we investigated the ability of CM derived from human Dental Pulp Stem Cells (hDPSC-CM) to support angiogenesis and tissue repair in a surgically injured medicinal leech model. The medicinal leech Hirudo verbana offers a valuable in vivo tool for studying wound healing and regeneration, due to its well-characterized anatomy, regenerative capabilities, and the evolutionary conservation of key immune and vascular pathways. METHODS: Adult leeches were surgically injured and treated with either FBS-free medium (CTRL) or hDPSC-CM. Animals were sacrificed at 48 h, 72 h, and 1 week post-treatment to evaluate tissue regeneration across experimental groups. Morphological, immunohistochemical, and molecular analyses were conducted. RESULTS: hDPSC-CM significantly accelerated wound healing, promoting extracellular matrix remodelling and muscle regeneration. The expression of key markers acid phosphatase, CD31, VEGF, Desmin, AIF-1, and COL Iα1, along with vessel counts, confirmed the immunomodulatory activity (38.5% overall decrease in macrophage number), enhanced angiogenesis (64.2% overall increase in CD31 expression), and improved muscle regeneration (222.5% overall increase in Desmin expression) induced by hDPSC-CM in the injured area. CONCLUSIONS: Our findings support the therapeutic potential of hDPSC-CM as a novel, cell-free strategy for enhancing wound healing and managing chronic conditions related to impaired tissue repair.