Synthesis and Characterization of an Alginate/Chitosan Scaffold for Stem Cell and Drug Co-Delivery

Endometrium-related infertility remains a major challenge in reproductive medicine because successful embryo implantation depends on adequate endometrial receptivity. Mesenchymal stem cells (MSCs) possess regenerative potential through their anti-inflammatory, anti-fibrotic, and proliferative properties, while their therapeutic efficacy can be enhanced using biomaterial scaffolds and pharmacological agents. This study evaluated pentoxifylline (PTX)-loaded alginate/chitosan (ALG/CS) scaffolds crosslinked with calcium chloride (CC) or genipin (GP) for endometrial regeneration. PTX (۲۰ mg/mL) was selected based on preliminary optimization, and human umbilical cord-derived MSCs (HUC-MSCs, passages ۳–۵) were seeded onto the scaffolds. All experiments were performed in triplicate (n = ۳). Scaffolds were characterized by Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), swelling, biodegradation, mechanical testing, and drug release analysis. An indirect transwell co-culture system was established to investigate the paracrine effects of HUC-MSCs on human endometrial stromal cells (HESCs). Statistical analyses were performed using two-way ANOVA following normality and homogeneity testing. GP-crosslinked scaffolds demonstrated significantly greater swelling capacity (۱۰.۹ ± ۰.۶ vs. ۷.۹ ± ۰.۵ at ۷۲ h, p = ۰.۰۰۴), higher PTX release (۶۸.۰۳ ± ۲.۱% vs. ۴۷.۳۹ ± ۱.۸%, p = ۰.۰۰۲), and improved mechanical strength (Young's modulus: ۶.۰ ± ۰.۴ vs. ۴.۲ ± ۰.۳ MPa, p = ۰.۰۰۱) compared with CC scaffolds. Controlled biodegradation and enhanced HUC-MSC adhesion were also observed in GP scaffolds. Furthermore, co-culture significantly increased HESC proliferation, with Ki۶۷-positive cells reaching ۶۸.۴ ± ۳.۲% compared with ۳۲.۱ ± ۲.۸% in the control group (p = ۰.۰۰۱). The superior performance of GP-crosslinked scaffolds is attributed to stable covalent crosslinking, which improves mechanical integrity and enables controlled degradation. Combined with the pro-angiogenic, anti-inflammatory, and antioxidative effects of PTX, this scaffold supports a favorable microenvironment for endometrial repair. These findings suggest that PTX-loaded GP-crosslinked ALG/CS scaffolds seeded with HUC-MSCs represent a promising strategy for enhancing endometrial regeneration and treating endometrium-related infertility.

کلیدواژه ها:

Endometrial regeneration ، Endometrial receptivity ، Human umbilical cord-derived mesenchymal stem cells ، Alginate/Chitosan scaffold ، pentoxifylline ، Genipin