Co-Encapsulation of Hepatocytes, Mesenchymal Stem Cells, and Growth Factor in Arginine-Glycine-Aspartate Functionalized Microbeads for Liver Disease

Acute liver failure is a life-threatening condition with limited treatment options, primarily liver transplantation, which is constrained by donor shortages and lifelong immunosuppression. This study presents a minimally invasive therapeutic approach using multifunctional microbeads co-encapsulating two cell types: immortalized hepatocytes and umbilical cord-derived mesenchymal stem cells, along with basic fibroblast growth factor-loaded poly(lactide-co-glycolide) microspheres. The alginate microbeads are functionalized with poly(ethylene glycol) and the arginine-glycine-aspartate tripeptide to enhance cell adhesion and are crosslinked via click chemistry for improved structural integrity. The bFGF-loaded PLGA microspheres were synthesized using a double-emulsion solvent evaporation method, achieving an average size of 4.25 ± 2.20 µm, a loading content of 0.078% and an entrapment efficiency of 3.52 ± 0.27%. Sustained bFGF release over 14 days (cumulative 2.39 ± 0.20 ng) enhanced hepatocyte proliferation, human mesenchymal stem cell differentiation and cell viability. Functional assessment demonstrated significantly improved hepatocyte performance, with microbeads producing 2032.53 ± 29.45 ng of albumin and 1057.00 ± 9.19 ng of alpha-fetoprotein over 14 days. Overall, this co-encapsulation strategy enhances hepatocyte regeneration, viability, function and offers a scalable therapeutic platform for ALF. Future studies should optimize the formulation and evaluate long-term efficacy <i>in vivo</i>.