A bioprocessing approach for osteogenic differentiation: combining electrical stimulation with the F1 protein fraction from natural latex of Hevea brasiliensis

The rising life expectancy and aging of bones have created a demand for new treatments for fractures and bone loss. Tissue engineering and development of scalable bioprocesses are emerging as a promising solution for tissue repair. This study aimed to evaluate in vitro proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs) from the bone marrow of Wistar rats subjected to electrical stimulation (ES) and cultured with F1 protein fraction from the latex of the Hevea brasiliensis rubber tree. Additionally, the study aimed to analyze the expression of genes related to osteogenesis. Cell viability (MTT) and osteogenic differentiation (Alizarin red) assays were performed using ES of 60 s, 150 s, and 300 s at 10 μA. The medium was supplemented with F1 at concentrations of 1%, 0.1%, and 0.01%. For gene expression analyses, cells were stimulated for 60 s and 300 s and supplemented with 0.001% and 0.0001% F1. All experimental conditions demonstrated cell viability above 70% with no cytotoxicity. Osteogenic differentiation exceeded 80%, with the ES-60 s group combined with 0.0001% F1 showing the highest efficacy. Expression of Bmp2 and Ltype genes increased with 60 s F1-0.001% and 0.0001%. Col1a1 gene expression was higher with F1-0.001% and ES-300 s, while Alp and Runx2 were prominent with F1-0.0001%. Camk2 showed increased expression with F1-0.001% at 60 s and 300 s. F1 protein fraction of latex has a significant biological effect on promoting the proliferation and osteogenic differentiation of MSCs, especially when combined with ES. This highlights its potential as a therapeutic option in regenerative medicine for bone repair.