A Biocompatible Nanoparticle Approach To Wound Healing: Betanin-Znonps Modulate TGF-Β/SMAD Signaling And Enhance Skin Regeneration

Abstract

Background: A balance between tissue regeneration, apoptosis and inflammation is necessary for effective wound healing. Promising approaches to improving this process include systems based on nanotechnology and bioactive chemicals produced from plants. Using zebrafish and PDL cell cultures, the wound healing abilities of betanin-derived zinc oxide nanoparticles (Betan-ZnONPs), with an emphasis in angiogenesis and apoptotic pathways.

Methods: Green synthesis was used to create Betan-ZnONPs,  which were then characterized by FTIR, XRD, SEM, and EDAX. Using the MTT assay, cytotoxicity was evaluated in PDL cells at doses of 10, 50, and 100 µg/mL. In order to assess cell migration at a non-toxic concentration (28.32 µg/mL), a scratch wound healing assay was used. Using adult zebrafish with dorsal skin injury, in vivo wound healing was investigated by comparing the treatment and control groups in day 7 and day 14. To investigate tissue regeneration and expression of apoptotic and angiogenic markers, histopathological examination and qPCR based gene expression profiling were conducted.

Results: The successful synthesis of crystalline, spherical Betan-ZnONPs with the proper functional group stabilization was validated by characterization. With a biocompatible concentration of 28.32 µg/mL, MTT analysis revealed dose-dependent cytotoxicity. In the scratch assay, treated cells migrated more readily. By day 14, the group that received Betan-ZnONP treatment showed better wound healing and re-epithelialization in zebrafish. In treated samples, histological investigation showed less inflammation and well-organized tissue regeneration. Increased apoptosis and angiogenesis were supported by gene expression analysis that revealed downregulation of Bcl-2 and over-expression of VEGF, HIF-1ɑ, Caspase-3, Caspase-8, Caspase-9.

Conclusion: Betan-ZnONPs significantly promote wound healing by modulating apoptotic and angiogenic pathways. These findings highlight their potential as a biocompatible, plant based nanotherapeutic for regenerative medicine.