NANO-bio interfaces : studying protien on polymer -derived nanoparticles for the development of bee venom-zno nanoparticles using collagen as a novel nanotherapeutic strategy for burn repair in rats

Abstract

This study aimed to biosynthesize a nanotherapeutic product for enhancing experimental wound healing in rats. Zinc oxide nanoparticles (ZnO NPs) were synthesized using natural collagen as a reducing and stabilizing agent and further bio-conjugated with bee venom to produce a nanomaterial with potential therapeutic applications. Using standard analytical methods and protocols, Collagen extracted from chicken feet exhibited characteristic UV–Visible absorption peaks between 210–280 nm and typical FTIR amide bands confirming its purity and protein structure. ZnO NPs-BV were successfully synthesized and characterized using UV–Visible spectroscopy, FTIR, scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) analysis, and X-ray diffraction (XRD). The rate of immobilized proteins was also quantified, and their biological activities were assessed, including antioxidant, anti-inflammatory, antibacterial, and ultraviolet (UV) protective effects. For the in vivo evaluation, 20 female Wistar albino rats were divided into four equal groups (n = 5): a healthy control group, an untreated burn group, and two treatment groups topically treated with 1% ZnO NPs-BV cream and 1% silver sulfadiazine cream, respectively. Cutaneous burns were induced under chloroform anesthesia using a 1.2 cm diameter electric soldering iron applied for 3 seconds. Treatments were applied every other day for three weeks, and wound healing was monitored macroscopically alongside the assessment of hematological, biochemical, and oxidative stress parameters, as well as histological examination of skin and spleen tissues. The results showed the formation of nanoparticles with a characteristic hexagonal crystalline structure and an average size of 33.4 nm, with effective immobilization of bee venom proteins. Results of in vitro biological evaluation demonstrated that ZnO NPs-BV possessed notable anti-inflammatory activity in both protein denaturation and RBC membrane stabilization assays. Antioxidant assays (DPPH and FRAP) indicated an important free radical scavenging and reducing power. Additionally, ZnO NPs-BV exhibited notable photoprotective activity, with performance comparable to that of the commercial sunscreen formulation. Antibacterial testing revealed concentration-dependent activity, with the strongest inhibition observed against Pseudomonas aeruginosa, and Staphylococcus aureus. For in vivo results, the experimental wounds applied to rats resulted in skin and spleen tissue abnormalities, abnormalities in hematological and biochemical parameters, as well as a disturbance in the oxidative stress balance.The group treated with ZnO NPs-BV demonstrated a marked acceleration in burn wound healing compared to the untreated group, evidenced by improved wound contraction and restoration of skin tissue architecture, along with reduced inflammatory markers and improved serum iron, albumin levels, and oxidative stress balance. Histological analysis further confirmed a clear improvement in the structural organization of both skin and spleen tissues. In conclusion, these findings suggest that ZnO NPs-BV exhibits a promising and safe therapeutic effect by reducing oxidative stress, enhancing tissue repair, accelerating burn wound healing, and restoring tissue function