Assessing cellular internalization and endosomal escape abilities of novel BUFII-Graphene oxide nanobioconjugates

Abstract

Cell-penetrating agents based on functionalized nanoplatforms have emerged as a promising approach for developing more efficient and multifunctional delivery vehicles for treating various complex diseases that require reaching different intracellular compartments. Our previous work has shown that achieving full cellular coverage and high endosomal escape rates is possible by interfacing magnetite nanoparticles with potent translocating peptides such as Buforin II (BUF-II). In this work, we extended such an approach to two graphene oxide (GO)-based nanoplatforms functionalized with different surface chemistries to which the peptide molecules were successfully conjugated. The developed nanobioconjugates were characterized via spectroscopic (FTIR, Raman), thermogravimetric, and microscopic (SEM, TEM, and AFM) techniques. Moreover, biocompatibility was assessed via standardized hemocompatibility and cytotoxicity assays in two cell lines. Finally, cell internalization and coverage and endosomal escape abilities were estimated with the aid of confocal microscopy analysis of colocalization of the nanobioconjugates with Lysotracker Green®. Our findings showed coverage values that approached 100% for both cell lines, high biocompatibility, and endosomal escape levels ranging from 30 to 45% and 12–24% for Vero and THP-1 cell lines. This work provides the first routes toward developing the next-generation, carbon-based, cell-penetrating nanovehicles to deliver therapeutic agents. Further studies will be focused on elucidating the intracellular trafficking pathways of the nanobioconjugates to reach different cellular compartments.

Invertase Immobilization on Magnetite Nanoparticles for Efficient Fructooligosaccharide Generation: A Comprehensive Kinetic Analysis and Reactor Design Strategy

“…The continuous operation was modeled for dilution rate (D) values ranging from 0 to 50 h −1 (0.02 to ∞ h in residence time (τ) equivalency). We solved a set of design balances defined by Equation (19) for each dilution rate value, repeatedly for each of the reactants and products considered in batch modeling. The resulting system of algebraic equations was solved using MATLAB's fsolve function.…”

“…In Equation (19), C i and C 0,i represent the final and inlet concentrations of species i, respectively. For each species i, the reaction rate r i is described by the right-hand side expression of the corresponding batch modeling differential equation (Equation (6) to Equation ( 13)).…”

“…In the CSTR model, based on the immobilized invertase model, seven additional equations were included relating to the new species involved in glucose oxidation. These equations were defined by Equation (19), with the reaction rate defined as the RHS expression in the corresponding differential equations for the new species represented in Equations ( 24)- (30). The glucose reaction rate was modified by inserting a new term addressing the new glucose-containing reaction.…”

“…The common nanostructured materials used in enzyme immobilization include silica, zirconia, gold, and iron oxide nanoparticles. Recently, polymeric (e.g., chitosan and gelatin) and carbon-based (e.g., graphene oxide and carbon quantum dots) nanomaterials have gained significant attention [19][20][21]. Among iron oxide nanoparticles, magnetite stands out due to its strong responsiveness to magnetic fields, ease of synthesis and functionalization, and low toxicity [18,21].…”