Gelatin Nanoparticles for Complexation and Enhanced Cellular Delivery of mRNA

Abstract

Messenger RNA (mRNA) is increasingly gaining interest as a modality in vaccination and protein replacement therapy. In regenerative medicine, the mRNA-mediated expression of growth factors has shown promising results. In contrast to protein delivery, successful mRNA delivery requires a vector to induce cellular uptake and subsequent endosomal escape to reach its end destination, the ribosome. Current non-viral vectors such as lipid- or polymer-based nanoparticles have been successfully used to express mRNA-encoded proteins. However, to advance the use of mRNA in regenerative medicine, it is required to assess the compatibility of mRNA with biomaterials that are typically applied in this field. Herein, we investigated the complexation, cellular uptake and maintenance of the integrity of mRNA complexed with gelatin nanoparticles (GNPs). To this end, GNPs with positive, neutral or negative surface charge were synthesized to assess their ability to bind and transport mRNA into cells. Positively charged GNPs exhibited the highest binding affinity and transported substantial amounts of mRNA into pre-osteoblastic cells, as assessed by confocal microscopy using fluorescently labeled mRNA. Furthermore, the GNP-bound mRNA remained stable. However, no expression of mRNA-encoded protein was detected, which is likely related to insufficient endosomal escape and/or mRNA release from the GNPs. Our results indicate that gelatin-based nanomaterials interact with mRNA in a charge-dependent manner and also mediate cellular uptake. These results create the basis for the incorporation of further functionality to yield endosomal release.

Biomaterials-Enhanced Intranasal Delivery of Drugs as a Direct Route for Brain Targeting

“…Similarly, nanocarriers provided with a lipid shell and a gelatin core are among the most widely exploited for IN drug delivery: they can be prepared in mild conditions and the lipid shell favors cell internalization ( Table 3 ) [ 57 , 58 ]. However, their stability could be limited by rapid in vivo disassembly and gelatin degradation by proteolytic enzymes [ 75 , 111 , 112 ]. Unfunctionalized lipid–gelatin nanoparticles have been generally used for IN drug delivery, however, their surface functionalization with targeting ligands could be achieved by the use of previously synthesized ligand-functionalized lipids [ 113 ].…”

Gelatin as It Is: History and Modernity

“…The formation of such structures is facilitated by a decrease in water concentration, shear stresses, and magnetic and electric fields [ 294 , 295 , 296 , 297 , 298 , 299 ]. In conclusion to this section, we should note that interest in the use of gelatin as a basis for the construction of nanomaterials is not decreasing, as evidenced by the works published already in the current century, in particular [ 300 , 301 , 302 , 303 , 304 , 305 , 306 , 307 , 308 , 309 , 310 , 311 , 312 , 313 , 314 , 315 , 316 , 317 , 318 , 319 , 320 , 321 , 322 , 323 , 324 , 325 ]. In fairness, it should also be noted that there are certain difficulties associated with the use of gelatin as a basis for creating functional materials due to its relatively low mechanical strength.…”

Gelatin Matrix as Functional Biomaterial for Immobilization of Nanoparticles of Metal-Containing Compounds

“…Thus, if the reaction of the formation of any substance proceeds in the gelatin matrix, the probability of its immobilization in the array (as well as in a thin layer) of this biopolymer seems to be quite high. In this regard, it is noteworthy that gelatin itself quite easily forms nanoparticles, both in the case when its molecules are neutral and in cases when they carry a positive or negative charge [ 71 ]. Images of these nanoparticles are shown in Figure 3 .…”

“…( A ) Zeta potential (19.0 ± 0.8), linear size (128 ± 18) nm; ( B ) zeta potential (2.9 ± 0.3), linear size (104 ± 23) nm; ( C ) zeta potential (–11.6 ± 0.8), linear size (133 ± 43) nm. (This figure was made by the author of the given article according to the data presented in [ 71 ]).…”