Synthesis, Characterization, and In Vivo Cytokinome Profile of IL-12-Loaded PLGA Nanospheres
We report the successful encapsulation and elution of recombinant murine IL-12 (rmIL-12) from poly(lactide-co-glycolic) acid (PLGA) nanospheres (IL-12-NS) synthesized using the double emulsion/solvent evaporation (DESE) technique with microsphere depletion through ultracentrifugation. Images obtaine...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2022-01-01
|
| Series: | Journal of Immunology Research |
| Online Access: | http://dx.doi.org/10.1155/2022/6993187 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832550784827392000 |
|---|---|
| author | Ryan A. Lacinski Justin E. Markel Jabeen Noore Hillary G. Pratt Brock A. Lindsey |
| author_facet | Ryan A. Lacinski Justin E. Markel Jabeen Noore Hillary G. Pratt Brock A. Lindsey |
| author_sort | Ryan A. Lacinski |
| collection | DOAJ |
| description | We report the successful encapsulation and elution of recombinant murine IL-12 (rmIL-12) from poly(lactide-co-glycolic) acid (PLGA) nanospheres (IL-12-NS) synthesized using the double emulsion/solvent evaporation (DESE) technique with microsphere depletion through ultracentrifugation. Images obtained with scanning electron microscopy (SEM) showcased a characteristic spherical shape with a mean particle diameter of 138.1±10.8 nm and zeta potential of −15.1±1.249 mV. These values suggest minimal flocculation when in solution, which was reflected in an in vivo biodistribution study that reported no observed morbidity/mortality. Encapsulation efficiency (EE) was determined to be 0.101±0.009% with average particle concentration obtained per batch of 1.66×109±4.45×108 particles/mL. Disparate zeta (ζ) potentials obtained from both protein-loaded and protein-unloaded batches suggested surface adsorption of protein, and confocal microscopy of BSA-FITC-loaded nanospheres confirmed the presence of protein within the polymeric shell. Furthermore, elution of rmIL-12 from IL-12-NS at a concentration of 500 million particles/mL was characterized using enzyme-linked immunosorbent assay (ELISA). When IL-12-NS was administered in vivo to female BALB/c mice through retroorbital injection, IL-12-NS produced a favorable systemic cytokine profile for tumoricidal activity within the peripheral blood. Whereas IFN-γ nadir occurred at 72 hours, levels recovered quickly and displayed positive correlations postburst out to 25 days postinjection. IL-12-NS administration induced proinflammatory changes while prompting minimal counterregulatory increases in anti-inflammatory IL-10 and IL-4 cytokine levels. Further, while IL-6 levels increased to 30 folds of the baseline during the burst phase, they normalized by 72 hours and trended negatively throughout the sill phase. Similar trends were observed with IL-1β and CXCL-1, suggesting a decreased likelihood of progression to a systemic inflammatory response syndrome-like state. As IL-12-NS delivers logarithmically lower amounts of IL-12 than previously administered during human clinical trials, our data reflect the importance of IL-12-NS which safely create a systemic immunostimulatory environment. |
| format | Article |
| id | doaj-art-2f31af1870fd473e806ab91dfe3438b3 |
| institution | Kabale University |
| issn | 2314-7156 |
| language | English |
| publishDate | 2022-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Immunology Research |
| spelling | doaj-art-2f31af1870fd473e806ab91dfe3438b32025-02-03T06:05:50ZengWileyJournal of Immunology Research2314-71562022-01-01202210.1155/2022/6993187Synthesis, Characterization, and In Vivo Cytokinome Profile of IL-12-Loaded PLGA NanospheresRyan A. Lacinski0Justin E. Markel1Jabeen Noore2Hillary G. Pratt3Brock A. Lindsey4Department of OrthopaedicsDepartment of OrthopaedicsDepartment of OrthopaedicsDepartment of SurgeryDepartment of OrthopaedicsWe report the successful encapsulation and elution of recombinant murine IL-12 (rmIL-12) from poly(lactide-co-glycolic) acid (PLGA) nanospheres (IL-12-NS) synthesized using the double emulsion/solvent evaporation (DESE) technique with microsphere depletion through ultracentrifugation. Images obtained with scanning electron microscopy (SEM) showcased a characteristic spherical shape with a mean particle diameter of 138.1±10.8 nm and zeta potential of −15.1±1.249 mV. These values suggest minimal flocculation when in solution, which was reflected in an in vivo biodistribution study that reported no observed morbidity/mortality. Encapsulation efficiency (EE) was determined to be 0.101±0.009% with average particle concentration obtained per batch of 1.66×109±4.45×108 particles/mL. Disparate zeta (ζ) potentials obtained from both protein-loaded and protein-unloaded batches suggested surface adsorption of protein, and confocal microscopy of BSA-FITC-loaded nanospheres confirmed the presence of protein within the polymeric shell. Furthermore, elution of rmIL-12 from IL-12-NS at a concentration of 500 million particles/mL was characterized using enzyme-linked immunosorbent assay (ELISA). When IL-12-NS was administered in vivo to female BALB/c mice through retroorbital injection, IL-12-NS produced a favorable systemic cytokine profile for tumoricidal activity within the peripheral blood. Whereas IFN-γ nadir occurred at 72 hours, levels recovered quickly and displayed positive correlations postburst out to 25 days postinjection. IL-12-NS administration induced proinflammatory changes while prompting minimal counterregulatory increases in anti-inflammatory IL-10 and IL-4 cytokine levels. Further, while IL-6 levels increased to 30 folds of the baseline during the burst phase, they normalized by 72 hours and trended negatively throughout the sill phase. Similar trends were observed with IL-1β and CXCL-1, suggesting a decreased likelihood of progression to a systemic inflammatory response syndrome-like state. As IL-12-NS delivers logarithmically lower amounts of IL-12 than previously administered during human clinical trials, our data reflect the importance of IL-12-NS which safely create a systemic immunostimulatory environment.http://dx.doi.org/10.1155/2022/6993187 |
| spellingShingle | Ryan A. Lacinski Justin E. Markel Jabeen Noore Hillary G. Pratt Brock A. Lindsey Synthesis, Characterization, and In Vivo Cytokinome Profile of IL-12-Loaded PLGA Nanospheres Journal of Immunology Research |
| title | Synthesis, Characterization, and In Vivo Cytokinome Profile of IL-12-Loaded PLGA Nanospheres |
| title_full | Synthesis, Characterization, and In Vivo Cytokinome Profile of IL-12-Loaded PLGA Nanospheres |
| title_fullStr | Synthesis, Characterization, and In Vivo Cytokinome Profile of IL-12-Loaded PLGA Nanospheres |
| title_full_unstemmed | Synthesis, Characterization, and In Vivo Cytokinome Profile of IL-12-Loaded PLGA Nanospheres |
| title_short | Synthesis, Characterization, and In Vivo Cytokinome Profile of IL-12-Loaded PLGA Nanospheres |
| title_sort | synthesis characterization and in vivo cytokinome profile of il 12 loaded plga nanospheres |
| url | http://dx.doi.org/10.1155/2022/6993187 |
| work_keys_str_mv | AT ryanalacinski synthesischaracterizationandinvivocytokinomeprofileofil12loadedplgananospheres AT justinemarkel synthesischaracterizationandinvivocytokinomeprofileofil12loadedplgananospheres AT jabeennoore synthesischaracterizationandinvivocytokinomeprofileofil12loadedplgananospheres AT hillarygpratt synthesischaracterizationandinvivocytokinomeprofileofil12loadedplgananospheres AT brockalindsey synthesischaracterizationandinvivocytokinomeprofileofil12loadedplgananospheres |