Nanoparticles of indinavir, ritonavir, efavirenz and atazanavir were manufactured using wet milling, homogenization and ultrasonication. These nanoformulations, collectively termed nanoformulated antiretroviral therapy (nanoART), assessed macrophage-based drug delivery. Monocyte-derived macrophage nanoART uptake, retention and sustained release were determined. These preliminary studies suggest the potential of nanoART for clinical use.
Total time
~3–5 days (manufacturing ~1–2 days; macrophage culture and uptake studies ~2–3 days)
Model organism
Human monocyte-derived macrophages (primary culture or THP-1 cells)
Steps
1
Manufacture nanoparticles by wet milling
Use NETZSCH MicroCer wet milling equipment to reduce bulk antiretroviral drug crystals (indinavir, ritonavir, efavirenz, atazanavir) to nanoparticle size.
▶ 03:21
2
Further reduce nanoART size by homogenization
Apply Avestin EmulsiFlex C5 homogenizer to milled nanoART suspensions to achieve uniform particle distribution and reduced mean diameter.
▶ 06:44
3
Optimize nanoparticle suspension by sonication
Use Cole Parmer ultrasonic processor to disperse and stabilize nanoART formulations, preventing aggregation.
▶ 07:52
4
Visualize and characterize nanoART morphology
Perform transmission electron microscopy and light scattering analysis to determine particle size, distribution, and morphology of final nanoformulations.
▶ 09:58
5
Culture macrophages and assess nanoART uptake
Differentiate monocytes to macrophages, expose to nanoART formulations, and measure intracellular drug uptake, retention kinetics, and sustained-release profiles.
▶ 10:56
6
Analyze and present representative results
Summarize nanoparticle characterization data, macrophage uptake/retention curves, and evidence supporting clinical potential of nanoART delivery.
▶ 15:37