The Response Of Doxorubicin Nanoparticles On Complex Tumor Microenvironments By Tailoring Multiple Physicochemical Properties

Background:Cellular internalization,delivery efficiency and therapeutic efficacy of nanoparticles vary according to the microenvironmental complexity for tumor types.Adjusting their physicochemical properties,such as size and surface properties,has significant potential for dealing with such complexities.Methods:The components of doxorubicin nanoparticles were analyzed by 1H NMR,and the particle size and morphology were observed by DLS and TEM respectively.The surface coating was analyzed by XPS,coagulation activity,and complement activation,and their targeted ligand content was detected by BCA assay.Flow cytometry,CCK-8 experiment,in vitro BBB permeability model were used to detect their cellular uptake,anti-proliferation and in vitro BBB permeability respectively.Intracranial glioma models and subcutaneous breast cancer models were constructed to detect their body distributions and anti-tumor effect.The mechanisms of action of doxorubicin nanoparticles were analyzed by immunohistochemistry and the safety in vivo was evaluated by HE staining.Results:We prepare four types of doxorubicin nanoparticles(DOX-D1,DOX-D2,DOX-W1 and DOX-W2 Nano)using simply changing reaction medium or reactant ratio.1H NMR confirmed that the components of doxorubicin nanoparticles were conjugated into heparin carrier successfully.DLS and TEM showed that DOX-D1 and DOX-D2 Nano exhibited different particle sizes,but DOX-W Nano presented similar particle sizes.And they could self-assemble into smaller particles in the blood-mimic conditions,the order of size was followed:DOX-D1>DOX-D2≈DOX-W Nano.XPS,coagulation activity,complement activation and BCA protein analysis indicated that DOX-D1 and DOX-D2 Nano exhibited the similar surface characteristics(surface coating and targeting ligand content),and DOX-W Nano presented similar surface properties,while DOX-W Nano has higher target ligand content than DOX-D Nano.Thus,the bioactivity in vitro and tumor microenvironment response of DOX-D1,DOX-D2 and DOX-W 1 are further investigated due to their different physicochemical properties.Cellular uptake and anti-proliferation experiments showed that DOX-W1 Nano had higher cell uptake and stronger anti-proliferation ability than DOX-D 1 and DOX-D2 Nano;in vitro BBB permeability experiments showed that DOX-D2 Nano had better BBB permeability than DOX-D 1 and DOX-W1 Nano;biodistribution and anti-tumor effect showed DOX-D 1 Nano could efficiently bypass BBB,deliver to intracranial glioma and inhibit the growth of glioma,while DOX-W1 Nano had excellent targeting and anti-tumor effect in subcutaneous tumor breast cancer.Immunohistochemistry showed that doxorubicin nanoparticles could inhibit the proliferation of tumor cells and reduce the generation of blood vessels.HE staining showed that doxorubicin nanoparticles had no organ toxicity in vivo.Conclusion:Tailoring multiple physicochemical properties of nanoparticles can play significant role in addressing tumor microenvironment complexity.