International Journal of Nanomedicine (Nov 2019)
Nanosome-Mediated Delivery Of Protein Kinase D Inhibitor Protects Chondrocytes From Interleukin-1β-Induced Stress And Apoptotic Death
Abstract
Hongsik Cho,1–3,* Fazal-Ur-Rehman Bhatti,1,3,* Karen A Hasty,1–3 Ae-Kyung Yi4 1Department of Orthopaedic Surgery and Biomedical Engineering, The University of Tennessee Health Science Center, Memphis, TN, USA; 2Department of Orthopaedic Surgery, Campbell Clinic, Memphis, TN, USA; 3151 Research Service, Veterans Affairs Medical Center, Memphis, TN, USA; 4Department of Microbiology, Immunology and Biochemistry, The University of Tennessee Health Science Center, Memphis, TN, USA*These authors contributed equally to this workCorrespondence: Hongsik ChoDepartment of Orthopaedic Surgery and Biomedical Engineering, The University of Tennessee Health Science Center, Research 151, VAMC, 1030 Jefferson Ave, Memphis TN 38104 USATel +1 901 523-8990 (Ext: 6456)Fax +1 901 577-7273Email [email protected] YiDepartment of Microbiology, Immunology and Biochemistry, The University of Tennessee Health Science Center, 858 Madison Ave., Suite 501C, Memphis, TN 38163, USATel +1 901 448-1775Fax +1 901 448-7360Email [email protected]: Inflammatory stress caused by protein kinase D (PKD) plays a critical role in damaging chondrocytes and extracellular matrix (ECM) during osteoarthritis (OA). The PKD inhibitor (PKDi) (CRT0066101) has been used to overcome inflammation in different cell types. However, the efficacy of a therapeutic drug can be limited due to off-target distribution, slow cellular internalization, and limited lysosomal escape. In order to overcome this issue, we developed nanosomes carrying CRT0066101 (PKDi-Nano) and tested their efficacy in vitro in chondrocytes.Methods: Chondrocytes were subjected to IL-1β-induced inflammatory stress treated with either PKDi or PKDi-Nano. Effects of treatment were measured in terms of cytotoxicity, cellular morphology, viability, apoptosis, phosphorylation of protein kinase B (Akt), and anabolic/catabolic gene expression analyses related to cartilage tissue.Results and Discussion: The effects of PKDi-Nano treatment were more pronounced as compared to PKDi treatment. Cytotoxicity and apoptosis were significantly reduced following PKDi-Nano treatment (P < 0.001). Cellular morphology was also restored to normal size and shape. The viability of chondrocytes was significantly enhanced in PKDi-Nano-treated cells (P < 0.001). The data indicated that PKDi-Nano acted independently of the Akt pathway. Gene expression analyses revealed significant increases in the expression levels of anabolic genes with concomitant decreases in the level of catabolic genes. Our results indicate that PKDi-Nano attenuated the effects of IL-1β via the nuclear factor kappa-light-chain enhancer of activated B cells (NF-κB) pathway.Conclusion: Taken together, these results suggest that PKDi-Nano can be used as a successful strategy to reduce IL1β-induced inflammatory stress in chondrocytes.Keywords: CRT0066101, liposome, cartilage, osteoarthritis, cytokine
