Zerumbone-Loaded Nanostructured Lipid Carrier Induces Apoptosis of Canine Mammary Adenocarcinoma Cells

Abstract

Canine mammary gland tumor (CMT) is the most common tumor in intact female dog. Zerumbone (ZER) has promising anticancer properties, but plagued with poor water solubility, poor absorption, bioavailability, and delivery to target tissues. To solubilize, ZER was loaded into nanostructured lipid carrier (NLC) to produce ZER-loaded NLC (ZER-NLC). The objectives of this study were to determine the antiproliferative effect and the mode of cell death induced by ZER-NLC and ZER on a canine mammary gland tumor (CMT) adenocarcinoma primary cell line. There was no significant difference (p>0.05) between ZER-NLC and ZER treatments in the inhibition of CMT cell proliferation; thus, the loading of ZER into NLC did not compromise the cytotoxic effect of ZER. Microscopically, ZER-NLC- and ZER-treated CMT cells showed apoptotic cell morphology. ZER-NLC and ZER treatments significantly downregulated the antiapoptotic Bcl-2 and upregulated the proapoptotic Bax gene expressions in CMT cells. Both ZER-NLC and ZER-treated CMT cells showed significant (p<0.0001) increases in caspase-8, -9, and -3/7 protein activities. In conclusion, ZER-NLC induced CMT cell death via regulation of Bcl-2 and Bax gene expressions and caspase activations, indicating the involvement of both the intrinsic and extrinsic pathways of apoptosis. This study provided evidences for the potential of ZER-NLC as an anticanine mammary gland adenocarcinoma chemotherapy.

Formulation and Nanotechnology-Based Approaches for Solubility and Bioavailability Enhancement of Zerumbone

“…It is completely soluble in dimethyl sulfoxide (DMSO) and ethanol but has extremely poor solubility in water (~1–1.5 mg/L at 25 °C) [ 34 , 58 ]. Zerumbone is highly lipophilic and the poor water solubility contributes to poor absorption, low oral bioavailability, and limited targeting to tissues and organs of interest [ 28 , 35 , 36 , 37 , 47 , 48 ]. The chemical structure and other physicochemical characteristics of zerumbone are presented in Figure 1 and Table 1 , respectively.…”

“…Zerumbone is known to exhibit numerous biological and pharmacological effects ( Figure 2 ) such as antitumor, antioxidant, antimicrobial, anti-nociceptive, hepatoprotective, immunomodulatory, anti-inflammatory, gastro-protective, and antiproliferative through modulation of various molecular targets ( Figure 3 ) and signaling mechanisms. Multiple reviews and extensive amount of studies have been performed detailing the molecular targets of zerumbone for various chronic diseases and have been reviewed in references elsewhere in great detail [ 27 , 28 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 43 , 45 , 46 , 47 , 49 , 56 , 59 , 60 ].…”

“…Most well-known natural compounds are polyphenols, flavonoids, and terpenes [ 26 ]. Zerumbone is one such natural compound, classified as a sesquiterpenoid that is extracted from the essential volatile oil of rhizomes from wild ginger, Zingiber zerumbet [ 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 ]. Zerumbone has been shown to possess strong antitumor [ 27 , 30 , 39 ], antioxidant [ 40 ], antimicrobial [ 41 ], anti-nociceptive [ 42 ], hepatoprotective [ 43 ], immunomodulatory [ 44 ], anti-inflammatory [ 33 , 45 , 46 ], and gastro-protective [ 40 ] activity.…”

“…However, despite promising in-vitro and in-vivo studies demonstrating the therapeutic utility of zerumbone, its preclinical and clinical development has been limited due to its poor water solubility, poor absorption, or low bioavailability, limiting its targeting into various tissues and organs of interest. Hence, to advance the therapeutic potential in various diseases and overcome the biopharmaceutical challenges of zerumbone, it is essential to develop advanced formulation and nanotechnology-based approaches [ 28 , 35 , 36 , 37 , 47 , 48 ].…”

“…Multiple reviews demonstrate the beneficial pharmacological effects of zerumbone for various diseases such as different types of cancer, wound healing, obesity, inflammatory bowel diseases [ 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 49 ]. However, to our knowledge, no review demonstrates the various formulation strategies developed to overcome the biopharmaceutical challenges related to zerumbone.…”