Toxicity and Safety Evaluation of Doxorubicin-Loaded Cockleshell-Derived Calcium Carbonate Nanoparticle in Dogs

Abstract

Doxorubicin (DOX) is a potent anticancer agent with cytotoxic effects which limit its clinical usage. This effect is due to its nonselective nature causing injury to the cells as a result of reactive free oxygen radical's release. Cockleshell-derived calcium carbonate nanoparticle (CS-CaCO3NP) is a pH-responsive carrier with targeted delivery potentials. This study aimed at evaluating the toxicity effects of repeated dose administration of DOX-loaded CS-CaCO3NP in healthy dogs. Fifteen dogs with an average body weight of 15 kg were randomized equally into 5 groups. Dogs were subjected to 5 doses at every 3-week interval with (i) normal saline, (ii) DOX, 30 mg/m2, and the experimental groups: CS-CaCO3NP-DOX at (iii) high dose, 50 mg/m2, (iv) clinical dose, 30 mg/m2, and (v) low dose, 20 mg/m2. Radiographs, electrocardiography, and blood samples were collected before every treatment for haematology, serum biochemistry, and cardiac injury assessment. Heart and kidney tissues were harvested after euthanasia for histological and ultrastructural evaluation. The cumulative dose of DOX 150 mg/m2 over 15 weeks revealed significant effects on body weight, blood cells, functional enzymes, and cardiac injury biomarkers with alterations in electrocardiogram, myocardium, and renal tissue morphology. However, the dogs given CS-CaCO3NP-DOX 150 mg/m2 and below did not show any significant change in toxicity biomarker as compared to those given normal saline. The study confirmed the safety of repeated dose administration of CS-CaCO3NP-DOX (30 mg/m2) for 5 cycles in dogs. This finding offers opportunity to dogs with cancer that might require long-term administration of DOX without adverse effects.

Cockle Shell-Derived Calcium Carbonate (Aragonite) Nanoparticles: A Dynamite to Nanomedicine

“…The safety of subcutaneous doses of this biogenic nanoparticle was tested on rats and a dose of 59 mg/m 2 and 590 mg/m 2 was considered safe as it showed negligible mild lesion and no mortality was recorded but some clinical signs and histopathological conditions were reported at a high dose of 5900 mg/m 2 and 29,500 mg/m 2 [16]. In addition, a repeated intravenous administration of Dox-loaded CSCaCO 3 NP in dogs revealed a cumulative dose of 150 mg/m 2 and below had no significant toxic effect suggesting a good safety margin of CSCaCO 3 NP-Dox at a dose of 30 mg/m 2 [21]. In addition, previous literature documented the ability of CSCaCO 3 NP to stimulate cellular differentiation leading to an increase in growth of osteoblast cells [11,132].…”

“…Great nanocarriers with homogenous shapes and size, surface morphology as well as distinctive compositions are derived to promote drug-system delivery applications notwithstanding the fact that homogenous spherically shaped CSCaCO 3 NP aragonite nanoparticles among others, has a promising great impact in nanomedicine due to its ability to enhance the therapeutic efficacy and pharmacokinetics of drugs [17]. In the past few decades, continuous synthesis and development of different nanoparticles has been in process in the quest to elicit the dose effect of drugs at a low concentration with minimal toxic effect [2,6,18,21,85,94,95]. These diverse nanoparticles facilitate several mechanisms of action of drugs in the targeted area of the body system.…”

“…In addition, the regulation of adequate therapeutic blood level of drugs at the minimum effective concentration and the minimum toxic concentration is achieved by the controlled kinetic release [153]. It was documented that CSCaCO 3 NP slowly decomposes in a physiologic state to produce Ca 2+ and CO 2 3− thereby releasing the encapsulated material from its inner matrix of which the by-products could be useful in bone remodelling, blood clotting and muscle contraction [21]. However, the exact fate of the control release mechanism of CaCO 3 of biogenic cockle shells origin in low pH remains unknown and yet to be documented.…”

“…It also possess a peculiar property of low thermodynamic stability thus, its size and shape are in concordance with the method and conditions of laboratory preparations [11]. In addition, numerous studies in recent years, were carried out on its toxicity to prove its wide safety margin both in vivo and in vitro [10,[16][17][18][19][20] A significant number of publications have appeared in recent years documenting the application of CSCaCO 3 NP of aragonite polymorph nanoparticles on different models of animals and cell lines for various therapeutic purposes with regard to their synthesis, functionalization, and both in vitro and in vivo toxicity studies [4,15,16,19,21,22]. Among all available documented literature on CSCaCO 3 NP with unique features so far, no review has been documented on the overview of this very nanoparticle of cockle shell biogenic origin.…”

Evaluation of In-Vitro Release Kinetic and Mechanisms of Curcumin Loaded-Cockle Shell-Derived Calcium Carbonate Nanoparticles

“…The applications of this biogenic inorganic carrier were reported in some pioneered works on a successful targeted delivery of drugs for cancer therapy and chemoprevention [23][24][25]. Notwithstanding the fact that its safety level was demonstrated using human breast cell line, osteoblastic and osteogenic cell lines for its biocompatibility [20,23,25,26] and in animal models [27,28]. Hence, the synthesis and use of biogenic inorganic cockle shell derived calcium carbonate for curcumin delivery with the ability to boost its therapeutic efficacy is essential.…”