Table 1 Summary of representative studies evaluating the biocompatibility and toxicity of liposomes
From: Nanotoxicological profiles of clinically approved nanoplatforms
Formulation composition | Preparation method | Liposome dose (administration route) | Animal/cell type | Cell/tissue target | Biocompatibility-/toxicity-related outcomes | References |
|---|---|---|---|---|---|---|
In vitro studies | ||||||
Rifampicin-loaded liposomes (RLip) made of soybean phosphatidylcholine; cholesterol and dicetyl phosphate or stearylamine | Thin-film hydration | Expressed in rifampicin concentration (0–25 µM) | Rat alveolar macrophage line NR 8383; human bronchial epithelial and small airway epithelial cells | Respiratory associated cells | RLip showed no marked cytotoxicity, with much higher cell viability than the free rifampicin incubated with the three cell lines over 24 h. There was no inflammatory response observed | Changsan et al. [22] |
Soybean lecithin liposomes co-loaded with rifampicin and the complex of isoniazid-phthalocyanine with gamma-cyclodextrin | Heating method, organic solvent-free | 0.1–1 mg/mL | HeLa cells and human peripheral lung fibroblasts and epithelial cells | Adenocarcinoma and normal respiratory cells | Following 24–48 h of incubation, no cytotoxicity was observed in the dark. Upon laser irradiation, highly loaded liposomes exhibited dose-dependent cytotoxicity | Nkanga et al. [101] |
Paclitaxel-loaded liposomes made of phosphatidylcholine, cholesterol and span 80 | Thin-film hydration | 0.5–5000 μg/mL | Lung cell line A549 | Cancer cells | Liposomes showed no cytotoxicity, while plain drug was cytotoxic following 24–48-h incubation | Utreja et al. [140] |
Rifapentine-loaded liposome made of hydrogenated soy phosphatidylcholine, cholesterol and Stearyl amine | Spray drying method | 20–100 μg/mL | Lung cell line A549 | Cancer cells | The formulation exhibited better cell viability than free drug following 24-h incubation | Patil-Gadhe et al. [108] |
Melittin-loaded liposomes containing poloxamer 188 | Not reported | Expressed in melittin concentration (2 μM) | HepG2 cells | Hepatic carcinoma cells | Empty liposomes showed no cytotoxicity while the melittin-liposomes were as cytotoxic as free melittin | Mao et al. [89] |
Cabazitaxel-liposomes made of egg phosphatidyl lipid-80, PEG-phosphoethanolamine and cholesterol | Thin-film hydration | Expressed in cabazitaxel concentration (10–20 µg/mL) | CT-26 and 4T1 cells | Mouse colon and breast cancer cells | Liposomal cabazitaxel showed lower cytotoxicity than the plain drug solution over 48 h of incubation | Yin et al. [157] |
Stearyl triphenylphosphonium liposome (STPP-L) and triphenylphosphonium-PEG-phosphatidylethanolamine liposome (TPP-PEG-L) | Thin-film hydration | 0–500 µg/mL | HeLa cells | Adenocarcinoma cells | Following 24-h incubation, TPP-PEG-L showed no cytotoxicity, while STPP-L was toxic (IC50 83 µg/mL), but to lesser extent than STPP + PEG-L (IC50 130 µg/mL) | Biswas et al. [18] |
Isoniazid-liposomes made of dipalmitoylphosphatidylcholine | Thin-film hydration | 0.1–1 mg/mL | L929 cells and human blood | Mouse fibroblast cells and human erythrocytes | No haemolysis was observed. Fibroblast morphology and monolayer confluence were unchanged after 24-h incubation | Chimote and Banerjee [24] |
Cationic liposomes containing phosphatidylethanolamine and positively charged cholesterol | Thin-film hydration | 5–65 µg/mL | Human kidney 293, liver carcinoma HepG2 and mouse fibroblast NIH3T3 cells | Kidney, liver and embryonic tissues | Liposomes showed much lower toxicity than lipofectin and polyethylenimine at concentrations required for optimal gene transfection | Joon Sig Choi et al. [63] |
Lipid-based Mitomycin C prodrug in PEG-coated liposome (hydrogenated soybean phosphatidylcholine and distearoyl phosphatidyl ethanolamine) | Thin-film hydration | Expressed in mitomycin C concentration (200–2000 nM) | Mouse carcinoma M109 cells | Murine lung tissues | In the absence of reducing agents (such as cysteine), liposome was fivefold to sixfold less cytotoxic than free mitomycin. No difference was observed on addition of the reducing agents | Gabizon et al. [40] |
Cationic lipids with a quaternary ammonium headgroup (CDA14) and a tri-peptide headgroup (CDO14) | Thin-film hydration | 15 µg/mL and 120 µg/mL | NCI-H460 cells | Human non-small cell lung cancer tissues | CDA14 induced more apoptosis than CDO14. CDA14 showed increased caspase-3/-9 activity with lower mitochondrial membrane potential and higher reactive oxygen species (ROS) levels | Cui et al. [28] |
Doxorubicin (Dox) loaded in liposomes made of distearoyl phosphatidylethanolamine-maleimide and cholesterol | Thin-film hydration | Expressed in doxorubicin concentration 1–10 µg/mL | 4T1 cell line | Breast tumour cells | Blank liposomes showed no effects on cells. Dox-loaded liposomes showed much better cytotoxic effects on tumour cells than free doxorubicin, which was due to enhanced cell uptake thanks to maleimide handles | Tang et al. [138] |
Integrated Nanotherapeutics Inc. proprietary anionic, neutral and cationic lipids | Not disclosed | 0–128 µg/mL | HL60, NB4, A549 and NIH3T3 cell lines | Tumour cell lines and healthy fibroblasts | Liposomes loaded with siRNAs did not affect viability both suspended and adherent cells at conventionally used concentrations | Syama et al. [135] |
mRNA mixed with positively and negatively charged lipids (DOTAP/POPS) | Thin-film hydration, extrusion, microfluidics | 0–48 µg/mL | Neuro-2a cells | Brain tissues | At 0.288 mg/mL positively charged liposomes led to 65% cell apoptosis, while no apoptosis was obvious for negatively charged liposomes; only 0.48 mg/mL caused 50% apoptosis. Mixing cationic with anionic lipids led to 80% cell viability | Wang et al. [144] |
In vivo studies | ||||||
Paclitaxel-loaded liposomes made of phosphatidylcholine, cholesterol and span 80 | Thin-film hydration | 10–200 mg/kg (IP) | Swiss albino mice | Liver, kidney, heart and spleen | No mortality, haematological, biochemical, histopathological, or weight changes were observed over 28 days | Utreja et al. [140] |
Rifapentine-loaded liposome made of hydrogenated soy phosphatidylcholine, cholesterol and Stearyl amine | Spray drying method | 1–10 mg/kg (Intratracheal) | Wistar rats | Lung | No changes in biochemical, histopathological parameters or body weight were observed at 1–5 mg/kg, while 10 mg/kg led to remarkable tissue toxicity and animal death | Patil-Gadhe et al. [108] |
Melittin-loaded liposomes containing poloxamer 188 | Not reported | 2–8 mg/kg (SC) | LM-3 xenograft tumour model | Hepatocellular carcinoma tissue | The liposomal melittin suppressed tumour growth with reduced side effects compared to the plain melittin | Mao et al. [89] |
Cabazitaxel-liposomes made of egg phosphatidyl lipid-80, PEG-phosphoethanolamine and cholesterol | Thin-film hydration | 5Â mg/kg (IV) | Balb/c mice | Erythrocyte, liver, spleen, kidney, heart and tumour tissues | Unlike the plain drug, cabazitaxel liposomes showed no significant haemolysis or histological lesions and stable body weight 48Â h post-injection | Yin et al. [157] |
Doxorubicin-liposomes made of PEG/phosphatidylcholine and cholesterol with Anti-CD19 mAb targeting ligand | Thin-film hydration | 10.4–13.6 mg/kg (IV) | SCID mice | Heart and blood | Drug-free liposomes showed no lethal toxicity. Targeted doxorubicin–liposomes increased mice life span to much higher levels than non-targeted ones, which were still better than plain doxorubicin | Allen et al. [4] |
Multivalent cationic liposome (Lipofectamineâ„¢), monovalent cationic DOTAP liposomes, neutral and negative liposomes | Heating method, organic solvent-free | 200Â nmol/mouse (intratracheal instillation) | Male Balb/c mice | Lung tissues | Lipofectamineâ„¢ induced greater toxic reactive oxygen intermediates than DOTAP liposomes, while neutral and negative liposomes showed no toxicity 24Â h post-intratracheal instillation | Dokka et al. [31] |
Lipid-based Mitomycin C prodrug in PEG-coated liposomes (hydrogenated soybean phosphatidylcholine and distearoyl phosphatidyl ethanolamine) | Thin-film hydration | 10 mg/kg/week (IV) | Female Balb/c mice | – | A single dose (10 mg/kg) of free mitomycin C exhibited drastic weight loss and toxic death, while the liposomes caused fatal toxicity only after three doses, and no weight loss was observed | Gabizon et al. [40] |
DOTAP cationic liposomes containing cholesterol | Thin-film hydration | 10, 25 or 100Â mg/kg/day (IV) | Male Wistar rats | Liver, kidney, lung, brain and spleen | Histological, haematological and chemico-clinical evaluations showed no significant changes after repeated doses. Dose-dependent genotoxicity (DNA strand breaks) was observed in the lung | Knudsen et al. [71] |
Doxorubicin (Dox) loaded in liposomes made of distearoyl phosphatidylethanolamine-maleimide and cholesterol | Thin-film hydration | Amount equivalent to 5Â mg/kg Dox (IV) | BALB/c mice | Heart tissues | The cardiac cavity of animals treated with Dox-loaded liposomes remained unchanged, while free doxorubicin animals exhibited several abnormalities (e.g. loose structure, myocardial vacuole degeneration, scattered inflammatory cell infiltration) | Tang et al. [138] |
PEGylated mitoxantrone liposomes from CSPC Zhongqi Pharmaceutical Technology (Shijiazhuang) Co., Ltd. | Not disclosed | 20Â mg/m2 (IV) | Randomized, open-label, active-controlled, single centre, phase II clinical trial with Chinese patients with advanced breast cancer | Breast | Liposomes exhibited lower incidence of cardiovascular disorders (13.3% vs. 20.0%) and increased cardiac troponin T (3.3% vs. 36.7%), but higher incidence of anaemia (76.7% vs. 46.7%), skin hyperpigmentation (66.7% vs. 3.3%) and fever (23.3% vs. 10.0%) than free mitoxantrone | Wang et al. [143] |
Cholesterol, egg phosphatidylcholine (Egg-PC) and 1,2-distaroyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)2000] and drug conjugate cholesterol-SN38 | Ethanol injection | 15Â mg/kg (IV) | ICR mice | Liver, heart, lung, spleen, kidney and ileum | The liposomal formulation of prodrug improved drug tolerability by alleviating bloody diarrhoea and liver damage, which were reported side effects of the prodrug | Shi et al. [126] |