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Evaluation of In Vivo Toxicity of Biological Nanoparticles
Julia Driscoll1, Irene K Yan1, Ramcharan Singh Angom2
1Department of Transplantation, Mayo Clinic, Jacksonville, Florida.
Abstract:
Biologically derived nanoparticles such as extracellular vesicles are promising candidates for therapeutic applications. In vivo toxicity of biological nanoparticles can result in tissue or organ damage, immunological perturbations, or developmental effects but cannot be readily predicted from in vitro studies. Therefore, an essential component of the preclinical assessment of these particles for their use as therapeutics requires screening for adverse effects and detailed characterization of their toxicity in vivo. However, there are no standardized, comprehensive methods to evaluate the toxicity profile of nanoparticle treatment in a preclinical model. Here, we first describe a method to prepare bovine milk-derived nanovesicles (MNVs). These MNVs are inexpensive to isolate, have a scalable production platform, and can be modified to achieve a desired biological effect. We also describe two vertebrate animal models, mice and zebrafish, that can be employed to evaluate the toxicity profile of biologically derived nanoparticles, using MNVs as an example. Treatment-induced organ toxicity and immunological effects can be assessed in mice receiving systemic injections of MNVs, and developmental toxicity can be assessed in zebrafish embryos exposed to MNVs in embryo water. Utilizing these animal models provides opportunities to analyze the toxicity profiles of therapeutic extracellular vesicles in vivo. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Preparation of milk-derived nanovesicles Basic Protocol 2: In vivo screening for organ toxicity and immune cell profiling using mice Basic Protocol 3: In vivo developmental toxicity screening using zebrafish.
Insights
This study presents a method for preparing milk-derived nanovesicles (MNVs) and utilizes mice and zebrafish models to screen for in vivo toxicity. These models offer a comprehensive approach to evaluating the safety of biologically derived nanoparticles for therapeutic use.
Area of Science:
- Biotechnology
- Nanomedicine
- Toxicology
Background:
- Biologically derived nanoparticles, like extracellular vesicles, show therapeutic promise but their in vivo toxicity is difficult to predict from in vitro studies.
- Preclinical assessment of nanoparticle therapeutics requires robust in vivo toxicity screening, yet standardized methods are lacking.
- Evaluating adverse effects and characterizing toxicity in vivo is crucial for the safe clinical translation of nanoparticle-based therapies.
Purpose of the Study:
- To develop a scalable method for preparing bovine milk-derived nanovesicles (MNVs).
- To establish and utilize vertebrate animal models (mice and zebrafish) for comprehensive in vivo toxicity assessment of biologically derived nanoparticles.
- To provide standardized protocols for evaluating organ toxicity, immunological effects, and developmental toxicity of nanoparticle treatments.
Main Methods:
- Isolation and preparation of milk-derived nanovesicles (MNVs).
- Systemic injection of MNVs into mice to assess organ toxicity and immune cell profiling.
- Exposure of zebrafish embryos to MNVs in embryo water to evaluate developmental toxicity.
Main Results:
- Demonstrated a method for inexpensive and scalable production of MNVs.
- Successfully employed mice and zebrafish models to screen for in vivo toxicity.
- Provided a framework for assessing organ, immune, and developmental toxicity of MNVs.
Conclusions:
- The described methods enable cost-effective and scalable production of MNVs.
- Mice and zebrafish serve as effective preclinical models for evaluating the in vivo toxicity of biologically derived nanoparticles.
- These protocols facilitate comprehensive safety assessments for therapeutic nanoparticle development.
