Bridging the In Vitro to In Vivo gap - Using the Chick Embryo Model to Accelerate Nanoparticle Validation and Qualification for In Vivo studies

Sep 21, 2026

Uploaded by Ben

The review addresses why many therapeutic nanoparticles fail to realize clinical potential: in vitro results often cannot predict in vivo performance. It presents the avian embryo and its chorioallantoic membrane (CAM) as an intermediate preclinical model that rapidly screens nanoparticle behavior under physiologically relevant conditions. The CAM’s vascular structure supports intravital imaging of fluorescent nanoparticles to quantify nano-bio interactions and evaluate stability, extravasation, biocompatibility, and distribution over time. The approach helps select candidate nanoparticles for rodent studies while enabling structure-property relationship development and reducing reliance on large mouse populations.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages25

Reading Time1 h 3 min

AccessFree to download

Introduction

Rationale for an intermediate preclinical model for bio-nano research

ABSTRACT

Bridging in vitro to in vivo using the chick embryo and CAM

Discussion

Assessing stability, extravasation, biocompatibility, distribution, and imaging

Conclusion

Emerging CAM techniques for future nano-bio studies

Made available for a limited time for personal research and study only License. [www.acsnano.org](www.acsnano.org) Bridging the In Vitro to In Vivo gap: Using the Chick Embryo Model to Accelerate Nanoparticle Validation and Qualification for In Vivo studies Kimberly S. Butler, C. Jeffrey Brinker, and Hon Sing Leong* articles . Downloaded via PURDUE UNIV on January 19, 2023 at 19:40:12 (UTC) . See [https://pubs.acs.org/sharingguidelines](https://pubs.acs.org/sharingguidelines) for options on how to legitimately share published | ACCESS | Metrics & More | | Article Recommendations | | *sı Supporting Information | | --- | --- | --- | --- | --- | --- | ABSTRACT: We postulate that nanoparticles (NPs) for use in therapeutic applications have largely not realized their clinical potential due to an overall inability to use in vitro results to predict NP performance in vivo. The avian embryo and associated chorioallantoic membrane (CAM) has emerged asan in vivo preclinical model that bridges the gap between in vitro and in vivo, enabling rapid screening of NP behavior under physiologically relevant conditions and providing a rapid, accessible, economical, and more ethical means of qualifying nanoparticles for in vivo use. The CAM is highly vascularized and mimics the diverging/converging vasculature of the liver, spleen, and lungs that serve as nanoparticle traps. Intravital imaging of fluorescently labeled NPs injected into the CAM vasculature enables immediate assessment and quantification of nano-bio interactions at the individual NP scale in any tissue of interest that is perfused with a microvasculature. In this review, we highlight how utilization of the avian embryo and its CAM as a preclinical model can be used to understand NP stability in blood and tissues, extravasation, biocompatibility, and NP distribution over time, thereby serving to identify a subset of NPs with the requisite stability and performance to introduce into rodent models and enabling the development of structure−property relationships and NP optimization without the sacrifice of large populations of mice or other rodents. We then review how the chicken embryo and CAM model systems have been used to accelerate the development of NP delivery and imaging agents by allowing direct visualization of targeted (active) and nontargeted (passive) NP binding, internalization, and cargo delivery to individual cells (of relevance for the treatment of leukemia and metastatic cancer) and cellular ensembles (e.g., cancer xenografts of interest for treatment or imaging of cancer tumors). We conclude by showcasing emerging techniques for the utilization of the CAM in future nano-bio studies. KEYWORDS: chick embryo, nanoparticle development, chorioallantoic membrane, nanoparticle bioavailability, drug development, intravital imaging, structure−function analysis INTRODUCTION: RATIONALE FOR AN INTERMEDIATE PRECLINICAL MODEL FOR BIO-NANO RESEARCH In 2018, MIRIBEL (minimum information reporting in bionano experimental literature) standards were established to stipulate how nanoparticles (NPs) intended for biomedical applications are to be characterized and how their in vitro interactions within any biological system are to be described and evaluated in the literature. 1 MIRIBEL defined a checklist of specific components to be reported, divided into three categories: material characterization, biological characterization, and details of experimental protocols with the intention of improving reproducibility, increasing quantitative comparisons of bio-nano materials, and facilitating meta-analyses and insilico modeling. In response to the publication of MIRIBEL in Nature Nanotechnology, the bio-nano research community was invited to provide suggestions to further these guidelines.2 We advocated for an ex ovo model system to “bridge the gap”between in vitro assessments of NPs stipulated by MIRIBELand in vivo rodent models. Our rationale was that, in spite o

FAQs

Why do nanoparticles often fail to translate from in vitro studies to clinical use?

In vitro assays typically cannot predict nanoparticle performance in vivo. Key clearance and uptake processes present in living organisms are not captured by in vitro investigations.

What role does the chick embryo chorioallantoic membrane (CAM) play in nanoparticle validation?

The CAM functions as an in vivo preclinical bridge between in vitro and in vivo. Its highly vascularized structure enables rapid screening under physiologically relevant conditions.

How does intravital imaging support nanoparticle assessment in the CAM model?

Fluorescently labeled nanoparticles injected into the CAM vasculature allow immediate assessment and quantification of nano-bio interactions at the individual nanoparticle scale in perfused tissues.

Sep 21, 2026

Uploaded by Ben