Nance Lab
Disease-Directed Engineering
Develop tunable, developmentally appropriate models of brain disease
The only way to simulate the complex environment of the brain is, well, the brain! We use living tissue models of the brain to study how disease, injury and treatment alter brain cells, vasculature, and extracellular spaces. Specifically, we have pioneered the use of organotypic whole hemisphere (OWH) brain slices from a range of species, including rat, ferret, mouse, rabbit, and OR-obtained human samples, and can study donor age, sex-dependent and brain-region dependent responses to various stimuli. Adapting from in vitro and in vivo techniques, we have published molecular, cellular, and functional assessments in slices to study changes in the cellular and extracellular space. The range of length and time scales possible for data generated from slice models allow us to apply data science and machine learning tools to uncover biological insights that are otherwise not obvious through standard data analysis methods. These slice platforms allow high throughput live tissue imaging in the acute and chronic windows after injury, or to study aspects of neurodevelopment, while retaining the 3D cytoarchitecture and in vivo physiological function of brain cells. A single brain can provide 20-30 slices, which also reduced the number of animal lives needed to study brain injury and treatment. In addition, we have developed microfluidic model systems of the blood-brain barrier (BBB) functions to study response to stimuli of multiple brain cell types to engage in a physiologically relevant 3-dimentional architecture in the presence of flow. Current project areas and recent publications are highlighted below.
Modeling hypoxia-ischemia ex vivo
Loss of oxygen and blood flow to the brain is a common cause of mortality around birth or brain damage after birth. We have developed living brain slice models of hypoxic-ischemia injury in the rat and ferret that mimic the injury seen in vivo and clinically in extremely preterm, preterm, and term equivalent infants. We have used these brain slice models to study the effect of developmental age, sex, and severity on regional cellular response and treatment response.
Modeling brain injury in low resource settings
There has long been a distinction in the data reported on the prevalence and associated mortality of neurological disease: incidence rates are higher and outcomes are worse in low resource settings (LRSs) compared to high resource settings (HRSs). Recent work in our lab focuses on developing ex vivo models of brain pathology in the LRS, which has more dominant white matter injury.
Microfluidic model of the blood-brain barrier (BBB)
The BBB describes the collection of endothelial cells, pericytes, astrocytes, and other cells which line vessels and capillaries in the brain. Endothelial cells also require perfusion as well as cell-cell interaction. Microfluidic modeling allows for incorporation of these cell types and flow rates that mimic blood flow in capillaries. We use a commercially available microfluidic device to investigate the effect of hypoxia and nutrient deprivation on BBB function, cell viability, and permeability in the presence of flow.
Modeling acute and traumatic brain injuries
OWH brain slice models can mimic disease hallmarks seen in acute and chronic neurological disease and be used to probe the mechanism of microenvironmental changes in a regionally dependent manner. Our work in this area has investigated chronic exposure to environmental toxins that disrupt mitochondrial function, 'double hit' models with multiple stimuli, and more recently modeling blast injury or concussive forces that result in traumatic brain injury.
Selected recent publications:
Butler B., Renney M., Bennett K., Charpentier G., Nance E.* A rotenone organotypic whole hemisphere slice model of mitochondrial abnormalities in the neonatal brain. J. Biological Engineering. (2024) Nov 14;18(1):67. doi: 10.1186/s13036-024-00465-w
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Floryanzia S., Lee S., Nance E. Isolation methods and characterization of primary rat neurovascular cells. J Biol Eng. (2024) Jul 11;18(1):39. doi: 10.1186/s13036-024-00434-3
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Floryanzia S.D. and Nance E*. Applications and considerations for microfluidic systems to model the blood-brain barrier. ACS Appl. Bio Mater. (2023) doi: 10.1021/acsabm.3c00364
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McKenna M., Filteau J., Sluis K., Chungyoun M., Butler B., Schimek N., Nance E.* Organotypic whole hemisphere brain slice models to study the effects of donor age and oxygen-glucose-deprivation on the extracellular properties of cortical and striatal tissue; J. Biological Engineering (2022); 16:14; doi: 10.1186/s13036-022-00293-w
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Wood T.R., Hildahl K., Helmbrecht H., Corry K.A., Moralejo D.H., Kolnik S.E., Prater K.E., Juul S.E., Nance E.* A ferret brain slice model of oxygen-glucose deprivation captures regional responses to perinatal injury and treatment associated with specific microglial phenotypes. Bioeng & Transl. Med (2021) e10265; doi: 10.1002/btm2.10265
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Liao R., Wood T.R., Nance E*. Superoxide dismutase reduces monosodium glutamate-induced injury in an organotypic whole hemisphere brain slice model of excitotoxicity. J Biol Eng (2020) Feb 4; 14:3. doi: 10.1186/s13036-020-0226-8
Design principles for nanotherapeutic delivery to the brain
We have identified general design principles for engineering therapeutics for delivery to the brain, with the goal that the design principles are adaptable to any nanoparticle system. We have shown that nanoparticles of sizes ranging from 10s of nanometers to larger than 100mm can overcome the BBB when it is impaired, or when the BBB is intact, by leveraging receptor transports , osmotically induced permeability, or with external non-invasive BBB permeability strategies. Our growing evidence also demonstrates that particles reaching the brain from systemic administration should have near-neutral or anionic net surface charge, and that cationic surface charges on nanoparticles result an inability to cross even in an impaired BBB. Within the brain parenchyma, we have shown that the estimated upper limit for rapid nanoparticle transport is around 120nm, with potential for larger sizes based on results by us and others that show effective spaces in the brain can range up to 1µm in size. We have also identified the importance of surface charge and functionalization in increasing nanoparticle penetration in the brain, with neutral or anionic net surface charges imparted by hydroxyl, methoxy, or carboxyl groups driving maximum diffusive ability in the brain. Thus far, we have applied these general design criteria to nanoparticles made from polymer, quantum dot, cellulose, peptoid, dendrimer, or extracellular vesicle materials. Current project areas and recent publications are highlighted below.​
Analyzing diffusion in the brain
Through application of multiple particle tracking, we can measure the real-time displacement of individual probes in the living brain. We can quantify Brownian motion and non-Brownian motion to determine what physical and chemical properties impact the ability of a molecule or particle to penetrate within the brain parenchyma. Our current efforts aim to decouple different physical and chemical properties from each other to determine which properties are the most significant contributors to a particle's ability to penetrate within the brain microenvironment.
Formulation parameters impact brain uptake
Stabilizers or emulsifiers are identified by the FDA as generally regarded as safe (GRAS) and are incorporated in the final formulation in very small quantities for many pharmaceutical products, including nanoparticles. We have shown that the type of emulsifier or stabilizer can influence particle distribution in the body, uptake in the brain, and distribution and cellular localization in the brain. Our ongoing work analyzes how the administration route and developmental age influence cell fate in the brain.
Nanoparticle agnostic analysis
We have worked with polymer, quantum dot, cellulose, peptoid, and dendrimer nanoparticles, and extracellular vesicles (a biologically produced nanoparticle) to study how design principles impact nanoparticle delivery to the brain. These particles can achieve cell specific uptake in the brain if they are less than 100nm in one dimension, near-neutral or net-negative in surface charge, surface functionalized with hydroxyl, methoxy, or carboxyl groups, and have a non-hydrophobic surface. We are currently exploring the role of nanoparticle shape, architecture, and rigidity on brain delivery for different nanoparticles.
Quantifying and predicting nanoparticle fate
Contradictory findings for individual brain cell-nanoparticle interactions persist in the field, likely due to variability of models used in each study. We use in vitro, ex vivo, and in vivo models to quantify nanoparticle fate in the brain. In addition, our analysis of nanoparticle behavior in the brain is data rich. We have leveraged data science and machine learning tools applied to our nanoparticle diffusion data to distinguish nanoparticle properties in living tissue, such as size and protein adsorption, and predict local viscosity and cell internalization.
Selected recent publications:
Schimek N., Wood T., Beck D.A.C., Toghani A., McKenna M., Nance E.* High fidelity predictions of diffusion in the brain microenvironment. Biophys J. (2024), Vol 123, Issue 22, 3935 – 3950. doi: 10.1016/j.bpj.2024.10.005
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Joseph A.* and Nance E.* Nanotherapeutics and the brain. Annual Reviews in Chemical & Biomolecular Engineering; (2022) 13:5.1-15.22, doi: 10.1146/annurev-chembioeng-092220-030853
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Joseph A., Motchoffo Simo G., Gao T., Alhindi N.2, Xu N., Graham D.J., Gamble L.J., Nance E.* Surfactants influence polymeric nanoparticle fate in the brain. Biomaterials (2021) 277: 121086; doi: 10.1016/j.biomaterials.2021.121086
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McKenna M., Shackelford D., Ferreira Pontes C., Ball B.K., Nance E.* Multiple particle tracking detects changes in brain extracellular matrix structure and predicts neurodevelopmental age. ACS Nano (2021) 15(5):8559-8573; doi: 10.1021/acsnano.1c00394
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Helmbrecht H., Joseph A., Zhang M., McKenna M., Nance E.* Governing transport principles for nanomedicine applications in the brain. Current Opinion in Chemical Engineering (2020) 30; 112-119 doi: 10.1016/j.coche.2020.08.010
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Joseph A., Liao R., Zhang M., Helmbrecht H., McKenna M., Filteau J., Nance E.* Nanoparticle-microglial interaction in the ischemic brain is modulated by injury duration and treatment. Bioengineering & Translational Medicine (2020) August; doi: 10.1002/btm2.10175
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Curtis C.D., McKenna M., Pontes C., Toghani D., Choe A., Nance E.* Predicting in situ nanoparticle behavior using multiple particle tracking and artificial neural networks. Nanoscale (2019) Nov 28; 11(46):22515-22530. doi: 10.1039/c9nr06327g
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Zhang M., Bishop B., Thompson N., Hildahl K., Dang B., Mironchuk O., Chen N., Aoki R., Holmberg V.*, Nance E.* Quantum dot cellular uptake and toxicity in the developing brain: implications for use as imaging probes. Nanoscale advances (2019) 1, 3424-3442. doi: 10.1039/C9NA00334G​​
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Curtis C., Toghani D., Wong B., Nance E.* Colloidal stability as a determinant of nanoparticle behavior in the brain. Colloids and Surfaces B: Biointerfaces (2018) 170, 673-682. doi: 10.1016/j.colsurfb.2018.06.050
Evaluating neurotherapeutics for improving brain health in newborns and children
Therapies intended for use in children can take up to 7 years longer to go from the first clinical trial in adults to the first trial in children; often, many approved adult therapeutics are used off-label for children. There is a significant gap in technological development for the neonatal and pediatric populations, particularly in technology that focuses on improving therapeutic outcomes for children and newborns with a range of conditions. Our research seeks to develop and evaluate therapeutic delivery systems for newborns and children, who have unique physiologies compared to adults. We focus specifically on engineering therapeutics that mitigate or attenuate ongoing injury in the brain, with the goal of improving neurological function and quality of life across the lifespan. We have ongoing collaborations that are developing therapeutics for ALS, Huntington's Disease, and other adult neurological conditions.
Nanoparticle pharmacokinetics in neonatal and pediatric populations
Most nanomedicine platforms are evaluated in adults. Off-label use of a drug can cause a higher risk of adverse drug reactions for children, especially for neonates, infants, and children younger than two years old, even when the original purpose of off-label drug use is to benefit these patients . The development of nanomedicine for pediatrics is challenged by the lack of pharmacokinetic (PK) data in the pediatric population, a gap in data even more significant for neonates. Our lab generates PK data for neonatal and pediatric populations in multiple species, and uses this data in collaboration with pharmacologists to do first-in-human predictions of how a drug or a nanomedicine will behave.
Ex vivo therapeutic screening
There is a need to screen promising therapeutics that may be neuroprotective for extremely preterm, preterm, and term brain injury. We use our brain slice culture models to screen individual drugs and combinations of drugs to evaluate their global and regional therapeutic effect via a variety of assessments, including global injury, regional cell death, inflammation and oxidative stress markers, protein and gene expression changes, and cell composition and morphological changes. We screen current FDA approved drugs, drugs being repurposed for our target patient populations and novel drugs just as nanotherapeutics.
Assessment of efficacy in animal models of perinatal brain injury
For therapeutics that show a beneficial effect in our ex vivo models, we next assess their therapeutic efficacy in clinically relevant models of brain injury. Our efforts largely focus on brain injury or disease that significantly impacts brain function early in life. We work with rat, ferret, and piglet models of perinatal and pediatric brain injury or disease. In addition to evaluating therapeutic biodistribution and safety, we also asses the neuroprotective effect of a therapeutic through evaluation of gross injury, regional neuropathology, cellular response, and gene expression changes.
Formulation stability and shelf-life assessment
There are many factors that influence the drug-delivery system behavior, stability, and shelf-life, all of which influence the integrity of the formulation for clinical use. Our formulations undergo rigorous experimental testing and scale-up evaluation. Formulations are subjected to cryoprotection and storage conditions, freeze thaw cycles, drug activity retention and colloidal stability analysis. Additional work is carried out to scale up formulations to amounts relevant to humans.
Selected recent publications:
Xu. N, Wixey J., Chand K., Wong M., Nance E*. Nano-formulated curcumin uptake and biodistribution in the fetal growth restricted newborn piglet brain. Drug Delivery & Translational Res. (2025) Mar 7; doi: 10.1007/s13346-025-01830-y
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Xu N., Wong M., Balistreri G., Nance E.* Neonatal pharmacokinetics and biodistribution of polymeric nanoparticles and effect of surfactant. Pharmaceutics (2023) 15(4): 1176; doi: 10.3390/pharmaceutics15041176
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Kolnik S., Corry K., Hildahl K., Filteau J., White O., Brandon O., Farid L., Shearlock AM, Mareljo D., Juul S., Nance E., Wood T.* Vitamin E decreases cytotoxicity and mitigates inflammatory and oxidative stress responses in a ferret organotypic brain slice model of neonatal hypoxia-ischemia. Developmental Neuroscience (2022) doi: 10.1159/000522485
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Nguyen, N.P., Helmbrecht H., Ye Z., Adebayo T., Hashi N., Doan M-A., Nance E.* Brain tissue derived extracellular vesicles for therapeutic in neonatal ischemic brain injury. Int Journal of Molec Sci. (2022) 23(2):620. doi: 10.3390/ijms23020620
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Nyambura C., Nance E., Pfaendtner J.*, Examining the Effect of Polymer Extension on Protein-Polymer Interactions That Occur during Formulation of Protein-Loaded Poly(lactic acid-co-glycolic acid)-polyethylene Glycol Nanoparticles. Polymers (2022) 14(21): 4730; doi: 10.3390/polym14214730
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Joseph A., Nyambura C., Bondurant D., Corry K., Beebout D., Wood T., Pfaendtner J., Nance E.* Formulation and efficacy of catalase-loaded nanoparticles for the treatment of neonatal hypoxic-ischemic encephalopathy. Pharmaceutics (2021) 13(8), 1131; doi: 10.3390/pharmaceutics13081131
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Liao R., Pon J., Chungyoun M., Nance E.* Enzymatic protection and biocompatibility screening of enzyme-loaded polymeric nanoparticles for neurotherapeutic applications. Biomaterials (2020) July; 257. doi: 10.1016/j.biomaterials.2020.120238
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Yellepeddi V*, Joseph A., Nance E.* Pharmacokinetics of nanotechnology-based formulations in pediatric populations. Adv. Drug. Del. Rev. (2019) Nov; 151:44-55. doi: 10.1016/j.addr.2019.08.008
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Joseph A., Wood T., Chen C-C., Corry K., Juul S., Snyder J., Parikh P., Nance E.* Curcumin-loaded brain penetrating nanoparticles for treatment of neonatal hypoxia-ischemia encephalopathy. Nano Research (2018) 11(10): 5670-5688. doi: 10.1007/s12274-018-2104-y