Paramagnetic Nanoparticle Formulations for Targeted Traumatic Brain Injury Treatment
Beeta Zamani - Parallel I
Author
09/26/2024
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Student’s name: Beeta Zamani
Home Institution: University of California, San Diego
NNCI Site: SDNI @ UC San Diego
REU Principal Investigator: Dr. Pedro Cabrales Arevalo – Jacobs School of Engineering, Shu Chien-Gene Lay Department of Bioengineering, UC San Diego
REU Mentor: Dr. Cynthia Muller - Jacobs School of Engineering, Shu Chien-Gene Lay Department of Bioengineering, UC San Diego
Abstract: Traumatic brain injuries (TBIs) are damages to the brain tissue that arise from external damage to the skull, and are a leading cause of death and disability in the United States. The severity of TBIs is largely correlated to incident injury severity, which results in localized damage that disrupts normal brain function. Primary injuries are observed in patients immediately after TBI incidence, and require immediate diagnosis and care. However, secondary injury can also arise weeks, months, or even years after, making TBI extremely dangerous. The urgent nature of this injury requires therapeutics that are fast-acting, localized, and have minimal side effects for best patient outcomes and prevention of future complications. This research carefully considers the cascading physiological effects of TBI to design a ferromagnetic nanoparticle for expedited treatment of TBIs. Targeted and localized delivery of nitric oxide is achieved with these ferromagnetic nanoparticles to restore blood flow to the injury site, which provides an innovative and non-invasive solution to treatment and reduction of TBI damage. This treatment was explored in rodents to carefully observe nanoparticle treatment efficacy, symptoms and side-effects after treatment, and performance in central nervous system function after administration post-TBI.
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- [00:00:00.200]Hi everyone, I'm Anita Tawani from UC San Diego, and my research over the summer has been focused on developing the
- [00:00:07.920]paramagnetic nanoparticle formulations for treatment of traumatic brain injuries.
- [00:00:14.280]So kind of getting into this, we're first going to look at what traumatic brain injuries are and the disease mechanism that leads them.
- [00:00:22.640]And from this, how we can develop different types of nanoparticles that can be used in an animal model to study traumatic brain injuries.
- [00:00:29.920]And then I'll go over my results, conclusions, and some future steps.
- [00:00:33.920]So firstly, traumatic brain injuries are actually the second most prevalent disability among Americans, according to the CDC.
- [00:00:41.160]And each year, 2.5 million individuals experience a TBI in the United States.
- [00:00:47.040]And what makes this disease really unique and really impactful is that it can be caused by any forceful head injury.
- [00:00:53.680]So this can be a fall, a car crash, a sports injury, or something personally school.
- [00:00:59.840]So given the nature of this disease, it can arise in many different ways.
- [00:01:04.080]You can have a blow to a different part of your head, and it can be to a different severity.
- [00:01:08.480]So this leads to us having primary versus secondary TBIs.
- [00:01:12.880]Every single traumatic brain injury will have a primary incidence.
- [00:01:16.640]So this will cause a cascade of physiological effects that I'll go over.
- [00:01:20.400]But only some of them will have secondary TBIs, which cause further injury due to prolonged effects.
- [00:01:27.920]And traumatic brain injuries
- [00:01:29.760]will ultimately result in physical and cognitive behavioral perception and sensory issues,
- [00:01:34.960]as you can see.
- [00:01:36.080]So it causes a very comprehensive cascade of symptoms that are dependent on the person
- [00:01:42.960]experiencing them.
- [00:01:43.840]So the pathogenesis of TBIs really depends on, again, the primary and secondary injury.
- [00:01:50.800]So in the primary injury, we immediately observe a cascade of cellular and molecular processes
- [00:01:56.400]after the initial flow.
- [00:01:58.080]And this can lead to hematoma,
- [00:01:59.680]contusions, or diffuse axonal injury.
- [00:02:02.560]However, only some individuals will experience secondary injury, which will lead to a breakdown
- [00:02:08.960]of the blood-brain barrier, which can cause brain infection, low blood pressure, hydrocephalus,
- [00:02:14.880]and more.
- [00:02:15.840]So we're really looking at a disease model that can progress weeks, months, or even years
- [00:02:21.280]later after the initial injury.
- [00:02:23.600]So based on this, and based on the very unique nature of TBI, depending on the person and
- [00:02:28.560]the situation,
- [00:02:29.600]there's a lot of different pathophysiological pathways that arise and a lot of mechanisms
- [00:02:34.560]of TBI.
- [00:02:35.600]This makes the disease really difficult to treat.
- [00:02:38.160]However, this also means that there are many therapeutic targets that we can achieve.
- [00:02:43.120]So looking into the methods that I took in my research to develop these paramagnetic
- [00:02:49.440]nanoparticle formulations.
- [00:02:51.440]So as you can see on the right, these are magnetic nanoparticles that can be loaded
- [00:02:56.320]with a drug, which is the blue star that you see in the core of this nanoparticle.
- [00:02:59.520]This can allow us to achieve targeted delivery and release of our drug by this magnetic drug
- [00:03:06.720]targeting mechanism.
- [00:03:07.760]So by administering these paramagnetic nanoparticles, we're able to also induce the magnetic field
- [00:03:15.360]at the TBI site and all the nanoparticles will accumulate in that site, release the
- [00:03:21.440]payload, and therefore have a therapeutic effect for TBI that we are trying to achieve.
- [00:03:27.120]So some really unique properties.
- [00:03:29.440]of this therapeutic that really set it apart from traditional methods of treatment are
- [00:03:34.480]the size scale.
- [00:03:35.600]So these nanoparticles I found in my research to be less than 100 nanometers in size, meaning
- [00:03:41.120]that they can pass through the blood-brain barrier without issue.
- [00:03:44.400]They're also paramagnetic, so they're attracted to the magnetic field, but after you remove
- [00:03:49.120]the magnetic field, they are no longer magnetized.
- [00:03:52.320]Therefore, we have targeted delivery and safe clearance, preventing any cytotoxic effects
- [00:03:57.840]from the drug remaining in that site.
- [00:03:59.360]In my study, I loaded these particles with an oxygen delivery agent, so in traumatic
- [00:04:06.880]brain injuries, in the tissue itself, there's a hypoxic environment.
- [00:04:11.200]By delivering these O2 agents, we're able to facilitate a return to the homeostatic
- [00:04:18.960]physiological function in the brain.
- [00:04:20.720]Ultimately, the key point is that these antiparticle drug formulations are not toxic, targeted
- [00:04:28.080]to the brain tissue.
- [00:04:29.280]They are able to overcome the blood-brain barrier and offer therapeutic TBI due to the
- [00:04:34.000]drug that flows through them.
- [00:04:36.000]Essentially, my research was focused on the synthesis of them, as you can see on the screen.
- [00:04:42.960]I determined the therapeutic dosage to be 10 milligrams per kilogram.
- [00:04:48.400]I'll go into that in more detail now.
- [00:04:50.400]I used an animal model in Wistar rats, about 350 to 400 grams in weight.
- [00:04:56.880]I ultimately had five
- [00:04:59.200]experimental groups, five control and five variable.
- [00:05:03.040]And then we were ultimately able to prepare the study in this manner.
- [00:05:08.320]So first, the rats were able to be instrumented with a catheter in the femoral artery.
- [00:05:15.200]And we had a two kilohertz sampling rate.
- [00:05:18.400]And we achieved measurements of arterial blood pressure and heart rate.
- [00:05:23.040]And then using the software, we were able to achieve all these other metrics as well.
- [00:05:29.120]So after instrumenting the rats, we also administered a cortical impact with surgery in the right
- [00:05:38.160]part of the brain, in the dura, we were able to impact the brain of the rats.
- [00:05:45.280]So this allowed us to achieve a very continuous model of TBI across the brain.
- [00:05:50.520]our groups. And ultimately, what I found from my study is that the TBI model was able to produce
- [00:05:58.490]successful and consistent induction of TBI. And this model requires you to have a specific
- [00:06:05.670]velocity and dwell time for the impactor, as you can see on this screen. So we had to also
- [00:06:12.290]determine those metrics as well, which were successful in producing consistent results for
- [00:06:17.430]TBI. And there were also no complications observed within this model. And that's kind of
- [00:06:23.810]the difficult part, right, in the study is not actually coming up with a nanoparticle, but making
- [00:06:28.730]sure your model is actually consistent and actually is trying to study what you want. So
- [00:06:34.190]that was really successful and allowed me to move into the study for the nanoparticle therapeutic
- [00:06:39.970]efficacy. So after achieving the data that you can see here, all of these different
- [00:06:46.190]tests, with the blood samples and with the five catheter assessment parameters, we were able to
- [00:06:53.430]find statistically significant results. So we saw statistically significant improvements in
- [00:06:59.790]recovery of arterial blood pressure and heart rate. So after TBI, these metrics dropped due
- [00:07:07.730]to the injury in the brain tissue, but we were able to observe statistically significant improvements.
- [00:07:12.770]And importantly, the targeting
- [00:07:16.170]of these nanoparticles was effectively achieved with this magnetic field induction. So that's
- [00:07:22.890]super important for us to know that we're actually able to IV administer the nanoparticle formulation,
- [00:07:27.990]bypass the blood-brain barrier, and have the magnetic field have all of the nanoparticles
- [00:07:33.910]accumulate in the specific part of the brain that we impacted. And this also brings into
- [00:07:39.170]consideration dosage. So as I mentioned, after some experimentation, I found the dosage to be
- [00:07:46.150]1,500 grams per kilogram, and this was effective in preventing ketotoxic effects, which are seen
- [00:07:52.670]if you have too high of a dosage, and premature cell uptake if you have too low dosage. So
- [00:07:57.830]ultimately, we were able to observe successful results in both of these aspects. And again,
- [00:08:04.630]what I was able to find is the optimal nanoparticle therapeutic dosage. We were able to
- [00:08:10.790]also see that the pyramid magnetic nanoparticle drug loading the O2 delivery agent can
- [00:08:16.130]statistically significantly act as a TBI therapeutic agent. So it can bypass the BVB
- [00:08:23.230]in the kidney and brain tissue, and it's also able to avoid toxic effects. So we're able to
- [00:08:28.850]have it be an effective therapeutic. And also, we were able to see statistically significant
- [00:08:34.190]differences in respiratory rate and blood pressure recovery over time in the rats. And
- [00:08:40.350]this ultimately indicates TBI recovery, which is significant for all of the preventive
- [00:08:46.110]conditions. Secondary effects that can arise when the primary injury is not treated properly.
- [00:08:51.790]So my next steps are to accumulate more animal study data, so just having more of the same
- [00:08:57.450]experimental groups. I can also confirm the nanoparticle release profile, since we can
- [00:09:02.050]see how these particles are actually being released, since they're magnetic. And there's
- [00:09:07.230]also microelectrics that we can apply to the affected part of the brain to see the local
- [00:09:12.710]concentration of the nanoparticles.
- [00:09:16.090]Ultimately, we would be looking at conducting more behavior tests, so having a live study
- [00:09:22.670]of the rats to see their TBI symptom evaluation over time. And ultimately, looking into diversifying
- [00:09:28.330]the TBI model and having the TBI into the brain.
- [00:09:32.810]And thank you so much for listening.
- [00:09:37.250]Awesome work. Thank you for sharing. I was wondering for the dosage, when you were changing
- [00:09:46.070]the dosage, did you change the actual amount of drug within each lipid nanoparticle? Or
- [00:09:50.850]did you change the amount of nanoparticles distributed?
- [00:09:54.370]So the way in which these nanoparticles were produced is that the synthesis was ultimately
- [00:10:02.310]allowing us to have a powder. So this powder is actually really unique. So our nanoparticles
- [00:10:08.670]come in this dry, solid form. And that means that whenever you need them, you can rehydrate
- [00:10:16.050]them in a sterile saline solution, which is a common practice for therapeutics. And
- [00:10:20.190]then they can be used in IV administration. So adjusting the dosage means that we rehydrate.
- [00:10:28.050]So in the same volume of saline, we would add more of the actual powder itself. So the
- [00:10:33.830]nanoparticles are already loaded with a specific amount of drug after synthesis. But since
- [00:10:38.630]they're shelf-stable in this dry form, we're able to adjust the dosage as needed, which
- [00:10:43.770]is helpful since you can do it for different weights.
- [00:10:46.030]Thank you. Yes?
- [00:10:47.030]That's the release mechanism for O2. Is it like whenever you magnetize it, does
- [00:10:53.110]it break the R?
- [00:10:54.110]So the way that it works is that since the actual drug itself is an oxygen delivery
- [00:11:01.550]-- is an oxygen delivery -- sorry, I have it written -- O2 delivery agent. So when we're
- [00:11:09.750]encapsulating it in a nanoparticle, the nanoparticle is being directed to the site of TBI, and
- [00:11:16.010]that's where you induce the magnetic field. And then in the magnetic field, it opens up
- [00:11:21.470]to release the payload, which is the drug itself. And the drug will cause the cascade
- [00:11:25.950]by -- it delivers oxygen to that site, and then it causes a cascade of molecular effects
- [00:11:30.970]that reverse the effects of TBI, since TBI causes a hypoxic environment. I hope that
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