Production and Evaluation of Lipid Nanoparticle as Drug Carrier for

Treatment of Alzheimer’s Disease to Enhance the Blood Brain

Barrier Penetration

S. Chitra, A. Sundar Raj, G. Yalini and K. Raghuram

Department of Biomedical Engineering, E.G.S Pillay Engineering College, Nagapattinam - 611002, Tamil Nadu, India

Keywords: Alzheimer's Disease, Lipid Nanoparticles, Blood‑Brain Barrier, Drug Delivery, Amyloid‑Beta, Tau Protein,

Neuroinflammation, Therapeutic Applications, siRNA, Sustained Release.

Abstract: Alzheimer's disease (AD) is a progressive neurodegenerative disorder that most frequently affects older

adults, resulting in cognitive impairment, memory loss, and behavioural changes. It is the most prevalent

cause of dementia and an increasing global health issue with aging populations. The pathophysiology of AD

is marked by amyloid-beta plaque deposition, tau protein tangle formation, and synaptic loss, leading to

neuronal dysfunction and cell death. Despite the fact that existing pharmacological therapies may correct

symptoms, they do not reverse or stop disease progression. One of the greatest obstacles to treating AD is the

blood-brain barrier (BBB), which shields the brain from the effective delivery of therapeutic compounds and

thus reduces the effectiveness of most drugs. This restriction necessitates novel drug delivery systems that

have the ability to cross the BBB and deliver drug therapeutic agents in adequate concentrations into the brain.

Lipid nanoparticles (LNPs) also present a potent solution to such a challenge. LNPs represent nanosized lipid-

based carrier systems that encapsulate hydrophobic and hydrophilic drugs, providing an environment of a

biocompatible platform for delivery of drugs as targeted therapy. The inherent characteristics of LNPs,

including their nanoscale dimensions, flexibility, and capacity to alter surface chemistry, make them capable

of interacting favourably with biological membranes and penetrating the BBB. This is a quality that makes

LNPs especially ideal for delivering therapeutic agents to the brain. The therapeutic uses of LNPs in the

treatment of AD are multifaceted, targeting the most important pathological characteristics of the disease,

including amyloid-beta plaques and tau protein tangles. LNPs may be used to deliver small molecules,

peptides, and antibodies to prevent amyloid-beta aggregation or facilitate clearance of amyloid-beta from the

brain, perhaps delaying disease. RNA therapies such as siRNA and antisense oligonucleotides are deliverable

through LNPs and can suppress production of amyloid-beta, serving as an alternate mechanism for

modification of disease. Overall, lipid nanoparticles are a promising drug delivery system for Alzheimer's

disease. Their capacity to penetrate the BBB, reach specific areas of the brain, and deliver therapeutic agents

in controlled release makes them an important resource for treating the intricacies of AD.

1 INTRODUCTION

1.1 Overview of Alzheimer's Disease

Alzheimer's disease (AD) is a chronic, progressive

neurodegenerative disease that causes cognitive

impairment, such as memory loss, compromised

reasoning, and problem-solving difficulty. It is the

most common cause of dementia, a syndrome of

profound loss of cognitive function, which has a

significant impact on an individual's capability to

perform activities of daily living. Alzheimer's disease

most commonly affects older individuals, and its

incidence rises with an aging population across the

globe. Estimates suggest that by the year 2050,

Alzheimer's disease will double the number of

individuals with the disease, and thus it will be an

enormous strain on the public health care systems

globally (Cummings et al., 2019). The disease is

accompanied by initial mild forgetfulness in its early

stage, but subsequently, the patient develops critical

memory loss, language problems, and confusion. In

its advanced stages, the patient loses the ability to

recognize close relatives, becomes bedridden, and

needs 24-hour care (Cunningham, C et al., 2020). At

168

Chitra, S., Raj, A. S., Yalini, G. and Raghuram, K.

Production and Evaluation of Lipid Nanoparticle as Drug Carrier for Treatment of Alzheimer’s Disease to Enhance the Blood Brain Barrier Penetration.

DOI: 10.5220/0013924500004919

Paper published under CC license (CC BY-NC-ND 4.0)

In Proceedings of the 1st International Conference on Research and Development in Information, Communication, and Computing Technologies (ICRDICCT‘25 2025) - Volume 5, pages

168-181

ISBN: 978-989-758-777-1

the molecular level, AD is defined by the presence of

amyloid-beta (Aβ) plaques deposits, tau protein

tangles, and neuroinflammation, which all lead to

neuronal dysfunction and cell death. The amyloid

plaques, composed of the aggregation of Aβ peptides,

block synaptic communication by disrupting

neuronal signalling (Cheng, S et al., 2020).

Meanwhile, tau tangles, due to the

hyperphosphorylation of tau proteins, disrupt

microtubule stability and intracellular transport,

further destroying neurons and inducing cognitive

impairment (Yuan et al., 2020). Despite years of

investigation into possible treatments, AD therapies

currently remain primarily symptomatic.

Acetylcholinesterase inhibitors like donepezil and

rivastigmine, which increase the concentration of

acetylcholine in the brain, have modest benefits in

improving performance on memory tasks and

reducing cognitive decline (Zhou et al., 2018).

Similarly, glutamate receptor antagonists like

memantine aim to block excitotoxicity through the

regulation of glutamate transmission, but these drugs

fail to treat the root causes of the disease (Cummings

et al., 2019). Thus, disease-modifying treatments that

not only alleviate symptoms but also reverse or slow

down AD progression are in dire need. A major

roadblock to identifying effective therapies is the

blood-brain barrier (BBB), a physical barrier that

impedes the delivery of therapeutic compounds into

the brain.

1.2 Blood-Brain Barrier (BBB) Issue

with Alzheimer's Therapy

The blood-brain barrier (BBB) is a selectively

permeable membrane between the circulatory system

and the brain. It is formed by tight junctions of

endothelial cells, pericytes, and astrocyte end-feet,

which collectively exclude potentially toxic

substances while allowing the entry of necessary

nutrients and gases (Zhou et al., 2018). The BBB is

an essential protective system for the brain, protecting

it from toxins, infection, and alterations in blood

composition. But the same factors that render the

BBB such a powerful protective system render very

significant obstacles to the delivery of drugs to the

brain. While the BBB allows small molecules such as

glucose and oxygen to pass across, it actively

excludes the passage of large molecules, including

most therapeutic drugs that might potentially be of

value in the treatment of neurological disorders such

as Alzheimer's (Zhou et al., 2018). In Alzheimer's, the

BBB poses another barrier: many drug candidates,

even amyloid-beta plaque, tau tangles, and

neuroinflammation candidates, are unable to cross the

BBB in therapeutic levels (Wang et al., 2022). As a

result, even potential candidates cannot reach their

target site in the brain, greatly hindering the

development of effective treatments. This is

compounded by the fact that AD pathophysiology

involves more than one molecular mechanism, such

as amyloid-beta deposition, tau

hyperphosphorylation, oxidative stress, and

neuroinflammation, all of which require targeted drug

delivery systems that can access specific regions of

the brain (Zhang et al., 2019). BBB penetration has

been a prominent area of study in drug delivery for

Alzheimer's disease. Different strategies have been

proposed, including the use of focused ultrasound for

reversible disruption of the BBB, receptor-mediated

delivery, and nanoparticle-based drug delivery

systems (Li et al., 2021). While some of these

strategies have seemed promising, they are typically

marred by problems of invasiveness, efficacy, and

safety. Of these strategies, lipid nanoparticles (LNPs)

have garnered a lot of attention as a promising

strategy to drug delivery to the brain. The unique

features of LNPs, including their nanoscale size,

biocompatibility, and capacity to encapsulate a range

of therapeutic agents, make them most suited to cross

the BBB and deliver drugs directly to the brain (Song

et al., 2020).

1.3 Use of Lipid Nanoparticles (LNPs)

in Drug Delivery

Lipid nanoparticles (LNPs) are nanocarriers that have

been extensively studied for their ability to deliver

therapeutic agents through the BBB and into the

brain. LNPs are typically composed of lipid

molecules in a formulation that is able to encapsulate

hydrophobic as well as hydrophilic drugs, thus

making them universal carriers of a range of

therapeutic compounds (Li et al., 2021). Lipid

composition of LNPs provides some benefits such as

biocompatibility, biodegradability, and reduced

toxicity, all of which play crucial roles in achieving

the successful delivery of drugs into the brain (Kalluri

et al., 2019). The strongest advantage of LNPs is also

their size, typically ranging between 20-100 nm. This

size helps LNPs move across the junctions of BBB

endothelial cells by mechanisms of endocytosis or

transcytosis (Li et al., 2021). The figure 1 shows the

Pictorial Representation of Lipid Nanoparticle (LNP)

for Alzheimer's Disease Treatment (Song et al.,

2020). The ability of LNPs to traverse the BBB and

deliver medicines to the brain in a specific manner is

useful particularly in diseases of the nerve system

Production and Evaluation of Lipid Nanoparticle as Drug Carrier for Treatment of Alzheimer’s Disease to Enhance the Blood Brain Barrier

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169

such as Alzheimer's disease, where selective delivery

is pivotal in order to achieve therapeutic performance.

Moreover, LNPs are engineered to provide a broad

selection of therapeutic species such as peptides,

proteins, nucleic acids, and small molecules of

applicability to therapy of various aspects of

Alzheimer's pathophysiology (Wang et al., 2022). In

addition to the ability to provide a variety of

therapeutic drugs, LNPs also have controlled and

sustained release. This is particularly critical in the

treatment of long-term conditions like Alzheimer's,

where stable drug levels in the brain over extended

timeframes can optimize therapeutic effects and

reduce dosing frequency (Song et al., 2020). With

controlled release of drug-encapsulated drugs, LNPs

can ensure that the drug is active in the brain for

extended durations without therapeutic failure caused

by ineffective drug levels.

Figure 1: Pictorial representation of lipid nanoparticle

(LNP) for Alzheimer's disease treatment (Song et al., 2020).

1.4 Therapeutic Applications of Lipid

Nanoparticles for the Treatment of

Alzheimer's Disease

Lipid nanoparticles (LNPs) have emerged to be

extremely promising in the treatment of several

therapeutic problems in Alzheimer's disease.

Amyloid-beta (Aβ), a peptide that aggregates to

deposit as plaques in the brain, disrupting synaptic

function and causing cognitive impairment, is one of

the principal therapeutic targets in AD. Aβ plaques

are one of the first and most spectacular

manifestations of AD pathology and are therefore a

significant target for drug treatments. Recent studies

have established the ability of LNPs to deliver

therapeutic cargoes, such as small molecule

inhibitors, antibodies, or RNA-based therapies, which

could prevent amyloid-beta aggregation or enable its

clearance from the brain (Yuan et al., 2020). These

approaches are intended to prevent amyloid-beta

toxicity on neurons and restoration of normal synaptic

function. Besides modulation of amyloid-beta

plaques, tau protein is another prime target of AD. Tau

form neurofibrillary tangles in neurons, leading to

intracellular transport disruption and

neurodegeneration. Tau tangles are also involved in

later stages of the disease and are believed to be at the

centre of cognitive impairment (Zhang et al., 2020).

LNPs can be used to deliver small molecule inhibitors

or RNA-based medicine that modulates tau

phosphorylation, aggregation, and clearance.

Through tau targeting, the researchers hope to

suppress or even reverse tau-mediated

neurodegeneration, offering a putative disease-

modifying treatment for Alzheimer's patients (Li et

al., 2021). They also suggest that neuroinflammation

is a key contributor to Alzheimer's pathogenesis.

Persistent brain inflammation exacerbates neuronal

damage and accelerates disease progression.

Activated microglia and inflammatory cytokines are

typically elevated in AD patient brains, which is

implicated in the neurodegenerative process (Kalluri

et al., 2020). LNPs can be utilized to deliver anti-

inflammatory medication to the brain, reducing

neuroinflammation and its harmful effects. This can

potentially slow disease progression and improve

cognitive function in Alzheimer's patients. The

flexibility of LNPs makes them amenable to deliver a

wide variety of therapeutic agents that can target

various aspects of Alzheimer's disease pathology.

Besides amyloid-beta, tau, and neuroinflammation,

LNPs can be designed to deliver drugs that modulate

neurotransmitter levels, enhance synaptic plasticity,

or provide neuroprotection against oxidative stress.

These multi-target approaches are likely to offer the

most effective way to treat Alzheimer's disease since

they can effectively address the multifactorial and

complex nature of the disease (Song et al., 2020).

2 LITERATURE REVIEW

2.1 Pathophysiology of Alzheimer's

Disease and Therapeutic

Challenges

Alzheimer's disease (AD) is a complex multifactorial

neurodegenerative disorder causing progressive

cognitive impairment, ultimately disabling the

individual's ability to perform activities of daily

living. The most prevalent form of dementia,

Alzheimer's disease affects millions of individuals

globally, and projections are that the number of cases

will swell exponentially within the next few decades

as the global population ages. AD typically begins

with the insidious and gradual onset of memory

impairment, accompanied by language deficits,

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spatial disorientation, and deficits in executive

function. The clinical course is one of slow

deterioration of cognitive function, with the earliest

and most prominent of symptoms being loss of

memory. The disease also impairs other cognitive

areas, including reasoning, problem-solving, and

decision-making.

At the pathological level, the two characteristic

features of AD are extracellular amyloid-beta (Aβ)

plaques and intracellular tau neurofibrillary tangles,

both of which disrupt normal neuronal function and

are responsible for the neurodegenerative process.

Amyloid plaques are composed of aggregates of

amyloid-beta peptide, which is the product of

aberrant cleavage of amyloid precursor protein (APP)

by enzymes like beta-secretase and gamma-secretase.

Such a build-up of plaques disrupts synaptic

transmission, which disrupts neural circuits, most

significantly in areas like the hippocampus and

cortex, which are essential to memory and cognition

functions (Cheng, S et al., 2020), (Yuan et al., 2020).

Alternatively, tau tangles resulting from tau protein

hyperphosphorylation also lead to neuronal

dysfunction. Tau is a microtubule-associated protein

that stabilizes the microtubule structure of the neuron

and promotes organelle and nutrient transport. In AD,

however, tau is abnormally hyperphosphorylated and,

in doing so, loses its microtubule association and

instead forms aggregates in the neuron to create

twisted tangles. These tangles disrupt neuronal

transport and lead to the cell death of affected

neurons, ultimately resulting in brain atrophy and the

resulting cognitive impairments (Yuan et al., 2020),

(Zhou et al., 2018).

Besides amyloid plaques and tau tangles,

neuroinflammation is another key mechanism in the

pathogenesis of AD. Neuroinflammation is the

activation of glial cells such as astrocytes and

microglia following neuronal damage. While glial

cells play a key role in ensuring homeostasis in the

brain, chronic glial cell activation is the reason behind

the release of pro-inflammatory cytokines and

reactive oxygen species, which also harm neurons

and promote neurodegeneration. Recent findings

have implicated the possibility of targeting

neuroinflammation as a potential way of reducing the

impact of AD and slowing the progression of the

disease (Zhou et al., 2018), (Wang et al., 2022).

While pathological mechanisms of AD are well

characterized, therapeutic interventions are

symptomatic. Currently approved drugs, e.g.,

acetylcholinesterase inhibitors (donepezil,

rivastigmine, and galantamine), act by increasing the

concentration of acetylcholine in the brain, a

neurotransmitter involved in learning and memory.

These drugs are not disease etiology curative but at

best modestly effective in slowing the rate of

cognitive decline. A second class of drugs, glutamate

modulators like memantine, decreases excitotoxicity

by modulating glutamate neurotransmission but, like

the first, is symptomatic only and does not alter the

course of the disease (Wang et al., 2022). The lack of

useful disease-modifying therapies is due to the

multifactorial and complicated etiology of AD, not

caused by a single but by the synergistic pathogenic

interaction of genetic, environmental, and lifestyle

factors. The reality of current drug discovery is

plagued with challenges in the ability to discover

molecular targets that can retard or halt disease

progression, and in the ability to provide assurance

that potential therapeutic agents can enter the brain.

The blood-brain barrier (BBB), a selective membrane

to protect the brain from toxic substances, is prone to

bar the effective delivery of drugs and biological

mediators, such as proteins, antibodies, and small

molecules. Therefore, the development of novel

therapeutic strategies to AD requires novel drug

delivery systems with the capability to traverse this

barrier (Zhang et al., 2019), (Song et al., 2020).

2.2 Drug Delivery and Blood-Brain

Barrier Issues

The blood-brain barrier (BBB) is a selective

semipermeable membrane that protects the CNS from

toxins and pathogens but does allow necessary

nutrients to pass through. While it serves a protective

role, however, the BBB is a significant barrier to the

delivery of drugs to the brain. BBB is made up of

pericytes, endothelial cells, and astrocytic end-feet,

which have tight junctions among them that limit the

diffusion of charged entities and large molecules from

the blood into the brain. Thus, many promising drugs

for the treatment of Alzheimer's disease and other

neurodegenerative disorders find it difficult to cross

the BBB in sufficient quantities to be effective (Song

et al., 2020), (Li et al., 2021).

Several strategies have been suggested to enhance

drug delivery through the BBB. One is to create drugs

that are sufficiently small or lipophilic to pass through

the BBB by passive diffusion. But this is typically not

sufficient, as many therapeutic compounds, like

antibodies, small molecules, and nucleic acids, are

too large or hydrophilic to pass through the BBB on

their own. A second strategy is to temporarily open up

the BBB via methods like focused ultrasound or

osmotic disruption so that drugs can travel more

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easily into the brain. But these methods are invasive,

and long-term safety is unknown (Li et al., 2021).

One of the most promising alternative approaches is

the application of drug delivery systems, such as

nanoparticles, liposomes, and viral vectors, in an

attempt to be designed to cross the BBB and deliver

therapeutic substances to the brain. Of these, lipid

nanoparticles (LNPs) have been of interest because

they have the potential to circumvent the BBB and

deliver a broad range of therapeutic molecules, such

as small molecules, proteins, and nucleic acids. The

small particle size of LNPs (range 20-100 nm)

ensures that they are able to traverse the BBB by

passive diffusion or receptor-mediated endocytosis.

LNPs can even be targeted by using targeting ligands

that direct them to areas in the brain in AD pathology

(Zhang et al., 2020), (Kalluri et al., 2019).

Lipid nanoparticles are also very promising because

they are biocompatible and biodegradable and thus

are ideally suited for long-term application in the

treatment of neurodegenerative disorders. The lipid

part of LNPs is typically derived from naturally

occurring lipids, such that the particles become safe

for use in humans. Further, the fact that LNPs can

entrap hydrophilic and hydrophobic drugs makes

them a simple drug delivery vehicle. They can be

employed to deliver a range of therapeutic drugs such

as small molecules, RNA therapeutics, and proteins to

the brain and provide an alternative non-invasive and

effective route of treatment (Yuan et al., 2020),

(Alzheimer et al. 2020).

2.3 Lipid Nanoparticles: Design and

Properties

Lipid nanoparticles (LNPs) are nanoscale drug

delivery systems composed primarily of lipid

components, used to encapsulate drugs and deliver

them to target tissues or organs. An LNP typically has

a lipid core to wrap around the drug load, and a

surfactant or excipient shell to stabilize the particle.

This composition allows LNPs to wrap a broad range

of therapeutic agents, from hydrophobic molecules to

small molecules, nucleic acids, and proteins.

Among the significant advantages of LNPs is that

they can cross the blood-brain barrier. Particle size in

the case of LNPs constitutes one of the most critical

parameters for BBB crossing. The nanoparticles of

the size range of 20-100 nm are more effective in

crossing the BBB because they possess the ability to

bind to endothelial cells and get internalized by

receptor-mediated endocytosis. Surface charge of the

LNPs is an important determinant for BBB crossing.

Cationic nanoparticles will be more likely to

penetrate the BBB at a higher rate due to electrostatic

attraction with negatively charged endothelial cells

lining the brain's blood vessels (Kalluri et al., 2019),

(Yuan et al., 2020).

Beyond their ability to traverse the BBB, LNPs may

be designed to produce therapeutic action at specific

areas of the brain that are impacted by Alzheimer's

disease. This can be achieved through surface

functionalization of the nanoparticles with targeting

ligands, which bind to the receptors that are

overexpressed in brain areas impacted by amyloid-

beta plaques or tau tangles. Targeting ligands such as

antibodies, peptides, or aptamers may be surface

functionalized onto the LNPs to make them more

targeted and selective towards specific brain areas

(Yuan et al., 2020), (Alzheimer et al. 2020). Besides

that, LNPs are highly biocompatible and

biodegradable, and hence can be employed in drug

delivery for an extended period of time without

inflicting any harm. The lipid components used in

LNPs are natural, and hence the risk of toxicity or

immunogenicity is minimized. Furthermore, the lipid

structure of LNPs is easily modifiable to improve

their drug delivery properties, such as stability,

release kinetics, and targeting capacity (Alzheimer et

al. 2020) (Wang, et al. 2020).

2.4 In Vivo Investigations of Lipid

Nanoparticles for Alzheimer's

Disease

Several preclinical studies have shown the

therapeutic potential of lipid nanoparticles in

delivering therapeutic agents to the brain and

improving cognitive function in animal models of

Alzheimer's disease. LNPs have been used to deliver

various therapeutic agents, including small molecule

inhibitors, antibodies, and RNA therapeutics, to the

brain. A study showed that LNPs could deliver anti-

amyloid-beta antibodies effectively to the animal

models, decreasing amyloid plaque burden and

memory performance. These results suggest that

LNPs can be used as an efficient drug delivery system

for amyloid-targeted therapy in AD (Zhang et al.,

2020), (Li et al., 2021). Apart from small molecules

and antibodies, LNPs have been used for delivery of

RNA therapeutics, such as siRNA and mRNA, in

Alzheimer's disease models. These RNA drugs can be

designed to selectively target genes that encode

amyloid-beta or tau to correct for the disease-causing

factors. For example, LNPs encapsulating siRNA for

tau have been effective in reducing tau pathology and

preventing neurodegeneration in preclinical AD

models. This treatment represents a novel

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intervention in the treatment of AD through

regulation of the molecular pathways of disease

pathology (Zhang et al., 2020), (Kalluri et al., 2020).

Another possible application of LNPs for AD is in

gene therapy. By introducing genetic material, such

as genes encoding therapeutic proteins, directly into

the brain, LNPs could potentially reverse the course

of AD and restore normal brain function. For

instance, LNPs have been used to deliver genes

encoding anti-inflammatory cytokines or

neurotrophic factors to promote neuronal survival and

suppress neuroinflammation. Such approaches also

have great promise for the development of disease-

modifying drugs that both offer symptomatic relief

and affect the underlying causes of AD pathology

(Zhang, et al. 2021).

3 MATERIALS AND

SPECIFICATIONS

3.1 High-Shear Homogenizer

High-shear homogenizer is a critical instrument in the

production of lipid nanoparticles (LNPs). It is

employed mainly for emulsification and dispersion of

the lipid-therapeutic agent blend to obtain uniform

particle size and composition. The homogenizer

functions by subjecting the lipid-API blend to

mechanical shear forces, which disperse it into

nanoscale droplets. The equipment is usually run at a

pressure of 500–2000 bar and a flow rate of 10–50

mL/min, depending on the production scale. The

high-speed rotor or impeller creates shear forces, and

the regulated flow rate provides for uniform

distribution of the droplets. The homogenizer should

be able to operate under sterile conditions to prevent

contamination during nanoparticle synthesis. Rotor-

stator mechanism prevents the lipid nanoparticles

from being aggregated, and ensures they are

homogenized to obtain the desired size range,

preferably 50 nm to 300 nm, for effective penetration

through the blood-brain barrier (BBB).

3.2 Sonicator

A sonicator employs ultrasonic sound waves to form

cavitation bubbles that impart shear forces to the

lipid-API blend, breaking the large lipid aggregates

into nanoparticles. Sonicators are normally run at

between 20-40 kHz frequencies with power outputs

ranging from 100–500 watts, depending on the

volume of the sample. The ultrasound waves are

passed via a probe or bath system, delivering the

mechanical power necessary for decreasing the

droplet size to the range of nanometres (usually 10 nm

to 200 nm). The sonication process is especially

effective in producing a smaller particle size and

enhancing the dispersion of the drug and lipid

components. For Alzheimer's drug delivery,

sonication helps in creating a stable suspension of

nanoparticles while ensuring the efficient

encapsulation of the active pharmaceutical ingredient

(API).

3.3 Rotary Evaporator

The rotary evaporator is utilized for the elimination

of organic solvents from the lipid-API mixture after

emulsification. It uses lower pressure to decrease the

solvent's boiling point to allow efficient evaporation

under lower temperatures while maintaining the

stability of both the lipids and the drug. Rotary

evaporators usually operate between a vacuum level

of 10–100 mbar, with temperatures regulated at 30–

50°C according to the used solvent (chloroform or

ethanol). The rotary evaporator runs at rotation rates

of 50-150 RPM, which gives the best mixing and

evaporation efficiency. The process ensures that trace

solvents, which may be harmful to patients, are

eliminated, and a stable colloidal suspension of lipid

nanoparticles is left for further analysis.

3.4 Dynamic Light Scattering (DLS)

Dynamic Light Scattering (DLS) is employed to

determine the size distribution and zeta potential of

lipid nanoparticles. The DLS method relies on the

phenomenon of light scattering by suspended

nanoparticles. The suspended particles induce light to

scatter, and the DLS instrument detects the rate of

change of the scattered light over time. The

nanoparticle size can be determined by measuring

these changes, and common nanoparticle sizes for

LNPs are between 10 nm and 300 nm. The zeta

potential, being an indicator of surface charge, plays

a central role in evaluating the stability of the

nanoparticle. A zeta potential value of greater than

±30 mV is normally essential for achieving stability

and inhibiting particle agglomeration. DLS systems

applied to the production of LNPs would generally

work at 173° angles for proper determination of

scattering intensity with sensitivity from 1 nm to a

few microns.

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3.5 Transmission Electron Microscopy

(TEM)

Transmission Electron Microscopy (TEM) is utilized

to study the structure and morphology of the lipid

nanoparticles. TEM offers high-resolution imaging in

the nanoscale, and by this means, particle shape, size,

and homogeneity are visualized. LNPs are generally

observed using TEM in a range of 50,000x to

1,000,000x magnification with an image resolution as

low as 1-2 nm. TEM is necessary to ensure that the

lipid nanoparticles are spherical or close to spherical

in shape, which is best for efficient drug delivery,

especially for crossing the blood-brain barrier.

Sample preparation for TEM is usually embedding

the nanoparticles in a resin and sectioning thin slices

to get good imaging.

3.6 UV-Vis Spectrophotometer

A UV-Vis Spectrophotometer is employed to measure

the encapsulation efficiency (EE) of the drug by the

lipid nanoparticles. The process is to record the

absorbance of the drug at a given wavelength, which

reflects the distinct absorption spectrum of the drug.

As an example, acetylcholinesterase inhibitors would

have a characteristic peak of absorbance between 230

nm and 300 nm. The encapsulation efficiency is

determined by comparing the drug's absorbance in the

supernatant (free drug) with the overall drug

concentration in the nanoparticle suspension. The

UV-Vis spectrophotometer is highly sensitive and can

detect drug concentrations as low as 1 µg/mL,

providing valuable data on the effectiveness of the

encapsulation process.

3.7 Lipids and Surfactants

The lipids used in the formulation of lipid

nanoparticles play a critical role in ensuring the

stability, solubility, and controlled release of the drug.

Lipid materials such as phosphatidylcholine, stearic

acid, and triglycerides are commonly employed.

Phosphatidylcholine (PC) is a phospholipid

employed to form a lipid bilayer, providing a more

stable nanoparticle structure. Stearic acid is a

saturated fatty acid that solidifies the matrix of the

nanoparticle, and triglycerides are employed to

impart fluidity and flexibility to the nanoparticle

structure. The selection of the lipid is based on the

required properties of the lipid nanoparticles,

including size, drug encapsulation efficiency, and the

rate of drug release. Surfactants are used to stabilize

lipid nanoparticles, minimize aggregation, and

enhance the dispersion of the lipid-API blend.

Surfactants such as polyethylene glycol (PEG)-ylated

lipids, chitosan, or biocompatible non-ionic

surfactants such as polyvinyl alcohol (PVA) can be

used. PEGylated lipids are especially useful for

enhancing the circulation time and biocompatibility

of nanoparticles. They also minimize opsonization

and immune recognition, thereby prolonging the half-

life of the nanoparticle in circulation. Surfactants also

lower the surface tension of the lipid nanoparticles,

which keeps them from aggregating or clumping

together to form larger clusters that can jeopardize

their stability and performance in drug delivery.

3.8 Drugs

The selection of drugs is paramount in the

formulation of lipid nanoparticle-based drug delivery

systems for Alzheimer's disease. Drugs like

acetylcholinesterase inhibitors (e.g., donepezil and

rivastigmine) are typically encapsulated in LNPs to

increase their bioavailability and therapeutic effects.

Neuroprotectants like curcumin or resveratrol are also

encapsulated to lower oxidative stress and hinder

neuronal injury. Gene therapy agents including small

interfering RNA (siRNA) or microRNA (miRNA)

targeting major proteins like amyloid precursor

protein (APP) or tau are being used more and more as

components of new therapies. These drugs are chosen

with great care depending on their capacity to

penetrate the blood-brain barrier and their suitability

for lipid nanoparticle formulation techniques.

3.9 Targeting Ligands and pH-

Sensitive Lipids

In order to enhance the specificity of the lipid

nanoparticles for the brain, targeting ligands are

usually attached to the surface of the nanoparticles.

Ligands such as transferrin (which binds to the

transferrin receptor on endothelial cells of the blood-

brain barrier) or cell-penetrating peptides (CPPs) can

facilitate the transport of the LNPs across the BBB.

The addition of these targeting ligands enhances the

precision of drug delivery, ensuring that therapeutic

agents are directed to the desired site in the brain

while minimizing off-target effects. pH-sensitive

lipids represent a distinctive family of lipids that

undergo physical changes when they sense shifts in

environmental pH, like that occurring in the acidic

microenvironment of neuroinflammation or amyloid

plaques in Alzheimer's disease. Such lipids facilitate

the triggered release of loaded drugs at specific brain

locations, promoting the site-specific therapeutic

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actions. These lipids are routinely added to

preparations of Alzheimer's treatments where

targeted drug release is desirable to achieve enhanced

patient response.

4 METHODOLOGY

Figure 2: Process flow of lipid nanoparticle (LNP) synthesis

for Alzheimer’s disease treatment (Source: Author).

The production of lipid nanoparticles (LNPs) for drug

delivery, particularly against Alzheimer's disease, is a

multi-stage process that seeks to entrap therapeutic

agents in a lipid matrix to counteract issues like

penetrating the blood-brain barrier (BBB) and

delivering controlled release of drugs. The figure 2

shows the Process Flow of Lipid Nanoparticle (LNP)

Synthesis for Alzheimer's Disease Treatment. The

following is a step-by-step detailed process of LNP

production and testing.

4.1 Lipid Solution Preparation and

Encapsulation of Active

Pharmaceutical Ingredient

Lipid solution preparation is the initial stage of LNP

production. This entails dissolving a blend of solid

and liquid lipids in an organic solvent.

Phosphatidylcholine (PC), stearic acid, and oleic acid

are usually employed lipids. The solid lipid, such as

stearic acid, contributes structural stability, whereas

the liquid lipid, oleic acid, maintains flexibility and

fluidity. The use of chloroform or ethanol dissolves

the lipids. The lipids are dissolved by placing them in

a round-bottom flask and subjecting them to gentle

heat (if necessary) to obtain a homogeneous lipid

solution. After preparing the lipid solution, the

therapeutic agent (API), e.g., acetylcholinesterase

inhibitors (e.g., donepezil, rivastigmine),

neuroprotective agents (e.g., curcumin, resveratrol),

or gene therapy vectors (siRNA or miRNA), is added.

The API is dissolved in a small amount of an aqueous

vehicle such as PBS (phosphate-buffered saline) or

water. This helps the hydrophobic parts of the API to

interact with the lipid phase. The lipid-API blend is

then mixed gently to mix the API with the lipid phase.

The hydrophobic regions of the lipid molecules get

associated with the hydrophobic regions of the API,

and the drug gets encapsulated.

4.2 Emulsification and High-Pressure

Homogenization

Emulsification is essential in the creation of the lipid

nanoparticle suspension. The lipid-API mixture is

combined with an aqueous phase to produce a stable

emulsion of nanoscale droplets. This is done by the

use of high-shear homogenization or sonication. The

lipid-API mixture is exposed to shear forces produced

by a high-shear homogenizer. This produces uniform

droplets ranging from 50 nm to 300 nm. The

homogenizer is used at pressures ranging from 500–

2000 bar with a flow rate of 10–50 mL/min. So

nicators are used as an alternative method to

introduce ultrasonic waves (20–40 kHz) that form

cavitation bubbles, and in effect disperse large

aggregates of lipids into smaller particles. The

emulsion is then subjected to high-pressure

homogenization or micro fluidization after the initial

emulsification to further minimize the size of the lipid

droplets to the nanoscale. The emulsion is pushed

through a narrow gap at high pressure (e.g., 500–2000

bar) by a high-pressure pump. The high shear forces

and turbulence rupture the droplets into uniform

nanoscale particles.

4.3 Solvent Evaporation

After the emulsion is brought down to nanoparticle

size, removal of the organic solvent from the lipid

phase (e.g., chloroform or ethanol) is the second step.

It is done under a rotary evaporator. A rotary

evaporator is run in reduced pressure (10–100 mbar)

to decrease the boiling point of the solvent to facilitate

rapid evaporation at 30–50°C. This process stabilizes

the lipids and API and removes the solvent. The flask

is shaken at 50–150 RPM to allow for effective

evaporation.

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4.4 Particle Size Optimization and

Dispersion

Following removal of the solvent, the lipid

nanoparticles are dispersed in a sterile aqueous

vehicle, usually PBS or saline, to form an equilibrium

colloidal suspension. The suspension is further

treated by sonication or high-shear homogenization to

obtain an optimized particle size. Dynamic Light

Scattering (DLS) is employed for the determination

of particle size distribution, which ranges from 10 nm

to 300 nm. The zeta potential of nanoparticles is

determined to determine the suspension stability. The

value of ±30 mV or greater confirms stable

nanoparticles with reduced aggregation possibility.

4.5 Characterization of Nanoparticles

and Encapsulation Efficiency (EE)

Characterization of lipid nanoparticles is important to

determine their physical and chemical characteristics.

Transmission Electron Microscopy (TEM) is utilized

for the observation of the nanoparticles' morphology

and size. The LNP particles would be ideally

spherical and uniform in size, and their resolution

power would be as high as 1-2 nm. Dynamic Light

Scattering (DLS) is utilized to identify the size

distribution and zeta potential of the nanoparticles.

This confirms that the particles are of the right size

and possess the proper surface charge for stability and

drug delivery.

The encapsulation efficiency is calculated to evaluate

the extent of the API that has been encapsulated

successfully in the lipid nanoparticles. The

suspension of nanoparticles is centrifuged to isolate

encapsulated and free drug. The free drug in the

supernatant is quantified by a UV-Vis

spectrophotometer to calculate the drug content. The

drug's absorbance at a given wavelength (e.g., 230-

300 nm for acetylcholinesterase inhibitors) is

determined to obtain the encapsulation efficiency

(EE).

4.6 In Vitro Drug Release Studies and

Final Product Testing

In vitro drug release studies are carried out after

verifying the encapsulation efficiency to examine the

release pattern of the entrapped drug. This is achieved

by replicating physiological conditions (37°C, PBS

buffer) to examine the controlled and sustained

release of the API. The release of the drug is tracked

over time, and the cumulative drug released is

quantified at various time intervals. This aids in

identifying whether the drug is released in a

controlled, sustained fashion, which is necessary for

successful treatment. The final lipid nanoparticle

product is evaluated for stability, bioavailability, and

targeting capacity. Physical and chemical stability of

the nanoparticles are examined over time under

different storage conditions. The capacity of the LNPs

to traverse the blood-brain barrier is determined using

animal or cellular models. If targeting ligands (e.g.,

transferrin or cell-penetrating peptides) are

incorporated into the nanoparticles, their capacity to

target brain cells specifically is determined.

4.7 Preparation of Lipid Nano Particles

as Drug Carrier

The manufacture of lipid nanoparticles (LNPs) as a

drug-delivery system to treat Alzheimer's disease is

an extremely detailed and systematic procedure with

the aim of encapsulating drug molecules within a

lipid matrix under conditions of stability and

controlled drug release. It is an essential process to

enable the bypass of the challenges to drug delivery

in the brain, especially in reaching across the blood-

brain barrier (BBB). The procedures in the

manufacture of LNPs start with lipid solution

preparation, where a mixture of liquid and solid

lipids, e.g., oleic acid and stearic acid, is dissolved

within an organic solvent such as chloroform or

ethanol. The main aim during this process is to obtain

good solubility and homogenization of the lipids.

Stearic acid is a solid lipid that stabilizes the

nanoparticle, while oleic acid is a liquid lipid that

imparts fluidity and flexibility to the particles.

Organic solvents have to be used in order to

effectively dissolve these lipids, whereby a

homogenous lipid solution is established to serve as

the core of the nanoparticle. Good preparation of the

lipid phase is important since any remaining

undissolved lipid or inefficient homogenization may

cause problems in the subsequent steps, like

inefficient encapsulation of the therapeutic molecule

or non-uniform nanoparticles formation. Once the

lipid solution is prepared, the next step is the

encapsulation of the therapeutic agent (API), such as

acetylcholinesterase inhibitors or small interfering

RNA (siRNA), which are commonly used in the

treatment of Alzheimer's disease. The API, in its

unadulterated form, is generally dissolved or

suspended in a limited volume of a proper solvent,

commonly an aqueous phase such as water or PBS,

prior to addition to the lipid blend. This is a sensitive

process that involves careful mixing to incorporate

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the API completely into the lipid phase to produce a

stable lipid-API mixture.

After the lipid-API mixture is prepared, the

subsequent process is emulsification. Emulsification

is the blending of two immiscible liquids, in this

instance the lipid-API mixture and an aqueous

medium (distilled water or PBS). The aim is to create

an emulsion of nanoscale droplets with the lipid-API

mixture dispersed in the aqueous medium. This is

usually attained via high-speed homogenization or

sonication. High-speed homogenizers or sonicators

apply mechanical energy to disrupt the lipid-API

phase into small droplets, hence forming an emulsion.

Sonication applies sound waves to create severe shear

forces that disrupt the lipid phase into small droplets.

These droplets are initially in the micrometre size

range but are then reduced down to nanometres in the

second step. Homogenization supplies mechanical

energy that also reduces the size of the droplets so that

the end product is of the required nanoparticle size.

Following this primary emulsification, the emulsion

is subjected to high-pressure homogenization or

micro fluidization, which further decreases the size of

the droplets. This involves the use of a high-pressure

pump to push the emulsion through a small gap,

creating high shear forces that shatter the droplets into

nanoscale particles. The mechanism of working of

high-pressure homogenization is based on fluid

dynamics such that the intense shear and turbulence

cause the lipid droplets to break up and achieve

uniformity and a nanoscale distribution of size.

Uniform distribution of size is the key to ensuring the

nanoparticles have consistency such that they work

optimally in terms of drug delivery, stability, and

bioavailability.

After the emulsion has been reduced to nanosized

droplets, the subsequent step is removal of the organic

solvent in which the lipids and API have been

dissolved. This is done through the use of a rotary

evaporator under lowered pressure for evaporation of

the solvent. The lowered pressure reduces the boiling

point of the solvent, which makes it easy to remove

the solvent quickly without destroying the lipid

nanoparticles or the therapeutic agent inside them.

The solvent evaporation principle is based on the

volatility of organic solvents such as ethanol or

chloroform and the ease with which they can be

removed under low pressure without leaving toxic

residues. This is an important step since residual

solvents may have adverse effects on the stability and

safety of the final nanoparticle product, particularly

for drug delivery applications. After the removal of

the solvent, the lipid nanoparticles are dispersed in a

sterile aqueous medium, like PBS or saline. This

process creates a stable colloidal suspension of lipid

nanoparticles. The suspension is then optimized in

terms of size through homogenization and sonication

parameters. The size of the nanoparticles is important

since smaller nanoparticles can easily penetrate the

blood-brain barrier and reach the site of action more

efficiently. The nanoparticles' size and surface

properties are determined by dynamic light scattering

(DLS), an analytical technique that quantifies light

scattering as the nanoparticles travel through a liquid

medium. DLS gives extensive information on the

nanoparticles' size distribution, thereby ensuring that

they are suitable for efficient drug delivery. The zeta

potential of the nanoparticles, or the surface charge,

is also quantified through measurement. Zeta

potential values above ±30 mV is usually indicative

of stable nanoparticles, as they tend to be less prone

to agglomeration or unstable clustering. High zeta

potential ensures that the particles will be well

dispersed in suspension, enhancing their stability and

functionality in vivo.

Morphological characterization of the nanoparticles

is done through Transmission Electron Microscopy

(TEM), which gives high-resolution images of the

nanoparticles at the nanoscale. TEM is especially

effective in identifying the shape, size, and

homogeneity of the nanoparticles. An ideal lipid

nanoparticle should be spherical and homogeneous in

size since this enhances its efficacy in drug delivery

and its ability to penetrate the blood-brain barrier. The

free drug is analysed in the supernatant with a UV-Vis

spectrophotometer, which detects the absorbance of

light by the drug. A high efficiency of encapsulation

is crucial, as it optimizes the therapeutic action by

making sure that most of the drug is efficiently

encapsulated in the nanoparticles to minimize the loss

of the active ingredient during processing.

Lastly, the release rates of the drug encapsulated are

examined using in vitro diffusion experiments. Here,

the drug release profile is followed over time,

generally in PBS buffer at 37°C, to mimic

physiological conditions. Drug release is evaluated to

check if the drug is released in a controlled, sustained

manner, which is necessary to maximize therapeutic

action and minimize side effects. A controlled release

guarantees that the drug is presented to the target site

for a longer duration of time, hence enhancing the

general effectiveness of the treatment. In summary,

the synthesis of lipid nanoparticles for Alzheimer's

disease treatment is a precise process that

encompasses several steps such as lipid preparation,

encapsulation of the API, emulsification, evaporation

of the solvent, and characterization. All these steps

are precisely regulated to achieve stability, size, and

Production and Evaluation of Lipid Nanoparticle as Drug Carrier for Treatment of Alzheimer’s Disease to Enhance the Blood Brain Barrier

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177

release rate of the nanoparticles that are required for

efficient delivery of the drug through the blood-brain

barrier.

5 RESULTS AND DISCUSSION

5.1 Encapsulation Efficiency and Drug

Lipid nanoparticles' (LNPs') efficacy as drug delivery

vehicles is largely determined by their encapsulation

efficiency (EE) and drug loading (DL). The

encapsulation efficiency (EE) of LNPs designed to

treat Alzheimer's disease ranged from 80% to 95% in

our experimental studies. Because it guarantees that

a considerable amount of the therapeutic chemical is

effectively integrated into the lipid nanoparticle

structure, reducing the requirement for high

medication dosages, this high EE is an important

accomplishment. Lowering the dosage can limit

systemic drug exposure, which lowers the risk of

adverse effects. the figure 3 shows theGraphical

representation of Encapsulation Efficiency and Drug

Table 1: Encapsulation efficiency and drug loading

(Source: Author).

Parameter Value Significance

Encapsulation

Efficiency

(EE)

87% High EE ensures

minimal drug loss and

reduced side effects.

Drug Loading

(DL)

84% Substantial drug payload

carried without

nanoparticle instability.

Drug Release

Rate

79% Allows for extended

therapeutic effect (24-48

hours).

Figure 3: Graphical representation of encapsulation

efficiency and drug loading (Source: Author).

Another crucial element in assessing the viability of

LNPs in practice, drug loading (DL), was also

optimized. Because aggregation can result in

decreased bioavailability and poor drug

administration, it is crucial to maintain the stability of

LNPs with high drug content without experiencing

severe aggregation. It is explained in the table 1

5.2 Stability and Release Profiles

Lipid nanoparticles' (LNPs') stability is essential to

guaranteeing their long-term efficacy and

dependability as a medication administration method.

The LNPs were kept at 4°C for a few weeks in order

to replicate storage conditions in our investigations,

and their stability was evaluated using a number of

metrics, such as size and zeta potential. The findings

showed that over the course of the investigation, the

nanoparticles exhibited little aggregation and retained

their size and zeta potential. This implies that the lipid

nanoparticles have outstanding long-term stability,

which is crucial to guaranteeing that the drug delivery

system doesn't experience physical alterations that

may diminish its effectiveness over time.

The LNPs showed a regulated, sustained release

profile in terms of drug release. The medication was

given gradually over the course of 24 to 48 hours,

guaranteeing that therapeutic concentrations were

sustained in the brain for a considerable amount of

time. When treating chronic illnesses like Alzheimer's

disease, where maintaining steady therapeutic levels

over time is essential to halting the course of the

disease and controlling symptoms, this sustained

release profile is extremely helpful. Controlled

medication release lessens the possibility of adverse

effects from high drug concentrations and eliminates

the need for frequent doses.

The table 2 shows a comprehensive analysis of the

stability, drug release profile, and blood-brain barrier

(BBB) penetration efficiency of lipid nanoparticles

(LNPs) with and without transferrin modification. It

points out that LNPs kept at 4°C are very stable,

having a consistent size of around 100 nm and the zeta

potential of -30 to -35 mV, demonstrating negligible

aggregation with time. The drug release profile

demonstrates a sustained and prolonged release of the

drug, with the concentrations rising from 0 μg/ml at 0

hours to 70 μg/ml at 48 hours, demonstrating

sustained long-term therapeutic effects with 80%

cumulative release for 48 hours. From the BBB

penetration perspective, LNPs without transferrin

modification demonstrate poor penetration (25%),

whereas transferrin-modified LNPs markedly

improve BBB penetration to 50%. Additionally, the

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transferrin-modified LNPs are highly efficient for

targeting, with 40% targeting the cortex and 35%

targeting the hippocampus, areas of greatest

importance for the treatment of Alzheimer's disease.

This detailed data proves the efficacy of transferrin-

modified LNPs for effective brain drug delivery and

long-term therapeutic effects.

Table 2: Stability, release profile and BBB penetration efficiency of lipid nanoparticle (Source: Author).

Metric

Condition/Para

mete

Data/Observatio

Key Insights

Stability of LNPs (Size) Storage at 4°C

Size of LNPs

(~100 nm)

LNPs maintain a consistent size (~100 nm)

over time, indicating excellent stability.

Stability of LNPs (Zeta

Potential)

Storage at 4°C

Zeta Potential

(-30 to -35 mV)

The LNPs retain their zeta potential over

time, indicating minimal aggregation and

maintaining stability.

Drug Release Profile

(Initial)

0 hours

Drug

Concentration:

/ml

No drug release at the initial time point (0

hrs).

Drug Release Profile (6 hrs) 6 hours

Drug

Concentration:

/ml

Gradual drug release, reaching 15 μg/ml

after 6 hours.

Drug Release Profile (12

hrs)

12 hours

Drug

Concentration:

/ml

Continued drug release, with concentration

reaching 30 μg/ml by 12 hours.

Drug Release Profile (24

hrs)

24 hours

Drug

Concentration:

50 μg/ml

Steady increase in drug concentration,

reaching 50 μg/ml at 24 hours,

demonstrating sustained release.

Drug Release Profile (48

hrs)

48 hours

Drug

Concentration:

70 μg/ml

Maximum drug concentration at 48 hours,

reflecting prolonged and sustained release.

Sustained Release Profile 24-48 hours

Cumulative

Release: 80%

80% cumulative drug release over 48 hours,

ensuring prolonged therapeutic efficacy.

BBB Penetration (Without

Transferrin Modification)

N/A

BBB

Penetration:

25%

Without transferrin modification, BBB

penetration is limited to 25%.

BBB Penetration (With

Transferrin Modification)

N/A

BBB

Penetration:

50%

Transferrin modification significantly

improves BBB penetration, reaching 50%.

Targeting to Cortex (With

Transferrin Modification)

N/A

Targeting

Efficiency: 40%

Transferrin-modified LNPs show a 40%

targeting efficiency to the cortex.

Targeting to Hippocampus

(With Transferrin

Modification)

N/A

Targeting

Efficiency: 35%

Transferrin-modified LNPs show 35%

targeting efficiency to the hippocampus,

crucial for Alzheimer's.

5.3 BBB Penetration and Targeting

Efficiency

Since the blood-brain barrier (BBB) keeps the

majority of therapeutic medicines from entering the

brain, lipid nanoparticles' (LNPs') capacity to

penetrate the BBB is essential for treating

neurological conditions like Alzheimer's. The lipid

nanoparticles showed notable penetration and

transport across the barrier in our tests utilizing in

vitro models of the blood-brain barrier. This was

accomplished by applying transferrin, a protein that

binds to transferrin receptors (TfR) found on the

BBB's endothelial cells, to the surface of the LNPs.

The table 3 shows the Comparison of Blood-Brain

Barrier (BBB) Penetration and Targeting Efficiency.

LNPs were able to enter the brain more easily thanks

to transferrin receptor-mediated endocytosis, which

guaranteed effective drug delivery to the intended

areas. Transferrin surface modification greatly

increased the LNPs' capacity to pass the blood-brain

barrier, increasing their therapeutic potential for

Alzheimer's disease treatment. The figure 4 shows the

Graphical representation of Blood-Brain Barrier

(BBB) Penetration and Targeting Efficiency (Source:

Author, (Zhou et al., 2018), (Wang et al., 2022),

(Zhang et al., 2020) We were able to improve

medication accumulation in particular brain areas that

are crucial for AD, such the cortex and hippocampus,

Production and Evaluation of Lipid Nanoparticle as Drug Carrier for Treatment of Alzheimer’s Disease to Enhance the Blood Brain Barrier

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179

which are important in memory and cognitive

processes, by improving the targeting effectiveness of

the LNPs. By ensuring that the medication reaches the

regions most impacted by Alzheimer's pathology, this

focused administration enhances treatment results.

Table 3: Comparison of Blood-Brain Barrier (BBB)

penetration and targeting efficiency (Source: Author, (Zhou

et al., 2018), (Wang et al., 2022), (Zhang et al., 2020).

Metric

Proposed

Methodology

(LNPs with

Transferrin

Surface

Modification

)

Existing

Methodolo

BBB

Penetration

(%)

50% 25% - 35%

Targeting to

Cortex (%)

40% 20% - 30%

Targeting to

Hippocampus

(

)

35% 15% - 25%

Figure 4: Graphical Representation of Blood-Brain Barrier

(BBB) penetration and targeting efficiency (Source:

Author, (Zhou et al., 2018), (Wang et al., 2022), (Zhang et

al., 2020).

6 CONCLUSIONS

The formulation of lipid nanoparticles (LNPs) as drug

delivery systems for treating Alzheimer's disease

(AD) is of special promise because they possess high

encapsulation efficiency (EE), regulated release of

drug, and an ability to permeate the blood-brain

barrier (BBB) efficiently. In this investigation, we

were able to establish that LNPs could be fine-tuned

to provide maximum therapeutic benefit by managing

the most influential parameters like encapsulation

efficiency, drug loading, stability, and targeting

efficiency. Encapsulation efficiency of the LNPs was

80% to 95%, with a remarkable 87% in the final

formulation, such that a high percentage of the

therapeutic agent is incorporated into the nanoparticle

structure, minimizing dosages of the drug and

reducing side effects caused by systemic

administration. The DL was also optimized to 84%,

such that a high drug payload can be achieved without

affecting nanoparticle stability. This drug loading is

high, which provides efficient delivery of therapeutic

molecules to the target location, maximizing the

overall therapeutic potential. Stability experiments

revealed that the LNPs were stable at 4°C for weeks,

with minimal aggregation and no change in size and

zeta potential, suggesting their stability for long-term

storage. Moreover, the sustained drug release profile,

with progressive release over 24 to 48 hours, is

perfect for the management of chronic diseases since

therapeutic drug levels are sustained for a long

duration of time, minimizing the risk of side effects

and enhancing patient compliance. The ability of

LNPs to cross the BBB is crucial for treating

neurological disorders like Alzheimer's, and our study

demonstrated that transferrin-modified LNPs

significantly penetrated the BBB in vitro through

transferrin receptor-mediated endocytosis. This

adjustment improved the LNPs' specificity to target

certain brain areas like the cortex and hippocampus,

which play important roles in memory and cognitive

processes, optimizing therapeutic effect by localizing

the drug where it is needed most. In summary, our

results demonstrate the promise of LNPs as a versatile

drug delivery system for Alzheimer's disease,

offering a solution with high encapsulation efficiency,

long-term release, and site-specific delivery to the

brain, and thus holding out hope for improved disease

control and, possibly, retardation of its progression.

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