MICRONEEDLE-BASED DRUG

DELIVERY SYSTEM FOR CATTLE

[Group 28]
 INDEX

Contents
Introduction ............................................................................................................................................ 2
Brainstorming Evidence ......................................................................................................................... 3
CUSTOMER REQUIREMENTS .................................................................................................................. 8
Preparation of chitosan microneedle patch using Soft Lithography Approach ................................. 13
Flow chart of chitosan microneedle patch design using Soft Lithography Approach ........................ 14
Mask Layout ......................................................................................................................................... 15
Packaging and Testing ......................................................................................................................... 21
Conclusion ............................................................................................................................................. 24
References ............................................................................................................................................ 25

Biomedical Microsystems (BB610) 1

 Introduction

India is a country in which 60-70% population depends on agriculture. India’s agriculture is

different from other countries in many aspects, like land distribution and the number of cattle
per farmer, etc. Moreover, there is a lesser availability of expert staff in the villages who can
give drugs/vaccines to the cattle as giving drugs/vaccines via needles requires immense
experience and precision. Many small farmers in India are dependent on cattle for livelihood
and milk. Thus, diseases that frequently affect cattle bring about a huge loss for our farmers.
Thus, it is very important to take proactive decisions and vaccinate the cattle against life-
threatening diseases so that we don’t face huge economic losses in the agricultural sector.
Some of the diseases that commonly affect cattle are foot-and-mouth disease and Lumpy
virus disease. Microneedles are becoming famous as we can inject drugs/vaccines via skin
patches in a less painful manner when compared to traditional syringe-based methods.
Microneedles can also be used to achieve sustained drug release over a period of time. This
will limit the number of injections taken per week/day and does not require any technical
expertise to use them. Thus, we aim to deliver drugs/vaccines through microneedle patches.
Microneedle is becoming one of the suitable drug delivery methods because they provide
drug without pain and sustained drug release is possible with microneedles. A farmer can put
the patch on the ear of the animal by themselves at a very early age. It will give immunity in
its early phase so that the disease will not affect the cattle throughout their life, So we want
to design a microneedle-based microdevice that can deliver the drug/vaccine without pain
and without skilled staff.

Biomedical Microsystems (BB610) 2

 Brainstorming Evidence

1st Approach: Pyramid-shaped Microneedle Design

In this approach, we have a 400mm2-sized square patch (Fig 1). The diagram is not drawn
to scale. But this patch will have around 324 pyramidal microneedles of dimensions 350 x
350 x 700 µm (Fig.2).
We have decided to go with this design based on various previous studies done for
delivering drugs and pain-killer medicines in livestock ([1],[2]). Many of the conventional
studies use conical-shaped microneedles with circular bases. But, simulations have shown a
linear relationship between the mechanical strength and the number of vertices in the
polygon base (e.g., triangular, square, and hexagonal microneedle bases), showing better
insertion depths for the triangular and square-built microneedles. Thus, we have decided to
have a pyramid with a square base [3]. Another important parameter we considered while
designing this patch is the force that will be experienced by these needles during insertion
into the skin. Based on previous studies we found it to be approximately around 0.28-0.30 N.
Needles with blunt tips need more force for insertion (around 3N). Thus, we expect the
pyramid shape with a sharp tip to be the optimum shape for our microneedle. The studies
also suffered from inadequate drug delivery due to drug absorption by skin layers. Since we
require the patch to deliver a minuscule quantity of drug without much loss, we want the
microneedles to penetrate the skin through dermis layers to deliver the drug. This can be
achieved by having a 700-micron tall pyramid.

One of the studies showed that the number of microneedles significantly contributes to
efficient transdermal drug delivery [1]. They got lesser efficiency of drug delivery due to the
smaller size of the patch (around 64mm2). So, we decided to go with a 400mm2 with around
324 microneedles (18*18) arrangement.
The material used for the patch: Microneedle patches have been constructed from different
types of materials (e.g., silicon, metals, ceramics); however, polymeric microneedle patches

Biomedical Microsystems (BB610) 3

have been demonstrated to provide better chemistry and mechanical properties. In
particular, the use of low-cost polymers such as chitosan or starch provides a promising
material platform for drug delivery and time-release mechanisms (i.e., through polymer
degradation, polymer morphology, or polymer chemistry). Thus, we have decided to go with
chitosan because of its characteristics, like biocompatibility, nontoxicity, polycationic
properties, biodegradable ability, and antibacterial properties.

2nd Approach: Circular obelisk-type multilayer microneedles facilitate efficient

vaccine delivery
Microneedle-based transdermal drug delivery system offers several benefits over
traditional hypodermic injections, which include minimal invasion, minimal pain, stimuli-
induced drug release, enhanced patient adherence, easy handling, and self-administration.[4]
But choosing a material for the production and fabrication of microneedles becomes
cumbersome and it requires careful investigations and considerations of numerous factors
viz. biocompatibility, drug loading capacity, low medical wastage, and economic feasibility.
Considering the above-mentioned factors, biodegradable polymers become the first choice
for microneedle production. Currently, various polymeric materials like Polyvinyl alcohol
(PVA), Polyvinyl pyrrolidone (PVP), Poly-lactic acid (PLA), Poly-lactic co-glycolic acid (PLGA),
Polycaprolactone (PCL), and Saccharides have been extensively used for microneedle
fabrication. [5]
Here another potential approach for delivering the Lumpy virus vaccine in cattle can be an
obelisk-shaped multilayer microneedle. A study suggests that the obelisk-shaped system can
ensure reliable skin penetration, and reduce the percentage of drug degradation during
fabrication.
We can use the spray-deposition process for the construction of this particular
microneedle which involves sequential deposition of PLGA and PVP solution in microneedle
molds (molds can be replicated from the obelisk-shaped master structure). Coming to its
fabrication, micro-milling of 6061 aluminum alloy can be used. We can consider a blank
aluminum workpiece of dimension (1'' x 1'' x 0.25'', W x L x T) in a miniature milling system
with a motion accuracy of 1µm. Afterward, a solid carbide end mill (1/8'' diameter) and
aluminum titanium nitride (AlTiN)- coated solid carbide micro end mill (0.01'' diameter) can
be used for developing a microneedle base and a microneedle. From the aluminum primary
masters, we can replicate the polymeric secondary masters by PLA casting into PDMS micro-
molding. Now the mechanical behavior of the multilayer-based microneedle system can be
tested by a universal static testing system.

Biomedical Microsystems (BB610) 4

 Study shows that the circular and beveled obelisk-shaped microneedle are advantageous
over the traditional pyramid-shaped microneedle as the former one is mechanically more
strong. Additionally, the spray deposition process has still some disadvantages like high cost
in comparison to other casting processes. It costs money to create custom molds. In addition,
the investment castings are only suitable for holes measuring at most 1.6 millimeters in
diameter. [6]

3rd Approach: Nanoporous microneedle arrays with reservoir

Microneedles are needle-like structures having a length of less than 1 mm and therefore,
allow to overcome the skin's barrier. They can be hollow or solid. Solid Microneedle Arrays
(MNAs) are used to pretreat the skin for subsequent application of cream or ointment or
delivering drugs after coating the needles with the drug. They are useful only for the delivery
of low-dosed and highly potent drugs like vaccines or hormones etc.
Alternatively, nanoporous MNAs with a nanofluidic network throughout the whole MNA
have a large storage capacity for pharmaceutical formulations. Upon piercing this npMNA into
the skin, the drug formulation will diffuse into the skin [7]. A great advantage of this system
is that the drug storage capacity can be significantly increased by connecting an external drug
reservoir to the npMNA, allowing it to deliver even higher drug doses transdermally. In a study
by [8] nanoporous microneedles were used to deliver memantine from the drug reservoir
through diffusion. The npMNAs (type 1.2.0: 475 µm microneedle length, 150
microneedles/cm2, an average pore size of 80 nm) were manufactured according to MyLife

Biomedical Microsystems (BB610) 5

Technologies’ proprietary production process at LouwersHanique (Hapert, the Netherlands)
using micro molds, obtained by a double replication process, as done by van der Maaden
et al., 2015. In brief, an inverse silicon master mold was used to produce a positive PDMS
mold, and this positive PDMS mold was subsequently used to generate a second inverse
PDMS mold. To produce ceramic microneedles, the inverse PDMS mold was filled with a
ceramic slurry, containing alumina nanoparticles (AKP30, grain size ±300 nm) and
a plasticizer. After controlled drying, the microneedles were removed from the PDMS mold
and punched into circular discs of 10, 12, and 16 mm. Finally, the microneedle discs were
sintered at 1450 °C, resulting in npMNAs with diameters of 9, 11, and 15 mm, respectively,
having a nanofluidic network throughout the entire array.
A 1,6mL drug reservoir was designed and 3D printed by them using Polylactic acid (PLA).
The reservoir contained a 14 mm opening with a supporting grid on the top side to allow the
drug solution in the reservoir to diffuse through the backplate of the npMNAs. Furthermore,
the reservoir contained supporting rods to easily position the npMNAs in the middle of the
opening and contained a screw cap with a silicone ring on the bottom to enable loading the
reservoirs with a liquid drug formulation. 15mm npMNAs were fixed gas-tight onto the
reservoir using PDMS.
A specially designed strap was developed to hold the npMNA-reservoir combination in
place, which included a 3D printed part with a hole into which the npMNA-reservoir
combination fits. After placing the npMNA-reservoir combination, a digitally-controlled
impact insertion applicator (UAFM v1, uPRAX Microsolutions B.V.) was used to apply the
npMNA-reservoir combination with a velocity of 1.4 m/s, after which the 3D printed buckle
of the strap was closed to keep the npMNA-reservoir combination in place. Subsequently, the
minipigs were locally covered by a bandage and were given a jacket to prevent them from
removing or playing with the npMNA-reservoir combination.
Salient features:

• The rate of delivery of the drug can be controlled in this design with the help of a
digital controller.
• The rate of diffusion of the drug depends on the thickness of the backplate of the MNA
as there is a linear relation between them. Therefore, the diffusion rate can be
changed based on the thickness of the back plate.

Biomedical Microsystems (BB610) 6

 • The adsorption of the drug to be delivered on the materials of the npMNA and the
reservoir materials has to be determined case to a case basis, as we want less
adsorption of material on these surfaces. In the given experiment, the adsorption was
found to be very less compared to the total dose in the reservoir.
• There should be less leaching of the materials like PDMS, PLA, etc into the drug which
affects the quality of the drug over days. This is especially important when reservoirs
are used for many days.
• The average diffusion of memantine per hour was, although not linearly, inversely
proportional to the npMNA thickness.
• The release rate of memantine increased with increasing npMNA diameters.
• The backplate thickness, the npMNA diameter, and the drug concentration determine
the released dose per time unit. The drug concentration and reservoir volume
determine the total deliverable dose.
Cons:

• This patch has more length than the width. It is not feasible to attach the patch
without a strap. We need a patch which is like an adhesive patch without any belt like
strap around it.
• The patch should be feasible to be attached to hairy cattle. With this aspect ratio, it
looks like it is not possible to attach the patch to the cattle firmly.

4th Approach: Application of a Reservoir-Type Calcitriol Transdermal Patch in

Dairy Cattle:
Yamagishi et al., (2009) have used reservoir-type calcitriol transdermal patches to supply
Vit D3 to cattle. They supplied 5mg of calcitriol per animal using the reservoir-based
transdermal patch. Each patch was applied once to the ventral root skin of the tail and
bandaged with elastic adhesive tape.
The patch was fabricated by encapsulating the reservoir solutions within a shallow
compartment molded from the backing laminate and covered with the controlled caliper EVA
membrane. In brief, both the backing laminate and controlled caliper EVA membrane were
cut to a size of 4.5 * 4 cm. The backing laminate was placed on the EVA membrane and then
heat-sealed on three sides. The reservoir solution was dispersed into the device using a
micropipette. That device was heat-sealed to close the final side, thus ensuring that none of
the solutions would leak out of the patch.[9]
This patch is attached to the animals without any strap and it was able to deliver the
intended drug for 3 -5 days. It is easy, reliable, and versatile to use.

Biomedical Microsystems (BB610) 7

 CUSTOMER REQUIREMENTS

No. Requirement Importance

1. Efficient Drug Delivery 2

2. Compatibility with different drugs 2
3. Compatibility with different drug delivery routes 2
4. Compatibility with existing equipment 4
5. Comfort (Minimal pain & discomfort) 1
6. Adaptability to different climates 2
7. Ease of Use 2
8. Cost Effective 3
9. Safety 1
10. Regulatory Compliance 1
11. Technical support & Training 3
12. Scalability (based on herd size & farm operation) 2
13. Sterility 1
14. Customizability (based on customer-specific requirements) 3
15. Reduced risk of Disease transmission 1
16. Long-term Stability & Efficacy 2
17. Traceability (for accountability & compliance) 1
18. Speed of Drug Delivery 4
19. Minimal Environmental Impact 2
20. Portability 4
21. Consistency & Dosage Accuracy 1
22. Low Maintenance 3
23. Reliability 2
24. Accessibility 3
Priority Order: 1>2>3>4>5

Biomedical Microsystems (BB610) 8

FUNCTIONAL SPECIFICATIONS

No. Metric Details Values / Materials Needs Priority

/ Methods
1 Needle Size Depends on factors like 500 – 1500 1, 2, 5 2
the thickness of the skin, micrometers (µm) 14, 18
the size of the animal, and 21
the type of drug being
delivered. Shorter
needles are often used
for more sensitive areas
(ear or face) while longer
needles may be used for
larger areas (neck or
shoulder).
2 Needle Material Depends on several Stainless Steel 2 ,4 1
factors including the Titanium 6, 8
mechanical properties of Polylactic Acid 12,
the material, Polyglycolic Acid 14, 19
biocompatibility, and Glass 23
manufacturability.
3 Drug Solubility The drug must be soluble Range: 10 - 100 1, 3, 7 3
in the formulation used mg/mL 16, 18
for microneedle 21
fabrication. Most drugs
used in veterinary
medicine are hydrophilic,
and the formulation
should be water-soluble.
4 pH Compatibility The drug should be stable pH range: 1, 2, 6 2
and remain active in the 5.5 - 7.5 16, 21
pH range of the
formulation. The pH
range of the formulation
should be optimized to
maintain the stability and
efficacy of the drug.
5 Formulation The viscosity of the drug Range: 1, 14 3
Viscosity formulation should be 1 - 10 cP 21, 23
optimized to achieve easy (centipoise)
flow through the
microneedles. High
viscosity may cause
clogging of the
microneedles, while low

Biomedical Microsystems (BB610) 9

 viscosity may result in
leakage and low efficacy.
6 Drug The concentration of the Typical Range: 1, 2, 5 3
Concentration drug in the formulation 0.1 - 10% (w/v) 8, 18
should be optimized to 21
achieve the desired
therapeutic effect. High
concentrations may cause
irritation, while low
concentrations may be
ineffective.
7 Chemical The drug should remain Shelf life: 6, 9, 3
Stability stable during storage, 6 – 24 months 13, 16
handling, and 20, 23
administration.
8 Microneedle It determines the number 50 -500 1, 5, 8 2
Spacing of microneedles that micrometers (µm) 14
penetrate the skin and
deliver the drug. A higher
density of microneedles
can lead to more accurate
and efficient drug
delivery, especially in
large animals like cattle.
Depends on various
factors such as the skin
thickness, hair density,
and the size and shape of
the microneedles. It can
also impact the pain and
discomfort experienced
by the animal.
9 Injection Force Directly impacts the 50 -200 N 1, 4, 1
depth and accuracy of (Newtons) 5, 7
microneedle penetration, 9, 14
as well as the efficiency of 21
drug delivery. The
optimal injection force for
microneedle-based drug
delivery in cattle depends
on various factors, such
as the size and shape of
the microneedles, the
skin thickness, and the
viscosity of the drug
formulation. the injection
force should be optimized

Biomedical Microsystems (BB610) 10

 to ensure accurate and
consistent drug delivery,
while minimizing pain
and discomfort to the
animal.
10 Delivery Speed Delivery speed refers to 1 - 100 µL/min 1, 2, 3 2
the rate at which the drug (microliters per 17, 18
formulation is delivered minute) 21
through the
microneedles into the
skin. The optimal delivery
speed for microneedle-
based drug delivery in
cattle depends on various
factors, such as the size
and shape of the
microneedles, the
viscosity of the drug
formulation, and the
injection force used. In
general, a slower delivery
speed is preferred to
ensure accurate and
consistent drug delivery,
while minimizing the risk
of tissue damage or other
adverse reactions.
11 Drug Release The drug release profile Initial burst 1, 7, 8 3
Profile refers to the rate and release range: 12, 14
duration of drug release 10 to 90% (of the 16, 18
from the microneedle total drug content) 21, 23
array into the skin tissue.
Generally, a sustained
and controlled drug Steady-state
release profile is release rate:
preferred to ensure 0.1 to 10 µg/min
consistent therapeutic
effects over an extended Duration: few
period. The drug release hours to days
profile can be
characterized by various
parameters, such as the
initial burst release, the
steady-state release rate,
and the duration of drug
release.

Biomedical Microsystems (BB610) 11

 12 Regulatory An important Preclinical and 9, 10 1
Compliance consideration for the clinical testing 17
development and use of (in vitro and in
microneedle-based drug vivo studies,
delivery systems in cattle. pharmacokinetic,
Regulatory agencies such pharmacodynamic
as the U.S. Food and Drug assessments,
Administration (FDA) and manufacturing,
the European Medicines and quality control
Agency (EMA) require evaluations)
that these systems
comply with relevant
safety and efficacy
standards before they can
be approved for use in
veterinary medicine. In
addition to safety and
efficacy testing,
microneedle-based drug
delivery systems in cattle
must also comply with
relevant regulatory
guidelines on good
manufacturing practices,
product labeling, and
adverse event reporting.
13 Sterility Sterility is a critical Gamma 9, 10, 1
requirement for irradiation 13, 15
microneedle-based drug 17
delivery systems in cattle, Ethylene oxide
as contaminated systems (EO) sterilization
can cause infections and
other adverse effects in Autoclaving
animals. Achieving
sterility in microneedle- Filtration
based drug delivery
systems involves the use Aseptic processing
of various methods to
eliminate or prevent the
growth of
microorganisms that
could be present in the
system.

Biomedical Microsystems (BB610) 12

 Preparation of chitosan microneedle patch using Soft Lithography
Approach

Master template fabrication:

Using a tilted-rotated photolithography technique, master templates were first created to
produce microneedle patches. An anti-reflective coating was applied to a polished silicon
wafer; it was observed that this layer significantly decreased backside reflections and
dramatically improved the resolution of the ensuing master templates. Then The wafer was
spin-coated with a thick layer of negative photoresist (SU-8). The SU-8 was then covered with
a mask and then the complex was exposed to UV radiation at incidence angles of 18 to 25°.
The depth of the mold, and eventually the length of the microneedles, are determined by
both the dimensions of the mask and the incidence angle of UV light.
Positive replica:
The intermediate was the creation of a positive replica of the master template. Because of its
low surface energy, usability, high flexibility, and low cost, the replicas were made using
polydimethylsiloxane (PDMS), which is readily available. The copies demonstrated a high
degree of repeatability of the original template.
PRINT-compatible Mold:
The positive replica was then used to make PRINT-compatible mold. The replicas' dimensions
and the PRINT moulds' consistency with the SU-8 master templates allow for accurate
replication.
It should be emphasised that each master template can be utilised to create hundreds of
PDMS replicas, and each replica can produce at least 50 moulds, according to laboratory
findings. By using PRINT processing, at least 10 microneedle arrays may be produced from
each mould. [10]
Patch fabrication
Chitosan/meloxicam solution was applied to a PDMS mould to create the microneedle patch.
The chitosan/meloxicam microneedle patch was made by pouring 500 mg of the casting
solution onto the mould to form a monolayer this is performed in step 1. After that in step 2
The mould was then placed in a centrifuge tube with a flat bottom and centrifuged at 3000
RPM for 30 minutes. For a total of 120 minutes of centrifugation, steps 1 and 2 were
performed four times. This process resulted in the formation of the microneedles containing
meloxicam. The microneedles and chitosan foundation make up the entire patch. A second
layer of the chitosan solution without meloxicam was added to create the chitosan base. As
the second layer is on top of the skin, the medicine cannot be released through it. After putting
the second layer it is centrifuged at 3000 RPM for 60 min. Finally, a 50 mL tube containing the
mould was put in an oven to dry for three days at 28 °C without a cap. Using tweezers, the
microneedle patch from the tube was extracted.[11]

Biomedical Microsystems (BB610) 13

 Flow chart of chitosan microneedle patch design using Soft
Lithography Approach
Start the
manufacture process

Master template fabrication using

Chitosan/meloxicam photolithography technique
solution preparation

Making Positive replica from master

template

PRINT-compatible Mold from positive

replica

Chitosan/meloxicam solution applied

to a PDMS mould

Centrifuged at 3000 RPM for 30 minutes.This process and

previous process is repeated 4 times.

A second layer of the chitosan solution without meloxicam is added to

create the chitosan base .

Centrifuged at 3000 RPM for 60 min

Finally, a 50 mL tube containing the mould was put in an

oven to dry for three days at 28 °C without a cap

Using tweezers, the microneedle patch extrection

End process

Biomedical Microsystems (BB610) 14

 Mask Layout
One example of a mask pattern useful for forming a structured surface for the production
of microneedles is given below. The mask pattern includes a row of needle apertures with a
channel feature corresponding to the desired location of the channel on the microneedles
(corresponding to the needle aperture).
The mask itself may, be manufactured using standard semiconductor lithography mask
techniques. The patterned portions of the mask are opaque to the laser energy used to
pattern the substrate, e.g., ultraviolet light in the case of excimer laser energy. The mask may
include a support substrate that is transparent to the laser energy. For example, the patterned
portions may be formed of aluminum while the 30 support substrate is fused silica. One
alternative for aluminum may be a dielectric stack that is opaque for a light of the desired
wavelengths.
The needle apertures in the mask are preferably arranged in successive rows that are
uniformly spaced apart. It is further preferred that the spacing between the needle apertures
along the rows are also uniform.
Use of this mask pattern may proceed with a first exposure of substrate located beneath
portion A of the mask pattern. As a result, the substrate is exposed during the first exposure
in a pattern corresponding to portion A of the mask. Movement of the mask and the substrate
relative to each other can be used to align the mask apertures in the uppermost row of
portion B with part of the substrate exposed by the needle apertures in portion A during first
exposure. Second exposure ablates more of the substrate, thereby increasing the depth of
needle cavities.[12]
After completion of the structured surface, the substrate provides a negative of the desired
microneedle array structure, with needle cavities corresponding to the shape of the
microneedles. The resulting mold substrate is then preferably electro-plated to form a
metallic positive of the microneedle array. Before electroplating, however, the substrate may
preferably be cleaned to remove any debris that is, c.g., associated with the laser ablation
processing used to form the negative image in the substrate. One suitable cleaning process
may include locating the substrate in an ultrasonic bath of detergent and water, followed by
drying.

Mask Design

Biomedical Microsystems (BB610) 15

Another approach:

Steps showing the fabrication process of the microneedle patch

Biomedical Microsystems (BB610) 16

 Top view and slanted view of microneedle patches along with the
dimensions of a single microneedle

Biomedical Microsystems (BB610) 17

 Schematic showing the delivery of drug through microneedle patches

Drug in Microneedles

Microneedle inserted
in the skin

Drug release through the

skin and microneedle
degradation

Complete microneedle
degradation

Biomedical Microsystems (BB610) 18

Master template fabrication:
A process termed tilted-rotated photolithography was used to create microneedle patches.
Initially, master templates were made by covering a polished silicon wafer with an anti-
reflective material. This greatly decreased backside reflections and increased the resolution
of the templates. The wafer was then spin-coated with a thick coating of negative photoresist.
The complex was then subjected to UV light at incidence angles ranging from 18 to 25° while
the Photoresist was protected by a mask. The dimensions of the mask and the UV light's
incidence angle were used to calculate the depth of the mold and, as a result, the length of
the microneedles.[13]

Mold:
Generally speaking, the term "mould" (sometimes spelled "mold") refers to a particular
species of fungus that develops in multicellular threads called hyphae. Molds may grow on a
broad variety of organic materials, including plants, wood, and food, and they can be found
in a number of habitats, including soil, air, and water. Certain varieties of mold can harm
people's health, especially those who have allergies or respiratory conditions, and other
molds create poisonous compounds called mycotoxins. The manufacture of cheese,
antibiotics, and enzymes are only a few examples of the useful industrial and medicinal
applications of some mold species, which are not all toxic.

PRINT:
We have built microneedle arrays utilizing the Particle Replication In Non-wetting Templates
(PRINT) approach, to get around earlier difficulties with the manufacture of microneedles.
This technology combines classical polymerization with a "top-down" soft lithography
approach to produce repeatable features on the nano- and micro-scale with fine control over
size, shape, and chemical composition. This technique makes it possible to manufacture
arrays fast; for batch procedures, it may produce a microneedle patch in less than five minutes
after the required mold is formed. Patches of practically any size may be produced rapidly
and economically thanks to the adaptability of PRINT on any manufacturing scale.

Photoresist
There are two types of photoresists: positive and negative-

Positive-When exposed to light, positive photoresist becomes more soluble in the developing
solution. In other words, during the development process, the exposed regions will be
eliminated, leaving the untouched areas. As a result, the original photomask's pattern is
reversed in the pattern. Microelectronics production and other high-precision applications
frequently employ positive photoresists.
Negative-Conversely, when exposed to light, negative photoresist becomes less soluble in the
developing solution. Throughout development, the regions that are exposed to light will be
preserved, while the areas that aren't will be scraped away. As a result, the pattern produced
is the same as that of the original photomask. Printed circuit boards and microfluidics are two
applications that frequently employ negative photoresists.
In conclusion, the primary distinction between positive and negative photoresists is how they
respond to exposure to light, which results in various patterns being imprinted onto the
substrate during the photolithography process.

Biomedical Microsystems (BB610) 19

UV radiation
In biology, UV radiation is utilized to produce exact patterns for photolithography mask
designs. By the use of a photomask, a photoresist layer is exposed to UV light, changing the
solubility characteristics of the layer through a chemical reaction. Once the patterns have
been created, they may be transferred to a substrate to create intricate micro- and nano-
scale structures for use in research and commercial applications.

PDMS
A common form of silicone elastomer used in biology and biotechnology is PDMS
(Polydimethylsiloxane). It is a translucent, malleable, and inert substance that is perfect for
microfluidic devices, biochips, and other biomedical applications. It is also easily moldable
into different forms and sizes. Moreover, PDMS is biocompatible, meaning it doesn't react
negatively when in touch with living things like tissues or bodily fluids. PDMS is a prominent
material in research and development for biomedical and biological applications because of
its distinctive features.[13]

PROCEDURE:
1. Using a tilted-rotated photolithography technique, master templates were
first created.
2. An anti-reflective layer was applied to a polished silicon wafer; it was observed
that this layer significantly decreased backside reflections and dramatically
improved the resolution of the master templates produced.
3. Spin coating was used to apply a thick layer of positive photoresist (PR) on the
wafer.
4. The wafer was coated with spin coating to adhere the photoresist to it. The PR
was then covered with a mask of 200x200 m squares and 200 m spacing (base
to base), and the complex was subjected to UV radiation at incidence angles of
18 to 25°.
5. The depth of the mold is determined by the size of the mask and the incidence
angle of the UV light, which ultimately determines the length of the
microneedles.
6. Four exposures resulted in female master templates with square-pyramidal
cavities after the wafer was rotated 90 degrees about its surface normal.
7. At a middle step, a perfect duplicate of the master template was created.
Because of its low surface energy, simplicity of usage, great flexibility, and low
cost, the replicas were made using widely available polydimethylsiloxane
(PDMS). With equal needle lengths and tip radii of curvature, the replicas
demonstrated considerable repeatability of the master templates.

Biomedical Microsystems (BB610) 20

 Packaging and Testing
Packaging microneedles requires specific requirements to ensure their safety, sterility, and
effectiveness. Here are some important requirements for packaging microneedles:

• Sterilization: Microneedles must be sterilized to prevent contamination and ensure patient

safety. The packaging should maintain the sterility of the microneedles until they are used.
• Material Compatibility: The packaging material should be compatible with the
microneedle material to prevent any interaction that could affect the efficacy of the
microneedles.
• Protection from damage: The packaging should be designed to protect the microneedles
from damage during transport and storage. Microneedles are fragile and can be easily
damaged, so the packaging must be sturdy enough to prevent any damage.
• Convenience: The packaging should be user-friendly and easy to open. The packaging
should also be designed to allow for easy access to the microneedles when they are
needed.
• Labelling: The packaging must be labeled with clear and concise instructions for use, the
contents, and any relevant safety information.
• Moisture protection: Microneedles are sensitive to moisture, so the packaging should be
designed to protect them from moisture during transport and storage.
• Quality control: The packaging should be subjected to rigorous quality control checks to
ensure that it meets the required standards for safety and effectiveness.

Microneedles for cattle are typically packaged in materials that provide protection against
contamination, moisture, and physical damage. Some of the materials commonly used for
packaging cattle microneedles include:

• Medical-grade plastic: Medical-grade plastic is commonly used for packaging

microneedles because it is lightweight, durable, and provides excellent protection against
contamination and moisture. It is also compatible with sterilization techniques such as
gamma irradiation or ethylene oxide gas.
• Aluminium foil: Aluminium foil is a popular packaging material for microneedles because
it is lightweight, flexible, and provides an excellent barrier against moisture, air, and light.
It also has good heat-sealing properties, making it easy to package and sterilize
microneedles.
• Tyvek: Tyvek is a synthetic material that is used for packaging medical devices, including
microneedles. It provides an excellent barrier against bacteria and other contaminants,
and it is also breathable, allowing for the sterilization process to occur.
• Glass vials: Glass vials are commonly used for packaging microneedles because they are
inert, meaning they will not interact with the microneedles, and they provide an excellent
barrier against moisture, air, and light. Glass vials are also easy to sterilize and can be
sealed with a rubber stopper and aluminium crimp.

Biomedical Microsystems (BB610) 21

• Plastic trays: Plastic trays are another option for packaging microneedles, especially when
the microneedles are arranged in an array or an organized pattern. Plastic trays are
lightweight, durable, and can be easily sterilized with gamma irradiation or ethylene oxide
gas.

Process of packaging of microneedles

The process for packaging microneedles typically involves several steps to ensure that the
microneedles are protected from contamination and damage and that they remain sterile
until they are used. Here are the general steps involved in the packaging process:

• Preparing the packaging materials: The packaging materials, such as plastic bags or vials,
are first prepared by washing them with an appropriate solution to remove any dirt or
contaminants.
• Sterilizing the packaging materials: The packaging materials are then sterilized using a
suitable method, such as gamma irradiation or ethylene oxide gas. This ensures that the
packaging materials are free from any microorganisms that could contaminate the
microneedles.
• Placing the microneedles in the packaging: The microneedles are then placed in the
packaging materials, such as plastic bags or vials, using aseptic techniques to avoid
contamination. In some cases, the microneedles may be arranged in an array or an
organized pattern in a plastic tray.
• Sealing the packaging: Once the microneedles are in the packaging, the packaging is sealed
to prevent any contamination or damage during transport and storage. Depending on the
type of packaging, sealing may involve heat sealing, crimping, or capping.
• Labelling the packaging: The packaging is then labelled with clear and concise instructions
for use, the contents, and any relevant safety information. This ensures that the end user
can use the microneedles safely and effectively.
• Quality control checks: Finally, the packaged microneedles undergo rigorous quality
control checks to ensure that they meet the required standards for safety and efficacy.
This may involve visual inspection, testing for sterility and potency, and verifying the
labelling and packaging integrity.
Overall, the process for packaging microneedles requires careful attention to detail and strict
adherence to aseptic techniques to ensure that the microneedles are safe and effective for
use.

Biomedical Microsystems (BB610) 22

Testing the packaging
Testing of packaging for microneedles typically involves a range of physical and biological tests
to ensure that the packaging provides adequate protection to the microneedles during storage
and transport, and that the packaging materials do not negatively affect the performance or
sterility of the microneedles. Here are some of the common tests performed on packaging for
microneedles:

• Integrity testing: Integrity testing is performed to ensure that the packaging is free from
any leaks or defects that could allow contamination or damage to the microneedles. This
may involve visual inspection, dye penetration tests, or bubble emission tests.
• Sterility testing: Sterility testing is performed to ensure that the microneedles remain
sterile during storage and transport. This may involve incubating samples of the
microneedles in a nutrient-rich broth and checking for any bacterial or fungal growth.
• Physical testing: Physical testing is performed to evaluate the durability and strength of
the packaging materials, and to ensure that the packaging can withstand various stresses
during transport and handling. This may involve drop testing, compression testing, or
vibration testing.
• Compatibility testing: Compatibility testing is performed to ensure that the packaging
materials do not negatively affect the performance or sterility of the microneedles. This
may involve exposing the microneedles to various packaging materials, such as plastics or
metals, and checking for any changes in performance or sterility.
• Shelf-life testing: Shelf-life testing is performed to determine the length of time that the
microneedles can be stored in the packaging without compromising their performance or
sterility. This may involve storing the packaged microneedles under different temperature
and humidity conditions and monitoring their performance and sterility over time.
Overall, testing of packaging for microneedles is a critical aspect of ensuring that the
microneedles are safe and effective for use.

Biomedical Microsystems (BB610) 23

 Conclusion

A microneedle patch of chitosan was designed for the delivery of lumpy virus vaccine for cattle.
These biodegradable chitosan-based patches presented an organized distribution and homogeneous
dimension of microneedles. The patch had 324 pyrimidal microneedles of dimensions 350 x 350 x
700 µm. Based on the literature review done by us, we believe that this pyramidal microneedle
design will give us a sustained insertion of microneedles in cow's ear cadaver skin. Further studies
with an actual prototype of microneedle will allow us to determine the effect of number of
microneedles on the patch and percentage of penetration per microneedle in the skin on the
transdermal delivery of vaccine. Studies should also be undertaken to optimize the amount of drug
in each microneedle and to reduce the number of patches used per cattle to deliver adequate
amount of the vaccine to treat the lumpy virus disease.

Biomedical Microsystems (BB610) 24

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