Introduction

Hydrogels constitute a three-dimensional network capable of absorbing significant amounts of

water, leading to swelling, thanks to the presence of hydrophilic groups such as -NH2, -COOH,
-OH, -CONH2, -CONH, and -SO3H. Typically formed by crosslinked polymer chains, hydrogels
can also arise from crosslinked colloidal clusters, resulting in flexible and soft structures due to
their water absorption properties. Chemical or physical crosslinking of natural or synthetic
polymer chains enables the design of hydrogels, which closely resemble living tissue owing to
their high water content, soft structure, and porosity[1,2].

The genesis of hydrogels for biomaterial applications traces back to the work of Wichterle and
Lim in 1960, who developed the synthetic poly-2-hydroxyethyl methacrylate (PHEMA)
hydrogel initially used as a filler for eye enucleation and contact lenses. Since then, hydrogels
have been increasingly explored, particularly in drug delivery and bioactive compound release
throughout the 1970s to the 1990s. Their utilization expanded into tissue engineering during the
1990s, initially confined to surface applications such as ocular treatments or wound care.

Tailoring the properties of hydrogels, including swelling-deswelling rates, stiffness,

degradability, and mesh size, can be achieved by adjusting the hydrophilic and hydrophobic
ratios, initiator or polymer concentrations, and reaction conditions such as time, temperature, and
container. Notably, the biomedical application scope of hydrogels has extended beyond surface
environments due to in situ gelation upon infection and stimuli responsiveness.

Over the past six decades, hydrogels have evolved to be implantable, injectable, and even
sprayable for various organs and tissues, gaining traction in environmental engineering, soft
robotics, and wastewater treatment. Despite these advancements, biomedical applications remain
a focal point, as evidenced by the substantial volume of research output in recent years.

In this review, we provide an overview of hydrogels, including their structures, classification,

and synthesis, before delving into recent biomedical applications. We emphasize their roles in 3D
cell cultures, drug delivery, wound dressings, and tissue engineering, highlighting their continued
relevance and potential in advancing healthcare solutions.

Hydrogels have emerged as a pivotal class of materials within the biomedical field, captivating
the attention of researchers, clinicians, and industry professionals alike. Their gel-like structure,
composed of crosslinked polymer chains with the ability to absorb and retain significant amounts
of water, underpins their remarkable versatility and utility across a broad spectrum of medical
applications. From drug delivery platforms to tissue engineering constructs and wound care
solutions,

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hydrogels offer a plethora of advantages rooted in their unique properties, including
biocompatibility, tunable mechanical characteristics, and controlled release capabilities.

At the heart of the appeal of hydrogels lies their ability to mimic the intricate architecture and
functionality of the extracellular matrix (ECM) found in natural tissues. By closely resembling
this native environment, hydrogels create a supportive milieu conducive to cellular adhesion,
proliferation, and differentiation[2].

This biomimetic quality not only facilitates the integration of hydrogel-based constructs with
host tissues but also enhances their therapeutic efficacy in regenerative medicine and tissue
engineering applications. Moreover, the ability to precisely engineer the chemical composition
and physical properties of hydrogels through tailored synthesis and crosslinking strategies
empowers researchers to fine-tune their behaviour and performance in vivo, thereby optimizing
their suitability for specific biomedical tasks.

In our comprehensive review, we aim to delve deeply into the latest advancements and emerging
trends in the utilization of hydrogels across a diverse array of biomedical contexts. Central to this
exploration is an examination of the fundamental principles governing the design and fabrication
of hydrogel-based systems. We will scrutinize the diverse range of materials and methodologies
employed to achieve desired properties and functionalities, considering factors such as
biocompatibility, mechanical strength, degradation kinetics, and responsiveness to external
stimuli. Additionally, we will delve into the complex interplay between hydrogels and biological

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environments, elucidating the mechanisms underlying cellular interactions, tissue integration,
and immunogenic responses[3].

Furthermore, we will confront head-on the current challenges and limitations impeding the
widespread adoption of hydrogels in clinical settings. Issues such as scalability, long-term
stability, and regulatory considerations represent significant barriers that must be addressed to
realize the full potential of hydrogel-based technologies. Through a rigorous analysis of these
obstacles and a forward-looking examination of potential solutions, we aim to chart a course
toward overcoming these hurdles and accelerating the translation of hydrogel-based innovations
from the laboratory to the bedside.

Ultimately, our review aspires to serve as an indispensable resource for researchers, clinicians,
and industry stakeholders seeking to harness the transformative power of hydrogels in addressing
pressing healthcare needs. By fostering interdisciplinary collaboration and knowledge exchange,
we envision a future where hydrogel-based solutions play a central role i revolutionizing modern
medicine, improving patient outcomes, and advancing human health and well-being on a global
scale[3,4].

Classification and Structure of Hydrogels

The classification and structural diversity of hydrogels are determined by a multitude of factors,
reflecting their complex nature and wide-ranging applications. These factors include their origin,
composition, response to environmental cues, methods of crosslinking, inherent properties,
configuration, and ionic characteristics, all of which collectively define their functionalities and
suitability for specific biomedical applications.

Hydrogels fundamentally consist of interconnected polymer chains, forming a three-dimensional

network capable of absorbing and retaining large quantities of water. Their sources can be
broadly categorized into natural, synthetic, or semi-synthetic polymers, each offering distinct
advantages and limitations[4].

Natural hydrogels
derive from biological sources and include materials such as cellulose, chitosan, collagen,
alginate, agarose, hyaluronic acid, gelatin, and fibrin. These materials possess inherent
biocompatibility, bioactivity, and biodegradability, making them ideal candidates for various
biomedical applications. However, natural hydrogels often exhibit weaker mechanical properties
and stability compared to their synthetic counterparts. Additionally, some components of natural
hydrogels may trigger allergic reactions in rare cases, necessitating caution when used in
sensitive populations.

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 Fig- protein-based natural hydrogel from the girdle of the ‘sea cockroach’

Synthetic hydrogels
synthesized through the polymerization of monomers, offer precise control over their chemical
composition and properties. Examples of synthetic polymers used in

Fig- The hydrogel consist of cross-linked chains, fixed ions and contain mobile ions.

hydrogel synthesis includes polyvinyl alcohol (PVA), polyethylene glycol (PEG), polyethylene
oxide (PEO), poly-2-hydroxyethyl methacrylate (PHEMA), poly-N-isopropyl acrylamide
(PNIPAM), polyacrylic acid (PAA), and polyacrylamide (PAAM).

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 These materials often exhibit enhanced mechanical strength and stability, making them suitable
for applications requiring robust materials. While some synthetic polymers may possess inherent
biocompatibility, careful consideration is needed to ensure their safety and compatibility with
biological systems.

Semi-synthetic hydrogels represent a hybrid approach, incorporating chemically modified natural

polymers or a combination of natural and synthetic polymers. Examples include
methacryloyl-modified gelatin (GelMA) and acrylate-modified hyaluronic acid (AcHyA), which
retain the bioactive properties of their natural counterparts while offering improved tunability
and functionality through chemical modifications. Additionally, semi-hydrogels can be
engineered using a combination of natural and synthetic polymers, allowing for tailored
properties and functionalities to meet specific biomedical needs[4].

In summary, the classification and structure of hydrogels encompass a diverse array of materials
and methodologies, each with unique advantages and challenges. Understanding these factors is
essential for harnessing the full potential of hydrogels in biomedical applications, ranging from
tissue engineering and drug delivery to wound healing and regenerative medicine. Continued
research and development in hydrogel technology hold promise for advancing healthcare
solutions and improving patient outcomes.

Fig- classification of hydrogel

Hydrogels exhibit diverse structures based on the composition of polymers they contain, which
can be categorized into homopolymers, copolymers, semi-interpenetrating networks
(semi-IPNs), or interpenetrating polymer networks (IPNs). Each of these structures offers distinct
properties and advantages[4]..

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Homopolymer hydrogels are composed of polymer chains originating from a single type of
monomer. In contrast, copolymer hydrogels consist of polymer chains derived from two or more
different monomer species. Copolymers can further be classified into block, alternate, or random
copolymers, depending on the arrangement of monomers within the polymer chains.

Both homopolymer and copolymer hydrogels feature a single type of polymer chain, providing
specific properties based on the monomeric composition. Semi-IPNs and IPN hydrogels,
however, involve multiple types of polymer chains, leading to enhanced structural complexity
and functionality.

Semi-IPN hydrogels incorporate a polymer network embedded within linear polymer chains,
with no additional crosslinking agent required. This configuration offers improved mechanical
strength and swelling properties compared to homopolymer and copolymer hydrogels due to the
presence of multiple polymer components[5].

IPN hydrogels, on the other hand, are formed by crosslinking two or more distinct polymer
networks together using a crosslinking agent. This results in a highly interconnected structure
with superior mechanical properties and swelling behavior compared to other hydrogel types.

Overall, semi-IPNs and IPNs present higher mechanical strength and swelling properties relative
to homopolymer and copolymer hydrogels, owing to their more complex structures involving
multiple polymer networks. This enhanced performance makes them particularly attractive for
various biomedical and industrial applications requiring robust and versatile materials.

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Hydrogels can exhibit various configurations, categorized as amorphous, crystalline, or
semi-crystalline. Amorphous hydrogels possess random network structures at the molecular
level, while crystalline hydrogels feature tightly packed polymer networks with a crystalline
order. Semi-crystalline hydrogels, introduced in 1994 through chemical crosslinking, incorporate
both amorphous and crystalline regions. Subsequent advancements have led to the development
of physically crosslinked semi-crystalline hydrogels via bulk and micellar polymerization. These
physical hydrogels undergo rapid transitions between solid and liquid states, offering
reversibility compared to their chemically crosslinked counterparts. This property renders them
suitable for applications such as injectable hydrogels and shape memory materials[4].

Hydrogels can be further classified based on their crosslinking method into chemical and
physical hydrogels. Chemical hydrogels feature permanent junctions formed by covalent
crosslinking, while physical hydrogels have transient junctions comprising physical interactions
like ionic interactions, hydrogen bonding, and crystallization. Physical hydrogels exhibit softer
mechanical properties due to weaker physical interactions but offer reversibility from liquid to
solid states.

Additionally, hydrogels can be categorized into nonionic, anionic, cationic, and ampholytic
groups based on their ionic charge. Anionic hydrogels carry negative charges, cationic hydrogels
bear positive charges, and ampholytic hydrogels contain both negative and positive charges.
These ionic hydrogels exhibit pH sensitivity and can form complexes with electronically charged
molecules, making them suitable for drug delivery applications.

Physical Response of Hydrogels

Hydrogels can respond to physical, chemical, or biomedical stimuli, adjusting their properties
such as deformation and self-assembly accordingly. Physical stimuli include temperature,
pressure, light, electric, and magnetic fields, while chemical stimuli encompass pH, ionic
strength, solvent composition, and molecular species. Biomedical stimuli involve responses to
antigens, ligands, and enzymes. Based on their responsiveness, hydrogels are classified as
conventional or smart hydrogels. Conventional hydrogels exhibit minor alterations in response to
environmental changes, while smart hydrogels dynamically adjust their properties to small
environmental changes[6].

The solid component of a hydrogel comprises a three-dimensional network of crosslinked

polymer chains, often referred to as a mesh at the molecular level. The crosslinking can occur via
covalent (chemical) or physical (junction/entanglement) linking. Supramolecular polymers, a
novel category, offer potential for biomaterial applications beyond conventional polymers. These

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polymers feature specific but nonpermanent interactions such as hydrogen bonding, π-π stacking,
and host-guest interactions, allowing for reversible polymeric properties in solution

Fig- As depicted in above figure, covalent bonds can form between two functional groups of
polymers either with or without the presence of covalent agents

Often facilitated by enzyme catalysis. On the other hand, physical junctions can arise from
electronic interactions between oppositely charged polymer groups, as well as through hydrogen
bonds or ionic interactions with ions in the surrounding solution. These bonding mechanisms
contribute to the formation and stability of hydrogel networks.

The hydrogel mesh, depicted in Figure 4, plays a crucial role in retaining water and exerting
elastic forces, primarily due to the swelling and release of water. Consequently, the mesh
structure maintains the integrity and solidity of hydrogels, ensuring their functionality. Meshes
also facilitate fluid exchange across the polymer network and serve as conduits for transporting
cells or drugs. The size of the mesh holes, known as mesh size, is a critical parameter that
influences drug release kinetics and is correlated with the linear distance between two
crosslinking points (ξ)[6].

Studies, such as that conducted by Demeter M et al., have utilized rheological analysis to
elucidate how the average molecular weight between crosslinking points (Mc) and crosslink
density (Ve) impact mesh size (ξ). Notably, there is a positive correlation between Mc and mesh
size, indicating that higher molecular weights lead to larger mesh sizes. Additionally, research by
Steinman NY et al. has demonstrated that hydrogel molecular weight is influenced by the

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polymer content, with different polyethylene glycol (PEG) molecular weights achievable at the
same concentration of crosslinking agent. This phenomenon underscores the infinite potential
molecular weight of hydrogels due to their three-dimensional network structure.

In summary, the intricate interplay between covalent and physical bonding mechanisms, coupled
with the structural properties of the hydrogel mesh, governs the functionality and performance of
hydrogel networks in various biomedical applications.

Characteristic of hydrogel
The water holding capacity and permeability stand out as crucial characteristics of hydrogels.
Upon contact with water, the polar hydrophilic groups within the hydrogel are the first to
hydrate, initiating the formation of primary bound water. Consequently, the network swells,
exposing hydrophobic groups that also interact with water molecules, leading to the formation of
hydrophobically-bound water, often referred to as "secondary bound water." The combination of
primary and secondary bound water constitutes the "total bound water."

Fig- water retention capacity of hydrogel at specific temp.

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As the network absorbs additional water, driven by osmotic forces of the network chains,
swelling occurs until an equilibrium swelling level is reached. This additional absorbed water,
termed "free water" or "bulk water," fills the spaces between the network chains or within larger
pores, macropores, or voids. However, the swelling process is counteracted by the presence of
covalent or physical cross-links, which generate an elastic network retraction force[7].
Ultimately, depending on the nature and composition of the hydrogel, the next step may involve
disintegration and/or dissolution if the network chains or cross-links are degradable.
Biodegradable hydrogels, containing labile bonds, offer advantages in applications such as tissue
engineering, wound healing, and drug delivery. These labile bonds, which can be present in the
polymer backbone or cross-links, can be broken under physiological conditions either
enzymatically or chemically, often through hydrolysis.

Biocompatibility stands as the third most critical property required of hydrogels. It entails
compatibility with the immune system of the hydrogel and its degradation products, ensuring
they are non-toxic. Ideally, these products should either metabolize into harmless substances or
be eliminated through renal filtration processes.

Hydrogels typically exhibit excellent biocompatibility due to their hydrophilic surfaces, which
possess low interfacial free energy when in contact with bodily fluids. This characteristic reduces
the tendency for proteins and cells to adhere to the surface of hydrogels. Additionally, the soft
and rubbery nature of hydrogels minimizes irritation to surrounding tissues.

Overall, ensuring biocompatibility is essential for the successful integration of hydrogels in

various biomedical applications, promoting safety and compatibility within biological systems.

The presence of cross-links between various polymer chains imparts viscoelastic and
occasionally purely elastic behavior to hydrogels, ultimately defining their structure (hardness),
elasticity, and contributing to their stickiness. Because of their high water content, hydrogels
exhibit a level of flexibility akin to natural tissue.

By controlling parameters such as polarity, surface properties, mechanical characteristics, and

swelling behavior, it is feasible to manipulate the chemistry of hydrogels. This control enables
tailoring hydrogels to specific applications, optimizing their performance for various biomedical
and industrial purposes[7,8].

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Stimuli responsive hydrogels
Hydrogels possess a unique capability to sense and respond to various stimuli present in their
surrounding environment, including factors such as temperature, pH levels, and the presence of
electrolytes. What sets these stimuli-sensitive hydrogels apart from their conventional
counterparts is their ability to undergo notable changes in volume when exposed to even minor
shifts in these environmental conditions.

Among these, temperature-sensitive hydrogels, often referred to as thermogels, stand out. These
hydrogels can dynamically adjust their swelling behavior in response to fluctuations in
temperature, showcasing a fascinating property known as positive thermo-sensitivity[7].

This means that certain polymers within these hydrogels contract or shrink when cooled below a
specific temperature threshold, known as the upper critical solution temperature (UCST).

Examples of stimuli-sensitive hydrogels span a diverse range of materials, including poly(vinyl

methyl ether) and poly(N-isopropyl acrylamide) gels, as well as those based on
kappa-carrageenan and calcium.

These hydrogels hold promise for various applications, including drug delivery systems, tissue
engineering, and biomedical devices, owing to their ability to adapt and respond to changing
environmental cues.

Recent advancements in the field have led to the development of biodegradable

temperature-sensitive polymers, such as polyesters, polyphosphazenes, polypeptides, and
chitosan. Similarly, pH/temperature-sensitive polymers,

like those based on sulfamethazine, poly(b-amino ester), poly(amino urethane), and

poly(amidoamine), have garnered significant attention for their potential biomedical
applications[8].

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 Characterisation
An easy way to quantify the presence of hydrogel in a system is to disperse the polymer in water
using a cylindrical vial and visually observe the formation of insoluble material. Visual
monitoring of the solution viscosity by turning the universal up-side down can also provide a
quick measure of the bulk viscosity[8].

Solubility
Normally the hydrogel content of a given material is estimated by measuring its insoluble part in
the dried sample after immersion in deionised water for 16 h or 48 h at room temperature. The
sample should be prepared at a dilute concentration (typically ~ 1%) to ensure that hydrogel
material is fully dispersed in water. The gel fraction is then measured as follows

G e l F r a c t i o n ( h y d r o g e l % ) = ( W d W i ) * 100

Where Wi is the initial weight of the dried sample and Wd is the weight of the dried insoluble
part of the sample after extraction with water.

Swelling measurement
The Japanese Industrial Standard K8150 method has been used to measure the swelling of
hydrogels. According to this method the dry hydrogel is immersed in deionised water for 48
hours at room temperature on a roller mixer. After swelling, the hydrogel is filtered by a stainless
steel net of 30 meshes (681 µm). The swelling is calculated as follows[8,9].

S w e l l i n g = W s − W d W d E3

Where, Ws is the weight of hydrogel in a swollen state and Wd is the weight of hydrogel in dry
state. The terms ‘swelling ratio’, ‘equilibrium degree of swelling’ (EDS) or ‘degree of swelling’
has been used for more or less similar measurements.

FTIR
FTIR (Fourier Transform Infrared Spectroscopy) is a useful technique for identifying chemical
structure of a substance. It is based on the principle that the basic components of a substance, i.e.
chemical bonds, usually can be excited and absorb infrared light at frequencies that are typical of
the types of the chemical bonds. The resulting IR absorption spectrum represents a fingerprint of
measured sample. This technique is widely used to investigate the structural arrangement in
hydrogel by comparison with the starting materials

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Scanning Electron Microscopy (SEM)
SEM can be used to provide information about the sample's surface topography, composition,
and other properties such as electrical conductivity. Magnification in SEM can be controlled over
a range of up to 6 orders of magnitude from about 10 to 500,000 times. This is a powerful
technique widely used to capture the characteristic ‘network’ structure in hydrogels[9].

Light scattering
Gel permeation chromatography coupled on line to a multi angle laser light scattering
(GPC-MALLS) is a widely used technique to determine the molecular distribution and
parameters of a polymeric system. Hydrogel in a polymeric system can be quantified using this
technique . This technique is widely used in quantifying the hydrogels of several hydrocolloids
such as gum arabic, gelatine and pullulan. It can be demonstrated how mass recovery data
obtained from GPC-MALLS correlate with actual amount of hydrogel obtained for dextran
radiation in solid state

fig.-Correlation between mass recovery data obtained from GPC-MALLS for dextran and
amount of hydrogel formed as a function of radiation dose.

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Other technique
The main methods used to characterise and quantify the amount of free and bound water in
hydrogels are differential scanning calorimetry (DSC) and nuclear magnetic resonance (NMR).
The proton NMR gives information about the interchange of water molecules between the
so-called free and bound states . The use of DSC is based on the assumption that only the free
water may be frozen, so it is assumed that the endotherm measured when warming the frozen gel
represents the melting of the free water, and that value will yield the amount of free water in the
hydrogel sample being tested. The bound water is then obtained by difference of the measured
total water content of the hydrogel test specimen, and the calculated free water content .
Thermo-gravimetric analysis , X-ray diffraction , sol-gel analysis etc. are also used to confirm
the formation of cross-linked network gel structures of hydrogel[9,10].

Synthesis of Hydrogels

The synthesis of hydrogels involves the formation of crosslinked polymer chains arranged in a
three-dimensional (3D) network structure. Crosslinking within hydrogels can occur either after
the polymer chain synthesis or concurrently with the growth of the polymer chain. Thus, the
process of synthesizing hydrogels typically initiates with monomers, prepolymers, or polymers,
as depicted in Figure 5. Regardless of the material used, crosslinking in hydrogels can be
achieved through physical or chemical means.

Physical crosslinking methods encompass a variety of mechanisms, including hydrogen bonding,

amphiphilic grafting, block polymer formation, crystallization, ionic interactions, and protein
interactions. Hydrogen bonding occurs between molecules containing functional groups such as
N-H, O-H, or F-H, enabling the formation of hydrogels from polymers possessing these
functional groups. Amphiphilic grafting and block polymer formation involve the self-assembly
of polymers in hydrophilic or hydrophobic solvents due to their amphiphilic nature.

Crystallization is another physical crosslinking method wherein polymer chains synthesize

hydrogels by adjusting their crystallization temperature. This process often involves freeze-thaw
cycles or heating-cooling procedures to induce crystallization. Ionic interactions occur through
the attraction between ionic groups, facilitating crosslinking within the hydrogel structure.

Protein interactions are unique physical crosslinking mechanisms where polymers modified with
proteins, through molecular medical technologies such as protein and genetic engineering,
contribute to hydrogel synthesis. These modified polymers can undergo crosslinking via
antibody-antigen interactions or through the inherent properties of the proteins, including
crystallization, functional groups, or hydrogen bonding.

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In summary, the synthesis of hydrogels involves a range of physical crosslinking methods that
offer versatility and control over the resulting hydrogel properties. These methods enable the
creation of hydrogel structures tailored to specific biomedical applications, from drug delivery
systems to tissue engineering scaffolds.

Physical cross-linking
also known as reversible gelation, has garnered increased attention due to its ease of production
and the absence of cross-linking agents. Unlike chemical cross-linking, which may interfere with
the integrity of substances to be entrapped,

physical cross-linking offers advantages such as simplicity and minimal disruption to entrapped
materials like cells and proteins. Furthermore, there is no need for the removal of cross-linking
agents before application, simplifying the process.

The formation of physically cross-linked hydrogels can be achieved through careful selection of
hydrocolloid type, concentration, and pH. This approach allows for a wide range of gel textures
to be obtained, making it an area of significant interest, particularly in industries such as food
production.

Researchers have explored various methods to obtain physically cross-linked hydrogels, which
are detailed in the literature. These methods offer diverse approaches to tailor the properties of
hydrogels for specific applications[10].

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 Fig- Schematic illustration of physical, chemical and double network crosslinking strategies in
hydrogels

Heating/cooling a polymer solution

Physically cross-linked gels can be formed by heating and subsequently cooling hot solutions of
gelatin or carrageenan. The gel formation process involves the formation of helices, their
association, and the creation of junction zones. Initially, carrageenan in a hot solution exists in a
random coil conformation above its melting transition temperature. However, upon cooling, it
undergoes a transformation.

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Rigid helical rods are formed when the carrageenan transforms into a helical structure upon
cooling. When salt ions such as K+ or Na+ are present, they screen the repulsion of the sulphonic
groups (SO3–), leading to the aggregation of double helices and the formation of stable gels.
Additionally, in some cases, hydrogels can be obtained by simply warming polymer solutions,
which induces block copolymerization. Examples include polyethylene oxide-polypropylene
oxide and polyethylene glycol-polylactic acid hydrogel[11].

Ionic interaction

fig.-Ionotropic gelation by interaction between anionic groups on alginate (COO-) with divalent
metal ions (Ca2+).

Ionic polymers can be cross-linked by the addition of di- or trivalent counterions. This method
underlies the principle of gelling a polyelectrolyte solution (e.g. Na+ alginate-) with a
multivalent ion of opposite charges (e.g. Ca2+ + 2Cl-) .

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Complex coacervation

fig.-Complex coacervation between a polyanion and a polycation.

Complex coacervate gels can be synthesized by combining a polyanion with a polycation. This
method operates on the principle that polymers carrying opposite charges are attracted to each
other, resulting in the formation of soluble or insoluble complexes, depending on the

concentration and pH of the solutions involved. An illustrative example involves coacervating

the polyanionic xanthan with the polycationic chitosan. Proteins that carry a positive charge
below their isoelectric point are inclined to interact with anionic hydrocolloids, leading to the
formation of polyion complex hydrogels, commonly known as complex coacervates.

Collagen hydrogels are categorized as natural hydrogels and hold particular significance due to
collagen's prevalence as the most abundant fibrous protein in the human body. Serving as a key
structural component of the extracellular matrix (ECM), collagen contributes to the elastic
strength of tissues and regulates crucial cellular processes such as adhesion, chemotaxis,
migration, and tissue development. Its importance extends to articular and bone tissue function.
Given collagen's biofunctionality within the human body, collagen hydrogels can effectively
emulate organic properties and characteristics.

Research indicates that collagen enhances cell growth, adhesion, and differentiation, particularly
in neural cells, and facilitates the formation of tissue-like structures, such as chondrocytes, for
co-culture applications involving neural cells and chondrocytes. Both in vivo and in vitro studies
highlight collagen's significance in maintaining chondrocyte phenotype and promoting
chondrogenesis, making it a focal point in 3D cell culture of chondrocytes[12].

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 Biomedical Applications of Hydrogels

3D cell culture-
Three-dimensional (3D) -cell cultures offer a valuable setting for cells to grow in vitro, allowing
them to expand in all directions. This contrasts with traditional 2D culture systems and provides
a more accurate representation of in vivo cell behaviour, as cells adopt a 3D structure akin to that
found in living tissue. The establishment of 3D cell cultures typically involves cultivating cells
on a scaffold that mimics the extracellular matrix (ECM), where cells are naturally embedded
and organized in a 3D manner. The ECM plays a crucial role in regulating cellular behaviour.
Hydrogels,

Fig. 3D cell culture

characterized by their 3D structure and water-absorbing capabilities, closely resemble the ECM,
making them ideal candidates for scaffolds in 3D cell cultures. Consequently, hydrogels have
garnered increasing interest for their application in supporting 3D cell cultures.

Hydrogels can be derived from natural, synthetic, or semi-synthetic polymers, each offering
distinct biochemical, physical, and mechanical properties conducive to 3D cell culture. Natural
hydrogels, in particular, exhibit excellent biocompatibility, contain endogenous factors, and
possess viscoelasticity and fibril structures akin to the ECM, making them well-suited for
sustaining cellular activity in 3D environments[13].

Collagen hydrogels are categorized as natural hydrogels and hold particular significance due to
collagen's prevalence as the most abundant fibrous protein in the human body. Serving as a key

19
structural component of the extracellular matrix (ECM), collagen contributes to the elastic
strength of tissues and regulates crucial cellular processes such as adhesion, chemotaxis,
migration, and tissue development. Its importance extends to articular and bone tissue function.
Given collagen's biofunctionality within the human body, collagen hydrogels can effectively
emulate organic properties and characteristics.

Research indicates that collagen enhances cell growth, adhesion, and differentiation, particularly
in neural cells, and facilitates the formation of tissue-like structures, such as chondrocytes, for
co-culture applications involving neural cells and chondrocytes. Both in vivo and in vitro studies
highlight collagen's significance in maintaining chondrocyte phenotype and promoting
chondrogenesis, making it a focal point in 3D cell culture of chondrocytes.

Studies of type I collagen, when used as a scaffold in 3D culture of rat chondrocytes, preserves
the chondrogenic phenotype. Moreover, investigations comparing type II collagen hydrogel
scaffolds to type I collagen reveal that type II collagen significantly enhances the chondrogenic
differentiation of human mesenchymal stem cells (hMSCs). Although type I and type II collagen
support chondrogenic phenotypes through distinct mechanisms, collagen hydrogel scaffolds
exhibit promising potential for cartilage tissue engineering.

Further studies indicates that rat mesenchymal stem cells (rMSCs) encapsulated within collagen
type I/II hydrogel scaffolds promote chondrogenic differentiation and offer enhanced cartilage
repair ability in a rat model of cartilage defects. These findings underscore the clinical relevance
and therapeutic potential of collagen hydrogels in cartilage repair applications[14].

The density of capillary networks plays a crucial role in providing essential nutrients to cells, a
key factor in the generation of natural tissue in vitro. Numerous studies have explored the use of
endothelial cells (ECs) in combination with natural or synthetic materials to reconstruct the
microvasculature of natural tissue in vitro. However, a significant drawback of this approach is
the unavoidable interference of fetal bovine serum (FBS), which limits its potential for future in
vivo applications.

In addressing this challenge, shown that collagen hydrogel scaffolds, under serum-free
conditions, can effectively promote the development of endothelial cell (EC) networks,
particularly when combined with human umbilical vein endothelial cells (HUVECs) and human
adipose tissue-derived stromal cells (hASCs). Despite this promising advancement, it's important
to note that the long-term performance of type I collagen may be compromised due to significant
shrinkage and weak mechanical properties.

To overcome these limitations, sientest introduced a novel approach by enhancing the

mechanical properties and addressing cell-mediated shrinkage of collagen hydrogels through

20
hybridization with silk fibroin. This collagen-silk fibroin hydrogel scaffold demonstrates
improved cell viability and growth of human mesenchymal stem cells (hMSCs) during long-term
3D cultures. This innovative strategy not only enhances the structural integrity of collagen
hydrogels but also supports cell viability and proliferation, thereby offering promising prospects
for tissue engineering applications.

Hyaluronic acid (HA) serves as a natural source of hydrogels and is a vital structural component
of the extracellular matrix (ECM) found abundantly in various tissues such as skin and cartilage.
Its presence facilitates cellular processes including survival, migration, angiogenesis,
differentiation, and neural regeneration by transducing intracellular signals. Studies have
revealed that HA hydrogel scaffolds can induce neural differentiation of human-induced
pluripotent stem cells (hiPSC-NPCs) and improve the cardiac differentiation efficiency of
hiPSCs when utilized in a microencapsulation approach. Furthermore, adjustable stiffness HA
hydrogel scaffolds have been synthesized to support the maintenance of stem cell characteristics
and promote direct cartilage differentiation. These findings underscore the potential of HA
hydrogels in stem cell therapy and tissue repair[15].

In addition to its constructive roles, HA is overexpressed in various types of cancers, where it

contributes to tumor progression and drug resistance. HA hydrogel scaffolds designed to mimic
the topography and mechanical properties of breast cancer tumors have been shown to enhance
the migration, invasion, and tumorigenicity of cancer cells compared to traditional 2D cultures.
Similarly, studies have demonstrated that HA hydrogel scaffolds promote the adhesion,
proliferation, and multidrug resistance protein activity of hepatocellular carcinoma cells,
reducing therapeutic effectiveness compared to 2D cultures. These findings highlight the
significance of HA hydrogels in cancer research.

Moreover, HA hydrogels have been leveraged to enhance the efficacy of 3D cell cultures by
combining them with various substances. For instance, microbially-transglutaminase-crosslinked
HA semi-interpenetrating networks with chondroitin have been shown to enhance the osteogenic
potential of dental pulp stem cells. Additionally, cell-adhesive peptide-RGD-modified HA
hydrogel scaffolds promote the differentiation of human neural stem/progenitor cells toward
oligodendrocytes and neurons, supporting long-term cell viability with minimal growth factors.
Furthermore, HA-collagen hydrogels have been developed to mimic the viscoelasticity and
fibrillary structure of the ECM, promoting cell spreading, fiber remodeling, and focal adhesion
of human mesenchymal stem cells in 3D cell culture settings.

Apart from HA, fibrin, another major ECM protein, has garnered attention for its bioactivities
including angiogenesis and cell-matrix interactions. Fibrin hydrogel constructs have been
utilized to mimic native ECM matrices, enhancing prevascular formation, cell viability, and
proliferation while promoting osteogenic differentiation and bone mineral deposition.

21
Furthermore, alginate, derived from brown algae, has been explored for 3D cell cultures, with
modifications enhancing its interaction with mammalian cells and supporting the differentiation
and formation of complex networks of neurons.

Synthetic hydrogels, such as polyvinyl alcohol (PVA) and polyethylene glycol (PEG) hydrogels,
offer robust mechanical strength and biocompatibility, making them suitable for 3D cell culture
scaffolds. PVA hydrogels have been shown to promote the expansion of hematopoietic stem cells
and the differentiation of spermatogonia stem cells, while PEG hydrogels have been employed
for tumor spheroid formation and stem cell differentiation studies. Additionally, semi-synthetic
hydrogels, combining natural and synthetic polymers, address the limitations of both natural and
synthetic hydrogels. These hydrogels create a cellular environment closer to in vivo conditions,
offering promising prospects in various medical applications, including stem cell therapy, tissue
repair, and drug screening. However, further research is needed to fully assess their
tissue-mimicking capabilities and performance.

Drug Delivery

Polymers stand out as highly promising materials for crafting drug delivery systems, offering
versatile options for various nanostructures such as polymeric micelles, vesicles, and hydrogels.
These nanostructures hold immense potential for efficient drug delivery. Particularly noteworthy
are smart hydrogels, which have garnered increased attention due to their stimuli-responsive
nature. This property enables the development of innovative targeted drug formulations and
allows for controlled drug release via non-intravenous routes, thereby mitigating the rapid onset
of opsonization upon contact with blood.

fig-hydrogels for controlled drug delivery

22
The key advantage of smart hydrogels lies in their capability to adapt their properties—including
mechanical strength, swelling capacity, hydrophilicity, and permeability of bioactive
molecules—in response to external stimuli such as temperature, pH, electromagnetic radiation,
magnetic fields, and biological cues. These smart hydrogels can be fabricated using either natural
or synthetic polymers. While natural hydrogels offer inherent biocompatibility, their mechanical
properties often pose challenges in maintaining consistency. However, this limitation can be
addressed through extensive chemical modification, albeit with significant processing
complexities[14].

In contrast, synthetic polymers offer greater flexibility in tuning their chemical and physical
properties, making them ideal candidates for smart hydrogel synthesis. Biodegradable and
hydrophilic synthetic polymers, in particular, emerge as highly competitive options for
fabricating smart hydrogels tailored for drug delivery applications. These synthetic polymers
confer smart hydrogels with desirable attributes such as low toxicity, minimal side effects, and
reduced blood material adhesion. Notably, the diminished blood material adhesion can
effectively slow down the opsonization process and mitigate phagocyte-mediated elimination.

Under fluctuations in temperature, thermo-responsive hydrogels (TRHs) undergo changes in

their matrix volume, leading to expansion or contraction, and alterations in the solubility,
conformation, and phase transition of the polymer. These hydrogels exhibit the unique ability to
maintain their gel-like structure within a specific temperature range. For instance, Zhao G et al.
observed that a prodrug, N-2-hydroxypropyl methacrylamide (HPMA)-copolymer-based
dexamethasone, transitions from a liquid state at 4 °C to a hydrogel state at 30 °C or higher by
increasing the content level of Dex. Upon intraarticular administration in rodent models of
inflammatory arthritis and osteoarthritis, this HPMA-Dex transforms into a hydrogel known as
ProGel-Dex, which can persist in the joint for more than 30 days and gradually release the
water-soluble polymeric prodrug. The released Free-Dex acts on phagocytic synoviocytes,
leading to persistent improvement in joint inflammation and pain. Importantly, the low molecular
weight of the water-soluble polymeric prodrug (6.8 kDa) ensures rapid renal clearance, thereby
reducing the risk of potential glucocorticoid-related side effects.

Similarly, Xing R et al. formulated topotecan-loaded lipid nanoparticles (TPT-SLNs) and

incorporated them into solutions of poloxamer 407 and poloxamer 188 to create
TPT-SLNs-TRHs, which exhibit gelation at temperatures above 31 °C. The researchers evaluated
the anticancer efficacy of TPT-SLNs-TRHs in a colorectal tumor xenograft rat model and
observed enhanced antitumor efficacy and reduced toxicity compared to pure TPT and
TPT-SLNs administration. Furthermore, the antitumor effect of TPT-SLNs-TRHs remained
stable for 28 days, with stability tests confirming no changes in particle size and incorporation
efficiency over six months. However, further research is needed to determine if the antitumor
effect of TPT-SLNs-TRHs can be sustained beyond 28 days.

23
Researchers are actively exploring the application of thermo-responsive hydrogels (TRHs) as
drug carriers for infectious diseases, such as HIV/AIDS. Lamivudine (3TC) and zidovudine
(AZT) are commonly used compounds for HIV treatment, but frequent dosing is required for
successful therapy. However, excessive dosing can lead to reduced adherence and treatment
failure. Witika B. A et al. utilized Pluronic® F-127 TRHs to encapsulate a nano co-crystal of
3TC and AZT (NCC-3TC-AZT). In vitro release studies demonstrated sustained release of 3TC
and AZT from NCC-3TC-AZT-TRHs over 168 hours, with no significant impact on cell viability
compared to individual drug treatments. While further investigation is needed to evaluate the
antiviral efficacy of NCC-3TC-AZT, these findings suggest the potential of TRHs as drug
carriers for AIDS treatment.

Moreover, sustained release strategies are being explored to prolong the residence time of
polymer-protein therapeutics in plasma and tissues, thereby enhancing efficacy. Huynh V et al.
utilized hydrophilic polyethylene glycol methyl ether methacrylate (PEGMA) polymers to
prepare antibody-conjugated TRHs, which can modulate protein release rates by adjusting the
PEGMA content. These TRHs dissolve slowly at 37 °C, releasing antibodies over a period of 13
days in vitro, suggesting their potential for sustained protein release[15].

pH-responsive hydrogels, which swell in response to changes in pH, offer promising applications
in drug delivery, particularly in oral administration targeting the stomach or intestine. Qing W et
al. developed a pH-responsive hydrogel by incorporating luteolin-conjugated methoxy PEG
(mPEG-LUL) with the chemotherapy drug bortezomib (BTZ) for colorectal cancer treatment.
This hydrogel sustained BTZ release at pH 5.5 for up to 50 hours and demonstrated significant
tumor reduction in vivo, highlighting its potential for targeted cancer therapy.

Additionally, photoresponsive hydrogels, which can alter their properties in response to light
energy, offer precise control over drug release. Ghani M et al. designed a photoresponsive
hydrogel using carboxylated spiropyran-modified silicone (SPCOOH)-based interpenetrating
networks (IPNs), enabling controlled release of doxycycline under UV light stimulation. This
hydrogel exhibited sustained drug release over 42 hours, suggesting its utility in controlled drug
delivery.

Furthermore, dual-responsive hydrogels, which respond to multiple stimuli, combine the

advantages of different responsive systems. Abedi F et al. developed a noncytotoxic
pH-/thermo-responsive hydrogel that simultaneously released doxorubicin (DOX) and curcumin
(CUR) under pH 5.8/40 °C conditions, demonstrating enhanced apoptosis in colon cancer cells
compared to individual drug treatments. These findings underscore the potential of
dual-responsive hydrogels for synergistic drug delivery.

24
In conclusion, various types of responsive hydrogels offer distinct advantages for drug delivery,
but each has its limitations and requires further evaluation for clinical translation.
Photoresponsive hydrogels, in particular, show promise due to their tunable properties under
light stimulation. However, additional research is needed to fully understand their in vivo
efficacy and safety[16].

Wound Dressings

fig-showing the development of hydrogels for wound healing applications

The skin, as the body's largest organ, serves as a vital protective barrier against external threats.
However, its constant exposure to physical, chemical, and thermal stressors makes it susceptible
to damage, resulting in wounds that disrupt its integrity. When such injuries occur, the body
initiates a complex series of events aimed at repairing the damaged tissues, a process known as
wound healing.

Wound healing can be broadly categorized into two types: acute and chronic. Acute wounds,
typically caused by sudden mechanical trauma like cuts, burns, or surgical incisions, follow a
well-defined timeline and generally heal completely within a few weeks. On the other hand,
chronic wounds are characterized by delayed or impaired healing, often persisting beyond the

25
expected healing period of 12 weeks. These wounds can arise from various factors, including
underlying health conditions like diabetes, compromised blood flow, or repeated tissue damage.

The process of wound healing unfolds in several distinct but overlapping phases. It begins with
hemostasis, where blood clotting mechanisms come into play to stop bleeding and prevent
further loss of blood. This is followed by the inflammatory phase, during which immune cells
migrate to the wound site to clear debris, neutralize pathogens, and release signalling molecules
that initiate tissue repair.

As the wound enters the proliferative phase, fibroblasts proliferate and migrate to the wound
area, synthesizing new extracellular matrix components like collagen and forming granulation
tissue. This tissue serves as a scaffold for the growth of new blood vessels, a process known as
angiogenesis, which helps deliver oxygen and nutrients to the healing site. Concurrently,
epithelial cells migrate over the wound surface to close the wound, a process called
re-epithelialization[26].

Finally, during the remodelling phase, the newly formed tissue undergoes maturation and
remodelling, characterized by the degradation of excess collagen and the strengthening of the
scar tissue. This phase may last for months to years, gradually restoring the skin's structural
integrity.

Effective wound management plays a critical role in supporting and accelerating the
wound-healing process. Wound dressings, in particular, serve as a primary intervention by
creating an optimal environment for healing. Ideally, a wound dressing should maintain
moisture, allow for gas exchange, absorb excess exudate, prevent infection, promote tissue
regeneration, and be well-tolerated by the body.

Hydrogels, which are three-dimensional networks of hydrophilic polymers capable of absorbing

and retaining large amounts of water, have emerged as promising candidates for wound
dressings. Their ability to maintain a moist wound environment, resemble the natural
extracellular matrix, and deliver therapeutic agents makes them attractive for wound healing
applications[17].

Recent advancements in hydrogel-based wound dressings have focused on incorporating

functional properties to address specific challenges in wound management. These properties may
include hemostatic activity to control bleeding, antibacterial effects to prevent infection,
angiogenic stimulation to promote blood vessel formation, anti-inflammatory properties to
reduce swelling and pain, and antioxidant capacity to counteract oxidative stress.

26
Natural polymers such as cellulose, chitosan, collagen, and hyaluronic acid have been
extensively explored for their biocompatibility and bioactive properties, making them popular
choices for hydrogel formulations. Additionally, synthetic polymers can be tailored to exhibit
desired characteristics such as controlled release of therapeutic agents or responsiveness to
environmental cues.

Furthermore, hydrogels loaded with bioactive compounds such as antibiotics, growth factors, or
antioxidants have been developed to enhance their therapeutic efficacy. These compounds can
help combat infections, stimulate tissue regeneration, and modulate the wound
microenvironment to support healing.

Overall, functional hydrogel-based wound dressings represent a promising approach to wound

management, offering versatile platforms for delivering targeted therapies and promoting[17].

Tissue Engineering

Tissue engineering holds immense promise for revolutionizing medical treatment by offering
solutions to address functional failures and severe tissue damage that traditional therapies
struggle to resolve. This interdisciplinary field combines principles of engineering, biology, and
materials science to develop innovative strategies for tissue repair and regeneration.

One of the key components of tissue engineering is the creation of scaffolds that mimic the
native extracellular matrix (ECM) found in living tissues. These scaffolds provide a supportive
framework for cells to adhere to, proliferate, and differentiate, ultimately facilitating the
formation of new functional tissue. Hydrogels, owing to their unique properties, have emerged as
particularly versatile and effective materials for constructing such scaffolds[18].

fig-Hydrogel based tissue engineering and its future

27
Hydrogels possess a remarkable ability to retain water and maintain a hydrated environment
similar to that found in living tissues, which is crucial for promoting cell viability and function.
Moreover, their mechanical properties can be tailored to match the specific requirements of
different tissue types, allowing for precise control over the microenvironment in which cells
grow and differentiate.

In tissue engineering, hydrogel scaffolds have shown tremendous potential for regenerating a
wide range of tissues, including nerves, cardiac tissue, cartilage, and bone. By providing a
supportive and biocompatible environment, hydrogels facilitate cell adhesion, proliferation, and
differentiation, leading to the formation of new tissue structures with improved functionality.

Furthermore, the development of advanced techniques such as 3D printing has enabled the
precise fabrication of hydrogel scaffolds with complex geometries, offering unprecedented
control over the architecture and properties of the resulting tissue constructs. This level of
precision allows researchers to tailor the scaffolds to match the intricate anatomical features of
different tissues, enhancing their compatibility and functionality[24].

While much progress has been made in preclinical studies demonstrating the effectiveness of
hydrogel-based tissue engineering approaches, translating these findings into clinical
applications presents additional challenges. Issues such as scalability, reproducibility, long-term
stability, and safety must be addressed to ensure the successful translation of hydrogel-based
therapies from the laboratory to the clinic.

Nevertheless, the remarkable versatility and efficacy of hydrogel-based tissue engineering

strategies offer tremendous promise for addressing a wide range of medical conditions associated
with tissue damage and dysfunction. With ongoing advancements in materials science,
biotechnology, and regenerative medicine, hydrogel-based approaches are poised to play a
central role in the future of medical treatment and tissue repair.

28
Buccal Delivery
The buccal drug delivery system has garnered significant interest due to the high permeability of
the buccal mucosal membrane, allowing for rapid absorption of pharmaceutical compounds into
the systemic circulation. An ideal polymer for buccal delivery should possess several key
characteristics, including excellent spreadability, swelling ability, rheological properties,
bioadhesivity in both dry and wet states, mechanical strength, non-toxicity, cost-effectiveness,
biocompatibility, and biodegradability[20].

Hydrogel-based formulations such as tablets, films, or bioadhesive patches have been

successfully developed to control the release and permeation of drugs across the buccal mucosa.
These formulations utilize hydrogels to modulate the release pattern of drugs based on hydration
and swelling processes. Commonly used hydrogel-based polymers for buccal delivery include
hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC),

hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), chitosan, polyacrylic

resins, polyvinyl alcohol (PVA), polyvinyl pyrrolidone, Kollicoat, maltodextrin, Lycoat NG 73,
and pullulan.

Buccal films, particularly those prepared from mucoadhesive polymers like

chitosan-carbopol/chitosan-gelation loaded with drugs such as miconazole nitrate, have
demonstrated excellent efficacy against Candida species in vitro.

These films offer flexibility, prolonged contact time, and enhanced protection to injured oral
mucosa. Similarly, bioadhesive patches fabricated from various ionic (e.g., sodium CMC and
chitosan) and non-ionic (e.g., PVA, HEC, HPMC) mucoadhesive hydrogel polymers have shown
promise for buccal drug delivery.

Nanocarriers loaded into hydrogels are utilized to improve drug residence time, bioavailability,
and protection from degradation in buccal delivery systems. Evaluation of mucoadhesion using
tensile strength and rotating cylinder tests has identified cellulose derivatives such as CMC,
HEC, and HPMC as promising options for buccal applications. These cellulose-based patches,
considered generally safe, can be used for various intraoral conditions[22].

The PROLOC™ bioadhesive drug delivery system, composed of starch-polyacrylic acid blends,
exhibits remarkable adhesion to hydrated mucous membranes and subsequently erodes
completely. Utilizing Proloc 15 polymer along with water-insoluble polymers like

29
Eudragit RL100/RS100, a buccal film for the effective delivery of almotriptan was developed,
demonstrating enhanced buccal permeation compared to oral suspension[23].

Furthermore, dental light-curable gelatin hydrogels containing antimicrobial peptides have

shown significant bioadhesivity to soft tissues like gingiva and hard surfaces like bone or
titanium implants. These multifunctional hydrogels exhibit cytocompatibility, biodegradability,
and tissue regenerative capabilities, making them promising candidates for various peri-implant
diseases.

fig-Enhanced buccal permeation demonstrated by the hydrogel-based mucoadhesive almotriptan

buccal film compared to oral suspension containing equivalent dose

30
 Conclusion
In conclusion, this review has provided an overview of the classification, structure, and recent
applications of hydrogels in various biomedical fields, including 3D cell cultures, drug delivery,
wound dressings, and tissue engineering. Hydrogels, owing to their unique properties such as
biocompatibility, tunable mechanical strength, and ability to mimic the extracellular matrix
(ECM), hold great promise for advancing medical treatments across multiple domains.

Semi-synthetic hydrogels represent a significant advancement in 3D cell culture technology,

offering the ability to optimize the drawbacks of both natural and synthetic hydrogels. These
hydrogels serve as excellent scaffolds for 3D cell cultures, closely resembling the ECM of living
tissues. Moreover, with the integration of 3D printing technology, these scaffolds show promise
in tissue engineering applications. However, further evaluation of their clinical efficacy is
warranted to fully realize their potential in tissue regeneration and repair.

In the realm of drug delivery, photoresponsive hydrogels have emerged as a promising approach,
offering precise control over drug release through light stimulation. While these hydrogels
demonstrate excellent properties for drug delivery under specific light conditions, the exploration
of alternative materials and delivery mechanisms may overcome limitations associated with
other responsive hydrogel systems.

Functional hydrogels have shown rapid advancements in wound healing applications, leveraging
their bioactive properties to accelerate the healing process. Multifunctional hydrogels, capable of
addressing the complex and dynamic nature of wound healing, hold promise as a strategic
approach to improve clinical outcomes. However, the commercial availability of hydrogels for
clinical applications in areas such as 3D scaffolds, drug delivery, and tissue engineering remains
limited.

Moving forward, a deeper understanding of the mechanical strength, elasticity, and biological
composition of the ECM in various cell and tissue types will be crucial for advancing the
medical applications of hydrogels. By leveraging this knowledge, researchers can develop
tailored hydrogel-based therapies with enhanced efficacy and safety profiles, ultimately
expanding the scope of hydrogel utilization in clinical treatments.

Furthermore, the development of hydrogel-based therapies will benefit from ongoing efforts to
explore novel formulations, fabrication techniques, and functionalization strategies. For instance,
the integration of bioactive molecules, growth factors, and nanoparticles into hydrogel matrices
holds potential for enhancing their therapeutic properties and targeting specific biological
processes. Moreover, the design of hybrid hydrogels, combining the advantages of different
polymer systems, offers opportunities to create customizable platforms with improved
performance characteristics.

31
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