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Graphene-Based Nanomaterials for Drug Delivery 245
allowing for regulated release in reaction to environmental stimuli, thanks to their special electrical features [75].
3.5 Nanoflakes of Graphene
Small, thin, and irregularly shaped graphene structures with lateral dimensions ranging from a few nanometers to micrometers are known as graphene nanoflakes. They can be created using bottom-up synthesis techniques or top-down procedures like mechanical exfoliation. Due to their large surface area, simplicity of function­alization, and biocompatibility, graphene nanoflakes have demonstrated potential as drug carriers. These nanoflakes are attractive candidates for intracellular drug delivery because of their irregular structure, which may enhance cellular uptake and drug release. For effective drug delivery, additional research is needed to maximize their drug-loading capability and enhance their biodegradability.
Promising prospects for drug delivery applications include graphene-based nano­materials such as GQDs, GO, graphene nanoribbons, oxidized graphene nanoribbons, and graphene nanoflakes. They are adaptable platforms for drug delivery s ystems thanks to their special qualities, which include a wide surface area, functionalization potential, and customizable electrical and optical properties. Utilizing these unique characteristics, scientists can create focused and regulated drug delivery systems that have the potential to completely change therapeutic approaches across a range of biological sectors [76]. To successfully translate these findings into practical appli­cations, more study is necessary to fully understand their biocompatibility, in vivo behavior, and long-term safety profiles.

4 Graphene Functionalized Materials for Drug Delivery

Over time, the discipline of medication administration has made incredible strides, changing how modern medicine is practiced. Graphene functionalized materials have become a viable platform with distinct features and a wide range of appli­cations among the many developments (Fig. 6). With a focus on their uses in bone tissue regeneration, neural regeneration, photodynamic and photothermal therapy, enhancing cellular and humoral immunity, antibacterial action against Gram-Positive and Gram-Negative bacteria, and other applications, this chapter examines the multi­faceted role of graphene functionalized materials in drug delivery. Targeting partic­ular tissues, organs, or cells while limiting negative effects on healthy tissues is the goal of drug delivery systems, which attempt to increase the therapeutic efficacy of drugs. Due to its outstanding biological, mechanical, electrical, and optical qualities, graphene, a single layer of carbon atoms organized in a two-dimensional lattice, has attracted a lot of interest. Researchers have modified graphene’s properties to fit
246 A. Mohamed Noor et al.
Fig. 6 Different applications of graphene functionalized for drug delivery
different drug delivery applications by functionalizing it with bioactive chemicals, peptides, or nanoparticles, unleashing its potential to transform medical treatments.

4.1 In Bone Tissue Regeneration

A crucial component of regenerative medicine is bone tissue regeneration, which has the potential to significantly improve the lives of millions of people with bone illnesses and injuries. Traditional methods of bone grafting have drawbacks, such as inadequate host tissue integration and possible immunological reactions. Graphene, graphene functionalized materials, and its derivatives offer a creative answer to these problems. Graphene derivatives have the capability to form compos­ites with diverse structures, thereby enhancing their properties. This, in turn, directly enhances the functionality of bone-related cells or indirectly promotes them through external stimuli. Graphene derivatives can facilitate the growth and proliferation of osteoblasts, induce osteogenic differentiation of stem cells by activating specific signal pathways, and up-regulate the expression of specific factors in macrophages, hence ultimately leading to bone regeneration. The remarkable mechanical strength and biocompatibility of graphene make it a prime choice for applications involving bone tissue regeneration. Researchers have added bioactive chemicals and growth factors to graphene to functionalize it, promoting osteogenesis and improving the integration of grafts with host tissue. These modified graphene scaffolds serve
Graphene-Based Nanomaterials for Drug Delivery 247
as precise drug delivery devices, dispensing growth hormones and osteogenic substances just where damage has occurred. As a result, bone regeneration is sped up, improving healing outcomes. Additionally, because of graphene’s electrical conduc­tivity, electroactive scaffolds can be created to promote bone tissue growth and nerve regeneration alongside bone healing [77, 78].

4.2 In Neural Regeneration

The intricate process of neural regeneration offers individuals with neurological diseases and traumas great hope [79]. The promotion of significant neuronal regen­eration and functional recovery is frequently difficult with conventional therapy. Graphene functionalized materials have become a fascinating approach to over­come these challenges. Because of graphene’s special qualities such as its electrical conductivity and capacity to promote brain cell growth, it is a desirable option for use in applications involving neural regeneration [80]. Researchers have successfully functionalized graphene with medicinal medicines and neurotrophic factors to help with neuronal regeneration and repair. In 2021, Qian et al. reported that graphene­based nanomaterials have little toxicity and can repair massive nerve damage in the central and peripheral nervous systems through dual control of Schwann cells and astroglia. By using 4% of graphene-based nanomaterials in the PCL matrix, it exerts low toxicity and displays a long-term effect of reparation. The toxicity still concerns researchers which may harm organs hence ensuing renal and hepatic failure, however, it is found that axon and myelin were repaired after the application of graphene­based scaffold implantation [81]. On the other hand, targeted medication delivery to injured brain tissues is made possible by these functionalized graphene carriers, which speeds up regeneration and promotes functional recovery. The restoration of sensory and motor functions in individuals with spinal cord injuries and other neuro­logical disorders has also shown promise for graphene-based neural interfaces and neuroprosthetic devices [82].

4.3 In Photodynamic and Photothermal Therapy

Non-invasive therapeutic techniques including photodynamic therapy (PDT) and photothermal t herapy (PTT) have drawn a lot of attention in the fight against cancer. These treatments rely on light-activated chemicals to selectively kill cancer cells while protecting healthy tissues. Materials with graphene functionalization have shown considerable promise for improving PDT and PTT effectiveness. Graphene is a prime candidate for photothermal therapy due to its outstanding light absorp­tion and photothermal conversion capabilities. In 2023, Lee et al. have reported that both reduced graphene oxide quantum dots (RGQDs) and hyaluronic acid
248 A. Mohamed Noor et al.
Fig. 7 Visible confocal fluorescence microscopy image of reduced graphene quantum dots and Hyaluronic acid graphene quantum dots within HeLa cells demonstrating both extracellular and intracellular photothermal therapy potential. Reproduced with permission from MDPI [83]
graphene quantum dots (HGQDs) display synergistic effect of promising biocompat­ible image-guided photothermal agents and drug delivery capabilities which further expand Graphene Quantum Dots applications into highly desired drug/PTT combo treatment methods. As shown in Fig. 7a, d, the green fluorescence of the 535 nm observed within HeLa cells using a lamp illumination at 460 ± 20 nm and Fig. 7b, e shows NIR imaging with 808 nm laser excitation. Figure 7c, f demonstrates the 1080 nm fluorescence spectra obtained within the cellular structures by RGQDs and HGQDs respectively, validating the intracellular presence of GQDs with less visible­overlapping and background-free measurement [83]. Graphene-based drug delivery systems can assemble in tumor tissues when functionalized with photosensitizers and targeting ligands. These functionalized graphene materials produce localized heat when exposed to near-infrared light, which causes cancer cells to be selec­tively destroyed [84]. Functionalized graphene can work as an effective photosensi­tizer carrier in photodynamic treatment. Reactive oxygen species are released by the functionalized graphene upon light activation, effectively eliminating cancer cells. The synergistic advantages of combining PDT and PTT in a single graphene-based nanocomposite have been astounding, increasing treatment efficacy while reducing side effects [ 85].

4.4 In Enhancing Cellular and Humoral Immunity

A potential strategy for treating a number of illnesses, including cancer and autoim­mune disorders, is immunotherapy. Exciting possibilities exist for improving cellular
Graphene-Based Nanomaterials for Drug Delivery 249
and humoral immunity thanks to graphene functionalized materials. With the use of immunomodulatory substances like cytokines or immunomodulatory peptides, researchers have functionalized graphene [86, 87]. As a result of these functional­ized graphene carriers’ potent immune response modulation, the body’s protection against infections is improved, and tissue regeneration is encouraged. Additionally, using functionalized graphene as a platform, immunomodulators can be precisely delivered to the troubled locations for treatment.
4.5 In Antibacterial Action Against Gram-Positive
and Gram-Negative Bacteria
An important global health concern is the emergence of antibiotic-resistant microor­ganisms. Graphene functionalized materials offer a potential remedy for efficiently battling viruses that are resistant to medication. Researchers have created antibacte­rial nanostructured graphene composites by functionalizing graphene with antimicro­bial peptides and nanoparticles. These modified graphene materials have the ability to damage bacterial membranes, which results in bacterial death. Antimicrobial medi­cations can be delivered to specific areas of the body utilizing graphene carriers, which limits harm to healthy tissues and lowers the chance of systemic toxicity [88]. Additionally, graphene’s special qualities, namely, its expansive surface area and high loading capacity, make it a desirable choice for antibacterial applications.

4.6 Miscellaneous

Graphene functionalized materials have demonstrated potential in numerous other drug delivery fields outside of the aforementioned applications. For instance, in order to effectively treat chronic inflammatory illnesses, researchers have looked into graphene-based carriers for the targeted delivery of anti-inflammatory phar­maceuticals. A potential advancement in the treatment of genetic problems is the delivery of small interfering RNA (siRNA) using functionalized graphene for use in gene therapy applications [89]. In addition, the problem of multidrug resistance in cancer treatment has been addressed by research into graphene functionalized materials to increase the effectiveness of conventional chemotherapy medications [90]. In the area of drug delivery, graphene functionalized materials have become a game-changing platform, transforming the management of numerous medical disor­ders. The special qualities and adaptability of graphene have opened up a wide range of potential for medical applications, including bone tissue regeneration, brain repair, cancer therapy, and immunomodulation. The development of graphene-based drug delivery systems is encouraging, but before they are widely used in clinical settings, significant issues with biocompatibility, long-term safety, and scalability
250 A. Mohamed Noor et al.
must be resolved. Table 1 summarizes graphene and graphene-based materials with their unique application and features. As research advances, graphene functional­ized materials have the potential to transform contemporary medicine and usher in a brand-new age of precise and effective medication delivery.

5 Challenges and Future Perspective

Due to their distinct and unrivaled structure and properties, graphene has recently attracted substantial attention in the field of medicine and gene delivery. These bioma­terials have made great strides in previous research, which highlights their enormous potential for biomedical applications. To fully explore their potential, additional preclinical research is still required. There is still a paucity of knowledge on what happens inside the body after loading graphene carriers with drugs, despite the focus on drug-carrying capability and bio-properties characterization in cell lines. More research is required on the interactions of graphene with cells and the efficacy of these carriers in vivo in animals, including studies on clearance processes, long-term cytotoxicity, tissue biodistribution, and intracellular uptake patterns. Understanding the behavior of graphene-based materials inside the body requires in-depth knowl­edge of their many characteristics and surface functions. Through the development of advanced graphene-based nanomaterials, gene transfection may be more effective. Although AIDS, neurological illnesses, and cardiovascular diseases are also impor­tant to pay attention to and investigate, the current trend in therapeutics research mostly focuses on cancer therapies. The goal of this assessment is to open the door for more in-depth investigation in this area.
The use of graphene and products based on it still entails inherent hazards despite efforts to lessen its toxicity. The European Scientific Committee on Emerging and Newly Identified Health Risks (SCENIHR) has designated these substances as hazardous agents. Critical elements like form, size, exposure type and duration, aggregation state, and concentration must be taken into account while researching nanomaterials for biomedical research in order to determine their safety. The toxi­city of the interaction between graphene nanoparticles and cells/tissues is still a major worry. For instance, the long-term interaction with tissues should be care­fully addressed when graphene-based devices are utilized as scaffolds in tissue engi­neering, demanding effective synthesis and purifying processes. Contaminants found in the samples can also be blamed for toxicity, emphasizing the value of meticu­lous cleaning procedures throughout production. Toxicology is also influenced by the morphological features of graphene, such as form and size. For determining graphene’s biocompatibility, one should consider factors such as its size, agglom­eration, and surface chemistry. Less toxicity has been observed in smaller flakes, highlighting the importance of graphene size in target cell internalization.
In conclusion, investigating graphene-based materials necessitates taking into account a variety of elements and traits, such as size, shape, functionalization, and functionalizing groups, as well as in-vivo concentration, pharmacokinetics,
Graphene-Based Nanomaterials for Drug Delivery 251
Ta bl e 1 Example of Graphene and graphene oxide-based materials with their promising advan­tages and properties. Reproduced with permission from WILEY Online Library [90]
Materials Applications Important features
Multifunctionalized GO Targeted cancer
Carboxymethyl cellulose-GO Targeted and sustained
GO Cancer therapy and
GO-hyaluronic acid-Arg-Gly-Asp peptide Targeted cancer
Magnetic GO-chitosan-PEG-N-Hydroxysuccinimide
polyvinylpyrrolidone­and β-cyclodextrin-modified GO
GO@soy phosphatidylcholine-folic acid nanohybrid
therapy and drug delivery
drug delivery
drug delivery system
therapy and anticancer drug delivery
Anticancer drug delivery system
Targeted anticancer drug delivery
Antitumor therapy and targeted drug delivery
· No noticeable toxic effects
· Higher drug stacking capability
· pH-responsive drug discharge features
· Particular target transport and effectual cell inhibition
· No noticeable toxicity with sustained and prolonged release of doxorubicin
· Incorporation of GO nanosheets highly improved the swelling capacity of hydrogels
· Sustained-release nanoformulation
· Improved suppression of cancer cell growth
· Low toxicity
· High drug loading
· Improved specificity and efficiency of anticancer drug delivery
· Good biocompatibility
· Low cytotoxicity
· pH-responsive controllable drug release behavior
· High drug loading potentials
· Low toxicity
· pH-dependent drug release
· No noticeable toxicity
· pH-dependable drug release
· Improved steadiness and good biocompatibility
(continued)
252 A. Mohamed Noor et al.
Ta bl e 1 (continued)
Materials Applications Important features
· Higher drug packing ability
· Effectual cellular uptake
· Regulated drug discharge
Chitosan-grafted-poly(methacrylic acid)/GOAnticancer drug
GO/chitosan oligosaccharide/ γ-polyglutamic acid
Superparamagnetic iron oxide-GO Smart nanotheranostics
Chitosan-carboxylated GO Gene delivery · High gene transferring
Modified GO Gene delivery · Low toxicity
GO/ethylene glycol-polycaprolactone Anticancer drug
GO-nanoscale hydroxyapatite Cancer therapy
delivery
Anticancer drug delivery
platform
delivery; tumor therapy
(chemotherapy and photothermal therapy)
· No detectable toxicity
· Significant biocompatibility
· High drug packing capacity
· pH-dependent drug delivery performance
· No detectable toxicity
· Simple delivery and controllable anticancer drug release behavior
· Good biocompatibility
· pH-dependable drug release
properties
· Improved release of DNA
· Suitable interaction with DNA and hydrophobic immune adjuvant
· Low cytotoxicity
· Improved biocompatibility and biodegradability
· High drug release and inhibition of tumor growth
· High biocompatibility
· High photothermal therapy activity
· Improved drug release behavior
(continued)
Graphene-Based Nanomaterials for Drug Delivery 253
Ta bl e 1 (continued)
Materials Applications Important features
· High drug loading capacity
Polymer G nano-aerogels Anticancer drug
Starch-G nanosheets Anticancer drug
Reduced-GO nanostructures Cancer therapy and
Reduced-GO nanostructures Anticancer drug
Nanoscale GO loaded with HN-1 (a tumor-targeted peptide)
D-mannose-mediated chitosan-functionalized GO nanosystems
delivery
delivery
anti-inflammatory effects
delivery
Anticancer drug delivery
Anticancer drug delivery
· High anticancer drug-releasing with pH-dependable behavior
· High anticancer drug loading capacity
· Sustained-release behavior
· Good biocompatibility
· Low toxicity with improved therapeutic efficacy
· Anti-proliferative activity with high efficacy
· Sustained pH-sensitive drug release
· Improved therapeutic efficacy
· High drug loading capacity
· High hemolytic toxicity to rabbit red blood cells
· High stability to the biological solution
· High tumor-targeting behavior
· pH-responsive drug release
· High cellular uptakes and cytotoxicity toward tumor cells
· Good biocompatibility
· Targeted and controlled delivery
· Intracellular discharge of marine algae-mediated anticancer drugs versus glioblastoma cancers (e.g., ulvan)
(continued)
254 A. Mohamed Noor et al.
Ta bl e 1 (continued)
Materials Applications Important features
5-Fluorouracil and curcumin loaded chitosan/reduced GO nanocomposites
Anticancer drug delivery
· Synergistic inhibitory effects against the growth of HT-29 colon cancerous cells
· Dual-drug loading properties
and pharmacodynamics. To ascertain the security and possible uses of graphene­based nanomaterials in medication delivery and other biomedical domains, thorough investigation and comprehension of these issues are required.

6 Conclusion

The book chapter offers a succinct summary of the many processes utilized to create graphene. Chemical reduction, CVD, mechanical and chemical exfoliation, epitaxial growth, and layer-by-layer assembly are a few of these techniques those utilized in the preparation of graphene and its derivatives. Regarding scalability, quality, and control over graphene characteristics, each process has specific benefits and draw­backs. Understanding these synthesis methods is essential for streamlining graphene production and modifying its characteristics for particular uses across a range of industries. The importance of examining various synthesis techniques is emphasized in this book chapter in order to fully harness the potential of graphene in advanced technologies, it is essential to explore its capabilities and applications.
Additionally, the major categories of graphene-based nanomaterials have been emphasized. These include graphene nanoflakes, graphene nanoribbons, GO, GQDs, and graphene nanoribbons. Each substance has unique qualities and production techniques that make them each strong contenders for applications in medication delivery. The properties of GQDs include fluorescence and quantum confinement effects, making them very relevant for many applications such as bioimaging and drug delivery. Controlled drug release systems can be easily functionalized using GO, which is generated from graphene. Targeted drug delivery may be possible with graphene nanoribbons and oxidized graphene nanoribbons with distinctive edge effects. Unusual-shaped graphene nanoflakes have potential for intracellular drug delivery. Understanding these materials makes it easier to use them as effectively as possible in various technological developments.
A brief summary of the uses of materials with graphene functionalized for drug delivery is also included in the book chapter. Researchers are looking into the effects of these chemicals in a variety of settings, including their potential use in photody­namic and photothermal therapy, in promoting cellular and humoral immunity, and in inhibiting the growth of Gram-positive and Gram-negative bacteria. Graphene’s