## 1 Introduction  
Many diseases that were previously thought to be incurable have now been successfully treated. Even though there are obstacles to overcome, the rapid growth of target-based anticancer drugs has provided the opportunity to selectively destroy tumor cells located deep within tissues [1]. Cancer therapies such as chemotherapy, hormonal therapy, and monoclonal antibody therapies are already in use in the clinic. However, they have significant limitations, such as relative ineffectiveness in certain types of cancers or serious side effects that disqualify them from being considered safe drugs. The development of novel target-based anticancer drugs allows for the selective targeting and eradication of cancerous cells [2; 3]. Advanced methods have been used, such as the selective accumulation of nanodrugs by modifying their physicochemical properties or the use of human stem cells. However, the use of these nanodrugs resulted in toxicity and was insufficiently effective considering the side effects caused using nanoparticles. Recent research has focused on developing a novel immunotherapeutic strategy that effectively directs immune cells to react against invasive cancer cells [4].  
For decades, antibiotics have been used to treat microbial infections. However, antibiotic resistance (AMR) has increasingly been reported locally and globally, leading to extremely high medical costs, reducing immunity, prolonging hospital stay, and high mortality rates [5]. A recent study from Saudi Arabia has reported the emergence and dissemination of colistin resistance among major Gram-negative pathogens, such as _Klebsiella pneumoniae_ and _Pseudomonas aeruginosa_[6]. Such a spread of colistin resistance represents a major challenge to public health given that colistin is usually used to treat infections caused by multidrug-resistant (MDR) bacteria, which can complicate the therapeutic options available for clinicians to manage patients. To combat this threat, it is crucial to develop better alternative agents that can reduce the resistance burden and enhance better antimicrobial activity [7].  
Biofilms are formed as an aggregate of at least one kind of microorganism that can develop on various surfaces by producing an extracellular polymeric matrix. The phenomenon of biofilm formation on the surface occurs in four stages. It is initially attached to the surface and may be reversible or irreversible after that maturation stage using the EOS matrix. This will develop micro- or macro-colonies. The maturation stage is very crucial, showing differences in susceptibility [8].  
Graphene and graphene oxide and their derivatives attract the scientific community's attention due to their remarkable physicochemical properties [9], mainly in biomedical applications where the graphene atom has a high surface area compared to weight; an important functional property to be considered [10]. To date, many scientists have investigated oxidation functional groups of graphene, which can be utilized to discover different compound materials, showing extraordinary improvement in biological, physical, and chemical characterization used in combination with different nanocomposites [11]. Along with the positive impact of graphene nanoformulations, some studies were reporting negative aspects of graphene that can lead to cell death due to inflammation, which is purely dependent on the type of treatment, duration of exposure, and concentration used [12]. Our main objective of the study was to understand the enhancement of nitrogen-doped graphene and its efficacy as an anticancer and antimicrobial potential towards tested cell lines and microorganisms in association with the physicochemical properties analyzed (Scheme 1).  
**Scheme 1.** Graphical representation of as-synthesized NDG, and the evaluation of their biofilm inhibition, antimicrobial, and anticancer activity.