Physiological Barriers Research Articles

Cardiac Arrest Pregnancy and Amniotic Fluid Embolism

Amniotic fluid embolism remains one of the most devastating conditions in obstetric practice with an incidence of approximately 1 in 40,000 deliveries and a reported mortality rate ranging from 20% to 60%. The pathophysiology involves an abnormal maternal response to fetal tissue exposure associated with breaches of the maternal-fetal physiologic barrier during parturition. This abnormal response triggers a cascade of events similar to the systemic inflammatory response syndrome, involving the activation of proinflammatory mediators. Maternal treatment in cases of cardiopulmonary arrest is mainly supportive, focusing on resuscitation efforts and stabilizing the mother’s condition. However, it is crucial to prioritize prompt delivery in order to improve the newborn’s outcome.

Overcoming the challenge: cell-penetrating peptides and membrane permeability

<p>Cell-penetrating peptides (CPPs) have emerged as a promising strategy for enhancing the membrane permeability of bioactive molecules, particularly in the treatment of central nervous system diseases. CPPs possess the ability to deliver a diverse array of bioactive molecules into cells using either covalent or non-covalent approaches, with a preference for non-covalent methods to preserve the biological activity of the transported molecules. By effectively traversing various physiological barriers, CPPs have exhibited significant potential in preclinical and clinical drug development. The discovery of CPPs represents a valuable solution to the challenge of limited membrane permeability of bioactive molecules and will continue to exert a crucial influence on the field of biomedical science.</p>

Pulmonary Delivery for miRs: Present and Future Potential

Administration through the respiratory tract can be advantageous, with high drug bioavailability, limited enzymatic activity, reduced dose requirements compared to oral, and potentially diminished side effects. Among the different types of drugs studied for pulmonary delivery, genetic material delivery has gained favorable scientific interest, using polymer-, lipid-, inorganic-, or vector-based nanocarriers. As pulmonary drug delivery has been associated with challenges, including physiological barriers and lung metabolism, the delivery of sensitive molecules such as nucleic acids can exacerbate these challenges. While short-interfering RNAs (siRNAs) have been extensively reported as suitable ribonucleic acid interference (RNAi) candidates for pulmonary delivery, discussion on micro-RNA (miR) pulmonary delivery is limited despite their significant therapeutic potential. Recently, these non-coding RNAs have been explored in targeted or non-targeted pulmonary administration against various diseases. This review addresses the information gap on miR-pulmonary delivery with updated and concentrated literature. We briefly discuss the barriers to lung administration, describe different functional nanocarriers for miR delivery, and provide an extensive literature update on the different miRs and their targeted diseases currently being studied.

Scarless wound healing programmed by core-shell microneedles

Effective reprogramming of chronic wound healing remains challenging due to the limited drug delivery efficacy hindered by physiological barriers, as well as the inappropriate dosing timing in distinct healing stages. Herein, a core-shell structured microneedle array patch with programmed functions (PF-MNs) is designed to dynamically modulate the wound immune microenvironment according to the varied healing phases. Specifically, PF-MNs combat multidrug-resistant bacterial biofilm at the early stage via generating reactive oxygen species (ROS) under laser irradiation. Subsequently, the ROS-sensitive MN shell gradually degrades to expose the MN core component, which neutralizes various inflammatory factors and promotes the phase transition from inflammation to proliferation. In addition, the released verteporfin inhibits scar formation by blocking Engrailed-1 (En1) activation in fibroblasts. Our experiments demonstrate that PF-MNs promote scarless wound repair in mouse models of both acute and chronic wounds, and inhibit the formation of hypertrophic scar in rabbit ear models.

Lipids as a key element of insect defense systems.

The relationship between insect pathogenic fungi and their insect hosts is a classic example of a co-evolutionary arms race between pathogen and target host: parasites evolve towards mechanisms that increase their advantage over the host, and the host increasingly strengthens its defenses. The present review summarizes the literature data describing the direct and indirect role of lipids as an important defense mechanism during fungal infection. Insect defense mechanisms comprise anatomical and physiological barriers, and cellular and humoral response mechanisms. The entomopathogenic fungi have the unique ability to digest the insect cuticle by producing hydrolytic enzymes with chitin-, lipo- and proteolytic activity; besides the oral tract, cuticle pays the way for fungal entry within the host. The key factor in insect resistance to fungal infection is the presence of certain types of lipids (free fatty acids, waxes or hydrocarbons) which can promote or inhibit fungal attachment to cuticle, and might also have antifungal activity. Lipids are considered as an important source of energy, and as triglycerides are stored in the fat body, a structure analogous to the liver and adipose tissue in vertebrates. In addition, the fat body plays a key role in innate humoral immunity by producing a range of bactericidal proteins and polypeptides, one of which is lysozyme. Energy derived from lipid metabolism is used by hemocytes to migrate to the site of fungal infection, and for phagocytosis, nodulation and encapsulation. One polyunsaturated fatty acid, arachidonic acid, is used in the synthesis of eicosanoids, which play several crucial roles in insect physiology and immunology. Apolipoprotein III is important compound with antifungal activity, which can modulate insect cellular response and is considered as important signal molecule.

Development of Phytochemical Delivery Systems by Nano-Suspension and Nano-Emulsion Techniques.

The awareness of the existence of plant bioactive compounds, namely, phytochemicals (PHYs), with health properties is progressively expanding. Therefore, their massive introduction in the normal diet and in food supplements and their use as natural therapeutics to treat several diseases are increasingly emphasized by several sectors. In particular, most PHYs possessing antifungal, antiviral, anti-inflammatory, antibacterial, antiulcer, anti-cholesterol, hypoglycemic, immunomodulatory, and antioxidant properties have been isolated from plants. Additionally, their secondary modification with new functionalities to further improve their intrinsic beneficial effects has been extensively investigated. Unfortunately, although the idea of exploiting PHYs as therapeutics is amazing, its realization is far from simple, and the possibility of employing them as efficient clinically administrable drugs is almost utopic. Most PHYs are insoluble in water, and, especially when introduced orally, they hardly manage to pass through physiological barriers and scarcely reach the site of action in therapeutic concentrations. Their degradation by enzymatic and microbial digestion, as well as their rapid metabolism and excretion, strongly limits their in vivo activity. To overcome these drawbacks, several nanotechnological approaches have been used, and many nanosized PHY-loaded delivery systems have been developed. This paper, by reporting various case studies, reviews the foremost nanosuspension- and nanoemulsion-based techniques developed for formulating the most relevant PHYs into more bioavailable nanoparticles (NPs) that are suitable or promising for clinical application, mainly by oral administration. In addition, the acute and chronic toxic effects due to exposure to NPs reported so far, the possible nanotoxicity that could result from their massive employment, and ongoing actions to improve knowledge in this field are discussed. The state of the art concerning the actual clinical application of both PHYs and the nanotechnologically engineered PHYs is also reviewed.

Mechanism of Synthesis and Effect of Autoantibodies: Implications for Therapy

The detailed mechanisms of autoantibody synthesis are different in each disease; however, the dysfunction of immune tolerance is attracting interest as the common mechanism in many autoantibody-associated diseases. Autoantibodies must pass through various physiological barriers, such as the blood-brain barrier, to approach their antigen in the central nervous system. The direct effects of autoantibodies on their antigens vary among antibodies. Exploring the detailed mechanism of synthesis and effect of autoantibodies would provide a more radical and effective therapeutic strategy.

Toward nanotechnology-enabled application of bilirubin in the treatment and diagnosis of various civilization diseases.

Bilirubin, an open chain tetrapyrrole, has powerful antioxidant, anti-inflammatory, immuno-suppressive, metabolic-modulating and anti-proliferative activities. Bilirubin is a natural molecule that is produced and metabolized within the human body, making it highly biocompatible and well suited for clinical use. However, the use of bilirubin has been hampered by its poor water solubility and instability. With advanced construction strategies, bilirubin-derived nanoparticles (BRNPs) have not only overcome the disadvantages of bilirubin but also enhanced its therapeutic effects by targeting damaged tissues, passing through physiological barriers, and ensuring controlled sustained release. We review the mechanisms underlying the biological activities of bilirubin, BRNP preparation strategies and BRNP applications in various disease models. Based on their superior performance, BRNPs require further exploration of their efficacy, biodistribution and long-term biosafety in nonhuman primate models that recapitulate human disease to promote their clinical translation.

Application of organic nanocarriers for intraocular drug delivery.

The application of intraocular drug delivery is usually limited due to special anatomical and physiological barriers, and the elimination mechanisms in the eye. Organic nano-drug delivery carriers exhibit excellent adhesion, permeability, targeted modification and controlled release abilities to overcome the obstacles and improve the efficiency of drug delivery and bioavailability. Solid lipid nanoparticles can entrap the active components in the lipid structure to improve the stability of drugs and reduce the production cost. Liposomes can transport hydrophobic or hydrophilic molecules, including small molecules, proteins and nucleic acids. Compared with linear macromolecules, dendrimers have a regular structure and well-defined molecular mass and size, which can precisely control the molecular shape and functional groups. Degradable polymer materials endow nano-delivery systems a variety of size, potential, morphology and other characteristics, which enable controlled release of drugs and are easy to modify with a variety of ligands and functional molecules. Organic biomimetic nanocarriers are highly optimized through evolution of natural particles, showing better biocompatibility and lower toxicity. In this article, we summarize the advantages of organic nanocarriers in overcoming multiple barriers and improving the bioavailability of drugs, and highlight the latest research progresses on the application of organic nanocarriers for treatment of ocular diseases.

Nanoparticle based intranasal drug delivery system of some herbal medicinal plants for Alzheimer’s disease

AbstractBackgroundDiseases of the Central Nervous System (CNS) such as epilepsy, schizophrenia, meningitis, migraine, Parkinson’s disease and Alzheimer’s disease require delivery of the drug to the brain for effective treatment but it remains problematic area to deal with, as the central nervous system (CNS) is isolated from the whole body by the blood–brain barrier (BBB). This tight physiological barrier limits the drug diffusion towards the brain parenchyma and is considered as the bottleneck in brain drug development. Now this can be circumvented by intraventricular or intracerebral administration. Intra Nasal delivery thus attributes to non‐invasive, essentially painless, high vascularity and the permeability of the nasal mucosa, which aids the drug in bypassing the aforementioned processes of degradation. Now the increasing realization of the side effects of allopathic medicines, coupled with the growing awareness about the medicinal benefits as well as therapeutic effect of herbal products is pushing up the demand for herbal extracts and herbal‐based supplements worldwide.MethodIn this study we designed a natural compound based Nanoformulations for drug delivery via nasal route to treat Alzheimer’s conditionResultAfter various preliminary studies we prepared standard extract loaded Nanoformulations with their particle size between the range 80 – 290nm and sustained release kinetics profile. Further, the toxicity analysis (morphological, cell viability, cytotoxicity assays) of these nanoformulations was performed on RPMI2650 (Human nasal septum) and NB41A3 (Mouse hippocampus) cell lines based on the route and site of drug delivery. Lastly, in vivo experiments were performed with AD (cognitive impairment) induced mice model. There was seemingly, a significant decrease in anti ß amyloid antibody positive neurons in case of optimised nanoformulations in comparison to the galantamine. It was also found to have reduced lesions caused due to the amyloid aggregation reflecting a noted improvement in dispersing and reducing the lesions in the hippocampal tissues.ConclusionIn conclusion, the present study demonstrates that the developed nanoformulations have potential therapeutic effects on improving the pathological state of AD, and might be taken further as a promising candidate for clinical research to evaluate its efficacy in humans.

Nanobiotechnology-based treatment strategies for malignant relapsed glioma.

Gliomas are the most aggressive and lethal tumors of the central nervous system, for which few therapeutic options exist. Surgical resection is the primary treatment for most gliomas; however, tumor recurrence is nearly inevitable. Emerging nanobiotechnology-based strategies have shown great prospects for early glioma diagnosis, physiological barrier traversing, postoperative regrowth suppression, and microenvironment remodeling. Herein, we focus on the postoperative scenario and summarize the key properties of the glioma microenvironment, especially its immune peculiarities. We elucidate the challenges of managing recurrent glioma. We also discuss the potential of nanobiotechnology in addressing the therapeutic challenges of recurrent glioma, including optimizing the design of drug delivery systems, enhancing intracranial accumulation, and restoring the anti-glioma immune response. The development of these technologies offers new opportunities for accelerating the drug development process and treating recurrent glioma.

Nanocarriers for ocular drug delivery: Recent advances and future opportunities.

Topical route of administration is very important and the most commonly used method of drug delivery for treatment of ocular diseases. However, due to unique anatomical and physiological barriers of eye, it is difficult to achieve the therapeutic concentration in the targeted tissue within the eye. To overcome the effect of these barriers in absorption and to provide targeted and sustained drug delivery, various advances have been made in developing safe and efficient drug delivery systems. Various formulation strategies for ocular drug delivery are used, like basic formulation techniques for improving availability of drugs, viscosity enhancers, and use of mucoadhesives for drug retention and penetration enhancers to promote drug transport to the eye. In this review, we present a summary of the current literature to understand the anatomical and physiological limitations in achieving adequate ocular bioavailability and targeted drug delivery of topically applied drugs and use of new techniques in formulating dosage forms in overcoming these limitations. The recent and future advances in nanocarrier-mediated drug delivery may have the potential to provide patient-friendly and noninvasive techniques for the treatment of diseases related to the anterior and posterior segments of the eye.

Local Delivery Strategies for Peptides and Proteins into the CNS: Status Quo, Challenges, and Future Perspectives.

Over the past decades, peptides and proteins have been increasingly important in the treatment of various human diseases and conditions owing to their specificity, potency, and minimized off-target toxicity. However, the existence of the practically impermeable blood brain barrier (BBB) limits the entry of macromolecular therapeutics into the central nervous systems (CNS). Consequently, clinical translation of peptide/protein therapeutics for the treatment of CNS diseases has been limited. Over the past decades, developing effective delivery strategies for peptides and proteins has gained extensive attention, in particular with localized delivery strategies, due to the fact that they are capable of circumventing the physiological barrier to directly introduce macromolecular therapeutics into the CNS to improve therapeutic effects and reduce systemic side effects. Here, we discuss various local administration and formulation strategies that have shown successes in the treatment of CNS diseases using peptide/protein therapeutics. Lastly, we discuss challenges and future perspectives of these approaches.

Sonocatalytic In Situ Induced Oxygen Storm Precision Enhanced Reactive Oxygen Therapy for Pancreatic Cancer

AbstractDeep‐buried tissue, extremely hypoxic blood supply, and complex tumor microenvironment are considered to be the hardest difficult barriers to the accurate treatment of pancreatic cancer. Herein, a novel Au@Co3O4 (AC)‐polyvinylpyrrolidone (AC‐PVP) sonocatalytic agent and radiation sensitizer with a multicore–shell structure are synthesized by a mild reduction method for reshaping the special physiological environment of pancreatic cancer. Oxygen storms can be generated under ultrasonic activation by in situ catalyzed H2O2 in tumor tissue. The oxygen storm improves the ability to produce superoxide anions by sonodynamic therapy as well as rapidly supplements the oxygen‐dependent on active oxygen such as auger electrons produced by high‐energy ray ionization during radiotherapy (RT), further enhancing the efficiency of RT. Interestingly, AC‐PVP can reduce the intracellular glutathione (GSH) level, thus preventing the generated reactive oxygen species from being consumed by GSH. More importantly, the glucose oxidase‐like of AC‐PVP can catalyze glucose in tumor tissue to produce H2O2, and continuously provide substrates for sonocatalytic to form self‐cycling oxygen production. This study offers a novel paradigm for achieving in situ precise self‐oxygenation to overcome the special physiological barrier of pancreatic cancer and efficient sonodynamic synergistic radiotherapy.

Dual Drug-Loaded Nanoliposomes Encapsulating Curcumin and 5-Fluorouracil with Advanced Medicinal Applications: Self-Monitoring and Antitumor Therapy.

Due to the presence of physiological barriers, it is difficult to achieve the desired therapeutic efficacy of drugs; thus, it is necessary to develop an efficient drug delivery system that enables advanced functions such as self-monitoring. Curcumin (CUR) is a naturally functional polyphenol whose effectiveness is limited by poor solubility and low bioavailability, and its natural fluorescent properties are often overlooked. Therefore, we aimed to improve the antitumor activity and drug uptake monitoring by simultaneously delivering CUR and 5-Fluorouracil (5-FU) in the form of liposomes. In this study, dual drug-loaded liposomes (FC-DP-Lip) encapsulating CUR and 5-FU were prepared by the thin-film hydration method; their physicochemical properties were characterized; and their biosafety, drug uptake distribution in vivo, and tumor cell toxicity were evaluated. The results showed that the nanoliposome FC-DP-Lip showed good morphology, stability, and drug encapsulation efficiency. It showed good biocompatibility, with no side effects on zebrafish embryonic development. In vivo uptake in zebrafish showed that FC-DP-Lip has a long circulation time and presents gastrointestinal accumulation. In addition, FC-DP-Lip was cytotoxic against a variety of cancer cells. This work showed that FC-DP-Lip nanoliposomes can enhance the toxicity of 5-FU to cancer cells, demonstrating safety and efficiency, and enabling real-time self-monitoring functions.

Vitamin A deficiency suppresses CEACAM1 to impair colonic epithelial barrier function via downregulating microbial-derived short-chain fatty acids

Vitamin A (VA) plays an essential role in modulating both the gut microbiota and gut barrier function. Short-chain fatty acids (SCFAs), as metabolites of the gut microbiota, protect the physiological intestinal barrier; however, they are compromised when VA is deficient. Thus, there is an urgent need to understand how and which SCFAs modulate colonic epithelial barrier integrity in VA deficiency (VAD). Herein, compared with normal VA rats (VAN), at the beginning of pregnancy, we confirmed that the colonic desmosome junction was impaired in the VAD group, and the amounts of acetate, propionate, and butyrate declined because of the decreased abundance of SCFA-producing bacteria (Romboutsia, Collinsella, and Allobaculum). The differentially expressed genes correlated with the gut barrier and the histone deacetylase complex between the VAD and VAN groups were enriched by RNA sequencing. In the VAD group, the expression levels of colonic CEA cell adhesion molecule 1 (CEACAM1) were down-regulated, and the levels of histone deacetylase 1 (HDAC1) and HDAC3 were up-regulated. Intriguingly, the above changes in the VAD groups were rescued by VA supplementation in the early postnatal period. Further study indicated that in Caco-2 cells, butyrate treatment significantly repressed the enrichment of HDAC3 on the promoter of the CEACAM1 gene to induce its expression. Our findings support that butyrate intervention can alleviate the impairment of colonic barrier function caused by VAD, and timely postnatal VA intervention may reverse the damage caused by VAD on gut barrier integrity during pregnancy.

Endoscopy and Antireflux Surgery: A Technical Review of Pre- and Postoperative Evaluation and Recognizing Patterns of Failure

Endoscopic evaluation after antireflux surgery is a challenge, due to the complexity of anatomy, variations in anatomical repairs, and the various patterns of failure. Studies have demonstrated that endoscopy reports of postoperative examinations are often inaccurate and inconsistent. The key to consistent high quality endoscopic examinations of the integrity of an antireflux surgery is a sound foundation in the native anatomical gastroesophageal junction and an understanding of the anatomy of the various postoperative configurations. This review will clarify the critical details necessary to perform a detailed endoscopic evaluation of the patient with suspected gastroesophageal reflux, and highlight key features to distinguish the intact repair from the dysfunctional one. It first explores the anatomical components of the native gastroesophageal junction and the unique geometric architecture that manifests in the physiologic reflux barrier. Then details the essential structures to evaluate and techniques to perform in an endoscopic examination prior to an antireflux surgery. It then systematically examines the altered anatomy and postoperative changes in endoscopic appearance after Nissen, Toupet, and Dor fundoplication, Collis gastroplasty, magnetic sphincter augmentation, and transoral incisionless fundoplication. Finally, to aid in the endoscopic diagnosis of specific dysfunction, it discusses the various patterns of failure after antireflux surgery, their characteristic endoscopic appearances, and the most useful adjunct testing modalities to augment the endoscopic examination when diagnosis is unclear.

Activation of nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 6 by Porphyromonas gingivalis regulates programmed cell death in epithelium

Background/purposeGingival epithelial cells form a physiological barrier against bacterial invasion. Programmed cell death (PCD) regulated by pathogen precognition receptors (PRRs) lead to tissue destruction and is closely related to inflammatory diseases. The purpose of this study was to investigate whether nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 6 (NLRP6) expresses in periodontal epithelium and induces PCD of epithelial cells infected by Porphyromonas gingivalis (P. gingivalis), therefore involves in periodontitis. Material and methodsThe expression of NLRP6 was detected in periodontal epithelium from human gingival sections and HaCaT cells stimulated by P. gingivalis. NLRP6 was over-expressed by adenovirus infection in HaCaT or knocked down by siRNA in P. gingivalis infected HaCaT, and the cell death was observed by transmission electron microscopy and flow cytometry analysis. In addition, qPCR and Western blot were performed to determine the expression of NLRP6 and the pyroptosis excutors, caspase-1 and gasdermin D. Enzyme-linked immunosorbent assay were performed to detect the secretion of IL-1β and IL-18. ResultsNLRP6 was up-regulated in both gingival epithelium of patients with periodontitis and P. gingivalis infected HaCaT. Over-expression of NLRP6 in HaCaT led to caspase-1 dependent pyroptosis. Interestingly, knockdown of NLRP6 with siRNA followed by P. gingivalis stimulation inhibited pyroptosis and induced apoptosis. ConclusionUp-regulation of NLRP6 by P. gingivalis in HaCaT led to pyroptosis, while knocking down NLRP6 inhibited pyroptosis and induced apoptosis, which indicated this PRR may play a crucial role in periodontitis by regulating PCD in periodontal epithelium.

ECOLOGICAL AND HYGIENIC ASSESSMENT OF QUALITY FLOWER OF TANACETUM VULGARE AGRO- AND URBANBIOCENOSES OF THE VORONEZH REGION

An important and little-researched aspect of the effect of human economic activity on medicinal plants is that in response to an increase in anthropo-genic load, additional synthesis of secondary metabolites, especially flavonoids, is induced, which play an important role in adapting plants to changing conditions. The purpose of the study was to study the accumulation of flavonoids and phenol carboxylic acids in the flowers of Tanacetum vulgare, col-lected in various agricultural and urbobiogeocenoses of the Voronezh region from the point of view of anthropogenic impact. As part of the study, 51 samples of Tanacetum vulgare flowers determined the content of heavy metals and arsenic and the sum of flavonoids and phenol carboxylic acids in terms of luteolin. By comparing the data on the content of heavy metals in the upper soil layers of the region and the content of these elements in the flowers of Tanacetum vulgare, it can be argued that there are significant physiological barriers that prevent the accumulation of ecotoxicants in the generative organs of the plant, which is especially noticeable for elements such as lead, mercury, arsenic, cadmium, cobalt and chromium. It turned out that ordinary pyjma is able to selectively concentrate copper and zinc. It was revealed that in the flowers of Tanacetum vulgare collected in agrobi-ogeocenoses and urbocenoses, the content of flavonoids and phenolcarboxylic acids is higher than in samples from protected areas, which is ex-plained by the fact that the key enzyme for the synthesis of flavonoids – phenylalaninammiacliase – has a pronounced stress inducibility. Under condi-tions of excessive toxic influence of pollutants, it is also possible to suppress the antioxidant system of plants.

Surface Area of Graphene Governs Its Neurotoxicity.

Due to their unique physicochemical properties, graphene and its derivatives are widely exploited for biomedical applications. It has been shown that graphene may exert different degrees of toxicity in in vivo or in vitro models when administered via different routes and penetrated through physiological barriers, subsequently being distributed within tissues or located within cells. In this study, in vitro neurotoxicity of graphene with different surface areas (150 and 750 m2/g) was examined on dopaminergic neuron model cells. SH-SY5Y cells were treated with graphene possessing two different surface areas (150 and 750 m2/g) in different concentrations between 400 and 3.125 μg/mL, and the cytotoxic and genotoxic effects were investigated. Both sizes of graphene have shown increased cell viability in decreasing concentrations. Cell damage increased with higher surface area. Lactate dehydrogenase (LDH) results have concluded that the viability loss of the cells is not through membrane damage. Neither of the two graphene types showed damage through lipid peroxidation (MDA) oxidative stress pathway. Glutathione (GSH) values increased within the first 24 and 48 h for both types of graphene. This increase suggests that graphene has an antioxidant effect on the SH-SY5Y model neurons. Comet analysis shows that graphene does not show genotoxicity on either surface area. Although there are many studies on graphene and its derivatives on their use with different cells in the literature, there are conflicting results in these studies, and most of the literature is focused on graphene oxide. Among these studies, no study examining the effect of graphene surface areas on the cell was found. Our study contributes to the literature in terms of examining the cytotoxic and genotoxic behavior of graphene with different surface areas.