Toxicity Screening Research Articles

CPA toxicity screening of cryoprotective solutions in rat hearts

In clinical practice, donor hearts are transported on ice prior to transplant and discarded if cold ischemia time exceeds ∼5 h. Methods to extend these preservation times are critically needed, and ideally, this storage time would extend indefinitely, enabling improved donor-to-patient matching, organ utilization, and immune tolerance induction protocols. Previously, we demonstrated successful vitrification and rewarming of whole rat hearts without ice formation by perfusion-loading a cryoprotective agent (CPA) solution prior to vitrification. However, these hearts did not recover any beating even in controls with CPA loading/unloading alone, which points to the chemical toxicity of the cryoprotective solution (VS55 in Euro-Collins carrier solution) as the likely culprit. To address this, we compared the toxicity of another established CPA cocktail (VEG) to VS55 using ex situ rat heart perfusion. The CPA exposure time was 150 min, and the normothermic assessment time was 60 min. Using Celsior as the carrier, we observed partial recovery of function (atria-only beating) for both VS55 and VEG. Upon further analysis, we found that the VEG CPA cocktail resulted in 50 % lower LDH release than VS55 (N = 4, p = 0.017), suggesting VEG has lower toxicity than VS55. Celsior was a better carrier solution than alternatives such as UW, as CPA + Celsior-treated hearts spent less time in cardiac arrest (N = 4, p = 0.029). While we showed substantial improvement in cardiac function after exposure to vitrifiable concentrations of CPA by improving both the CPA and carrier solution formulation, further improvements will be required before we achieve healthy cryopreserved organs for transplant.

Imperative Role of Natural Product Chemistry in Cosmeceutical R&D - Phytonanocosmeceuticals

Natural product chemistry has always been attractive for drug and cosmetic industries as natural products can address these industries very well. By consumer demand, the cosmetic industry is looking for innovative, safer, more effective, and environmentally friendly products. In this sense, a relatively new concept of cosmetics has emerged under “cosmeceuticals or dermocosmetics/medcosmetics”. Cosmeceuticals are briefly defined as a subclass of cosmetics that contain drug active substances or bioactive natural products with enhanced efficacy for therapeutic or cosmetic purposes. They are also described as a combination of cosmetics and pharmaceutics or medical-grade cosmetics, which particularly enhance skin penetration and the restorative effect of active ingredients in cosmetic formulations. On the other hand, nanotechnology has become another exciting area in cosmetics as nanoformulations enhance skin penetration. Therefore, we have been working on research and development of novel phyto-based cosmeceuticals via extensive screening studies on plant extracts and pure natural substances using in vitro (enzyme inhibition, etc), in silico (molecular docking and toxicity screening), and cell-based assays. In this regard, an anti-acne formulation based on a number of plant extracts tested against Propionibacterium acnes has been developed by our group. Besides, an antimicrobial formulation as an oral spray for mouth defense is currently a commercial product. We have been studying nanofiber formulations loaded with plant extract for wound healing. All our ongoing studies on discovering novel active natural ingredients for cosmeceutical purposes have so far yielded three patents and four patent applications and commercialized a final product. In the present mini review, examples and details of our group’s recent findings on phyto-based cosmeceuticals and nanoformulations will be underlined.

PROFIL FITOKIMIA DAN ANALISIS TOKSISITAS Ficus racemosa L. MENGGUNAKAN METODE BRINE SHRIMP (Artemia salina) LETHALITY TEST

A traditional plant, Ficus racemosa L., known as the ara plant in West Nusa Tenggara, is potentially beneficial as a drug candidate, which is one option to develop the biodiversity of natural ingredients in Indonesia. The extract was produced by maceration method with ethanol 96% solvent. Furthermore, they partitioned using various solvents, n-hexane, ethyl acetate, and chloroform. The presence or absence of secondary metabolite is determined by the phytochemical screening test. Meanwhile, the Brine Shrimp Lethality Test (BSLT) is a toxicity screening to determine the LC50 of extract and their partition with various concentrations used were 50, 100, 250, 500, and 1000 ppm. The metabolite secondary flavonoids, alkaloids, and saponins are confirmed. The analysis showed that LC50 of extract was 218,34 ppm; n-hexane fraction was 413,57 ppm; ethyl acetate fraction was 309.51 ppm; and chloroform fraction was 314.60 ppm; all had toxic potential.

3D hepatic organoid production from human pluripotent stem cells

Hepatic organoids might provide a golden opportunity for realizing precision medicine in various hepatic diseases. Previously described hepatic organoid protocols from pluripotent stem cells rely on complicated multiple differentiation steps consisting of both 2D and 3D differentiation procedures. Therefore, the spontaneous formation of hepatic organoids from 2D monolayer culture is associated with a low-throughput production, which might hinder the standardization of hepatic organoid production and hamper the translation of this technology to the clinical or industrial setting. Here we describe the stepwise and fully 3D production of hepatic organoids from human pluripotent stem cells. We optimized every differentiation step by screening for optimal concentrations and timing of differentiation signals in each differentiation step. Hepatic organoids are stably expandable without losing their hepatic functionality. Moreover, upon treatment of drugs with known hepatotoxicity, we found hepatic organoids are more sensitive to drug-induced hepatotoxicity compared with 2D hepatocytes differentiated from PSCs, making them highly suitable for in vitro toxicity screening of drug candidates. The standardized fully 3D protocol described in the current study for producing functional hepatic organoids might serve as a novel platform for the industrial and clinical translation of hepatic organoid technology.

A Behavioural and Histological Approach to Dose Dependent Chronic Toxicity Screening of Cyclotide in Zebrafishes

A Behavioural and Histological Approach to Dose Dependent Chronic Toxicity Screening of Cyclotide in Zebrafishes

Simultaneous screening of membrane permeability and toxicity for discovery of new cryoprotectants

Simultaneous screening of membrane permeability and toxicity for discovery of new cryoprotectants

Toxicity Screening of Surface Waters in Zaghen Restoration Area with Toxkit Microbiotests

Abstract In this study, we report the findings regarding the toxicity of surface waters collected in March 2023 from five sampling stations in the Zaghen restoration area, located within the Danube Delta Biosphere Reserve, Romania. The assessment of acute toxicity was conducted using two microbiotests: the Thamnotoxkit F and the Daphtoxkit FTM magna. Even if the acute toxicity effects are less than 50% and do not allow the calculation of a toxicity score for the water samples, the experiments show a different vulnerability of the two zooplanktonic species to the presence of heavy metals in the water with mortality being higher in Thamnocephalus platyurus (20%) than in Daphnia magna (15%).

New melatonin derivatives with donepezil fragment in Alzheimer’s disease as neuroprotectors

AbstractBackgroundAlzheimer’s disease is characterized by progressive, irreversible degeneration of acetylcholine‐producing neurons, cognitive impairment, and the accumulation of neurofibrillary tangles and amyloid plaques. The natural decline in melatonin levels with aging has been associated with insomnia and depression as well as the development of neurodegenerative disorders such as AD. Furthermore, the complexity of AD pathology has encouraged the design of molecules that are able to interact with two or more complementary targets, with the expectation that these multitarget ligands (MTL) may represent important advantages in the treatment of the disease. This study aimed to find new ChE inhibitors with improved toxicity.Method In vitro toxicity screening of the newly compounds on human neuroblastoma SH‐SY5Y, mouse neuroblastoma cell line Neuro‐2a and mouse fibroblast cells CCL‐1. The viability of the cells was assessed by MTT assay. The neuroprotective properties of the compounds were determined in a model of H2O2‐induced oxidative damage in SH‐SY5Y cells. For comparison, effects donepezil, melatonin and galanthamine were determined.ResultMost of the compounds have IC50 values over 100 µM. Compound 3f possess statistically significant protective effects. It should be noted that the neuroprotective effects were more pronounced than those of donepezil and similar to melatonin and rasagiline.ConclusionThe new series of melatonin derivatives with donepezil fragment exhibit low toxicity and a good in vitro safety profile. Further, in vitro protection study proved efficient prevention against oxidative stress in H2O2‐induced injury in SH‐SY5Ycells, especially by compound 3f and was proven as the most perspective for further investigations.Acknowledgments: The financial support by the National Science Fund of Bulgaria (Grant KP‐06‐Н63/11 14.12.2022) is gratefully acknowledged.

A CPA toxicity screening of cryoprotective solutions in rat hearts

A CPA toxicity screening of cryoprotective solutions in rat hearts

Toxicological Assessment of Higher Olefins in OECD TG 422 Repeated Dose and Reproductive /Developmental Toxicity Screening Tests in Han Wistar Rats.

Higher olefins (HO) are used primarily as intermediates in the production of other chemicals, such as polymers, fatty acids, plasticizer alcohols, surfactants, lubricants, amine oxides, and detergent alcohols. The potential toxicity of five HO (i.e., 1-Octene, Nonene, Decene, Hexadecene, and 1-Octadecene) with carbon ranging from C8 to C18 was examined in a combined repeated dose and reproduction/developmental toxicity screening study (OECD TG 422). These five HO were administered to Han Wistar rats by gavage at 0 (controls), 100, 300, and 1000mg/kg bw/day. As a group of substances, adaptive changes in the liver (liver weight increase without pathological evidence), as well as increased kidney weight in male rats, were observed in HO with carbon numbers from C8 to C10. The overall systemic no observed adverse effect level (NOAEL) for all HO was determined at 1000mg/kg bw/day. In the reproductive/developmental toxicity assessment, offspring viability, size, and weights were reduced in litters from females treated with Nonene at 1000mg/kg bw/day. The overall no observed effects level (NOEL) for reproductive toxicity was considered to be 300mg/kg bw/day for Nonene and 1000mg/kg bw/day for the other four HO, respectively. These data significantly enrich the database on the toxicity of linear and branched HO, allowing comparison with similar data published on a range of linear and branched HO. Comparisons between structural class and study outcome provide further supportive data in order to validate the read-across hypothesis as part of an overall holistic testing strategy.

National service is free, not cancer disease: Financial toxicity in a public universal coverage healthcare system.

32 Background: Italian National Health Service provide regional control on clinical pathways to be cost-free. ASL Napoli 1 centralized model of Palliative Care Unit ensure free multidisciplinary medical and nursing assistance, beyond aids and devices. Since healthcare costs are rising and private spending is becoming necessary, our aim is screening financial toxicity in the most densely populated area in Europe. Methods: From January to April 2023 we performed simple random sampling to select 27 home care patients. From the richest neighborhoods to poorest ones, patients answered the COST-FACIT questionnaire. We collected information about medical history, sociodemographic characteristics (employment status or retired, family composition, if benefiting from state allowances) and, above all, out-of-pocket costs. Results: Data highlight that wealthiest and middle-class patients are afraid about out of control medical expenses and their satisfaction about economic status is declining. Patients from a lower socioeconomic background are deeply concerned about not being able to work and not being of financial support to often single income families. Score analysis reveals significant variation among the different items, and the average score is 12.4. Highest scores count 28 and 29, respectively, suggesting that these patients are more able to manage the costs of treatment or access financial resources to cope with medical expenses. Three scores of 0 suggest higher financial toxicity in the realm of palliative oncology care. These patients face greater difficulty in handling treatment costs, encounter barriers in accessing financial resources, or experience high expenses. Patients have highlighted that purchase of non-reimbursable drugs is the most onerous out-of-pocket cost in home care setting. Conclusions: Our centralized Palliative Care Unit organizational model coordinates by adapting purchasing and providing holistic sanitary approach beyond patients' economic abilities. Our centralized Palliative Care Unit provides health services to endure equity across all the city. However, score analysis suggests a range financial suffering for home care patients. Low average score 12.4 suggests how much financial suffering in palliative care impacts, even in a model of centralization of health care with global coverage. So, screening for financial toxicity plays a key role in directing health policies on pharmaceutical expenditure.

Implementation of financial toxicity screening and a novel financial navigation program.

290 Background: “Financial Toxicity” is the financial burden and stress imposed on a patient due to a disease and its associated treatment. About 50% of patients with gynecologic cancer experience financial toxicity. The aim of the study is to evaluate implementation of financial toxicity screening and a novel financial navigation program for patients with gynecologic oncology. Methods: Patients presenting for initial consultation with a gynecologic oncology provider from July 2022 to January 2023 were included. A new financial navigation program began in July 2022. On October 17, 2022, a financial toxicity screening question was added to new patient intake forms. We quantified patient referrals to the navigator before and after screening implementation, along with needs identified, action taken by the navigator and institutional support services utilized. Results: We included 458 new patients, 49% before and 51% after introduction of screening. The majority were white (61%) and had private insurance (59%) followed by Medicare (31%); median age was 59 (IQR 47-69). Following screening, a higher proportion of patients were referred to the financial navigator (15%) than before screening (4%, p<0.001). Additionally, referrals to other hospital social support services decreased from 9% before screening to 3% after (p<0.001). Uterine and cervical cancers were more prevalent in the 29 patients (12%) who screened positive (31% and 14% respectively) compared to those who screened negative (17% and 1%) or did not respond (24% and 4%). Referrals to the navigator included 59% of those who screened positive, 8% who screened negative and 10% who did not respond. Over six months, 62 patients (new and follow-up) were referred to the navigator. The most common need was transportation (21%) followed by financial assistance (16%) and emotional support (16%). The navigator assisted 11% of patients with transportation and referred 13% to financial counseling/billing and 27% to other resources. Conclusions: Implementing a financial toxicity screening question on intake forms was feasible, and increased identification of patients with financial toxicity and referrals to our financial navigator. Further investigation is needed to understand screening barriers and evaluate the program.

Critical assessment of the endocrine potential of Linalool and Linalyl acetate: proactive testing strategy assessing estrogenic and androgenic activity of Lavender oil main components.

The acyclic linear monoterpenes Linalool (Lin) and Linalyl acetate (LinAc) occur in nature as major constituents of various essential oils such as lavender oils. A potential endocrine activity of these compounds was discussed in literature including premature thelarche and prepubertal gynecomastia due to lavender product use. This study aims to follow-up on these critical findings reported by testing Lin and LinAc in several studies in line with current guidance and regulatory framework. No relevant anti-/ER and AR-mediated activity was observed in recombinant yeast cell-based screening tests and guideline reporter gene in vitro assays in mammalian cells. Findings in the screening test suggested an anti-androgenic activity, which could not be confirmed in the respective mammalian cell guideline assay. Mechanistic guideline in vivo studies (Uterotrophic and Hershberger assays) with Lin did not show significant dose related changes in estrogen or androgen sensitive organ weights and a guideline reproductive toxicity screening study did not reveal evident effects on sex steroid hormone sensitive organ weights, associated histopathological findings and altered sperm parameters. Estrous cycling and mating/fertility indices were not affected and no evident Lin-related steroid hormone dependent effects were found in the offspring. Overall, the initial concerns from literature were not confirmed. Findings in the yeast screening test were aberrant from follow-up guideline in vitro and in vivo studies, which underlines the need to apply careful interpretation of single in vitro test results to support a respective line of evidence and to establish a biologically plausible link to an adverse outcome.

Digital symptom assessment tools: the next frontier in financial toxicity screening.

Digital symptom assessment tools: the next frontier in financial toxicity screening.

Generation of Fibrotic Liver Organoids Using Hepatocytes, Primary Liver Sinusoidal Endothelial Cells, Hepatic Stellate Cells, and Macrophages.

Liver organoids generated with single or multiple cell types have been used to investigate liver fibrosis development, toxicity, pathogenesis, and drug screening. However, organoid generation is limited by the availability of cells isolated from primary tissues or differentiated from various stem cells. To ensure cell availability for organoid formation, we investigated whether liver organoids could be generated with cell-line-based Huh-7 hepatocellular carcinoma cells, macrophages differentiated from THP-1 monocytes, and LX-2 hepatic stellate cells (HSCs) and primary liver sinusoidal endothelial cells (LSECs). In liver organoids, hepatocyte-, LSEC-, macrophage-, and HSC-related gene expression increased relative to that in two-dimensional (2D)-cultured Huh-7/LSEC/THP-1/LX-2 cells without Matrigel. Thioacetamide (TAA) increased α-smooth muscle actin expression in liver organoids but not in 2D-cultured cells, whereas in TAA-treated organoids, the expression of hepatic and LSEC markers decreased and that of macrophage and HSC markers increased. TAA-induced fibrosis was suppressed by treatment with N-acetyl-L-cysteine or tumor-necrosis-factor-stimulated gene 6 protein. The results showed that liver toxicants could induce fibrotic and inflammatory responses in liver organoids comprising Huh-7/LSEC/macrophages/LX-2 cells, resulting in fibrotic liver organoids. We propose that cell-line-based organoids can be used for disease modeling and drug screening to improve liver fibrosis treatment.

Biosorption of methylene blue in aqueous solution using structurally modified rice husk and its notable compatibility, biosafety potential – A sustainable approach towards the management of hazardous dyes

In vitro batch system was adopted to determine the dye removal and uptake efficacy by providing various optimum condition which includes initial dye concentration, adsorbent dosage, contact time, pH, temperature. Surface modified rice husk was screened for biocompatibility or biosafety potential using phytotoxicity assessment, brine shrimp toxicity, hemocompatibility and mutagenicity screening. Optimum condition for the dye adsorption includes 1750 mg of adsorbent dosage, 25 mg/L of initial dye concentration, 100 min of contact time, 45 oC temperature, 7.5 pH Adsorbed dye in aqueous solution thus obtained under optimum condition was not shown any phytotoxic effect on the seedling index of Vigna mungo, Vigna radiata and Brassica niger, enzymatic anti-oxidants level, biochemical composition like total protein, total carbohydrate, total chlorophyll, qualitative protein expression pattern. No impact on the hatching, survival rate, morphology, anti-oxidants profile of brine shrimp -Artemia salina that exposed to surface modified rice husk treated dye in aqueous solution revealed best biocompatibility. No sign of hemolysis on the peripheral blood that treated with adsorbed dye in aqueous solution also confirmed the biocompatibility. No induction of cytotoxicity, and apoptosis in the RAW cell line that exposed to treated dye in aqueous solution was also confirmed the best biocompatibility. Mutagenicity screening with bacterial strain Salmonella typhimurium His- bacterial detection system adopting Ames test showed absence of revertant bacterial colonies indicated the best biosafety or biocompatibility. The present study suggests the possible utilization of rice husk treated methylene blue in aqueous solution for agriculture or irrigation purpose with high biocompatibility and biosafety.

Toxicity evaluation of zinc oxide nanoparticles green synthesized using papaya extract in zebrafish.

In green synthesis of zinc oxide nanoparticles (ZnO NPs), the use of papaya extract as a capping and reducing agent shows promise for potential applications of these particles in biomedicine. However, toxicity evaluation is necessary to ensure the safety of humans and the environment. The zebrafish model is used to assess toxicity with embryo developmental observation as it is a rapid, simple method for screening of toxicity. The objective of the present study was to assess the toxicological characteristics of ZnO NPs produced from papaya extract using a zebrafish model. The preparation of plant extracts from papaya using two solvents (water and methanol) and characterization of bioactive compounds in the extracts were reported. ZnO NPs were synthesized from both plant extracts and characterized with scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy. Toxicity evaluation was conducted on zebrafish embryos for 96 h. ZnO NPs synthesized from aqueous and methanol extracts had mean crystallite diameters of 13 and 12 nm, respectively. Mortality, hatching rate and malformation of zebrafish embryos were assessed at different concentrations of ZnO NPs. Both NPs showed high mortality rates at high concentrations, with 100 (aqueous) and 20 mg/l (methanol extract) being lethal for all embryos. Concentrations <10 mg/l for both synthesized ZnO NPs had similar results to the negative control, indicating a safe dosage for embryos. The hatching rate and malformation were also affected, with higher concentrations of NPs causing a delayed hatching rate and malformation in pericardial and yolk sac edema. Whole embryo mRNA expression of immune-associated genes, including IL-1 and -10 and TNF-α, was upregulated following lethal concentration 50 (LC50) ZnO NP exposure. ZnO NPs synthesized from papaya extract (both in aqueous and methanol environments) had a dose- and time-dependent embryonic toxicity effect. Hence, the present study demonstrated initial toxicity screening of ZnO NPs synthesized from plant extract.

Evaluation of Alectinib Metabolic Stability in HLMs Using Fast LC-MS/MS Method: In Silico ADME Profile, P450 Metabolic Lability, and Toxic Alerts Screening.

Alectinib, also known as Alecensa®, is prescribed for the therapeutic treatment of individuals diagnosed with metastatic non-small cell lung cancer (NSCLC) who have a specific genetic mutation referred to as anaplastic lymphoma kinase (ALK) positivity. The Food and Drug Administration granted regular approval to alectinib, a drug developed by Hoffmann-La Roche, Inc. (Basel, Switzerland)/Genentech, Inc. (South San Francisco, CA, USA), on 6 November 2017. The screening of the metabolic stability and identification of hazardous alarms within the chemical structure of ALC was conducted using the StarDrop software package (version 6.6), which incorporates the P450 metabolic module and DEREK software (KB 2018 1.1). The primary aim of this investigation was to develop a high-throughput and accurate LC-MS/MS technique for the quantification of ALC in the metabolic matrix (human liver microsomes; HLMs). The aforementioned methodology was subsequently employed to assess the metabolic stability of ALC in HLMs through in vitro tests, with the obtained results further validated using in silico software. The calibration curve of the ALC showed a linear correlation that exists within the concentration range from 1 to 3000 ng/mL. The LC-MS/MS approach that was recommended exhibited accuracy and precision levels for both inter-day and intra-day measurements. Specifically, the accuracy values ranged from -2.56% to 3.45%, while the precision values ranged from -3.78% to 4.33%. The sensitivity of the established approach was proved by its ability to adhere to an LLOQ of 0.82 ng/mL. The half-life (t1/2) and intrinsic clearance (Clint) of ALC were estimated to be 22.28 min and 36.37 mL/min/kg, correspondingly, using in vitro experiments. The ALC exhibited a moderate extraction ratio. The metabolic stability and safety properties of newly created derivatives can be enhanced by making modest adjustments to the morpholine and piperidine rings or by substituting the substituent, as per computational software. In in silico ADME prediction, ALC was shown to have poor water solubility and high gastrointestinal absorption along with inhibition of some cytochrome P450s (CYP2C19 and CYP2C9) without inhibition of others (CYP1A2, CYP3A4, and CYP2D6) and P-glycoprotein substrate. The study design that involves using both laboratory experiments and different in silico software demonstrates a novel and groundbreaking approach in the establishment and uniformization of LC-MS/MS techniques for the estimation of ALC concentrations, identifying structural alerts and the assessment of its metabolic stability. The utilization of this study strategy has the potential to be employed in the screening and optimization of prospective compounds during the drug creation process. This strategy may also facilitate the development of novel derivatives of the medicine that maintain the same biological action by targeted structural modifications, based on an understanding of the structural alerts included within the chemical structure of ALC.

Enhancing cardiomyocytes contraction force measuring in drug testing: Integration of a highly sensitive single-crystal silicon strain sensor into SU-8 cantilevers

The development of efficient tools for predicting drug-induced cardiotoxicity in the preclinical phase would greatly benefit the drug development process. This study presents an SU-8 cantilever integrated with a single-crystal silicon strain sensor to enhance force sensitivity in toxicity screening methods based on changes in the contraction force of cardiomyocytes. The proposed cantilever device enables real-time measurements of cardiomyocytes contraction force with high sensitivity, thereby facilitating the assessment of drug cardiotoxicity. The experimental results obtained herein demonstrate the responsiveness of the proposed platform in detecting forces smaller than 0.02 μN with a force sensitivity that is nearly 17 times higher than those of conventional metal-based strain sensors. Moreover, the integration of strain sensors demonstrates the potential for manufacturing cantilever arrays that can be used in high-throughput screening applications. The developed methodology successfully facilitates in vitro culturing of cardiomyocytes and allows for continuous monitoring of their contraction force. The practical applicability of the proposed platform is further validated through cardiotoxicity analysis. The cultured cardiomyocytes are treated with two cardiovascular drugs, namely verapamil (an L-type calcium channel blocker) and isoproterenol (a sympathomimetic drug targeting β1 and β2 adrenergic receptors), to analyze the drug induced effects in the cardiomyocytes.

Impact of the Physical Cellular Microenvironment on the Structure and Function of a Model Hepatocyte Cell Line for Drug Toxicity Applications.

It is widely recognised that cells respond to their microenvironment, which has implications for cell culture practices. Growth cues provided by 2D cell culture substrates are far removed from native 3D tissue structure in vivo. Geometry is one of many factors that differs between in vitro culture and in vivo cellular environments. Cultured cells are far removed from their native counterparts and lose some of their predictive capability and reliability. In this study, we examine the cellular processes that occur when a cell is cultured on 2D or 3D surfaces for a short period of 8 days prior to its use in functional assays, which we term: "priming". We follow the process of mechanotransduction from cytoskeletal alterations, to changes to nuclear structure, leading to alterations in gene expression, protein expression and improved functional capabilities. In this study, we utilise HepG2 cells as a hepatocyte model cell line, due to their robustness for drug toxicity screening. Here, we demonstrate enhanced functionality and improved drug toxicity profiles that better reflect the in vivo clinical response. However, findings more broadly reflect in vitro cell culture practises across many areas of cell biology, demonstrating the fundamental impact of mechanotransduction in bioengineering and cell biology.