Real-time monitoring of cortical brain activity in response to acute pain using wide-area Ca2+ imaging.

Previous human and rodent studies indicated that nociceptive stimuli activate many brain regions that is involved in the somatosensory and emotional sensation. Although these studies have identified several important brain regions involved in pain perception, it has been a challenge to observe neural activity directly and simultaneously in these multiple brain regions during pain perception. Using a transgenic mouse expressing G-CaMP7 in majority of astrocytes and a subpopulation of excitatory neurons, we recorded the brain activity in the mouse cerebral cortex during acute pain stimulation. Both of hind paw pinch and intraplantar administration of formalin caused strong transient increase of the fluorescence in several cortical regions, including primary somatosensory, motor and retrosplenial cortex. This increase of the fluorescence intensity was attenuated by the pretreatment with morphine. The present study provides important insight into the cortico-cortical network during pain perception.

Investigating the Influence of Morphine and Cocaine on the Mesolimbic Pathway Using a Novel Microimaging Platform.

Dopamine (DA)'s relationship with addiction is complex, and the related pathways in the mesocorticolimbic system are used to deliver DA, regulating both behavioral and perceptual actions. Specifically, the mesolimbic pathway connecting the ventral tegmental area (VTA) and the nucleus accumbens (NAc) is crucial in regulating memory, emotion, motivation, and behavior due to its responsibility to modulate dopamine. To better investigate the relationship between DA and addiction, more advanced mapping methods are necessary to monitor its production and propagation accurately and efficiently. In this study, we incorporate dLight1.2 adeno-associated virus (AAV) into our latest CMOS (complementary metal-oxide semiconductor) imaging platform to investigate the effects of two pharmacological substances, morphine and cocaine, in the NAc using adult mice. By implanting our self-fabricated CMOS imaging device into the deep brain, fluorescence imaging of the NAc using the dLight1.2 AAV allows for the visualization of DA molecules delivered from the VTA in real time. Our results suggest that changes in extracellular DA can be observed with this adapted system, showing potential for new applications and methods for approaching addiction studies. Additionally, we can identify the unique characteristic trend of DA release for both morphine and cocaine, further validating the underlying biochemical mechanisms used to modulate dopaminergic activation.

Multi-Region Microdialysis Imaging Platform Revealed Dorsal Raphe Nucleus Calcium Signaling and Serotonin Dynamics during Nociceptive Pain.

In this research, we combined our ultralight micro-imaging device for calcium imaging with microdialysis to simultaneously visualize neural activity in the dorsal raphe nucleus (DRN) and measure serotonin release in the central nucleus of the amygdala (CeA) and the anterior cingulate cortex (ACC). Using this platform, we observed brain activity following nociception induced by formalin injection in the mouse's hind paw. Our device showed that DRN fluorescence intensity increased after formalin injection, and the increase was highly correlated with the elevation in serotonin release in both the CeA and ACC. The increase in calcium fluorescence intensity occurred during the acute and inflammatory phases, which suggests the biphasic response of nociceptive pain. Furthermore, we found that the increase in fluorescence intensity was positively correlated with mouse licking behavior. Lastly, we compared the laterality of pain stimulation and found that DRN fluorescence activity was higher for contralateral stimulation. Microdialysis showed that CeA serotonin concentration increased only after contralateral stimulation, while ACC serotonin release responded bilaterally. In conclusion, our study not only revealed the inter-regional serotonergic connection among the DRN, the CeA, and the ACC, but also demonstrated that our device is feasible for multi-site implantation in conjunction with a microdialysis system, allowing the simultaneous multi-modal observation of different regions in the brain.

The Role of Neurons in Human Health and Disease.

Neurons are the functional units of the nervous system [...].

Prenatal nicotine exposure alters gene expression profiles of neurons in the sub-regions of the VTA during early postnatal development.

Brain growth occurs during the first 2 weeks of postnatal development in rats. This developmental period is equivalent to the third trimester of human gestation. Dendritic arborization, axonal growth, and gliogenesis are observed along with a strong maturation of neurotransmission during this critical development period. Furthermore, nicotine exposure during early development causes deficiencies in sensory and cognitive processing in adults. In this study, we further investigated the gene expression of neuron groups and the influence of perinatal nicotine exposure on gene expressions of neurons within the sub-regions of the ventral tegmental area (VTA) in 1 week, 2 week and 3-week-old rat pups. We exposed pregnant rats to nicotine perinatally on gestational day 7 through postnatal day 14. Pups are exposed to nicotine during pregnancy and through breastfeeding to investigate its effect in rat pups during early neuronal development. Real time PCR was used to find the relative expressions of gamma-aminobutyric acid (GABA), dopamine, and glutamate neuron markers within the three sub-regions of the VTA including the parabrachial pigmented nucleus (PBP), parainterfascicular (PIF), and paranigral nucleus (PN). Our results indicated that during early maturation, the dopamine marker tyrosine hydroxylase (TH) showed a consistently increased significance in PN sub-region compared to PIF and PBP. These results suggest that following perinatal nicotine exposure, VTA dopamine neurons, especially within the PN sub-region, are significantly excited starting from birth.

Investigating the Modulation of the VTA Neurons in Nicotine-Exposed Pups during Early Maturation Using Optogenetics.

Advancing the understanding of the relationship between perinatal nicotine addiction and the reward mechanism of the brain is crucial for uncovering and implementing new treatments for addiction control and prevention. The mesolimbic pathway of the brain, also known as the reward pathway, consists of two main areas that regulate dopamine (DA) and addiction-related behaviors. The ventral tegmental area (VTA) releases DA when stimulated, causing the propagation of neuronal firing along the pathway. This ends in the release of DA into the extracellular space of the nucleus accumbens (NAc), which is directly modulated by the uptake of DA. Much research has been conducted on the effects of nicotine addiction, but little research has been conducted concerning nicotine addiction and the mesolimbic pathway regarding maturation due to the small brain size. In this study, we apply our novel microstimulation experimental system to rat pups that have been perinatally exposed to nicotine. By using our self-fabricated photo-stimulation (PS) device, we can stimulate the VTA and collect dialysate, which is then used to estimate DA released into the NAc. The proposed platform has demonstrated the potential to monitor neural pathways as the pups mature.

Processes for Designing Innovative Biomedical Hardware to Use in Space and on Earth.

The new era of space exploration is increasing the astronaut's number and diversity in low orbit and beyond. The influx of such a diverse crew population will also increase the need for medical technologies to ensure safe and productive missions. Such a need represents a unique opportunity to innovate and develop diagnostics and treatment tools to meet future needs. Historically, terrestrial regulatory oversight of biomedical design processes was considered separate from spaceflight regulatory processes because it did not address spaceflight constraints. These constraints challenge the creative development of unique solutions for use in space. Translation between healthcare innovation in spaceflight to healthcare on Earth and vice versa requires understanding the commonalities, unique needs and constraints. This manuscript provides a framework for comparing Earth-space design processes and a perspective on the best practices to improve healthcare equity and health outcomes.

Targeted Sensitization of Glioblastoma Multiforme Using AAAPT Technology.

Glioblastoma Multiforme (GBM) is the most malignant type of all brain tumors. Current GBM treatment options include surgery, followed by radiation and chemotherapy. However, GBM can become resistant to therapy, resulting in tumor recurrence. GBM cells develop resistance to treatments by either downregulating cell death pathways (CD95) or upregulating cell survival pathways (NF-κB (p65)). Healthy tissues can be affected by the increased therapeutic dose. Therefore, it is important to develop a method that can only target GBM tumor cells, thereby reducing the non-specific uptake which will reduce the side effects. Here we demonstrate an application of novel priori activation of apoptosis pathways of tumor technology (AAAPT), which has been used to demonstrate the effect of targeted tumor sensitizers to make chemotherapy work at lower doses in breast, lung and prostate cancers. Treatment of GBM spheroids with AAAPT in 3D PEGDA microwells, showed an increase in cell death, an upregulation of cell death pathways, and a downregulation of cell survival pathways, in comparison to Temozolomide (TMZ), an oral alkylating agent, which is a commonly used chemotherapy in the treatment of GBM. The dose of AAAPT sensitizers may provide a promising method to increase treatment efficacy and reduce off-target toxicity, as an alternative to existing methods which cause significant off-target damage.

Automatic Upper-Limb Brunnstrom Recovery Stage Evaluation via Daily Activity Monitoring.

Motor function assessment is crucial for post-stroke rehabilitation. Conventional evaluation methods are subjective, heavily depending on the experience of therapists. In light of the strong correlation between the stroke severity level and the performance of activities of daily living (ADLs), we explored the possibility of automatically evaluating the upper-limb Brunnstrom Recovery Stage (BRS) via three typical ADLs (tooth brushing, face washing and drinking). Multimodal data (acceleration, angular velocity, surface electromyography) were synchronously collected from 5 upper-limb-worn sensor modules. The performance of BRS evaluation system is known to be variable with different system parameters (e.g., number of sensor modules, feature types and classifiers). We systematically searched for the optimal parameters from different data segmentation strategies (five window lengths and four overlaps), 42 types of features, 12 feature optimization techniques and 9 classifiers with the leave-one-subject-out cross-validation. To achieve reliable and low-cost monitoring, we further explored whether it was possible to obtain a satisfactory result using a relatively small number of sensor modules. As a result, the proposed approach can correctly recognize the stages of all 27 participants using only three sensor modules with the optimized data segmentation parameters (window length: 7s, overlap: 50%), extracted features (simple square integral, slope sign change, modified mean absolute value 1 and modified mean absolute value 2), the feature optimization method (principal component analysis) and the logistic regression classifier. According to the literature, this is the first study to comprehensively optimize sensor configuration and parameters in each stage of the BRS classification framework. The proposed approach can serve as a factor-screening tool towards the automatic BRS classification and is promising to be further used at home.

Investigating miRNA-mRNA Interactions and Gene Regulatory Networks From VTA Dopaminergic Neurons Following Perinatal Nicotine and Alcohol Exposure Using Bayesian Network Analysis.

MicroRNAs play an important role in gene regulation for many biological systems, including nicotine and alcohol addiction. However, the underlying mechanism behind miRNAs and mRNA interaction is not well characterized. Microarrays are commonly used to quantify the expression levels of mRNAs and/or miRNAs simultaneously. In this study, we performed a Bayesian network analysis to identify mRNA and miRNA interactions following perinatal exposure to nicotine and/or alcohol. We utilized three sets of microarray data to predict the regulation relationship between mRNA and miRNAs. Following perinatal alcohol exposure, we identified two miRNAs: miR-542-5p and miR-874-3p, that exhibited a strong mutual influence on several mRNA in gene regulatory pathways, mainly Axon guidance and Dopaminergic synapses. Finally, we confirmed our predicted addiction pathways based on the Bayesian network analysis with the widely used Kyoto Encyclopedia of Genes and Genomes (KEGG)-based database and identified comparable relevant miRNA-mRNA pairs. We believe the Bayesian network can provide insight into the complexity biological process related to addiction and can potentially be applied to other diseases.

Investigating the Influence of GABA Neurons on Dopamine Neurons in the Ventral Tegmental Area Using Optogenetic Techniques.

Dopamine (DA) is the key regulator of reward behavior. The DA neurons in the ventral tegmental area (VTA) and their projection areas, which include the prefrontal cortex (PFC), nucleus accumbens (NAc), and amygdala, play a primary role in the process of reward-driven behavior induced by the drugs of addiction, including nicotine and alcohol. In our previous study, we developed a novel platform consisting of micro-LED array devices to stimulate a large area of the brain of rats and monkeys with photo-stimulation and a microdialysis probe to estimate the DA release in the PFC. Our results suggested that the platform was able to detect the increased level of dopamine in the PFC in response to the photo-stimulation of both the PFC and VTA. In this study, we used this platform to photo-stimulate the VTA neurons in both ChrimsonR-expressing (non-specific) wild and dopamine transporter (DAT)-Cre (dopamine specific) mice, and measured the dopamine release in the nucleus accumbens shell (NAcShell). We measured the DA release in the NAcShell in response to optogenetic stimulation of the VTA neurons and investigated the effect of GABAergic neurons on dopaminergic neurons by histochemical studies. Comparing the photo-stimulation frequency of 2 Hz with that of 20 Hz, the change in DA concentration at the NAcShell was greater at 20 Hz in both cases. When ChrimsonR was expressed specifically for DA, the release of DA at the NAcShell increased in response to photo-stimulation of the VTA. In contrast, when ChrimsonR was expressed non-specifically, the amount of DA released was almost unchanged upon photo-stimulation. However, for nonspecifically expressed ChrimsonR, intraperitoneal injection of bicuculline, a competitive antagonist at the GABA-binding site of the GABAA receptor, also significantly increased the release of DA at the NAcShell in response to photo-stimulation of the VTA. The results of immunochemical staining confirm that GABAergic neurons in the VTA suppress DA activation, and also indicate that alterations in GABAergic neurons may have serious downstream effects on DA activity, NAcShell release, and neural adaptation of the VTA. This study also confirms that optogenetics technology is crucial to study the relationship between the mesolimbic dopaminergic and GABAergic neurons in a neural-specific manner.

Predicting Feeding Conditions of Premature Infants Through Non-Nutritive Sucking Skills Using a Sensitized Pacifier.

Due to the lack of enough physical or suck central pattern generator (SCPG) development, premature infants require assistance in improving their sucking skills as one of the first coordinated muscular activities in infants. Hence, we need to quantitatively measure their sucking abilities for future studies on their sucking interventions. Here, we present a new device that can measure both intraoral pressure (IP) and expression pressure (EP) as ororhithmic behavior parameters of non-nutritive sucking skills in infants. Our device is low-cost, easy-to-use, and accurate, which makes it appropriate for extensive studies. To showcase one of the applications of our device, we collected weekly data from 137 premature infants from 29 week-old to 36 week-old. Around half of the infants in our study needed intensive care even after they were 36 week-old. We call them full attainment of oral feeding (FAOF) infants. We then used the Non-nutritive sucking (NNS) features of EP and IP signals of infants recorded by our device to predict FAOF infants' sucking conditions. We found that our pipeline can predict FAOF infants several weeks before discharge from the hospital. Thus, this application of our device presents a robust and inexpensive alternative to monitor oral feeding ability in premature infants.

Successful International EMBS Conference on Biomedical Engineering and Innovation Held in Hungary.

The 1st IEEE Engineering in Medicine and Biology Society (EMBS) and the University of Pécs International Conference on Biomedical Engineering and Innovation (ICBEI) was held on 24-26 October 2022, in Pécs, Hungary. The conference opened with welcome talks by Prof. Attila Miseta, the rector of the University of Pécs (UP), Prof. Miklos Nyitrai, the dean of the UP, and Prof. Metin Akay, the president of IEEE EMBS, who delivered the first keynote talk at the conference. Then, we enjoyed three more exceptional keynote talks by Dr. Yasemin Akay, Dr. Gabor Forgacs, and Dr. Istvan Ulbert (Figure 1). With the participation of 230+ people, nine keynote lectures, and nine health care companies holding exhibitions, the conference was exceptional and well organized by Dr. Luca Toth, Dr. Adam Schiffer, and Dr. Peter Maroti, M.D., Ph.D., from the Centre for Biomedical Engineering and Innovation (Figure 2). The conference organizing team did a fantastic job. The attendance by several medical school students, the participation of health care companies (Figure 3), the hackathon competition, and several workshops were the highlights of the event. The first day of the conference concluded with a special wine tasting sponsored by the University of Pécs Wine Research Institute. Then, the conference organizing committee appreciation dinner hosted by Luca Tóth, Adam Schiffer, and Péter Maróti in the historical city of Pécs was indeed an exceptional social and cultural event (Figure 4).

Investigating the effects of chronic perinatal alcohol and combined nicotine and alcohol exposure on dopaminergic and non-dopaminergic neurons in the VTA.

The ventral tegmental area (VTA) is the origin of dopaminergic neurons and the dopamine (DA) reward pathway. This pathway has been widely studied in addiction and drug reinforcement studies and is believed to be the central processing component of the reward circuit. In this study, we used a well-established rat model to expose mother dams to alcohol, nicotine-alcohol, and saline perinatally. DA and non-DA neurons collected from the VTA of the rat pups were used to study expression profiles of miRNAs and mRNAs. miRNA pathway interactions, putative miRNA-mRNA target pairs, and downstream modulated biological pathways were analyzed. In the DA neurons, 4607 genes were differentially upregulated and 4682 were differentially downregulated following nicotine-alcohol exposure. However, in the non-DA neurons, only 543 genes were differentially upregulated and 506 were differentially downregulated. Cell proliferation, differentiation, and survival pathways were enriched after the treatments. Specifically, in the PI3K/AKT signaling pathway, there were 41 miRNAs and 136 mRNAs differentially expressed in the DA neurons while only 16 miRNAs and 20 mRNAs were differentially expressed in the non-DA neurons after the nicotine-alcohol exposure. These results depicted that chronic nicotine and alcohol exposures during pregnancy differentially affect both miRNA and gene expression profiles more in DA than the non-DA neurons in the VTA. Understanding how the expression signatures representing specific neuronal subpopulations become enriched in the VTA after addictive substance administration helps us to identify how neuronal functions may be altered in the brain.

Deep Learning Classification of Systemic Sclerosis Skin Using the MobileNetV2 Model.

Goal: Systemic sclerosis (SSc) is a rare autoimmune, systemic disease with prominent fibrosis of skin and internal organs. Early diagnosis of the disease is crucial for designing effective therapy and management plans. Machine learning algorithms, especially deep learning, have been found to be greatly useful in biology, medicine, healthcare, and biomedical applications, in the areas of medical image processing and speech recognition. However, the need for a large training data set and the requirement for a graphics processing unit (GPU) have hindered the wide application of machine learning algorithms as a diagnostic tool in resource-constrained environments (e.g., clinics). Methods: In this paper, we propose a novel mobile deep learning network for the characterization of SSc skin. The proposed network architecture consists of the UNet, a dense connectivity convolutional neural network (CNN) with added classifier layers that when combined with limited training data, yields better image segmentation and more accurate classification, and a mobile training module. In addition, to improve the computational efficiency and diagnostic accuracy, the highly efficient training model called "MobileNetV2," which is designed for mobile and embedded applications, was used to train the network. Results: The proposed network was implemented using a standard laptop (2.5 GHz Intel Core i7). After fine tuning, our results showed the proposed network reached 100% accuracy on the training image set, 96.8% accuracy on the validation image set, and 95.2% on the testing image set. The training time was less than 5 hours. We also analyzed the same normal vs SSc skin image sets using the CNN using the same laptop. The CNN reached 100% accuracy on the training image set, 87.7% accuracy on the validation image set, and 82.9% on the testing image set. Additionally, it took more than 14 hours to train the CNN architecture. We also utilized the MobileNetV2 model to analyze an additional dataset of images and classified them as normal, early (mid and moderate) SSc or late (severe) SSc skin images. The network reached 100% accuracy on the training image set, 97.2% on the validation set, and 94.8% on the testing image set. Using the same normal, early and late phase SSc skin images, the CNN reached 100% accuracy on the training image set, 87.7% accuracy on the validation image set, and 82.9% on the testing image set. These results indicated that the MobileNetV2 architecture is more accurate and efficient compared to the CNN to classify normal, early and late phase SSc skin images. Conclusions: Our preliminary study, intended to show the efficacy of the proposed network architecture, holds promise in the characterization of SSc. We believe that the proposed network architecture could easily be implemented in a clinical setting, providing a simple, inexpensive, and accurate screening tool for SSc.

Astrocytes Decreased the Sensitivity of Glioblastoma Cells to Temozolomide and Bay 11-7082.

Glioblastoma multiforme (GBM) is the most common malignant type of astrocytic tumors. GBM patients have a poor prognosis with a median survival of approximately 15 months despite the "Stupp" Regimen and high tumor recurrence due to the tumor resistance to chemotherapy. In this study, we co-cultured GBM cells with human astrocytes in three-dimensional (3D) poly(ethylene glycol) dimethyl acrylate (PEGDA) microwells to mimic the tumor microenvironment. We treated 3D co- and mono-cultured cells with Temozolomide (TMZ) and the nuclear factor-κB (NF-κB) inhibitor Bay 11-7082 and investigated the combined effect of the drugs. We assessed the expressions of glial fibrillary acidic protein (GFAP) and vimentin that play a role in the tumor malignancy and activation of the astrocytes as well as Notch-1 and survivin that play a role in GBM malignancy after the drug treatment to understand how astrocytes induced GBM drug response. Our results showed that in the co-culture, astrocytes increased GBM survival and resistance after combined drug treatment compared to mono-cultures. These data restated the importance of 3D cell culture to mimic the tumor microenvironment for drug screening.

Investigating the influence of perinatal nicotine and alcohol exposure on the genetic profiles of dopaminergic neurons in the VTA using miRNA-mRNA analysis.

Nicotine and alcohol are two of the most commonly used and abused recreational drugs, are often used simultaneously, and have been linked to significant health hazards. Furthermore, patients diagnosed with dependence on one drug are highly likely to be dependent on the other. Several studies have shown the effects of each drug independently on gene expression within many brain regions, including the ventral tegmental area (VTA). Dopaminergic (DA) neurons of the dopamine reward pathway originate from the VTA, which is believed to be central to the mechanism of addiction and drug reinforcement. Using a well-established rat model for both nicotine and alcohol perinatal exposure, we investigated miRNA and mRNA expression of dopaminergic (DA) neurons of the VTA in rat pups following perinatal alcohol and joint nicotine-alcohol exposure. Microarray analysis was then used to profile the differential expression of both miRNAs and mRNAs from DA neurons of each treatment group to further explore the altered genes and related biological pathways modulated. Predicted and validated miRNA-gene target pairs were analyzed to further understand the roles of miRNAs within these networks following each treatment, along with their post transcription regulation points affecting gene expression throughout development. This study suggested that glutamatergic synapse and axon guidance pathways were specifically enriched and many miRNAs and genes were significantly altered following alcohol or nicotine-alcohol perinatal exposure when compared to saline control. These results provide more detailed insight into the cell proliferation, neuronal migration, neuronal axon guidance during the infancy in rats in response to perinatal alcohol/ or nicotine-alcohol exposure.

NF-κB inhibitor with Temozolomide results in significant apoptosis in glioblastoma via the NF-κB(p65) and actin cytoskeleton regulatory pathways.

Glioblastoma (GBM) is the most malignant brain tumor characterized by intrinsic or acquired resistance to chemotherapy. GBM tumors show nuclear factor-κB (NF-κB) activity that has been associated with tumor formation, growth, and increased resistance to therapy. We investigated the effect of NF-κB inhibitor BAY 11-7082 with Temozolomide (TMZ) on the signaling pathways in GBM pathogenesis. GBM cells and patient-derived GBM cells cultured in 3D microwells were co-treated with BAY 11-7082 and TMZ or BAY 11-7082 and TMZ alone, and combined experiments of cell proliferation, apoptosis, wound healing assay, as well as reverse-phase protein arrays, western blot and immunofluorescence staining were used to evaluate the effects of drugs on GBM cells. The results revealed that the co-treatment significantly altered cell proliferation by decreasing GBM viability, suppressed NF-κB pathway and enhanced apoptosis. Moreover, it was found that the co-treatment of BAY 11-7082 and TMZ significantly contributed to a decrease in the migration pattern of patient-derived GBM cells by modulating actin cytoskeleton pathway. These findings suggest that in addition to TMZ treatment, NF-κB can be used as a potential target to increase the treatment's outcomes. The drug combination strategy, which is significantly improved by NF-κB inhibitor could be used to better understand the underlying mechanism of GBM pathways in vivo and as a potential therapeutic tool for GBM treatment.

Investigating the influence of perinatal nicotine exposure on genetic profiles of neurons in the sub-regions of the VTA.

Chronic nicotine exposure during pregnancy has been shown to induce physiological and anatomical alterations in offspring. Previously, we investigated the complexity of dopamine (DA) neuron firing in the sub-regions of the ventral tegmental area (VTA) following perinatal nicotine exposure. Using approximate entropy, we found that within the middle sub-region, the parainterfascicular nucleus (PIF), there was higher complexity indicating more random neural firing and a less homogeneous neuron population. Therefore, we sought to investigate the neuron populations within the sub-regions of the VTA following perinatal nicotine exposure. We used real time PCR in order to find the relative quantity of glutamate to γ-aminobutyric acid (GABA), DA, and glutamate neurons within three sub-regions: the parabrachial pigmented nucleus (PBP), parainterfascicular nucleus (PIF), and paranigral nucleus (PN). Our results showed that the PIF region of the VTA contained a more diverse population of neurons resulting in a more complex system. In addition, we found that DA neurons are more activated in PN sub-region of the VTA, which mediates the rewarding effects of drugs including nicotine. Lastly, using immunohistochemistry, we observed an overall decrease in DA neurons following perinatal nicotine exposure.

Temozolomide in Combination With NF-κB Inhibitor Significantly Disrupts the Glioblastoma Multiforme Spheroid Formation.

Glioblastoma multiforme (GBM) is the most common malignant primary brain tumor, accounting for 50% of all cases. GBM patients have a five-year survival rate of merely 5.6% and a median overall survival of 14.6 months with the "Stupp" regimen, 20.9 months with tumor treatment fields (TTF, OptuneR) in patients who participated in clinical trials, and 11 months for all GBM patients prior to TTF use. Objective: Our group recently developed a brain cancer chip which generates tumor spheroids, and provides large-scale assessments on the response of tumor cells to various concentrations and combinations of drugs. This platform could optimize the use of tumor samples derived from GBM patients to provide valuable insight on the tumor growth and responses to drug therapies. To minimize any sample loss in vitro, we improved our brain cancer chip system by adding an additional laminar flow distribution layer, which reduces sample loss during cell seeding and prevents spheroids from escaping from the microwells. Methods: In this study, we cultured 3D spheroids from GBM cell lines and patient-derived GBM cells in vitro, and investigated the effect of the combination of Temozolomide and nuclear factor-κB inhibitor on tumor growth. Results: Our study revealed that these drugs have synergistic effects in inhibiting spheroid formation when used in combination. Conclusions: These results suggest that the brain cancer chip enables large-scale, inexpensive and sample-effective drug screening to 3D cancer tumors in vitro, and could be applied to related tissue engineering drug screening studies.