The high metabolic demands of the brain require an efficient vascular system to be coupled with neural activity to supply adequate nutrients and oxygen.This supply is coordinated by the action of neurons,glial and vas...The high metabolic demands of the brain require an efficient vascular system to be coupled with neural activity to supply adequate nutrients and oxygen.This supply is coordinated by the action of neurons,glial and vascular cells,known collectively as the neurovascular unit,which temporally and spatially regulate local cerebral blood flow through a process known as neurovascular coupling.In many neurodegenerative diseases,changes in functions of the neurovascular unit not only impair neurovascular coupling but also permeability of the blood-brain barrier,cerebral blood flow and clearance of waste from the brain.In order to study disease mechanisms,we need improved physiologicallyrelevant human models of the neurovascular unit.Advances towards modeling the cellular complexity of the neurovascular unit in vitro have been made using stem-cell derived organoids and more recently,vascularized organoids,enabling intricate studies of non-cell autonomous processes.Engineering and design innovations in microfluidic devices and tissue engineering are progressing our ability to interrogate the cerebrovasculature.These advanced models are being used to gain a better understanding of neurodegenerative disease processes and potential therapeutics.Continued innovation is required to build more physiologically-relevant models of the neurovascular unit encompassing both the cellular complexity and designed features to interrogate neurovascular unit functionality.展开更多
The aim of this work was to develop an in vitro model to study mechanical compression effects on cartilage. A pressure-controlled compression device was used in this study. Cartilage explants obtained from human knee ...The aim of this work was to develop an in vitro model to study mechanical compression effects on cartilage. A pressure-controlled compression device was used in this study. Cartilage explants obtained from human knee were compressed at 1MPa/1Hz for 7 hours (30 min ON, 30 min OFF) under normoxia (5% CO2, 21% O2) or hypoxia (5% CO2, 5% O2). Cell viability was analyzed while nitric oxide (NO) and glycosaminoglycans (GAG) release was assayed in culture media. Mechanical stimulation increased NO production and GAG release by human cartilage explants under normoxia and hypoxia culture. In normoxia and hypoxia conditions, mechanical stimulation alters human OA cartilage metabolism. There is also, an increase in matrix degradation after compression, as shown by levels of GAG found in culture media. This study put in evidence the importance of mechanical compression in the progression of the osteoarthritis and present and in vitro model for mechanobiological and pharmacological studies.展开更多
Background:Risk factors for the etiology of post-weaning diarrhea,a major problem in swine industry associated with enormous economic losses,remain to be fully elucidated.In concordance with the ethical concerns raise...Background:Risk factors for the etiology of post-weaning diarrhea,a major problem in swine industry associated with enormous economic losses,remain to be fully elucidated.In concordance with the ethical concerns raised by animal experiments,we developed a new in vitro model of the weaning piglet colon(MPigut-IVM)including a mucin bead compartment to reproduce the mucus surface from the gut to which gut microbes can adhere.Results:Our results indicated that the MPigut-IVM is able to establish a representative piglet archaeal and bacterial colon microbiota in terms of taxonomic composition and function.The MPigut-IVM was consequently used to investigate the potential effects of feed deprivation,a common consequence of weaning in piglets,on the microbiota.The lack of nutrients in the MPigut-IVM led to an increased abundance of Prevotellaceae and Escherichia-Shigella and a decrease in Bacteroidiaceae and confirms previous in vivo findings.On top of a strong increase in redox potential,the feed deprivation stress induced modifications of microbial metabolite production such as a decrease in acetate and an increase in proportional valerate,isovalerate and isobutyrate production.Conclusions:The MPigut-IVM is able to simulate luminal and mucosal piglet microbiota and represent an innovative tool for comparative studies to investigate the impact of weaning stressors on piglet microbiota.Besides,weaning-associated feed deprivation in piglets provokes disruptions of MPigut-IVM microbiota composition and functionality and could be implicated in the onset of post-weaning dysbiosis in piglets.展开更多
Neurodegeneration is a catastrophic process that develops progressive damage leading to functional andstructural loss of the cells of the nervous system and is among the biggest unavoidable problems of our age.Animalm...Neurodegeneration is a catastrophic process that develops progressive damage leading to functional andstructural loss of the cells of the nervous system and is among the biggest unavoidable problems of our age.Animalmodels do not reflect the pathophysiology observed in humans due to distinct differences between the neuralpathways,gene expression patterns,neuronal plasticity,and other disease-related mechanisms in animals andhumans.Classical in vitro cell culture models are also not sufficient for pre-clinical drug testing in reflecting thecomplex pathophysiology of neurodegenerative diseases.Today,modern,engineered techniques are applied to developmulticellular,intricate in vitro models and to create the closest microenvironment simulating biological,biochemical,and mechanical characteristics of the in vivo degenerating tissue.In THIS review,the capabilities and shortcomings ofscaffold-based and scaffold-free techniques,organoids,and microfluidic models that best reflect neurodegeneration invitro in the biomimetic framework are discussed.展开更多
Traumatic brain injuries are serious clinical incidents associated with some of the poorest outcomes in neurological practice.Coupled with the limited regenerative capacity of the brain,this has significant implicatio...Traumatic brain injuries are serious clinical incidents associated with some of the poorest outcomes in neurological practice.Coupled with the limited regenerative capacity of the brain,this has significant implications for patients,carers,and healthcare systems,and the requirement for life-long care in some cases.Clinical treatment currently focuses on limiting the initial neural damage with longterm care/support from multidisciplinary teams.Therapies targeting neuroprotection and neural regeneration are not currently available but are the focus of intensive research.Biomaterial-based interventions are gaining popularity for a range of applications including biomolecule and drug delive ry,and to function as cellular scaffolds.Experimental investigations into the development of such novel therapeutics for traumatic brain injury will be critically underpinned by the availability of appropriate high thro ughput,facile,ethically viable,and pathomimetic biological model systems.This represents a significant challenge for researchers given the pathological complexity of traumatic brain injury.Specifically,there is a concerted post-injury response mounted by multiple neural cell types which includes microglial activation and astroglial scarring with the expression of a range of growth inhibito ry molecules and cytokines in the lesion environment.Here,we review common models used for the study of traumatic brain injury(ranging from live animal models to in vitro systems),focusing on penetrating traumatic brain injury models.We discuss their relative advantages and drawbacks for the developmental testing of biomaterial-based therapies.展开更多
The human gut microbiota is widely considered to be a metabolic organ hidden within our bodies,playing a crucial role in the host’s physiology.Several factors affect its composition,so a wide variety of microbes resi...The human gut microbiota is widely considered to be a metabolic organ hidden within our bodies,playing a crucial role in the host’s physiology.Several factors affect its composition,so a wide variety of microbes residing in the gut are present in the world population.Individual excessive imbalances in microbial composition are often associated with human disorders and pathologies,and new investigative strategies to gain insight into these pathologies and define pharmaceutical therapies for their treatment are needed.In vitro models of the human gut microbiota are commonly used to study microbial fermentation patterns,community composition,and host-microbe interactions.Bioreactors and microfluidic devices have been designed to culture microorganisms from the human gut microbiota in a dynamic environment in the presence or absence of eukaryotic cells to interact with.In this review,we will describe the overall elements required to create a functioning,reproducible,and accurate in vitro culture of the human gut microbiota.In addition,we will analyze some of the devices currently used to study fermentation processes and relationships between the human gut microbiota and host eukaryotic cells.展开更多
Liver injury is a common cause of drug approval withdrawal during drug development,pre-clinical research,and clinical treatment.If not properly treated,patients with severe liver injury can suffer from acute liver fai...Liver injury is a common cause of drug approval withdrawal during drug development,pre-clinical research,and clinical treatment.If not properly treated,patients with severe liver injury can suffer from acute liver failure or even death.Thus,utilization of the convenient in vitro hepatotoxicity assessment model for early detection of drug-induced hepatotoxicity is vital for drug development and safe personalized medication.Biomaterials(e.g.,hydrogels,nanofibers,decellularized liver matrix)and bioengineering technologies(e.g.,microarrays,micropatterns,3D printing,and microfluidics)have been applied for in vitro hepatotoxicity assessment models.This review summarizes the structure and functions of the liver as well as the components of in vitro hepatotoxicity assessment models.In addition,it highlights the latest advances in developing hepatotoxicity models with the ultimate goal of further clinical translation.展开更多
Introduction Cancer is an attractive target of gene therapy and currently represents the disease in most clinical trials[1]. Strategies for cancer gene therapy include: (1) stimulation of immune responses to tumor cel...Introduction Cancer is an attractive target of gene therapy and currently represents the disease in most clinical trials[1]. Strategies for cancer gene therapy include: (1) stimulation of immune responses to tumor cells,(2) delivery of specific enzymes展开更多
The introduction of living cells to manufacturing process has enabled the engineering of complex biological tissues in vitro.The recent advances in biofabrication with extremely high resolution(e.g.at single cell leve...The introduction of living cells to manufacturing process has enabled the engineering of complex biological tissues in vitro.The recent advances in biofabrication with extremely high resolution(e.g.at single cell level)have greatly enhanced this capacity and opened new avenues for tissue engineering.In this review,we comprehensively overview the current biofabrication strategies with single-cell resolution and categorize them based on the dimension of the single-cell building blocks,i.e.zero-dimensional single-cell droplets,one-dimensional single-cell filaments and two-dimensional single-cell sheets.We provide an informative introduction to the most recent advances in these approaches(e.g.cell trapping,bioprinting,electrospinning,microfluidics and cell sheets)and further illustrated how they can be used in in vitro tissue modelling and regenerative medicine.We highlight the significance of single-cell-level biofabrication and discuss the challenges and opportunities in the field.展开更多
Wheat flour,as the most important source of food globally,is one of the most common causative agents of food allergy.This study aimed to investigate the effects of fermentation on wheat protein digestibility and aller...Wheat flour,as the most important source of food globally,is one of the most common causative agents of food allergy.This study aimed to investigate the effects of fermentation on wheat protein digestibility and allergenicity.Protein digestibility were evaluated using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and enzyme-linked immunosorbent assay.The effect of protein on intestinal permeability was investigated by Caco-2 cell monolayers.Co-culture fermentation with Pediococcus acidilactici XZ31 and yeast leads to improvement in digestibility of wheat protein compared to single strain fermentation.Fermentation leads to a decrease in albumin/globulin antigenicity and an increase in gluten R5 reactivity,with the most significant changes in the co-culture group.Digestion strengthen the decrease of protein antigenicity and counteracts the difference in antigenicity induced by fermentation between groups.However,pretreatment with P.acidilactici XZ31 reduces the amount of allergens across Caco-2 monolayer and attenuates the gluten-induced increase in permeability of Caco-2 cell monolayer by reducing actin polymerization and villous atrophy.Co-culture fermentation reduces gluten-induced cell monolayer damage to a greater extent than P.acidilactici XZ31 monoculture.These results gives valuable insight into the effects of P.acidilactici XZ31 fermentation on the allergenicity and toxicity of wheat proteins,which contribute to promoting the application of multi-strain leavening agent in hypoallergenic and gluten-free wheat products.展开更多
Microglia, the main driver of neuroinflammation, play a central role in the initiation and exacerbation of various neurodegenerative diseases and are now considered a promising therapeutic target. Previous studies on ...Microglia, the main driver of neuroinflammation, play a central role in the initiation and exacerbation of various neurodegenerative diseases and are now considered a promising therapeutic target. Previous studies on in vitro human microglia and in vivo rodent models lacked scalability, consistency, or physiological relevance, which deterred successful therapeutic outcomes for the past decade. Here we review human blood monocyte-derived microglia-like cells as a robust and consistent approach to generate a patient-specific microglia-like model that can be used in extensive cohort studies for drug testing. We will highlight the strength and applicability of human blood monocyte-derived microglia-like cells to increase translational outcomes by reviewing the advantages of human blood monocyte-derived microglia-like cells in addressing patient heterogeneity and stratification, the basis of personalized medicine.展开更多
Brain metastases are a leading cause of cancer-related mortality.However,progress in their treatment has been limited over the past decade,due to an incomplete understanding of the underlying biological mechanisms.Emp...Brain metastases are a leading cause of cancer-related mortality.However,progress in their treatment has been limited over the past decade,due to an incomplete understanding of the underlying biological mechanisms.Employing accurate in vitro and in vivo models to recapitulate the complexities of brain metastasis offers the most promising approach to unravel the intricate cellular and physiological processes involved.Here,we present a comprehensive review of the currently accessible models for studying brain metastasis.We introduce a diverse array of in vitro and in vivo models,including cultured cells using the Transwell system,organoids,microfluidic models,syngeneic models,xenograft models,and genetically engineered models.We have also provided a concise summary of the merits and limitations inherent to each model while identifying the optimal contexts for their effective utilization.This review serves as a comprehensive resource,aiding researchers in making well-informed decisions regarding model selection that align with specific research questions.展开更多
The SOCS1/JAK2/STAT3 axis is strongly associated with tumor growth and progression,and participates in cytokine secretion in many diseases.However,the effects of the SOCS1/JAK2/STAT3 axis in experimental subarachnoid ...The SOCS1/JAK2/STAT3 axis is strongly associated with tumor growth and progression,and participates in cytokine secretion in many diseases.However,the effects of the SOCS1/JAK2/STAT3 axis in experimental subarachnoid hemorrhage remain to be studied.A subarachnoid hemorrhage model was established in rats by infusing autologous blood into the optic chiasm pool.Some rats were first treated with JAK2/STAT3 small interfering RNA(Si-JAK2/Si-STAT3)or overexpression plasmids of JAK2/STAT3.In the brains of subarachnoid hemorrhage model rats,the expression levels of both JAK2 and STAT3 were upregulated and the expression of SOCS1 was downregulated,reaching a peak at 48 hours after injury.Simultaneously,the interactions between JAK2 and SOCS1 were reduced.In contrast,the interactions between JAK2 and STAT3 were markedly enhanced.Si-JAK2 and Si-STAT3 treatment alleviated cortical neuronal cell apoptosis and necrosis,destruction of the blood-brain barrier,brain edema,and cognitive functional impairment after subarachnoid hemorrhage.This was accompanied by decreased phosphorylation of JAK2 and STAT3 protein,decreased total levels of JAK2 and STAT3 protein,and increased SOCS1 protein expression.However,overexpression of JAK2 and STAT3 exerted opposite effects,aggravating subarachnoid hemorrhage-induced early brain injury.Si-JAK2 and Si-STAT3 inhibited M1-type microglial conversion and the release of pro-inflammatory factors(inducible nitric oxide synthase,interleukin-1β,and tumor necrosis factor-α)and increased the release of anti-inflammatory factors(arginase-1,interleukin-10,and interleukin-4).Furthermore,primary neurons stimulated with oxyhemoglobin were used to simulate subarachnoid hemorrhage in vitro,and the JAK2 inhibitor AG490 was used as an intervention.The in vitro results also suggested that neuronal protection is mediated by the inhibition of JAK2 and STAT3 expression.Together,our findings indicate that the SOCS1/JAK2/STAT3 axis contributes to early brain injury after subarachnoid hemorrhage both in vitro and in vivo by inducing inflammatory responses.This study was approved by the Animal Ethics Committee of Anhui Medical University and the First Affiliated Hospital of University of Science and Technology of China(approval No.LLSC-20180202)on March 1,2018.展开更多
Biomedical field has been seeking a feasible standard drug screening system consisting of 3D tumor model array for drug researching due to providing sufficient samples and simulating actual in vivo tumor growth situat...Biomedical field has been seeking a feasible standard drug screening system consisting of 3D tumor model array for drug researching due to providing sufficient samples and simulating actual in vivo tumor growth situation,which is still a challenge to rapidly and uniformly establish though.Here,we propose a novel drug screening system,namely 3D tumor array chip with“layer cake”structure,for drug screening.Accurate gelatin methacryloyl hydrogel droplets(~0.1μL)containing tumor cells can be automatically deposited on demand with electrohydrodynamic 3D printing.Transparent conductive membrane is introduced as a chip basement for preventing charges accumulation during fabricating and convenient observing during screening.Culturing chambers formed by stainless steel and silicon interlayer is convenient to be assembled and recycled.As this chip is compatible with the existing 96-well culturing plate,the drug screening protocols could keep the same as convention.Important properties of this chip,namely printing stability,customizability,accuracy,microenvironment,tumor functionalization,are detailly examined.As a demonstration,it is applied for screening of epirubicin and paclitaxel with breast tumor cells to confirm the compatibility of the proposed screening system with the traditional screening methods.We believe this chip will potentially play a significant role in drug evaluation in the future.展开更多
Background:Pudilan Xiaoyan Oral Liquid(PDL),a famous traditional Chinese formula for treating acute and chronic inflammation.To evaluate the broad-spectrum antiviral effect of Pudilan Xiaoyan Oral Liquid,and provide a...Background:Pudilan Xiaoyan Oral Liquid(PDL),a famous traditional Chinese formula for treating acute and chronic inflammation.To evaluate the broad-spectrum antiviral effect of Pudilan Xiaoyan Oral Liquid,and provide a basis for clinical medication.Methods:Its inhibitory effect on different respiratory viruses was observed by cytopathic test.The potential mechanism of the anti-influenza effect was determined by neuraminidase activity.In order to observe the therapeutic effect of PDL on viral pneumonia caused by different respiratory viruses.The viral pneumonia model was established by nasal infection with different respiratory viruses,and then PDL was given Therapeutic and prophylactically to evaluate its pharmacodynamic activity in vivo.Results:The results of in vitro experiments showed that PDL had different inhibitory effects on cytopathic effects caused by different respiratory viruses.And it has obvious inhibitory effect on the neuraminidase activity of influenza A virus,which indicates that it exerts anti-influenza virus effect by inhibiting neuraminidase activity of influenza virus.The results in vivo showed that PDL exhibited an inhibitory effect on pulmonary index(PI)and effectively reduced the degree of lesions in the lungs.The lethal rate of mice was significantly decreased while survival time of mice was dramatically increased by PDL treatment in comparison to infection control,respectively.Conclusions:Our study demonstrates that PDL had a significant protection and treatment effect for respiratory virus infection in vitro and in vivo.展开更多
Adult-onset brain cancers,such as glioblastomas,are particularly lethal.People with glioblastoma multiforme(GBM)do not anticipate living for more than 15 months if there is no cure.The results of conventional treatmen...Adult-onset brain cancers,such as glioblastomas,are particularly lethal.People with glioblastoma multiforme(GBM)do not anticipate living for more than 15 months if there is no cure.The results of conventional treatments over the past 20 years have been underwhelming.Tumor aggressiveness,location,and lack of systemic therapies that can penetrate the blood–brain barrier are all contributing factors.For GBM treatments that appear promising in preclinical studies,there is a considerable rate of failure in phaseⅠandⅡclinical trials.Unfortunately,access becomes impossible due to the intricate architecture of tumors.In vitro,bioengineered cancer models are currently being used by researchers to study disease development,test novel therapies,and advance specialized medications.Many different techniques for creating in vitro systems have arisen over the past few decades due to developments in cellular and tissue engineering.Later-stage research may yield better results if in vitro models that resemble brain tissue and the blood–brain barrier are used.With the use of 3D preclinical models made available by biomaterials,researchers have discovered that it is possible to overcome these limitations.Innovative in vitro models for the treatment of GBM are possible using biomaterials and novel drug carriers.This review discusses the benefits and drawbacks of 3D in vitro glioblastoma modeling systems.展开更多
Fibrotic tumors,such as pancreatic ductal adenocarcinoma(PDAC),are characterized for high desmoplastic reaction,which results in high intra-tumoral solid stress leading to the compression of blood vessels.These microa...Fibrotic tumors,such as pancreatic ductal adenocarcinoma(PDAC),are characterized for high desmoplastic reaction,which results in high intra-tumoral solid stress leading to the compression of blood vessels.These microarchitectural alterations cause loss of blood flow and poor intra-tumoral delivery of therapeutics.Currently,there is a lack of relevant in vitro models capable of replicating these mechanical characteristics and to test anti-desmoplastic compounds.Here,a multi-layered vascularized 3D PDAC model consisting of primary human pancreatic stellate cells(PSCs)embedded in collagen/fibrinogen(Col/Fib),mimicking tumor tissue within adjunct healthy tissue,is presented to study the fibrosis-induced compression of vasculature in PDAC.It is demonstrated how the mechanical and biological stimulation induce PSC activation,extracellular matrix production and eventually vessel compression.The clinical relevance is confirmed by correlating with patient transcriptomic data.Furthermore,the effects of gradual vessel compression on the fluid dynamics occurring within the channel is evaluated in silico.Finally,it is demonstrated how cancer-associated fibroblast(CAF)-modulatory therapeutics can inhibit the cell-mediated compression of blood vessels in PDAC in vitro,in silico and in vivo.It is envisioned that this 3D model is used to improve the understanding of mechanical characteristics in tumors and for evaluating novel anti-desmoplastic therapeutics.展开更多
Background:The ingestion of golden thread contaminated with heavy metals through the food chain leads to detrimental effects to human health.During digestion,not all of the heavy metals could be released to the gastro...Background:The ingestion of golden thread contaminated with heavy metals through the food chain leads to detrimental effects to human health.During digestion,not all of the heavy metals could be released to the gastrointestinal tract and readily to be absorbed by human body.Thus,bioaccessibility is an important issue in health risk assessments.Aims and Objectives:The aims and objectives of this study were to investigate the bioaccessibility of Cd in golden thread and assess the associated health risks based on the exposure to bioaccessible Cd.Materials and Methods:Inductively coupled plasma mass spectrometry(ICP-MS)has been applied to determine the Cd content in golden thread.Physiologically based extraction test(PBET)digestion was performed in the in vitro/Caco2 cell model to investigate the bioaccessibility of Cd in golden thread.Furthermore,the target hazard quotient(THQ)was used to assess the risks of the total and the bioaccessible content of Cd in golden thread.Results:The results revealed that the total Cd content in six batches of golden thread ranged from 3.203 to 5.723 mg/kg.After uptake by Caco2 cells,the bioaccessibility of Cd ranged from 42.36%to 59.73%.The results of the risk assessment indicated that prior to uptake by Caco2 cells,the THQ values of Cd for all batches of golden thread were greater than 1.However,after uptake by Caco2 cells,the THQ values of Cd in all samples were less than 1,thus suggesting that the risks were at a safe level.Conclusion:This study was the first to perform health risk assessment with bioaccessible heavy metals present in traditional Chinese medicine by PBET digestion using an in vitro/Caco2 cell model,thus enabling us to obtain more accurate and objective results while allowing us to avoid unnecessary government intervention and to establish more reasonable limit standards for heavy metals.展开更多
Decidualization is a special type of differentiation of endometrial stromal cells into secretory decidualized cells,which is closely related to the occurrence of menstruation and establishment of pregnancy.Decidualiza...Decidualization is a special type of differentiation of endometrial stromal cells into secretory decidualized cells,which is closely related to the occurrence of menstruation and establishment of pregnancy.Decidualization abnormalities can cause female infertility and abortion,and the decidualization modelin vitro is an important tool for studying relevant mechanisms.This article summarizes severalin vitro decidualization models in recent research from three aspects,including the selection of model cells and culture systems,evaluation of decidualization markers,and induction schemes.These models can be appropriately selected and applied in specific endometrium-related disease models,such as endometriosis,recurrent pregnancy loss,and preeclampsia.展开更多
The pathogenic cascade of Alzheimer’s disease(AD)characterized by amyloid-β protein accumulation is still poorly understood,partially owing to the limitations of relevant models without in vivo neural tissue microen...The pathogenic cascade of Alzheimer’s disease(AD)characterized by amyloid-β protein accumulation is still poorly understood,partially owing to the limitations of relevant models without in vivo neural tissue microenvironment to recapitulate cell-cell interactions.To better mimic neural tissue microenvironment,three-dimensional(3D)core-shell AD model constructs containing human neural progenitor cells(NSCs)with 2% matrigel as core bioink and 2% alginate as shell bioink have been bioprinted by a co-axial bioprinter,with a suitable shell thickness for nutrient exchange and barrier-free cell interaction cores.These constructs exhibit cell self-clustering and-assembling properties and engineered reproducibility with long-term cell viability and self-renewal,and a higher differentiation level compared to 2D and 3D MIX models.The different effects of 3D bioprinted,2D,and MIX microenvironments on the growth of NSCs are mainly related to biosynthesis of amino acids and glyoxylate and dicarboxylate metabolism on day 2 and ribosome,biosynthesis of amino acids and proteasome on day 14.Particularly,the model constructs demonstrated Aβ aggregation and higher expression of Aβ and tau isoform genes compared to 2D and MIX controls.AD model constructs will provide a promising strategy to facilitate the development of a 3D in vitro AD model for neurodegeneration research.展开更多
基金supported by the Weston Brain Institute Rapid Response Grant,No.RR182093(to JR).
文摘The high metabolic demands of the brain require an efficient vascular system to be coupled with neural activity to supply adequate nutrients and oxygen.This supply is coordinated by the action of neurons,glial and vascular cells,known collectively as the neurovascular unit,which temporally and spatially regulate local cerebral blood flow through a process known as neurovascular coupling.In many neurodegenerative diseases,changes in functions of the neurovascular unit not only impair neurovascular coupling but also permeability of the blood-brain barrier,cerebral blood flow and clearance of waste from the brain.In order to study disease mechanisms,we need improved physiologicallyrelevant human models of the neurovascular unit.Advances towards modeling the cellular complexity of the neurovascular unit in vitro have been made using stem-cell derived organoids and more recently,vascularized organoids,enabling intricate studies of non-cell autonomous processes.Engineering and design innovations in microfluidic devices and tissue engineering are progressing our ability to interrogate the cerebrovasculature.These advanced models are being used to gain a better understanding of neurodegenerative disease processes and potential therapeutics.Continued innovation is required to build more physiologically-relevant models of the neurovascular unit encompassing both the cellular complexity and designed features to interrogate neurovascular unit functionality.
文摘The aim of this work was to develop an in vitro model to study mechanical compression effects on cartilage. A pressure-controlled compression device was used in this study. Cartilage explants obtained from human knee were compressed at 1MPa/1Hz for 7 hours (30 min ON, 30 min OFF) under normoxia (5% CO2, 21% O2) or hypoxia (5% CO2, 5% O2). Cell viability was analyzed while nitric oxide (NO) and glycosaminoglycans (GAG) release was assayed in culture media. Mechanical stimulation increased NO production and GAG release by human cartilage explants under normoxia and hypoxia culture. In normoxia and hypoxia conditions, mechanical stimulation alters human OA cartilage metabolism. There is also, an increase in matrix degradation after compression, as shown by levels of GAG found in culture media. This study put in evidence the importance of mechanical compression in the progression of the osteoarthritis and present and in vitro model for mechanobiological and pharmacological studies.
文摘Background:Risk factors for the etiology of post-weaning diarrhea,a major problem in swine industry associated with enormous economic losses,remain to be fully elucidated.In concordance with the ethical concerns raised by animal experiments,we developed a new in vitro model of the weaning piglet colon(MPigut-IVM)including a mucin bead compartment to reproduce the mucus surface from the gut to which gut microbes can adhere.Results:Our results indicated that the MPigut-IVM is able to establish a representative piglet archaeal and bacterial colon microbiota in terms of taxonomic composition and function.The MPigut-IVM was consequently used to investigate the potential effects of feed deprivation,a common consequence of weaning in piglets,on the microbiota.The lack of nutrients in the MPigut-IVM led to an increased abundance of Prevotellaceae and Escherichia-Shigella and a decrease in Bacteroidiaceae and confirms previous in vivo findings.On top of a strong increase in redox potential,the feed deprivation stress induced modifications of microbial metabolite production such as a decrease in acetate and an increase in proportional valerate,isovalerate and isobutyrate production.Conclusions:The MPigut-IVM is able to simulate luminal and mucosal piglet microbiota and represent an innovative tool for comparative studies to investigate the impact of weaning stressors on piglet microbiota.Besides,weaning-associated feed deprivation in piglets provokes disruptions of MPigut-IVM microbiota composition and functionality and could be implicated in the onset of post-weaning dysbiosis in piglets.
文摘Neurodegeneration is a catastrophic process that develops progressive damage leading to functional andstructural loss of the cells of the nervous system and is among the biggest unavoidable problems of our age.Animalmodels do not reflect the pathophysiology observed in humans due to distinct differences between the neuralpathways,gene expression patterns,neuronal plasticity,and other disease-related mechanisms in animals andhumans.Classical in vitro cell culture models are also not sufficient for pre-clinical drug testing in reflecting thecomplex pathophysiology of neurodegenerative diseases.Today,modern,engineered techniques are applied to developmulticellular,intricate in vitro models and to create the closest microenvironment simulating biological,biochemical,and mechanical characteristics of the in vivo degenerating tissue.In THIS review,the capabilities and shortcomings ofscaffold-based and scaffold-free techniques,organoids,and microfluidic models that best reflect neurodegeneration invitro in the biomimetic framework are discussed.
基金funded by awards from the EPSRC Doctoral Training Centre in Regenerative Medicine and an NHS bursary。
文摘Traumatic brain injuries are serious clinical incidents associated with some of the poorest outcomes in neurological practice.Coupled with the limited regenerative capacity of the brain,this has significant implications for patients,carers,and healthcare systems,and the requirement for life-long care in some cases.Clinical treatment currently focuses on limiting the initial neural damage with longterm care/support from multidisciplinary teams.Therapies targeting neuroprotection and neural regeneration are not currently available but are the focus of intensive research.Biomaterial-based interventions are gaining popularity for a range of applications including biomolecule and drug delive ry,and to function as cellular scaffolds.Experimental investigations into the development of such novel therapeutics for traumatic brain injury will be critically underpinned by the availability of appropriate high thro ughput,facile,ethically viable,and pathomimetic biological model systems.This represents a significant challenge for researchers given the pathological complexity of traumatic brain injury.Specifically,there is a concerted post-injury response mounted by multiple neural cell types which includes microglial activation and astroglial scarring with the expression of a range of growth inhibito ry molecules and cytokines in the lesion environment.Here,we review common models used for the study of traumatic brain injury(ranging from live animal models to in vitro systems),focusing on penetrating traumatic brain injury models.We discuss their relative advantages and drawbacks for the developmental testing of biomaterial-based therapies.
基金supported by the BIOMEMBRANE project (M-ERA.net 2 project 4246)the KERAPACK project (MANUNET MNET 17/NMAT-0060)+2 种基金the PRA_2018_68 (grant supported by the University of Pisa)MIT-UNIPI project (grant supported by the University of Pisa and the MIT)the support of the Additive Manufacturing Cross-Lab of the Department of Information Engineering of the University of Pisa
文摘The human gut microbiota is widely considered to be a metabolic organ hidden within our bodies,playing a crucial role in the host’s physiology.Several factors affect its composition,so a wide variety of microbes residing in the gut are present in the world population.Individual excessive imbalances in microbial composition are often associated with human disorders and pathologies,and new investigative strategies to gain insight into these pathologies and define pharmaceutical therapies for their treatment are needed.In vitro models of the human gut microbiota are commonly used to study microbial fermentation patterns,community composition,and host-microbe interactions.Bioreactors and microfluidic devices have been designed to culture microorganisms from the human gut microbiota in a dynamic environment in the presence or absence of eukaryotic cells to interact with.In this review,we will describe the overall elements required to create a functioning,reproducible,and accurate in vitro culture of the human gut microbiota.In addition,we will analyze some of the devices currently used to study fermentation processes and relationships between the human gut microbiota and host eukaryotic cells.
基金supports from General Program from the National Natural Science Foundation of China(No.31871016)the National Key Research and Development Program(2016YFC1101302)from the Ministry of Science and Technology of China.
文摘Liver injury is a common cause of drug approval withdrawal during drug development,pre-clinical research,and clinical treatment.If not properly treated,patients with severe liver injury can suffer from acute liver failure or even death.Thus,utilization of the convenient in vitro hepatotoxicity assessment model for early detection of drug-induced hepatotoxicity is vital for drug development and safe personalized medication.Biomaterials(e.g.,hydrogels,nanofibers,decellularized liver matrix)and bioengineering technologies(e.g.,microarrays,micropatterns,3D printing,and microfluidics)have been applied for in vitro hepatotoxicity assessment models.This review summarizes the structure and functions of the liver as well as the components of in vitro hepatotoxicity assessment models.In addition,it highlights the latest advances in developing hepatotoxicity models with the ultimate goal of further clinical translation.
基金supported by a predoctoral fellowship from the National Institutes of Health and a research grant from the National Science Foundation
文摘Introduction Cancer is an attractive target of gene therapy and currently represents the disease in most clinical trials[1]. Strategies for cancer gene therapy include: (1) stimulation of immune responses to tumor cells,(2) delivery of specific enzymes
基金support from the National Natural Science Foundation of China(No.52105306,32211530075)New Faculty Start-up Funding provided by Tsinghua University(012-53330200421).
文摘The introduction of living cells to manufacturing process has enabled the engineering of complex biological tissues in vitro.The recent advances in biofabrication with extremely high resolution(e.g.at single cell level)have greatly enhanced this capacity and opened new avenues for tissue engineering.In this review,we comprehensively overview the current biofabrication strategies with single-cell resolution and categorize them based on the dimension of the single-cell building blocks,i.e.zero-dimensional single-cell droplets,one-dimensional single-cell filaments and two-dimensional single-cell sheets.We provide an informative introduction to the most recent advances in these approaches(e.g.cell trapping,bioprinting,electrospinning,microfluidics and cell sheets)and further illustrated how they can be used in in vitro tissue modelling and regenerative medicine.We highlight the significance of single-cell-level biofabrication and discuss the challenges and opportunities in the field.
基金supported by the National Key Research and Development Program of China(2019YFC1605000)National Natural Science Foundation of China(31872904)。
文摘Wheat flour,as the most important source of food globally,is one of the most common causative agents of food allergy.This study aimed to investigate the effects of fermentation on wheat protein digestibility and allergenicity.Protein digestibility were evaluated using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and enzyme-linked immunosorbent assay.The effect of protein on intestinal permeability was investigated by Caco-2 cell monolayers.Co-culture fermentation with Pediococcus acidilactici XZ31 and yeast leads to improvement in digestibility of wheat protein compared to single strain fermentation.Fermentation leads to a decrease in albumin/globulin antigenicity and an increase in gluten R5 reactivity,with the most significant changes in the co-culture group.Digestion strengthen the decrease of protein antigenicity and counteracts the difference in antigenicity induced by fermentation between groups.However,pretreatment with P.acidilactici XZ31 reduces the amount of allergens across Caco-2 monolayer and attenuates the gluten-induced increase in permeability of Caco-2 cell monolayer by reducing actin polymerization and villous atrophy.Co-culture fermentation reduces gluten-induced cell monolayer damage to a greater extent than P.acidilactici XZ31 monoculture.These results gives valuable insight into the effects of P.acidilactici XZ31 fermentation on the allergenicity and toxicity of wheat proteins,which contribute to promoting the application of multi-strain leavening agent in hypoallergenic and gluten-free wheat products.
基金supported by grants from NHMRC (APP1125796)The Col Bambrick Memorial MND Research Grant+2 种基金The NTI MND Research Grantthe FightMND Foundationsupported by an NHMRC Senior Research Fellowship APP1118452。
文摘Microglia, the main driver of neuroinflammation, play a central role in the initiation and exacerbation of various neurodegenerative diseases and are now considered a promising therapeutic target. Previous studies on in vitro human microglia and in vivo rodent models lacked scalability, consistency, or physiological relevance, which deterred successful therapeutic outcomes for the past decade. Here we review human blood monocyte-derived microglia-like cells as a robust and consistent approach to generate a patient-specific microglia-like model that can be used in extensive cohort studies for drug testing. We will highlight the strength and applicability of human blood monocyte-derived microglia-like cells to increase translational outcomes by reviewing the advantages of human blood monocyte-derived microglia-like cells in addressing patient heterogeneity and stratification, the basis of personalized medicine.
基金This work was supported by the National Natural Science Foundation of China(No.82072787)the Department of Science and Technology of Guangdong Province International Cooperation Project(No.2022A0505050012)+1 种基金the Tertiary Education Scientific Research Project of Guangzhou Municipal Education Bureau(No.202235421)the Guangzhou Basic and Applied Basic Research Scheme(No.G23151016).
文摘Brain metastases are a leading cause of cancer-related mortality.However,progress in their treatment has been limited over the past decade,due to an incomplete understanding of the underlying biological mechanisms.Employing accurate in vitro and in vivo models to recapitulate the complexities of brain metastasis offers the most promising approach to unravel the intricate cellular and physiological processes involved.Here,we present a comprehensive review of the currently accessible models for studying brain metastasis.We introduce a diverse array of in vitro and in vivo models,including cultured cells using the Transwell system,organoids,microfluidic models,syngeneic models,xenograft models,and genetically engineered models.We have also provided a concise summary of the merits and limitations inherent to each model while identifying the optimal contexts for their effective utilization.This review serves as a comprehensive resource,aiding researchers in making well-informed decisions regarding model selection that align with specific research questions.
基金This study was supported by the National Natural Science Foundation of China,No.81500375(to XQK)the Fundamental Research Funds for the Central Universities,No.WK9110000112(to YW)+1 种基金the Anhui Provincial Natural Science Foundation of China,No.1508085QH184(to YW)Shandong Provincial Natural Science Foundation of China,No.ZR2015HQ001(to XQK).
文摘The SOCS1/JAK2/STAT3 axis is strongly associated with tumor growth and progression,and participates in cytokine secretion in many diseases.However,the effects of the SOCS1/JAK2/STAT3 axis in experimental subarachnoid hemorrhage remain to be studied.A subarachnoid hemorrhage model was established in rats by infusing autologous blood into the optic chiasm pool.Some rats were first treated with JAK2/STAT3 small interfering RNA(Si-JAK2/Si-STAT3)or overexpression plasmids of JAK2/STAT3.In the brains of subarachnoid hemorrhage model rats,the expression levels of both JAK2 and STAT3 were upregulated and the expression of SOCS1 was downregulated,reaching a peak at 48 hours after injury.Simultaneously,the interactions between JAK2 and SOCS1 were reduced.In contrast,the interactions between JAK2 and STAT3 were markedly enhanced.Si-JAK2 and Si-STAT3 treatment alleviated cortical neuronal cell apoptosis and necrosis,destruction of the blood-brain barrier,brain edema,and cognitive functional impairment after subarachnoid hemorrhage.This was accompanied by decreased phosphorylation of JAK2 and STAT3 protein,decreased total levels of JAK2 and STAT3 protein,and increased SOCS1 protein expression.However,overexpression of JAK2 and STAT3 exerted opposite effects,aggravating subarachnoid hemorrhage-induced early brain injury.Si-JAK2 and Si-STAT3 inhibited M1-type microglial conversion and the release of pro-inflammatory factors(inducible nitric oxide synthase,interleukin-1β,and tumor necrosis factor-α)and increased the release of anti-inflammatory factors(arginase-1,interleukin-10,and interleukin-4).Furthermore,primary neurons stimulated with oxyhemoglobin were used to simulate subarachnoid hemorrhage in vitro,and the JAK2 inhibitor AG490 was used as an intervention.The in vitro results also suggested that neuronal protection is mediated by the inhibition of JAK2 and STAT3 expression.Together,our findings indicate that the SOCS1/JAK2/STAT3 axis contributes to early brain injury after subarachnoid hemorrhage both in vitro and in vivo by inducing inflammatory responses.This study was approved by the Animal Ethics Committee of Anhui Medical University and the First Affiliated Hospital of University of Science and Technology of China(approval No.LLSC-20180202)on March 1,2018.
基金This work was sponsored by the National Nature Science Foundation of China(No.U1609207)the National Key Research and Development Program of China(2018YFA0703000)the Science Fund for Creative Research Groups of the National Natural Science Foundation of China(No.51521064).
文摘Biomedical field has been seeking a feasible standard drug screening system consisting of 3D tumor model array for drug researching due to providing sufficient samples and simulating actual in vivo tumor growth situation,which is still a challenge to rapidly and uniformly establish though.Here,we propose a novel drug screening system,namely 3D tumor array chip with“layer cake”structure,for drug screening.Accurate gelatin methacryloyl hydrogel droplets(~0.1μL)containing tumor cells can be automatically deposited on demand with electrohydrodynamic 3D printing.Transparent conductive membrane is introduced as a chip basement for preventing charges accumulation during fabricating and convenient observing during screening.Culturing chambers formed by stainless steel and silicon interlayer is convenient to be assembled and recycled.As this chip is compatible with the existing 96-well culturing plate,the drug screening protocols could keep the same as convention.Important properties of this chip,namely printing stability,customizability,accuracy,microenvironment,tumor functionalization,are detailly examined.As a demonstration,it is applied for screening of epirubicin and paclitaxel with breast tumor cells to confirm the compatibility of the proposed screening system with the traditional screening methods.We believe this chip will potentially play a significant role in drug evaluation in the future.
基金supported by the National Natural Science Foundation of China(No.81774204)Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences(No.CI2021A04608)。
文摘Background:Pudilan Xiaoyan Oral Liquid(PDL),a famous traditional Chinese formula for treating acute and chronic inflammation.To evaluate the broad-spectrum antiviral effect of Pudilan Xiaoyan Oral Liquid,and provide a basis for clinical medication.Methods:Its inhibitory effect on different respiratory viruses was observed by cytopathic test.The potential mechanism of the anti-influenza effect was determined by neuraminidase activity.In order to observe the therapeutic effect of PDL on viral pneumonia caused by different respiratory viruses.The viral pneumonia model was established by nasal infection with different respiratory viruses,and then PDL was given Therapeutic and prophylactically to evaluate its pharmacodynamic activity in vivo.Results:The results of in vitro experiments showed that PDL had different inhibitory effects on cytopathic effects caused by different respiratory viruses.And it has obvious inhibitory effect on the neuraminidase activity of influenza A virus,which indicates that it exerts anti-influenza virus effect by inhibiting neuraminidase activity of influenza virus.The results in vivo showed that PDL exhibited an inhibitory effect on pulmonary index(PI)and effectively reduced the degree of lesions in the lungs.The lethal rate of mice was significantly decreased while survival time of mice was dramatically increased by PDL treatment in comparison to infection control,respectively.Conclusions:Our study demonstrates that PDL had a significant protection and treatment effect for respiratory virus infection in vitro and in vivo.
文摘Adult-onset brain cancers,such as glioblastomas,are particularly lethal.People with glioblastoma multiforme(GBM)do not anticipate living for more than 15 months if there is no cure.The results of conventional treatments over the past 20 years have been underwhelming.Tumor aggressiveness,location,and lack of systemic therapies that can penetrate the blood–brain barrier are all contributing factors.For GBM treatments that appear promising in preclinical studies,there is a considerable rate of failure in phaseⅠandⅡclinical trials.Unfortunately,access becomes impossible due to the intricate architecture of tumors.In vitro,bioengineered cancer models are currently being used by researchers to study disease development,test novel therapies,and advance specialized medications.Many different techniques for creating in vitro systems have arisen over the past few decades due to developments in cellular and tissue engineering.Later-stage research may yield better results if in vitro models that resemble brain tissue and the blood–brain barrier are used.With the use of 3D preclinical models made available by biomaterials,researchers have discovered that it is possible to overcome these limitations.Innovative in vitro models for the treatment of GBM are possible using biomaterials and novel drug carriers.This review discusses the benefits and drawbacks of 3D in vitro glioblastoma modeling systems.
文摘Fibrotic tumors,such as pancreatic ductal adenocarcinoma(PDAC),are characterized for high desmoplastic reaction,which results in high intra-tumoral solid stress leading to the compression of blood vessels.These microarchitectural alterations cause loss of blood flow and poor intra-tumoral delivery of therapeutics.Currently,there is a lack of relevant in vitro models capable of replicating these mechanical characteristics and to test anti-desmoplastic compounds.Here,a multi-layered vascularized 3D PDAC model consisting of primary human pancreatic stellate cells(PSCs)embedded in collagen/fibrinogen(Col/Fib),mimicking tumor tissue within adjunct healthy tissue,is presented to study the fibrosis-induced compression of vasculature in PDAC.It is demonstrated how the mechanical and biological stimulation induce PSC activation,extracellular matrix production and eventually vessel compression.The clinical relevance is confirmed by correlating with patient transcriptomic data.Furthermore,the effects of gradual vessel compression on the fluid dynamics occurring within the channel is evaluated in silico.Finally,it is demonstrated how cancer-associated fibroblast(CAF)-modulatory therapeutics can inhibit the cell-mediated compression of blood vessels in PDAC in vitro,in silico and in vivo.It is envisioned that this 3D model is used to improve the understanding of mechanical characteristics in tumors and for evaluating novel anti-desmoplastic therapeutics.
基金financially supported by the 13th Five-Year National Significant New Drugs Creation Feature Subjects grant(2018ZX09735006)by the Project for Medicine and Medical Instruments Review and Approval System Reform grant(ZG2016-1)
文摘Background:The ingestion of golden thread contaminated with heavy metals through the food chain leads to detrimental effects to human health.During digestion,not all of the heavy metals could be released to the gastrointestinal tract and readily to be absorbed by human body.Thus,bioaccessibility is an important issue in health risk assessments.Aims and Objectives:The aims and objectives of this study were to investigate the bioaccessibility of Cd in golden thread and assess the associated health risks based on the exposure to bioaccessible Cd.Materials and Methods:Inductively coupled plasma mass spectrometry(ICP-MS)has been applied to determine the Cd content in golden thread.Physiologically based extraction test(PBET)digestion was performed in the in vitro/Caco2 cell model to investigate the bioaccessibility of Cd in golden thread.Furthermore,the target hazard quotient(THQ)was used to assess the risks of the total and the bioaccessible content of Cd in golden thread.Results:The results revealed that the total Cd content in six batches of golden thread ranged from 3.203 to 5.723 mg/kg.After uptake by Caco2 cells,the bioaccessibility of Cd ranged from 42.36%to 59.73%.The results of the risk assessment indicated that prior to uptake by Caco2 cells,the THQ values of Cd for all batches of golden thread were greater than 1.However,after uptake by Caco2 cells,the THQ values of Cd in all samples were less than 1,thus suggesting that the risks were at a safe level.Conclusion:This study was the first to perform health risk assessment with bioaccessible heavy metals present in traditional Chinese medicine by PBET digestion using an in vitro/Caco2 cell model,thus enabling us to obtain more accurate and objective results while allowing us to avoid unnecessary government intervention and to establish more reasonable limit standards for heavy metals.
基金The work was supported by grant from the National Natural Science Foundation of China(81771587).
文摘Decidualization is a special type of differentiation of endometrial stromal cells into secretory decidualized cells,which is closely related to the occurrence of menstruation and establishment of pregnancy.Decidualization abnormalities can cause female infertility and abortion,and the decidualization modelin vitro is an important tool for studying relevant mechanisms.This article summarizes severalin vitro decidualization models in recent research from three aspects,including the selection of model cells and culture systems,evaluation of decidualization markers,and induction schemes.These models can be appropriately selected and applied in specific endometrium-related disease models,such as endometriosis,recurrent pregnancy loss,and preeclampsia.
基金supported from the Key Research and Development Projects of Chinese People’s Liberation Army(Grant No.BWS17J036)Natural Science Foundation of China(Grant No.32000956)China Postdoctoral Science Foundation(Grant No.2020M670294).
文摘The pathogenic cascade of Alzheimer’s disease(AD)characterized by amyloid-β protein accumulation is still poorly understood,partially owing to the limitations of relevant models without in vivo neural tissue microenvironment to recapitulate cell-cell interactions.To better mimic neural tissue microenvironment,three-dimensional(3D)core-shell AD model constructs containing human neural progenitor cells(NSCs)with 2% matrigel as core bioink and 2% alginate as shell bioink have been bioprinted by a co-axial bioprinter,with a suitable shell thickness for nutrient exchange and barrier-free cell interaction cores.These constructs exhibit cell self-clustering and-assembling properties and engineered reproducibility with long-term cell viability and self-renewal,and a higher differentiation level compared to 2D and 3D MIX models.The different effects of 3D bioprinted,2D,and MIX microenvironments on the growth of NSCs are mainly related to biosynthesis of amino acids and glyoxylate and dicarboxylate metabolism on day 2 and ribosome,biosynthesis of amino acids and proteasome on day 14.Particularly,the model constructs demonstrated Aβ aggregation and higher expression of Aβ and tau isoform genes compared to 2D and MIX controls.AD model constructs will provide a promising strategy to facilitate the development of a 3D in vitro AD model for neurodegeneration research.
