Therapeutic options for neurological disorders currently remain limited. The intrinsic complexity of the brain architecture prevents potential therapeutics from reaching their cerebral target, thus ...limiting their efficacy. Recent advances in microfluidic technology and organ-on-chip systems have enabled the development of a new generation of in vitro platforms that can recapitulate complex in vivo microenvironments and physiological responses. In this context, microfluidic-based in vitro models of the blood–brain barrier (BBB) are of particular interest as they provide an innovative approach for conducting research related to the brain, including modeling of neurodegenerative diseases and high-throughput drug screening. Here, we present the most recent advances in BBB-on-chip devices and examine validation steps that will strengthen their future applications.
Microfluidic-based blood–brain barrier-on-chip (μBBB) technology is a powerful approach to study the physiological function of the BBB in vitro and to facilitate drug discovery targeting brain disorders.Mimicry of the complexity of multiple cell crosstalk and thin extracellular matrix as basal membrane are essential but challenging. Different biomaterials and chip designs have been explored in the fabrication of μBBBs.Other key features such as shear stress, cell type/origin, and cell co-culture spatial configuration must be carefully controlled and selected. Appropriate BBB permeability assays and parameters (e.g., TEER measurement, small molecule drugs, and fluorescent probes) should be standardized and compared with in vivo data.μBBBs hold great potential in disease modeling, drug discovery, neurotoxicity screening, and personalized medicine applications.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
BACKGROUND and AIM: The regenerative and anti-inflammatory potential of mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) is well established. Bronchopulmonary dysplasia (BPD) is a ...lung pathology developed by premature babies. In a rat model of BPD, our study found that MSC-EVs could prevent oxidative stress and fibrosis, the two distinct signs of the disease. To clarify the mechanism of action of MSC-EVs, in vitro experiments were assessed to test the efficacy of Good Manufacturing Practice-grade (GMP-grade) MSC-EVs against oxidative stress and fibrosis.
The fibrosis assay considered the use of primary macrophages isolated from rat bone marrow. After 7 days of TGFβ1 treatment, αSMA and CD90 expressions were analysed by flow cytometry to test the anti-fibrotic ability of MSC-EVs in comparison with a positive control represented by TGFβ receptor I inhibitor. Cells were analysed for profibrotic markers by qRT-PCR, immunofluorescence, and flow cytometry. Human alveolar epithelial cells collected from healthy donors were used for oxidative stress assay. Cells were damaged by hydrogen peroxide and rotenone. The efficacy of MSC-EVs in restoring oxidative damage was investigated in comparison with vitamin C over 10 days. Immunofluorescence was used to analyse DNA oxidation and cell proliferation.
MSC-EVs inhibited αSMA induction and collagen deposition in macrophages more strongly than TGFβ receptor I inhibitors in a dose-dependent manner. Furthermore, MSC-EVs were able to protect alveolar epithelial cells from DNA oxidation in a dose-responsive manner. MSC-EV was a better scavenger than vitamin C. MSC-EVs functioned by downregulating cell proliferation and subsequently repairing DNA damage.
MSC-EVs can counteract the development of fibrosis and oxidative stress in BPD possibly interfering in TGFβ activation.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Systems-oriented approaches involving integrated in vitro and in silico models are emerging as a basis for future risk assessment strategies to address the current challenge in chemical risk ...assessment of increased numbers of registered chemicals with no toxicological data. Liver toxicity poses a major concern in risk assessment as the liver is exposed to most xenobiotic compounds and their bioactive metabolites, making the liver susceptible to chemically induced liver diseases. Nonalcoholic fatty liver disease (NAFLD) is a rising global burden with an estimated prevalence of about 25% in the general population. Risk factors for NAFLD include metabolic disorders, genetics, drugs, and environmental exposures. To integrate in vitro data into computational models and use in vitro to in vivo extrapolation to systematically assess the risk of chemicals to induce NAFLD, sophisticated human cell models are needed. Reviewed here are characteristics and limitations of 2D and 3D monoculture and coculture human liver cell models, including microphysiological systems, in which NAFLD-relevant end points were addressed together with their results. The aim of this review is to inform selection of experimental test systems depending on the toxicological question to be addressed and to promote the further development of in vitro approaches that address existing technological gaps.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Gynaecological cancers originate within the female reproductive system and are classified according to the site in the reproductive system where they arise. However, over 50 % of these malignancies ...are categorized as rare, encompassing 30 distinct histological subtypes, which complicates their diagnosis and treatment. The focus of this review is to give an overview of established in vitro models for the investigation of rare gynaecological cancers, as well as an overview of available online databases that contain detailed descriptions of cell line characteristics. Cell lines represent the main models for the research of carcinogenesis, drug resistance, pharmacodynamics and novel therapy treatment options. Nowadays, classic 2D cell models are increasingly being replaced with 3D cell models, such as spheroids, organoids, and tumoroids because they provide a more accurate representation of numerous tumour characteristics, and their response to therapy differs from the response of adherent cell lines. It is crucial to use the correct cell line model, as rare tumour types can show characteristics that differ from the most common tumour types and can therefore respond unexpectedly to classic treatment. Additionally, some cell lines have been misclassified or misidentified, which could lead to false results. Even though rare gynaecological cancers are rare, this review will demonstrate that there are available options for investigation of such cancers in vitro on biologically relevant models.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Absorption, distribution, metabolism and excretion (ADME) studies represent a fundamental step in the early stages of drug discovery. In particular, the absorption of orally administered drugs, which ...occurs at the intestinal level, has gained attention since poor oral bioavailability often led to failures for new drug approval.
In this context, several in vitro preclinical models have been recently developed and optimized to better resemble human physiology in the lab and serve as an animal alternative to accomplish the 3Rs principles. However, numerous models are ineffective in recapitulating the key features of the human small intestine epithelium and lack of prediction potential for drug absorption and metabolism during the preclinical stage.
In this review, we provide an overview of in vitro models aimed at mimicking the intestinal barrier for pharmaceutical screening. After briefly describing how the human small intestine works, we present i) conventional 2D synthetic and cell-based systems, ii) 3D models replicating the main features of the intestinal architecture, iii) micro-physiological systems (MPSs) reproducing the dynamic stimuli to which cells are exposed in the native microenvironment. In this review, we will highlight the benefits and drawbacks of the leading intestinal models used for drug absorption and metabolism studies.
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•Poor oral drug bioavailability is the primary cause of failures for new drugs approval.•Intestinal in vitro models include 2D traditional monolayers, complex 3D systems and emerging fluid-dynamic platforms.•Multicellular models display a good correlation between in vitro permability data and the fractions absorbed in humans.•Microphysiological systems closely recapitulate the intestinal physiological cues but still need to be standardized.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Commercialization of Organoids Choudhury, Deepak; Ashok, Aswathi; Naing, May Win
Trends in molecular medicine,
March 2020, 2020-Mar, 2020-03-00, 20200301, Volume:
26, Issue:
3
Journal Article
Peer reviewed
Organoids have been successfully exploited for drug screening, disease modeling, pathogenesis, and regenerative medicine. Herein, we discuss the progress achieved in the commercialization of ...organoids in the last few years. We further elaborate on the concept of organoid biobank and highlight ethical and regulatory issues surrounding organoid research and commercialization.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The influence of microorganisms growing as sessile communities in a large number of human infections has been extensively studied and recognized for 30-40 years, therefore warranting intense ...scientific and medical research. Nonetheless, mimicking the biofilm-life style of bacteria and biofilm-related infections has been an arduous task. Models used to study biofilms range from simple in vitro to complex in vivo models of tissues or device-related infections. These different models have progressively contributed to the current knowledge of biofilm physiology within the host context. While far from a complete understanding of the multiple elements controlling the dynamic interactions between the host and biofilms, we are nowadays witnessing the emergence of promising preventive or curative strategies to fight biofilm-related infections. This review undertakes a comprehensive analysis of the literature from a historic perspective commenting on the contribution of the different models and discussing future venues and new approaches that can be merged with more traditional techniques in order to model biofilm-infections and efficiently fight them.
Responses of the indigenous human gut commensal microbiota to iron are poorly understood because of an emphasis on in vitro studies of pathogen iron sensitivity. In a study of iron supplementation in ...healthy humans, we identified gradual microbiota shifts in some participants correlated with bacterial iron internalization. To identify direct effects due to taxon-specific iron sensitivity, we used participant stool samples to derive diverse in vitro communities. Iron supplementation of these communities caused small compositional shifts, mimicking those in vivo, whereas iron deprivation dramatically inhibited growth with irreversible, cumulative reduction in diversity and replacement of dominant species. Sensitivity of individual species to iron deprivation in axenic culture generally predicted iron dependency in a community. Finally, exogenous heme acted as a source of inorganic iron to prevent depletion of some species. Our results highlight the complementarity of in vivo and in vitro studies in understanding how environmental factors affect gut microbiotas.
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•Iron supplementation elicits small, person-specific effects on healthy gut microbiomes•Iron deprivation gradually leads to irreversible changes in community structure•Taxon-specific sensitivity to iron deprivation can predict changes in a community•Heme buffers effects of iron deprivation by acting as a source of inorganic iron
In vitro communities enable characterization of perturbation-specific effects on community structure outside the complex host environment. By pairing a human study of the effects of iron supplementation on the gut microbiome with in vitro community experiments, we provide a foundation for further characterization of iron metabolism in healthy human gut communities.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Nonalcoholic steatohepatitis (NASH) is a severe form of liver injury that can be caused by a variety of stimuli and has a significant mortality rate. A common technique to induce in vitro steatosis ...involves culturing primary human hepatocytes (PHH) in fatty acid‐enriched media. This study compared the lipidome of PHH cultured in fatty acid‐enriched media to hepatocytes from patients with NASH and healthy controls. Hepatocytes from NASH patients displayed increased total cellular abundance of glycerolipids and phospholipids compared to healthy control hepatocytes. PHH cultured in fatty acid‐enriched media demonstrated increased glycerolipids. However, these culture conditions did not induce elevated phospholipid levels. Thus, culturing PHH in fatty acid‐enriched media has limited capacity to emulate the environment of hepatocytes in NASH patients.
The lipidome of hepatocytes from nonalcoholic steatohepatitis (NASH) patients displays increased abundance of glycerolipids and phospholipids compared to healthy controls. Primary human hepatocytes (PHH) cultured in fatty acid‐enriched media, a common technique for inducing in vitro steatosis, only increased glycerolipids. Thus, culturing PHH in fatty acid‐enriched media has limited capacity to emulate the environment of hepatocytes in NASH patients.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
In vitro human osteoarthritis (OA)-mimicking models enabling pathophysiological studies and evaluation of emerging therapies such as cartilage tissue engineering are of great importance.
We describe ...the development and characterization of a human OA osteochondral organ culture. We also apply this model for evaluation of the phenotype maintenance of a human MSC derived engineered cartilage, as an example of emerging therapeutics, under long term exposure to the OA-mimicking environment. We also test the sensitivity of the model to a series of external factors and a potential disease-modifying agent, in terms of chondrogenic phenotype maintenance of the engineered cartilage, under OA-mimicking environment.
Excised joint tissues from total knee replacement surgeries were carved into numerous miniaturized and standardized osteochondral plugs for subsequent OA organ culture. The organ cultures were characterized in detail before being co-cultured with a tissue engineered cartilage. The chondrogenic phenotype of the tissue engineered cartilage co-cultured in long term up to 8 weeks under this OA-mimicking microenvironment was evaluated. Using the same co-culture model, we also screened for a number of biomimetic environmental factors, including oxygen tension, the presence of serum and the application of compression loading. Finally, we studied the effect of a matrix metalloprotease inhibitor, as an example of potential disease-modifying agents, on the co-cultured engineered cartilage.
We demonstrate that cells in the OA organ culture were viable while both the typical chondrogenic phenotype and the characteristic OA phenotype were maintained for long period of time. We then demonstrate that upon co-culture with the OA-mimicking organ culture, the engineered cartilage initially exhibited a more fibrocartilage phenotype but progressively reverted back to the chondrogenic phenotype upon long term co-culture up to 8 weeks. The engineered cartilage was also found to be sensitive to all biomimetic environmental factors screened (oxygen tension, serum and compression). Moreover, under the effect of a MMP inhibitor, the chondrogenic phenotype of engineered cartilage was better maintained.
We demonstrated the development of a human OA osteochondral organ culture and tested the feasibility and potential of using this model as an in vitro evaluation tool for emerging cartilage therapies.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK, ZRSKP
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