Synthesis of Cell-Adhesive Anisotropic Multifunctional Particles by Stop Flow Lithography and Streptavidin–Biotin Interactions

Cell-adhesive particles are of significant interest in biotechnology, the bioengineering of complex tissues, and biomedical research. Their applications range from platforms to increase the efficiency of anchorage-dependent cell culture to building blocks to loading cells in heterogeneous structures to clonal-population growth monitoring to cell sorting. Although useful, currently available cell-adhesive particles can accommodate only homogeneous cell culture. Here, we report the design of anisotropic hydrogel microparticles with tunable cell-adhesive regions as first step toward micropatterned cell cultures on particles. We employed stop flow lithography (SFL), the coupling reaction between amine and N-hydroxysuccinimide (NHS) and streptavidin–biotin chemistry to adjust the localization of conjugated collagen and poly-l-lysine on the surface of microscale particles. Using the new particles, we demonstrate the attachment and formation of tight junctions between brain endothelial cells. We also demonstrate the geometric patterning of breast cancer cells on particles with heterogeneous collagen coatings. This new approach avoids the exposure of cells to potentially toxic photoinitiators and ultraviolet light and decouples in time the microparticle synthesis and the cell culture steps to take advantage of the most recent advances in cell patterning available for traditional culture substrates.National Institutes of Health (U.S.) (GM092804)National Science Foundation (U.S.) (CMMI-1120724 and DMR-1006147)Samsung Scholarship Foundatio

Human brain organoids in Alzheimer’s disease

Alzheimer’s disease (AD) is a progressive neurological disorder that typically involves neuronal damage leading to the deterioration of cognitive and essential body functions in aging brains. Major signatures of AD pathology include the deposition of amyloid plaques and neurofibrillary tangles, disruption of the blood-brain barrier, and induction of hyper-activated proinflammation in the brain, leading to synaptic impairment and neuronal loss. However, conventional pharmacotherapeutic modalities merely alleviate symptoms, but do not cure AD, partly because drug screening has used model systems with limited accuracy in terms of reflecting AD pathology in human brains. In this regard, several AD organoids have received substantial attention as alternatives to AD animal models. In this review, we summarize the key characteristics required for the generation of a pathologically relevant AD brain organoid. We also overview major experimental organoid models of AD brains, such as spheroids, three-dimensional (3D) bioprinted constructs, and 3D brain-on-chips, and discuss their strengths and weaknesses for AD research. This review will provide valuable information that will inspire future efforts to engineer authentic AD organoids for the study of AD pathology and for the discovery of novel AD therapeutic strategies

Therapeutic nanoplatforms and delivery strategies for neurological disorders

Abstract The major neurological disorders found in a central nervous system (CNS), such as brain tumors, Alzheimer’s diseases, Parkinson’s diseases, and Huntington’s disease, have led to devastating outcomes on the human public health. Of these disorders, early diagnostics remains poor, and no treatment has been successfully discovered; therefore, they become the most life-threatening medical burdens worldwide compared to other major diseases. The major obstacles for the drug discovery are the presence of a restrictive blood–brain barrier (BBB), limiting drug entry into brains and undesired neuroimmune activities caused by untargeted drugs, leading to irreversible neuronal damages. Recent advances in nanotechnology have contributed to the development of novel nanoplatforms and effective delivering strategies to improve the CNS disorder treatment while less disturbing brain systems. The nanoscale drug carriers, including liposomes, dendrimers, viral capsids, polymeric nanoparticles, silicon nanoparticles, and magnetic/metallic nanoparticles, enable the effective drug delivery penetrating across the BBB, the aforementioned challenges in the CNS. Moreover, drugs encapsulated by the nanocarriers can reach further deeper into targeting regions while preventing the degradation. In this review, we classify novel disease hallmarks incorporated with emerging nanoplatforms, describe promising approaches for improving drug delivery to the disordered CNS, and discuss their implications for clinical practice

An Air Particulate Pollutant Induces Neuroinflammation and Neurodegeneration in Human Brain Models

10.1002/advs.202101251Advanced Science821210125

Engineered Human Intervertebral Disc Model Inducing Degenerative Microglial Proinflammation

Degeneration of the intervertebral disc (IVD) is a major contributor to low back pain (LBP). IVD degeneration is characterized by abnormal production of inflammatory cytokines secreted by IVD cells. Although the underlying molecular mechanisms of LBP have not been elucidated, increasing evidence suggests that LBP is associated particularly with microglia in IVD tissues and the peridiscal space, aggravating the cascade of degenerative events. In this study, we implemented our microfluidic chemotaxis platform to investigate microglial inflammation in response to our reconstituted degenerative IVD models. The IVD models were constructed by stimulating human nucleus pulposus (NP) cells with interleukin-1β and producing interleukin-6 (129.93 folds), interleukin-8 (18.31 folds), C-C motif chemokine ligand-2 (CCL-2) (6.12 folds), and CCL-5 (5.68 folds). We measured microglial chemotaxis (p < 0.05) toward the conditioned media of the IVD models. In addition, we observed considerable activation of neurodegenerative and deactivation of protective microglia via upregulated expression of CD11b (p < 0.001) and down-regulation of CD206 protein (p < 0.001) by soluble factors from IVD models. This, in turn, enhances the inflammatory milieu in IVD tissues, causing matrix degradation and cellular damage. Our findings indicate that degenerative IVD may induce degenerative microglial proinflammation, leading to LBP development

Oral Pathogenic Bacteria-Inducing Neurodegenerative Microgliosis in Human Neural Cell Platform

Porphyromonas gingivalis is a gram-negative bacterium found in the human oral cavity and is responsible for the development of chronic periodontitis as well as neurological diseases, including Alzheimer’s disease (AD). Given the significance of the roles of P. gingivalis in AD pathogenesis, it is critical to understand the underlying mechanisms of P. gingivalis-driven neuroinflammation and their contribution to neurodegeneration. Herein, we hypothesize that P. gingivalis produces secondary metabolites that may cause neurodegeneration through direct or indirect pathways mediated by microglia. To test our hypothesis, we treated human neural cells with bacterial conditioned media on our brain platforms and assessed microgliosis, astrogliosis and neurodegeneration. We found that bacteria-mediated microgliosis induced the production of nitric oxide, which causes neurodegeneration assessed with high pTau level. Our study demonstrated the elevation of detrimental protein mediators, CD86 and iNOS and the production of several pro-inflammatory markers from stimulated microglia. Through inhibition of LPS and succinate dehydrogenase in a bacterial conditioned medium, we showed a decrease in neurodegenerative microgliosis. In addition, we demonstrated the bidirectional effect of microgliosis and astrogliosis on each other exacerbating neurodegeneration. Overall, our study suggests that the mouth-brain axis may contribute to the pathogenesis of AD

Physicochemical properties and bioavailability of naturally formulated fat-soluble vitamins extracted from agricultural products for complementary use for natural vitamin supplements

The purpose of the current study was to evaluate the physicochemical properties, digestive stability, storage stability, and intestinal absorption of formulated natural vitamins (FNV) by mixing fat-soluble vitamins extracted from agricultural products with their synthetic vitamin (SYNV) counterparts using a 6 to 4 ratio (w:w, dry weight). The FNV A, D, E, and K were evenly dispersed without crystal growth in the dispersion specifications for the functional tablet foods. The FNV A, D, E, and K had 89, 73, 65, and 36% of the digestive recovery, respectively, which was comparable to that of the SYNV. FNV D, E, and K were retained over 77%, but rapidly decreased to 15% after 6 months during accelerated storage at 25 30 and 35℃. The comparable radical scavenging capacity was found between the FNV and the SYNV. Results from the current study suggest that fat-soluble vitamins extracted from agricultural products could be reasonable complementary use for natural vitamin supplements

Development of Physiologically relevant Human Brain Models by Using Brain Decellularized Extracellular Matrix

Introduction: In-vitro human brain models have proven to be effective for platforms for drug screening as they reconstruct pathophysiological features found in brain diseases. Conventional culture systems, however, rely on synthetic or natural bioinks derived from tissues, other than brains, and therefore, they lack most of necessary components in the native microenvironment. In this regard, decellularized extracellular matrices have recently emerged as promising bioinks to combat this limitation. To facilitate more accurate pathophysiological study and drug screening, a recently developed our porcine derived brain decellularized extracellular matrix (BdECM) was combined with Matrigel, one of the gold standard hydrogels, and employed to our validated microfluidic human brain model in order to recapitulate microenvironment found in human brains. Materials and Methods: The BdECM was synthesized through decelluarization of porcine brains. The mixture of BdECM and Matrigel hybrid hydrogel was incorporated into ourthe recently developed our microfluidic models consisting of neurons, astrocytes and the microglia. To validate the model, we assessed microglial immune response by performing immunocytochemistry for CD11B and CD206. Furthermore, Tto quantitatively measure potential cytotoxicity of BdECM, lactate dehydrogenase (LDH) assay was performed. Results and Discussion : The gelation kinetics of BdECM hydrogel was tested at concentrations of 0.5, 1, 2, and 4% of BdECM. The results showed that BdECM was capable of achieving a storage modulus from 100 to 200Pa, which is comparable to the modulus found in human brain tissues. This indicates the inclusion of BdECM in the cell culture hydrogel may further improve the physical properties of conventional hydrogel, such as Matrigel. To check the physiological relevance of the combination of Matrigel and BdECM, we assessed the cytotoxicity of the mixture and found that the cytotoxicity of Matrigel and the mixture was 1.9 and 2.3% respectively indicating there was no significant increase in cytotoxicity with the addition ofthere was no significant cell death compared to Matrigel, as seen in Figure 1 BdECM. In addition,Even though the permeability of BdECM+Matrigel (0.0104 ± 0.0051 cm/s) was comparable tolower that of Matrigel (0.013 cm/s and 0.00678 ± 0.01395 cm/s respectively), the limited nutrient transport through the gel did not affect the cell viability. In addition, no immune response was found when the microglia cultured on BdECM, determined by microglial activation markers, such as CD11B and CD206. Conclusions: Our results showed that BdECM hydrogel will provide relevant stiffness to native brain and the hybrid of BdECM and Matrigel provided microenvironments for brain cells without causing significant cell death and immune response. We are expecting that this gel recipe will offer a promising cell culture platform for in vitro brain models, which closely recapitulate human brain.1