using-multilayered-hydrogel-bioink-three-dimensional-bioprinting-for

Using Multilayered Hydrogel Bioink in Three-Dimensional Bioprinting for Homogeneous Cell Distribution

During the extrusion-based three-dimensional bioprinting process, liquid-like bioinks with low viscosity can protect cells from membrane damage induced by ...

construction-human-aorta-smooth-muscle-cell-organ-on-chip-model-for

Construction of a Human Aorta Smooth Muscle Cell Organ-On-A-Chip Model for Recapitulating Biomechanical Strain in the Aortic Wall

Conventional two-dimensional cell culture techniques and animal models have been used in the study of human thoracic aortic aneurysm and dissection ...

Fudan University Pudong Medical Center

Fudan University

Although there are numerous claims of remote group participation leading to the synthesis of the expected glycosides with improved anomeric geometry outcome in glycosylation, there is still a lack of enough strong evidence and this has led to a long-term debate, particularly for equatorial 4-O group participation. In this work, we were able to synthesize and isolate a stable seven-membered trichlorooxazepine ring bridging intermediate with a high yield by employing a 2-azido-2-deoxy-glucopyranosyl donor, which provides strong evidence to support the putative participation of the equatorial 4-O group in glycosylation.

Evidence of robust participation by an equatorial 4-O group in glycosylation on a 2-azido-2-deoxy-glucopyranosyl donor.

A versatile strategy for the synthesis of 6″-functionalized α-GalCers by using NAP ether group for permanent hydroxyl protection was developed, which provide the flexibility necessary for the incorporation of a wide range of functional groups in target molecules including alkyne, azide, thiol that are intolerant to Pd-catalyzed hydrogenolysis as well as other functionalities like carboxylic acid and amine. This strategy is also adaptable to other glycoconjugate synthesis especially those containing clickable tags and unsaturated functionalities.

Rapid access to 6″-functionalized α-galactosyl ceramides by using 2-naphthylmethyl ether as the permanent protecting group.

Starting from readily available vanillin and α-D-acetobromo glucose, two natural acylated phenolic glycosides vitexnegheteroin A and ovatoside D were synthesized for the first time in 4 steps with overall yields of 54% and 65%, respectively. The key steps involve the directly regioselective O-6 acylation of vanillin β-D-glucopyranoside with acyl chlorides, and simultaneous removal of the allyl protecting groups on the phenolic acid moiety and reduction of the aldehyde in the aglycon moiety by using Pd(PPh)-NaBH system in one pot.

Concise total synthesis of acylated phenolic glycosides vitexnegheteroin A and ovatoside D.

Cancer immunotherapy fights against cancer by modulating the immune response and is delivering encouraging results in clinical treatments. However, it is challenging to achieve durable response in all cancer patients during treatment due to the diversity and dynamic nature of immune system as well as inter- and intratumor heterogeneity. A comprehensive assessment of system immunity and tumor microenvironment is crucial for effective and safe cancer therapy, which can potentially be resolved by single-cell proteomic analysis. Single-cell proteomic technologies enable system-wide profiling of protein levels in a number of single cells within the immune system and tumor microenvironment, and thereby provide direct assessment of the functional state of the immune cells and tumor-immune interaction that could be used to evaluate efficacy of immunotherapy and to improve clinical outcome. In this chapter, we summarized current single-cell proteomic technologies and their applications in cancer immunotherapy.

Single-Cell Proteomics for Cancer Immunotherapy.

One of the challenges for extrusion bioprinting using low-viscosity bioinks is the fast gravity-driven sedimentation of cells. Cells in a hydrogel bioink that features low viscosity tend to settle to the bottom of the bioink reservoir, and as such, their bioprintability is hindered by association with the inhomogeneous cellularized structures that are deposited. This is particularly true in cases where longer periods are required to print complex or larger tissue constructs. Increasing the bioink's viscosity efficiently retards sedimentation but gives rise to cell membranolysis or functional disruption due to increased shear stress on the cells during the extrusion process. Inspired by the rainbow cocktail, we report the development of a multilayered modification strategy for gelatin methacryloyl (GelMA) bioink to manipulate multiple liquid interfaces, providing interfacial retention to retard cell sedimentation in the bioink reservoir. Indeed, the interfacial tension in our layer-by-layer bioink system, characterized by the pendant drop method, was found to be exponentially higher than the sedimental pull (Δ = ∼10 N) of cells, indicating that the interfacial retention is crucial for preventing cell sedimentation across the adjacent layers. It was demonstrated that the encapsulated cells displayed better dispersibility in constructs bioprinted using the multilayered GelMA bioink system than that of pristine GelMA where the index of homogeneity of the cell distribution in the multilayered bioink was 4 times that of the latter.

Hydrogel Bioink with Multilayered Interfaces Improves Dispersibility of Encapsulated Cells in Extrusion Bioprinting.

The organ-on-a-chip model mimics the structural and functional features of human tissues or organs and has great importance in translational research. For vessel-on-a-chip model, conventional fabrication techniques are unable to efficiently imitate the intimal-medial unit of the vessel wall. Bioprinting technology, which can precisely control the organization of cells, biomolecules, and the extracellular matrix, has the potential to fabricate three-dimensional (3D) tissue constructs with spatial heterogeneity. In this study, we applied the gelatin-methacryloyl-based bioprinting technology to print 3D construct containing endothelial cells (ECs) and smooth muscle cells (SMCs) on a microfluidic chip. Compared with traditional culture system, EC-SMC coculturing chip model upregulated αSMA and SM22 protein expression of the SMC to a greater degree and maintains the contractile phenotype of the SMC, which mimics the natural vascular microenvironment. This strategy enabled us to establish an in vitro vascular model for studies of the physiologic and pathologic process in vascular wall.

Bioprinting a 3D vascular construct for engineering a vessel-on-a-chip.

A major goal of polydimethylsiloxane (PDMS) microfabrication is to develop a simple and inexpensive method for rapid fabrication. Despite the recent advancements in this field, facile PDMS microfabrication on non-planar surfaces remains elusive. Here we report a facile method for rapid prototyping of PDMS microdevices viaPLAT (microscale plasma-activated templating) on non-planar surfaces through micropatterning of hydrophilic/hydrophobic (HL/HB) interface by flexible polyvinyl chloride (PVC) hollow-out mask. This mask can be easily prepared with flexible PVC film through a cutting crafter and applied as pattern definer during the plasma treatment for microscale HL/HB interface formation on different substrates. The whole process requires low inputs in terms of time as well as toxic chemicals. Inspired by liquid molding, we demonstrated its use for rapid prototyping of PDMS microstructures. Following the proof-of-concept study, we also demonstrated the use of the flexible hollow-out mask to facilitate cell patterning on curved substrates, which is difficult to realize with conventional methods. Collectively, our work utilizes flexible and foldable PVC film as mask materials for facile microscale HL non-planar surface modification to establish a useful tool for PDMS prototyping and cell patterning.

Rapid prototyping of PDMS microdevices viaPLAT on nonplanar surfaces with flexible hollow-out mask.

Bicuspid aortic valve (BAV) is the most common congenital heart anomaly and is prone to cause complications, such as valvular stenosis and thoracic aortic dilation. There is currently no reliable way to predict the progression rate to thoracic aortic aneurysm. Here, we aimed to characterize the proteomic landscape in the plasma of stenotic BAV patients and provide potential biomarkers to predict progressive aortic dilation.

Plasma proteomic profiling reveals biomarkers associated with aortic dilation in patients with bicuspid aortic valve.

Bicuspid aortic valve (BAV) is the most common congenital cardiovascular disease in general population and is frequently associated with the development of thoracic aortic aneurysm (TAA). There is no effective strategy to intervene with TAA progression due to an incomplete understanding of the pathogenesis. Insufficiency of NOTCH1 expression is highly related to BAV-TAA, but the underlying mechanism remains to be clarified.

Aorta smooth muscle-on-a-chip reveals impaired mitochondrial dynamics as a therapeutic target for aortic aneurysm in bicuspid aortic valve disease.

To assure complete tumor removal, frozen section analysis is the most common procedure for intraoperative pathological assessment of resected tumor margins. However, during one operation, multiple biopsies may be sent for examination, but only few of them are made into cryosections because of the complex preparation protocols and time-consuming pathological analysis, which potentially increases the risk of overlooking tumor involvement. Here, we propose a fluorescence-based pre-screening strategy that allows high-throughput, convenient, and fast gross assessment of resected tumor margins. A dual-activatable cationic fluorescent molecular rotor was developed to specifically illuminate live tumor cells' cytoplasm by emitting two different fluorescence signals in response to elevations in hypoxia-induced nitroreductase (a biochemical marker) and cytoplasmic viscosity (a biophysical marker), two characteristics of cancer cells. The ability of the fluorescent molecular rotor in detecting tumor cells was evaluated in mouse and human specimens of multiple tissues by comparing with hematoxylin and eosin staining. Importantly, the fluorescent molecular rotor achieved 100 % specificity in discriminating lung and liver cancers from normal tissue, allowing pre-screening of the tumor-free surgical margins and promoting clinical decision. Altogether, this type of fluorescent molecular rotor and the proposed strategy may serve as a new option to facilitate intraoperative assessment of resected tumor margins.

A pre-screening strategy to assess resected tumor margins by imaging cytoplasmic viscosity and hypoxia.

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in millions of deaths and seriously threatened public health and safety. Despite COVID-19 vaccines being readily popularized worldwide, targeted therapeutic agents for the treatment of this disease remain very limited. Here, we studied the inhibitory activity of the scutellarein and its methylated derivatives against SARS-CoV-2 main protease (M) by the fluorescence resonance energy transfer (FRET) assay. Among all the methylated derivatives we studied, 4'-O-methylscutellarein exhibited the most promising enzyme inhibitory activity in vitro, with the half-maximal inhibitory concentration value (IC) of 0.40 ± 0.03 μM. Additionally, the mechanism of action of the hits was further characterized through enzyme kinetic studies and molecular docking. Overall, our results implied that 4'-O-methylscutellarein could be a primary lead compound with clinical potential for the development of inhibitors against the SARS-CoV-2 M.

Discovery of 4'-O-methylscutellarein as a potent SARS-CoV-2 main protease inhibitor.

Thoracic aortic aneurysm (TAA) is the permanent dilation of the thoracic aortic wall that predisposes patients to lethal events such as aortic dissection or rupture, for which effective medical therapy remains scarce. Human-relevant microphysiological models serve as a promising tool in drug screening and discovery.

ArticleTotal : 2	Year
Publication title Cited by 4	2020
Publication title Cited by 1	2022

Article	Year
Evidence of robust participation by an equatorial 4-O group in glycosylation on a 2-azido-2-deoxy-glucopyranosyl donor. Chemical communications (Cambridge, England)\| PubMed ID: 28239733	2017
Rapid access to 6″-functionalized α-galactosyl ceramides by using 2-naphthylmethyl ether as the permanent protecting group. Bioorganic & medicinal chemistry letters\| PubMed ID: 28274628	2017
Concise total synthesis of acylated phenolic glycosides vitexnegheteroin A and ovatoside D. Carbohydrate research\| PubMed ID: 29522916	2018
Single-Cell Proteomics for Cancer Immunotherapy. Advances in cancer research\| PubMed ID: 29941105	2018
Hydrogel Bioink with Multilayered Interfaces Improves Dispersibility of Encapsulated Cells in Extrusion Bioprinting. ACS applied materials & interfaces\| PubMed ID: 31378063	2019
Bioprinting a 3D vascular construct for engineering a vessel-on-a-chip. Biomedical microdevices\| PubMed ID: 31875940	2019
Rapid prototyping of PDMS microdevices viaPLAT on nonplanar surfaces with flexible hollow-out mask. Biofabrication\| PubMed ID: 33418543	2021
Plasma proteomic profiling reveals biomarkers associated with aortic dilation in patients with bicuspid aortic valve. Annals of translational medicine\| PubMed ID: 34430623	2021
Aorta smooth muscle-on-a-chip reveals impaired mitochondrial dynamics as a therapeutic target for aortic aneurysm in bicuspid aortic valve disease. eLife\| PubMed ID: 34486519	2021
A pre-screening strategy to assess resected tumor margins by imaging cytoplasmic viscosity and hypoxia. eLife\| PubMed ID: 34633289	2021
Discovery of 4'-O-methylscutellarein as a potent SARS-CoV-2 main protease inhibitor. Biochemical and biophysical research communications\| PubMed ID: 35303682	2022
Patient-derived microphysiological model identifies the therapeutic potential of metformin for thoracic aortic aneurysm. EBioMedicine\| PubMed ID: 35636318	2022

Shiqiang Yan

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