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modeling gut neuro-epithelial connections in a novel microfluidic device

microsystems & Nanoengineering 2023年第6期9卷 235-246页

作者： Jose m.de Hoyos-Vega Xi Yu alan m.gonzalez-suarez Sisi Chen Arnaldo mercado-Perez Eugene Krueger Jeric Hernandez Yaroslav Fedyshyn Brooke R.Druliner David R.Linden Arthur Beyder Alexander Revzin Department of Physiology and Biomedical Engineering Mayo ClinicRochesterMNUSA Enteric Neuroscience Program(ENSP) Mayo ClinicRochesterMNUSA Division of Gastroenterology and Hepatology Department of MedicineMayo ClinicRochesterMNUSA

The intestinal lumen is filled with diverse chemical and physical stimuli.Intestinal epithelial cells sense these stimuli and signal to enteric neurons which coordinate a range of physiologic processes required for normal digestive tract function.Yet,the neuro-epithelial connections remain poorly resolved,in part because the tools for orchestrating interactions between these cellular compartments are lacking.We describe the development of a two-compartment microfluidic device for co-culturing enteric neurons with intestinal epithelial cells.The device contains epithelial and neuronal compartments connected by microgrooves.The epithelial compartment was designed for cell seeding via injection and confinement of intestinal epithelial cells derived from human intestinal organoids.We demonstrated that organoids planarized effectively and retained epithelial phenotype for over a week.In the second chamber we dissociated and cultured intestinal myenteric neurons including intrinsic primary afferent neurons(IPANs)from transgenic mice that expressed the fluorescent protein tdTomato.IPANs extended projections into microgrooves,surrounded and frequently made contacts with epithelial cells.The density and directionality of neuronal projections were enhanced by the presence of epithelial cells in the adjacent compartment.Our microfluidic device represents a platform that may,in the future,be used to dissect structure and function of neuro-epithelial connections in the gut and other organs(skin,lung,bladder,and others)in health and disease.

关键词： fluid bladder organs

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Bioactive hydrogel microcapsules for guiding stem cell fate decisions by release and reloading of growth factors

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Bioactive materials 2022年第9期7卷 1-14页

作者： Kihak Gwon Hye Jin Hong alan m.gonzalez-suarez michael Q.Slama Daheui Choi Jinkee Hong Harihara Baskaran Gulnaz Stybayeva Quinn P.Peterson Alexander Revzin Department of Physiology and Biomedical Engineering Mayo ClinicRochesterMN55902USA Department of Chemical and Biomolecular Engineering Yonsei UniversitySeoul03722Republic of Korea Department of Chemical and Biomolecular Engineering Case Western Reserve UniversityClevelandOH44106USA

Human pluripotent stem cells(hPSC)hold considerable promise as a source of adult cells for treatment of diseases ranging from diabetes to liver failure.Some of the challenges that limit the clinical/translational impact of hPSCs are high cost and difficulty in scaling-up of existing differentiation protocols.In this paper,we sought to address these challenges through the development of bioactive microcapsules.A co-axial flow focusing microfluidic device was used to encapsulate hPSCs in microcapsules comprised of an aqueous core and a hydrogel shell.Importantly,the shell contained heparin moieties for growth factor(GF)binding and release.The aqueous core enabled rapid aggregation of hPSCs into 3D spheroids while the bioactive hydrogel shell was used to load inductive cues driving pluripotency maintenance and endodermal differentiation.Specifically,we demonstrated that one-time,1 h long loading of pluripotency signals,fibroblast growth factor(FGF)-2 and transforming growth factor(TGF)-β1,into bioactive microcapsules was sufficient to induce and maintain pluripotency of hPSCs over the course of 5 days at levels similar to or better than a standard protocol with soluble GFs.Furthermore,stem cell-carrying microcapsules that previously contained pluripotency signals could be reloaded with an endodermal cue,Nodal,resulting in higher levels of endodermal markers compared to stem cells differentiated in a standard protocol.Overall,bioactive heparin-containing core-shell microcapsules decreased GF usage five-fold while improving stem cell phenotype and are well suited for 3D cultivation of hPSCs.

关键词： Droplet microfluidics Bioactive core-shell microcapsule Growth factor release 3D stem cell culture Stem cell differentiation

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Function of hepatocyte spheroids in bioactive microcapsules is enhanced by endogenous and exogenous hepatocyte growth factor

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Bioactive materials 2023年第10期28卷 183-195页

作者： Kihak Gwon Daheui Choi Jose mde Hoyos-Vega Harihara Baskaran alan m.gonzalez-suarez Seonhwa Lee Hye Jin Hong Kianna m.Nguyen Ether Dharmesh Go Sugahara Yuji Ishida Takeshi Saito Gulnaz Stybayeva Alexander Revzin Department of Physiology and Biomedical Engineering Mayo ClinicRochesterMNUSA Department of Chemical and Biomolecular Engineering Case Western Reserve UniversityClevelandOHUSA Biomedical Engineering Saint Louis UniversitySt.LouisMOUSA University of Southern Californi1 Keck School of MedicineDepartment of MedicineDivision of Gastrointestinal and Liver DiseasesLos AngelesCAUSA Research and Development Department PhoenixBioCo.LtdKagamiyam1Higashi-Hiroshim1Hiroshim1Japan USC Research Center for Liver Diseases Los AngelesCAUSA

The ability to maintain functional hepatocytes has important implications for bioartificial liver development,cell-based therapies,drug screening,and tissue engineering.Several approaches can be used to restore hepatocyte function in vitro,including coating a culture substrate with extracellular matrix(ECm),encapsulating cells within biomimetic gels(Collagen-or matrigel-based),or co-cultivation with other cells.This paper describes the use of bioactive heparin-based core-shell microcapsules to form and cultivate hepatocyte spheroids.These microcapsules are comprised of an aqueous core that facilitates hepatocyte aggregation into spheroids and a heparin hydrogel shell that binds and releases growth factors.We demonstrate that bioactive microcapsules retain and release endogenous signals thus enhancing the function of encapsulated hepatocytes.We also demonstrate that hepatic function may be further enhanced by loading exogenous hepatocyte growth factor(HGF)into microcapsules and inhibiting transforming growth factor(TGF)-β1 signaling.Overall,bioactive microcapsules described here represent a promising new strategy for the encapsulation and maintenance of primary hepatocytes and will be beneficial for liver tissue engineering,regenerative medicine,and drug testing applications.

关键词： Droplet microfluidics Bioactive microcapsules Hepatocyte spheroid Heparin Hepatocyte growth factor

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A versatile microfluidic device for multiple ex vivo/ in vitro tissue assays unrestrained from tissue topography

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microsystems & Nanoengineering 2020年第1期6卷 806-819页

作者： Jose m.de Hoyos-Vega alan m.gonzalez-suarez Jose L.Garcia-Cordero Unidad monterrey Centro de Investigacion y de Estudios Avanzados del IPNVia del conocimiento 201Parque PIITApodacaNL 66628Mexico

Precision-cut tissue slices are an important in vitro system to study organ function because they preserve most of the native cellular microenvironments of organs,including complex intercellular connections.However,during sample manipulation or slicing,some of the natural surface topology and structure of these tissues is lost or damaged.Here,we introduce a microfluidic platform to perform multiple assays on the surface of a tissue section,unhindered by surface topography.The device consists of a valve on one side and eight open microchannels located on the opposite side,with the tissue section sandwiched between these two structures.When the valve is actuated,eight independent microfluidic channels are formed over a tissue section.This strategy prevents cross-contamination when performing assays and enables parallelization.Using irregular tissues such as an aorta,we conducted multiple in vitro and ex vivo assays on tissue sections,including short-term culturing,a drug toxicity assay,a fluorescence immunohistochemistry staining assay,and an immune cell assay,in which we observed the interaction of neutrophils with lipopolysaccharide(LPS)-stimulated endothelium.Our microfluidic platform can be employed in other disciplines,such as tissue physiology and pathophysiology,morphogenesis,drug toxicity and efficiency,metabolism studies,and diagnostics,enabling the conduction of several assays with a single biopsy sample.

关键词： metabolism topography versatile

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A microfluidic platform for cultivating ovarian cancer spheroids and testing their responses to chemotherapies

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microsystems & Nanoengineering 2020年第1期6卷 315-326页

作者： Neda Dadgar alan m.gonzalez-suarez Pouria Fattahi Xiaonan Hou John S.Weroha Alexandre Gaspar-maia Gulnaz Stybayeva Alexander Revzin Department of Physiology and Biomedical Engineering Mayo ClinicRochesterMN 55905USA Department of medical Oncology Mayo ClinicRochesterMN 55905USA Department of Laboratory medicine and Pathology Mayo ClinicRochesterMN 55905USA

There is increasing interest in utilizing in vitro cultures as patient avatars to develop personalized treatment for cancer.Typical cultures utilize matrigel-coated plates and media to promote the proliferation of cancer cells as spheroids or tumor explants.However,standard culture conditions operate in large volumes and require a high concentration of cancer cells to initiate this process.Other limitations include variability in the ability to successfully establish a stable line and inconsistency in the dimensions of these microcancers for in vivo drug response measurements.This paper explored the utility of microfluidics in the cultivation of cancer cell spheroids.Six patient-derived xenograft(PDX)tumors of high-grade serous ovarian cancer were used as the source material to demonstrate that viability and epithelial marker expression in the microfluidic cultures was superior to that of matrigel or large volume 3D cultures.To further demonstrate the potential for miniaturization and multiplexing,we fabricated multichamber microfluidic devices with integrated microvalves to enable serial seeding of several chambers followed by parallel testing of several drug concentrations.These valve-enabled microfluidic devices permitted the formation of spheroids and testing of seven drug concentrations with as few as 100,000 cancer cells per device.Overall,we demonstrate the feasibility of maintaining difficul-to-culture primary cancer cells and testing drugs in a microfluidic device.This microfluidic platform may be ideal for drug testing and personalized therapy when tumor material is limited,such as following the acquisition of biopsy specimens obtained by fine-needle aspiration.

关键词： testing fluid cancer

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Predicting outcomes after kidney transplantation: Can Pareto’s rules help us to do so?

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World Journal of Transplantation 2024年第1期14卷 9-11页

作者： Fernando m.gonzalez Francisca gonzalez Cohens Department of Nephrology Faculty of MedicineUniversidad de ChileSantiago 7500922Chile Web Intelligence Centre Faculty of Physics and Mathematical SciencesSantiago 7500922Chile

Kidney transplantation is the best option for kidney replacement therapy,even considering that most of the times the grafts do not survive as long as their recipients.In the Khalil et al's experience,published in this issue of the Journal,they analyze their second kidney graft survival and describe those significant predictors of early loss.This editorial comments on the results and put in perspective that most of the times,long-term graft survival could be inadvertently jeopardized if the immunosuppressive therapy is reduced or withdrawn for any reason,and that it could happen frequently if the transplant physician intends to innovate with the clinical care without proper evidence-based data.

关键词： Kidney transplantation Graft survival Acute rejection Interstitial fibrosis and tubular atrophy Immunosuppression

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modeling mitochondria,where are the numbers?

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Neural Regeneration Research 2024年第7期19卷 1435-1436页

作者： Adrian m.Davies alan G.Holt University Center Shrewsbury ShrewsburyShropshireUK

models and simulations are particularly useful for exploring ideas and concepts in the biological sciences that are expe rimentally impracticable.in silico methods are also gaining acceptance with regulatory authorities as an alternative to animal testing.For example,the Environmental Protection Agency aims to eliminate animal testi ng by 2035.

关键词： eliminate authorities acceptance

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Excess dietary Lys reduces feed intake, stimulates jejunal CCK secretion and alters essential and non‑essential blood AA profile in pigs

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Journal of Animal Science and Biotechnology 2024年第3期15卷 1336-1346页

作者： maximiliano muller Elout Van Liefferinge alan Tilbrook Robert van Barneveld Eugeni Roura Centre for Nutrition and Food Sciences Queensland Alliance for Agriculture and Food InnovationThe University of QueenslandBrisbaneQueensland 4072Australia Laboratory of Animal Nutrition and Animal Product Quality(LANUPRO) Department of Animal Sciences and Aquatic EcologyGhent University339000 GhentFlandersBelgium Centre for Animal Science Queensland Alliance for Agriculture and Food Innovation and the School of Veterinary ScienceThe University of QueenslandBrisbaneQueensland 4072Australia SunPork Group BrisbaneQueensland 4009Australia

Background Commercial diets are frequently formulated to meet or exceed nutrient levels including those of lim-iting essential amino acids(AA)covering potential individual variations within the herd.However,the provision of dietary excess of AA,such as Lys,may lead to reduced appetite and growth in pigs.The mechanisms modulat-ing these responses have not been extensively investigated.This study evaluated the effect of Lys dietary excesses on performance and satiety biomarkers in post weaning pigs.methods Twenty-four pigs aged 21 d and weighing 6.81±0.12kg(mean±SEm)were individually housed and offered 1 of 4 dietary treatments for 3weeks:a diet containing a standardized ileal digestible Lys reaching 100%(T0),120%(T1),150%(T2)or 200%(T3)of the NRC(2012)requirements.At the end of the experiment,blood samples from the cephalic vein of the T0 and T3 groups were obtained for AA analysis.In addition,primary intestinal cultures from T0 pigs were used,following their humane killing,to evaluate the effect of Lys on gut hormone secretion and AA sensors gene expression under ex vivo conditions.Results Feed intake was linearly reduced(Pmetabotropic glutamate receptor 1 and the solute carrier family 7 member 2(Pmal jejunum tissues incubated with Lys at 20mmol/L when compared to the control(Lys 0mmol/L).Plasma Lys and Glu(Pmpared to T0 pigs.In contrast,plasma levels of His,Val,Thr,Leu(Pm that excess dietary Lys inhibits hunger in pigs.moreover,the results provide evidence of pre-and post-absorptive mechanisms modulating these responses.Lys dietary excesses should be nar-rowed,when possible,to avoid negative effects of the AA on appetite in pigs.

关键词： Amino acid Blood Cholecystokinin Feed intake Lysine Pig Satiety

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Critical care specialists,the missing link in organ procurement for transplantation

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World Journal of Critical Care medicine 2024年第2期13卷 1-6页

作者： Francisca Del Rocio gonzalez Cohens Fernando m.gonzalez Web Intelligence Centre Faculty of Physics and Mathematical SciencesUniversidad de ChileSantiago 8370397Chile Department of Nephrology Faculty of MedicineUniversidad de ChileSantiago 7500922Chile

The procurement process for organ donation begins with the identification of potential organ donors in emergency or critical care units(CCU),followed by their clinical evaluation,diagnostic procedures,and therapeutic interventions,mostly conducted in CCUs.It concludes with the request for organ donation and,if accepted,the retrieval of organs.Despite most interventions occurring in detection units,there has been a neglect of the strategic role played by critical care specialists(CCS)in managing and caring for brain-dead or near-brain-death patients.Questions arise:Are they willing to undertake this responsibility?Do they fully comprehend the nature of organ procurement?Are they aware of the specific interventions required to maintain possible organ donors in optimal physiological condition?Our objective is to examine the role of CCS in organ procurement and propose ways to enhance it,ultimately aiming to increase and enhance organ donation rates.

关键词： Organ procurement Transplantation Brain death Cardiac death Organ donation

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Generation of lossy mode resonances(LmR)using perovskite nanofilms

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Opto-Electronic Advances 2024年第2期7卷 33-40页

作者： Dayron Armas Ignacio R.matias m.Carmen Lopez-gonzalez Carlos Ruiz Zamarreño Pablo Zubiate Ignacio del Villar Beatriz Romero Electrical Electronic and Communications Engineering DepartmentPublic University of NavarraPamplona 31006Spain Institute of Smart Cities(ISC) Public University of NavarraPamplona 31006Spain Experimental Science and Technology School Rey Juan Carlos UniversityMostoles 28933Spain

The results presented here show for the first time the experimental demonstration of the fabrication of lossy mode resonance(LmR) devices based on perovskite coatings deposited on planar waveguides. Perovskite thin films have been obtained by means of the spin coating technique and their presence was confirmed by ellipsometry, scanning electron microscopy, and X-ray diffraction testing. The LmRs can be generated in a wide wavelength range and the experimental results agree with the theoretical simulations. Overall, this study highlights the potential of perovskite thin films for the development of novel LmR-based devices that can be used for environmental monitoring, industrial sensing, and gas detection, among other applications.

关键词： perovskite thin films slab waveguide lossy mode resonance

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