Yongyao Tan

Tongji Hospital

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Guigang Li

Huazhong University of Science and Technology

Lingjuan Xu

Tongji Hospital

Guanyu Su

Huazhong University of Science and Technology

Wei Wang

Huazhong University of Science and Technology

Yunming Wang

Huazhong University of Science and Technology

Xiao Zhou

Huazhong University of Science and Technology

Rong Liu

Huazhong University of Science and Technology

Chunxiu You

Huazhong University of Science and Technology

Huamin Zhou

Huazhong University of Science and Technology

Xiao Zhou

Huazhong University of Science and Technology

Cooperative Institutions

Huazhong University of Science and Technology

Tongji Hospital

Shanghai Jiao Tong University

Chinese Academy of Sciences

Sun Yat-sen University

University of Chinese Academy of Sciences

Zhejiang Provincial People's Hospital

Lanzhou University

Chinese Academy of Medical Sciences & Peking Union Medical College

Tsinghua University

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Colletotrichum Keratitis: An Important Fungal Infection of Nine Human Eyes

Guigang Li Wei Wang Hongya Gong yang xiaoxuan Lingjuan Xu

Colletotrichum is a relatively uncommon cause of human infection, and previous findings on Colletotrichum keratitis have been scarce. This research describes the clinical characteristics and treatment outcomes of 9 cases of Colletotrichum keratitis discovered in our hospital using next-generation sequencing (NGS). Among 78 patients with NGS-proven fungal keratitis, 9 (11.5%) were infected with Colletotrichum species. NGS revealed that 3 patients were infected with C. truncatum, 3 with C. gloeosporioides, and 3 with C. fructicola. Seven patients had a history of corneal plant trauma, one patient was infected by mosquitoes, and one patient had an unknown origin. Seven patients underwent penetrating keratoplasty, and two underwent lamellar keratoplasty. Eight patients healed after keratoplasty, but one required evisceration due to recurrence of fungal infection. NGS allows for more precise diagnosis and enhances awareness of Colletotrichum keratitis, which is not as rare as previously reported.

Fungal Keratitis

Colletotrichum

10.2139/ssrn.4686275

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Colletotrichum Keratitis: an important fungal infection of nine human eyes

Diagnostic Microbiology and Infectious Disease (2024)

Wei Wang Hongya Gong Xiaoxuan Yang Lingjuan Xu Xiaoyu Huang

Fungal Keratitis

Colletotrichum

10.1016/j.diagmicrobio.2024.116540

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Isolation and characterization of rabbit limbal niche cells

Experimental Eye Research (2024)

Guanyu Su Xiaojie Guo Lingjuan Xu Bihui Jin Yongyao Tan

Limbal niche cells (LNCs) are one of the most important supporting cells for corneal epithelial stem cells (CES), however, research on LNCs has been mostly limited to humans and rats previously. To expand the research work into the rabbit animal model, one of the most often used animals in stem cell study, this study was carried out for the in vitro isolation and identification of rabbit LNCs. Rabbit LNCs were isolated by collagenase A digestion method and single cells were obtained, the cells were then seeded on 5% Matrigel-coated plastic surface and cultured in modified embryonic stem cell medium (MESCM). Three biological replicates of the isolating and characterization were recorded from New Zealand White rabbits aged from 2.5 months to 5 months. LNC markers (VIM/CD90/CD105/SCF/PDGFRβ) were analyzed using tyramide signal amplification (TSA) staining, immunohistochemical staining (IHC), western blotting (WB), and real-time reverse transcription polymerase chain reaction (qPCR). TSA staining suggested that VIM was highly expressed in rabbit limbus stroma, which was confirmed by WB, and P63α was expressed in the basal limbus epithelium. Pan-CK and CK12 were highly expressed in the central corneal epithelium but lightly expressed in the limbal epithelium. The WB result indicated that PDGFRβ and VIM expressions in rabbit-LNCs P4 were higher than in P1 and P7. In addition, rabbit corneal epithelium highly expressed Paired Box 6 (PAX6) and Epidermal growth factor-like domain 6(EGFL6). For the three repeat experiments, the cell expansion activity of rabbit-LNC was highest at P4. Rabbit-LNCs were passaged from P0 to P7, and the number of cell doublings (NCD) of P4 for the three repeat experiments was 2.816, 2.737, and 2.849. qPCR showed that high mRNA expression levels of VIM, CD90, CD105, SCF, and PDGFRβ in rabbit-LNCs P4. In conclusion, rabbit-LNCs could be successfully isolated by the collagenase A digestion method as used in human tissue. There were similar characteristics between rabbit and human LNCs (VIM+/CD90+/CD105+/SCF+/PAX6+/PDGFRβ+).

Matrigel

CD90

10.1016/j.exer.2024.109838

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Basic Fibroblast Growth Factor Supports the Function of Limbal Niche Cells via the Wnt/β-Catenin Pathway

Journal of Ocular Pharmacology and Therapeutics (2024)

Bihui Jin Guanyu Su Xiao Zhou Lingjuan Xu Wei Wang

To test the effects and underlying mechanisms of basic fibroblast growth factor (bFGF) on the limbal niche cell (LNC) function

10.1089/jop.2024.0042

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Limbal Niche Cells in Three-Dimensional Matrigel Induced Dedifferentiation of Mature Corneal Epithelial Cells toward a Progenitor State

bioRxiv (Cold Spring Harbor Laboratory) (2020)

Hui Zhu Wei Wang Lingjuan Xu Menglin Jiang Yongyao Tan

ABSTRACT Purpose To investigate the possibility and the key factors of stably committed mature corneal epithelial cells dedifferentiate into corneal epithelial stem cells in vitro . Methods Mature cornea epithelia cell (MCEC) sheets or limbal epithelial progenitor cell (LEPC) sheets were isolated from central corneas or limbal segments by Dispase II and further digested with 0.25% trypsin/1 mM EDTA (T/E) to yield single cells. Limbal niche cells (LNC) were isolated from the limbal stroma by collagenase A and expanded on 5% Matrigel coated plastic. Single MCECs were seeded on 50% Matrigel with or without LNC culturing for 10 days, regarding as three-dimensional MCEC (3D-MCEC) group or three-dimensional MCEC+LNC (3D-MCEC+LNC) group. Expression of CK12, p63α, PCK, Vimentin were analyzed with immunofluorescence staining. Results The expression of mature cornea epithelial marker (CK12) in MCEC was higher than that in LEPC (P=0.020) but epithelial stem cell marker (p63α) was lower than that in LEPC (P=0.000). When seeded in 3D Matrigel, single MCEC cells could form spheres within 72 hours, and the expression of CK12 reduced (P=0.005) and the expression of p63α also reduced to zero (P=0.000) compared to MCEC. Serial passages of LNC which were expanded in coated Matrigel could form spheres in 3D Matrigel. After mixing MCECs with LNC, rounder spheres emerged within 24 hours which consisted of both epithelia cells (PCK + /Vim - ) and LNC (PCK - /Vim + ). Moreover, epithelia cells in 3D-MCEC+LNC group expressed less CK12 and more p63α than those in MCEC group (P=0.043, 0.000). Besides, the diameter of spheres in 3D-MCEC+LNC group were larger than that in 3D-MCEC group (P=0.000). Conclusion Human LNC and three-dimensional Matrigel could induce the dedifferentiation of mature corneal epithelial cells into corneal epithelial stem cells.

Matrigel

Dispase

10.1101/2020.06.01.128629

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Thermoplastic PHB-Reinforced Chitosan Piezoelectric Films for Biodegradable Pressure Sensors

ACS Applied Bio Materials (2024)

Shuang Sun Yongyao Tan Qikuan Cheng Yuchen Cai Jiaqi Zheng

Flexible and wearable pressure sensors have attracted significant attention in the fields of smart medicine and human health monitoring. Nevertheless, the design and fabrication of degradable disposable pressure sensors still face urgent challenges. Herein, we fabricated poly(3-hydroxybutyrate) (PHB)-reinforced chitosan (CS) piezoelectric films for intelligent sensors through a simple, low-cost, and environmentally friendly roll-forming method. The results show that PHB doping successfully increased the effective piezoelectric coefficient of the chitosan-based film from 40.12 to 49.38 pm/V (a 23% increase). Simultaneously, the pressure sensor based on the CS/PHB film exhibited excellent response sensitivity (484 mV/kPa) and a wide linear response range (0-130 kPa), which could be used as haptic sensors and motion monitoring sensors for the fast response to human motion signals. Additionally, the CS/PHB film could be completely degraded within 18 days in a natural soil environment, demonstrating outstanding degradability. Therefore, chitosan-based piezoelectric films with excellent biodegradability and piezoelectric characteristics have been successfully fabricated in this work, which will promote the innovative development of green chitosan-based electronic devices and disposable pressure sensors.

10.1021/acsabm.4c00966

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Isolation and Identification of Limbal Niche Cells

Journal of Visualized Experiments (2023)

Guanyu Su Wei Wang Lingjuan Xu Rong Liu Yongyao Tan

Here we report a standard procedure for the isolation and identification of limbal niche cells (LNCs). Limbus tissue obtained from an eye bank was used for LNCs isolation. The tissue was divided into 12 pieces under aseptic conditions and digested for 18 h at 37 °C in the cell culture incubator using collagenase A to obtain cell clusters with LNCs and limbal epithelial progenitor cells. The cell clusters were further digested for 15 min at 37 °C using 0.25% trypsin-EDTA to obtain single cells and then cultured in modified embryonic stem cell medium (MESCM) on a plastic surface coated with 5% Matrigel. Cells were passaged upon 70% confluence, and LNCs were identified using immunofluorescence, real-time quantitative PCR (qPCR), and flow cytometry. Primary LNCs were isolated and passaged more than 12 times. The proliferation activity of LNCs from P4 to P6 was the highest. LNCs expressed higher stem cell markers than BMMSCs (SCF, Nestin, Rex1, SSEA4, CD73, CD90, MSX1, P75NTR, and PDGFRβ). Furthermore, results showed that P4 LNCs uniformly expressed VIM, CD90, CD105, and PDGFRβ, but not Pan-CK, which could be used as a marker for the identification of LNCs. Flow cytometric analysis showed that approximately 95%, 97%, 92%, and 11% of LNCs expressed CD73, CD90, CD105, and SCF respectively, while they were 68%, 99%, 20%, and 3% in BMMSCs. The standard process for LNC isolation and identification could provide a reliable laboratory basis for the widespread use of LNCs.

CD90

Limbal stem cell

Cluster of differentiation

Stem cell marker

Matrigel

Immunofluorescence

10.3791/65618

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Citations (2)

Limbal Bio‐Engineered Tissue Employing 3D Nanofiber‐Aerogel Scaffold to Facilitate LSCs Growth and Migration

Macromolecular Bioscience (2022)

Yongyao Tan Dan Chen Yunming Wang Wei Wang Lingjuan Xu

Constrained by the existing scaffold inability to mimic limbal niche, limbal bio-engineered tissue constructed in vitro is challenging to be widely used in clinical practice. Here, a 3D nanofiber-aerogel scaffold is fabricated by employing thermal cross-linking electrospinned film polycaprolactone (PCL) and gelatin (GEL) as the precursor. Benefiting from the cross-linked (160 °C, vacuum) structure, the homogenized and lyophilized 3D nanofiber-aerogel scaffold with preferable mechanical strength is capable of refraining the volume collapse in humid vitro. Intriguingly, compared with traditional electrospinning scaffolds, the authors' 3D nanofiber-aerogel scaffolds possess enhanced water absorption (1100-1300%), controllable aperture (50-100 µm), and excellent biocompatibility (optical density value, 0.953 ± 0.021). The well-matched aperture and nanostructure of the scaffolds with cells enable the construction of limbal bio-engineered tissue. It is foreseen that the proposed general method can be extended to various aerogels, providing new opportunities for the development of novel limbal bio-engineered tissue.

Gelatin

Polycaprolactone

Electrospinning

Biocompatibility

10.1002/mabi.202100441

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Citations (8)

JoVE Video Dataset

Guanyu Su Wei Wang Lingjuan Xu Rong Liu Yongyao Tan

10.3791/65618-v

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Speaking-Induced Charge-Laden Face Masks with Durable Protectiveness and Wearing Breathability

ACS Applied Materials & Interfaces (2022)

Dan Chen Lianwei Tang Yunming Wang Yongyao Tan Yue Fu

Face masks, which serve as personal protection equipment, have become ubiquitous for combating the ongoing COVID-19. However, conventional electrostatic-based mask filters are disposable and short-term effective with high breathing resistance, causing respiratory ailments and massive consumption (129 billion monthly), intensifying global environmental pollution. In an effort to address these challenges, the introduction of a piezoelectric polymer was adopted to realize the charge-laden melt-blown via the melt-blowing method. The charge-laden melt-blown could be applied to manufacture face masks and to generate charges triggered by mechanical and acoustic energy originated from daily speaking. Through an efficient and scalable industrial melt-blown process, our charge-laden mask is capable of overcoming the inevitable electrostatic attenuation, even in a high-humidity atmosphere by long-wearing (prolonging from 4 to 72 h) and three-cycle common decontamination methods. Combined with outstanding protective properties (PM2.5 filtration efficiency >99.9%), breathability (differential pressure <17 Pa/cm2), and mechanical strength, the resultant charge-laden mask could enable the decreased replacement of masks, thereby lowering to 94.4% of output masks worldwide (∼122 billion monthly) without substituting the existing structure or assembling process.

Face masks

Differential pressure

Filtration (mathematics)

10.1021/acsami.2c01077

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Citations (16)