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Article
| Open AccessSynergy between the anthocyanin and RDR6/SGS3/DCL4 siRNA pathways expose hidden features of Arabidopsis carbon metabolism
TRANSPARENT TESTA19 (TT19) encodes a glutathione S-transferase which functions in anthocyanin stabilization and vacuolar transport. Here, by tt19 suppressor screening, the authors show that RDR6/SGS3/DCL4 siRNA pathway constituents synergistically interact with components of the flavonoid pathway to control carbon metabolism.
- Nan Jiang
- , Aimer Gutierrez-Diaz
- & Erich Grotewold
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Article
| Open AccessIntegrating molecular markers into metabolic models improves genomic selection for Arabidopsis growth
An increase in genomic selection (GS) accuracy can accelerate genetic gain by shortening the breeding cycles. Here, the authors introduce a network-based GS method that uses metabolic models and improves the prediction accuracy of Arabidopsis growth within and across environments.
- Hao Tong
- , Anika Küken
- & Zoran Nikoloski
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Article
| Open AccessInvestigating the dynamics of microbial consortia in spatially structured environments
The spatial organisation of microbial communities is caused by the interplay of biotic and abiotic factors. Here the authors design a microfluidic platform to quantify the spatiotemporal parameters influencing diffusion-mediated interactions, and use this device to investigate information transmission and metabolic cross-feeding in synthetic microbial consortia.
- Sonali Gupta
- , Tyler D. Ross
- & Ophelia S. Venturelli
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Article
| Open AccessLITESEC-T3SS - Light-controlled protein delivery into eukaryotic cells with high spatial and temporal resolution
The type III secretion system (T3SS) of bacteria can be used to inject cargo into eukaryotic cells but its lack of target specificity is a disadvantage. Here the authors place the T3SS under the regulation of light by engineering optogenetic switches into the dynamic cytosolic T3SS component SctQ.
- Florian Lindner
- , Bailey Milne-Davies
- & Andreas Diepold
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Article
| Open AccessTarget-responsive vasoactive probes for ultrasensitive molecular imaging
Noninvasive detection of molecular targets in living subjects could provide valuable insights into healthy function and disease. Here, the authors develop vasoactive imaging probes which allow wide-field in vivo mapping of nanomolar-level molecular species in rat brain.
- Robert Ohlendorf
- , Agata Wiśniowska
- & Alan Jasanoff
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Article
| Open AccessORF Capture-Seq as a versatile method for targeted identification of full-length isoforms
Most human protein-coding genes are expressed as multiple isoforms. Here the authors present ORF Capture-seq that uses cloned ORFs as probes to capture and sequence full length transcript sequences. This enables highly sensitive characterization of eukaryotic transcriptomes.
- Gloria M. Sheynkman
- , Katharine S. Tuttle
- & Marc Vidal
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Article
| Open AccessMulti-omic single-cell snapshots reveal multiple independent trajectories to drug tolerance in a melanoma cell line
Detailed understanding of how cancer cells transition from a drug sensitive to a tolerant state is lacking. Here, using single cell proteomic and metabolic data the authors uncover that isogenic BRAF mutant melanoma cells can take two distinct paths to become tolerant to BRAF inhibition.
- Yapeng Su
- , Melissa E. Ko
- & James R. Heath
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Article
| Open AccessSize and surface charge characterization of nanoparticles with a salt gradient
Exosomes are used as disease biomarkers, but their characterization in biological samples is challenging. Here the authors achieve simultaneous characterization of size and zeta potential of individual nanoparticles and particle mixtures at physiological salinity conditions, exploiting a salt gradient in a capillary channel.
- Martin K. Rasmussen
- , Jonas N. Pedersen
- & Rodolphe Marie
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Article
| Open AccessLight-powered Escherichia coli cell division for chemical production
Manipulation of genes controlling microbial shapes can affect bio-production. Here, the authors employ an optogenetic method to realize dynamic morphological engineering of E. coli replication and division and show the increased production of acetoin and poly(lactate-co-3-hydroxybutyrate).
- Qiang Ding
- , Danlei Ma
- & Xiulai Chen
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Article
| Open AccessLight-triggered release of conventional local anesthetics from a macromolecular prodrug for on-demand local anesthesia
Local anesthetic sustained release systems suffer from untriggered rapid drug release upon application. Here the authors overcome this issue by covalently linking tetracaine to a polymer gel via a photo-cleavable linkage, enabling light-triggered and repeatable drug release.
- Wei Zhang
- , Tianjiao Ji
- & Daniel S. Kohane
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Article
| Open AccessMinimal lactazole scaffold for in vitro thiopeptide bioengineering
Lactazole A is a thiopeptide from Streptomyces lactacystinaeus, encoded by a compact 9.8 kb biosynthetic gene cluster. Here, the authors show a platform for in vitro biosynthesis of lactazole A via a combination of a flexible in vitro translation system with recombinantly produced lactazole biosynthetic enzymes.
- Alexander A. Vinogradov
- , Morito Shimomura
- & Hiroyasu Onaka
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Article
| Open AccessMagnetically driven active topography for long-term biofilm control
Biofilm formation is a major problem in indwelling medical devices. Here, the authors report on the development of a magnetically responsive micro pillar surface for the controlled prevention and removal of biofilms which also increased sensitivity to antibiotics.
- Huan Gu
- , Sang Won Lee
- & Dacheng Ren
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Article
| Open AccessA single-cell transcriptomic landscape of primate arterial aging
Arterial degeneration, closely associated with cardiovascular diseases, is driven by aging-related vascular cell-specific transcriptomics changes. This study provides a single-cell transcriptomic atlas for senile aortic and coronary arteries and underscores FOXO3A-based the transcriptional network in vasoprotection during aging.
- Weiqi Zhang
- , Shu Zhang
- & Jing Qu
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Article
| Open AccessComprehensive aptamer-based screening identifies a spectrum of urinary biomarkers of lupus nephritis across ethnicities
Developing noninvasive diagnostic biomarkers for lupus nephritis (LN) diagnosis is an important clinical goal. Here the authors identify urinary proteins correlated with active LN and disease severity, which differ across ethnicities but collectively outperform the current clinical method.
- Samantha Stanley
- , Kamala Vanarsa
- & Chandra Mohan
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Article
| Open AccessPhotoactivatable Cre recombinase 3.0 for in vivo mouse applications
Previous versions of photoactivatable Cre recombinase (PA-Cre) suffered from unintentional recombination in dark conditions. Here, the authors develop an improved version of PA-Cre, called PA-Cre 3.0, which shows reduced leakiness and improved efficiency upon activation, and make mouse lines that express PA-Cre 3.0 conditionally.
- Kumi Morikawa
- , Kazuhiro Furuhashi
- & Masayuki Yazawa
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Article
| Open AccessStress-induced expression is enriched for evolutionarily young genes in diverse budding yeasts
Fermentation parameters of industrial processes are often not the ideal growth conditions for industrial microbes. Here, the authors reveal that young genes are more responsive to environmental stress than ancient genes using a new gene age assignment method and provide targeted genes for metabolic engineering.
- Tyler W. Doughty
- , Iván Domenzain
- & John P. Morrissey
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Article
| Open AccessControlled packing and single-droplet resolution of 3D-printed functional synthetic tissues
Precise patterning of lipid-stabilised aqueous droplets is a key challenge in building synthetic tissue designs. Here, the authors show how the interactions between pairs of droplets direct the packing of droplets within 3D-printed networks, enabling the formation of synthetic tissues with high-resolution features.
- Alessandro Alcinesio
- , Oliver J. Meacock
- & Hagan Bayley
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Article
| Open AccessModel-driven generation of artificial yeast promoters
A small set of promoters is used for most genetic construct design in S. cerevisiae. Here, the authors develop a predictive model of promoter activity trained on a data set of over one million sequences and use it to design large sets of high-activity promoters.
- Benjamin J. Kotopka
- & Christina D. Smolke
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Article
| Open AccessHighly parallel and efficient single cell mRNA sequencing with paired picoliter chambers
Single-cell RNA-seq can reveal accurate and precise cell types and states. Here the authors present an scRNA-seq platform, Paired-seq, which uses differential flow resistance to achieve 95% cell utilisation efficiency for improved cell-free RNA removal and gene detection.
- Mingxia Zhang
- , Yuan Zou
- & Chaoyong Yang
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Article
| Open AccessA drug-inducible sex-separation technique for insects
Sorting insects based on sex is error prone and frequently labour intensive. Here, the authors present a drug-inducible sex separation system based on sex-specific rescue from antibotic toxicity.
- Nikolay P. Kandul
- , Junru Liu
- & Omar S. Akbari
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Article
| Open AccessSynthetic hybrids of six yeast species
Many industrial organisms are the result of recent or ancient allopolypoidy events. Here the authors iteratively combine the genomes of six yeast species to generate a viable hybrid.
- David Peris
- , William G. Alexander
- & Chris Todd Hittinger
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Article
| Open AccessArtifact-free and high-temporal-resolution in vivo opto-electrophysiology with microLED optoelectrodes
Artifact-free opto-electrophysiology is key for precise modulation and monitoring of individual neurons at high spatio-temporal resolution. The authors present a method for eliminating stimulation artifacts in high-density micro-LED optoelectrodes for accurate functional mapping of local circuits.
- Kanghwan Kim
- , Mihály Vöröslakos
- & Euisik Yoon
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Article
| Open AccessCytosine base editors with minimized unguided DNA and RNA off-target events and high on-target activity
Cytosine base editors have been reported to induce off-target mutations in DNA and RNA. Here the authors identify next-generation CBEs with reduced guide-independent off-target editing profiles and retain high on-target editing activity.
- Yi Yu
- , Thomas C. Leete
- & Nicole M. Gaudelli
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Article
| Open AccessTargeted delivery of nanomaterials with chemical cargoes in plants enabled by a biorecognition motif
Targeted delivery of nanomaterials to subcellular compartments could allow precision engineering of plant function. Here, Santana et al. show that quantum dots functionalized with a rationally-designed targeting peptide are preferentially delivered to chloroplasts where they can be used to tune organellar redox status.
- Israel Santana
- , Honghong Wu
- & Juan Pablo Giraldo
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Article
| Open AccessChemical modifications of adenine base editor mRNA and guide RNA expand its application scope
Cas9 base editors are promising tools for correcting pathogenic single nucleotide mutations. Here the authors chemically modify mRNA encoding the editor and the gRNA to enhance editing and broaden its application.
- Tingting Jiang
- , Jordana M. Henderson
- & Wen Xue
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Article
| Open AccessEnhanced succinic acid production by Mannheimia employing optimal malate dehydrogenase
Malate dehydrogenase (MDH) is one of the key enzymes for succinic acid (SA) bioproduction. Here, the authors report biochemical and structural analyses of various MDHs to reveal amino acids influencing the specific activity and susceptibility to substrate inhibition, and achieve industrial-level SA production.
- Jung Ho Ahn
- , Hogyun Seo
- & Sang Yup Lee
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Article
| Open AccessDevelopment of a thermophilic coculture for corn fiber conversion to ethanol
Corn fiber is a difficult feedstock to utilize due to its recalcitrant hemicellulose. Here, the authors characterize the recalcitrant structures, isolate a new bacterium to consume the hemicellulose, identify its enzymes, and show the benefit with increased conversion of corn fiber to ethanol.
- Dhananjay Beri
- , William S. York
- & Christopher D. Herring
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Article
| Open AccessMultiplex secretome engineering enhances recombinant protein production and purity
Host cell proteins can contaminate biotherapeutics and compromise and degrade their quality. Here the authors use modelling and CRISPR to delete secreted host proteins in CHO cells, leading to improved monoclonal antibody production and purity.
- Stefan Kol
- , Daniel Ley
- & Nathan E. Lewis
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Article
| Open AccessA fully orthogonal system for protein synthesis in bacterial cells
Ribosome engineering is an emerging powerful approach for synthetic protein synthesis. Here the authors invert the Ribo-T system, using the engineered ribosome to translate the proteome while the native ribosome translates specific mRNA.
- Nikolay A. Aleksashin
- , Teresa Szal
- & Alexander S. Mankin
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Article
| Open AccessPerturbing proteomes at single residue resolution using base editing
Base editors allow for the precise modification of genes. Here the authors use Target-AID to systematically test 17,000 sites across the yeast genome.
- Philippe C. Després
- , Alexandre K. Dubé
- & Christian R. Landry
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Article
| Open AccessTransformable hybrid semiconducting polymer nanozyme for second near-infrared photothermal ferrotherapy
Due to tumour microenvironment, Fenton reactions have low therapeutic efficiency. Here the authors report on the application of NIR-II hybrid semiconducting nanozymes for combined photothermal therapy and enhanced ferrotherapy with photoacoustic imaging and show application in vivo in tumour models.
- Yuyan Jiang
- , Xuhui Zhao
- & Ruiping Zhang
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Article
| Open AccessImaging small molecule-induced endosomal escape of siRNA
Therapeutic siRNA becomes trapped in endosomes, limiting its efficacy. Here the authors use fluorescently-tagged galectin-9 as a biosensor for membrane damage to monitor endosomal escape of cholesterol-conjugated siRNA following treatment of small molecule membrane-destabilising drugs.
- Hampus Du Rietz
- , Hampus Hedlund
- & Anders Wittrup
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Article
| Open AccessSynthetic biology based construction of biological activity-related library of fungal decalin-containing diterpenoid pyrones
Combining genome mining and heterologous expression in a genetically tractable host can lead to bioactive natural products discovery and production. Here, the authors employ this strategy for new decalin-containing diterpenoid pyrenes production by expressing native, extended, and shunt pathways in Aspergillus oryzae.
- Kento Tsukada
- , Shono Shinki
- & Teigo Asai
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Article
| Open AccessAn in vivo platform to select and evolve aggregation-resistant proteins
Protein aggregation remains a significant challenge for manufacturing of protein biopharmaceuticals. Here, the authors demonstrate the use of directed evolution and an assay for in vivo innate protein aggregation-propensity to generate aggregation-resistant scFv fragments.
- Jessica S. Ebo
- , Janet C. Saunders
- & David J. Brockwell
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Perspective
| Open AccessDeveloping a new class of engineered live bacterial therapeutics to treat human diseases
The role microbes play in human health and the ability of synthetic biology to engineer microbial properties opens up new ways of treating disease. In this perspective, the authors describe the design and development of these living therapeutics.
- Mark R. Charbonneau
- , Vincent M. Isabella
- & Caroline B. Kurtz
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Article
| Open AccessDNA punch cards for storing data on native DNA sequences via enzymatic nicking
Current synthetic DNA-based data storage systems have high recording costs, read-write latency and error-rates that make them uncompetitive compared to traditional digital storage. The authors use nicks in native DNA to encode data in parallel and create access sites for in-memory computations.
- S. Kasra Tabatabaei
- , Boya Wang
- & Olgica Milenkovic
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Article
| Open AccessPooled CRISPRi screening of the cyanobacterium Synechocystis sp PCC 6803 for enhanced industrial phenotypes
Developing cyanobacteria as CO2-neutral cell factories relies on the knowledge of the regulation mechanisms for growth and metabolism. Here, the authors develop an inducible CRISPRi gene repression library in Synechocystis sp. PCC 6803 and screens genes potentially affecting growth and L-lactate tolerance and production.
- Lun Yao
- , Kiyan Shabestary
- & Elton P. Hudson
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Article
| Open AccessStem cell-derived polarized hepatocytes
To model hepatocyte function accurately in vitro, it is necessary to generate and maintain a polarized epithelium. Here, the authors describe a protocol to generate polarized human pluripotent stem cell-derived hepatocyte-like cells (HLCs) to model enteric virus production and drug secretion in vitro.
- Viet Loan Dao Thi
- , Xianfang Wu
- & Charles M. Rice
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Article
| Open AccessAntibacterial effects of nanopillar surfaces are mediated by cell impedance, penetration and induction of oxidative stress
Synthetic nanopillars resembling the wing surface of certain insects display antibacterial activities. Here, Jenkins et al. show that these activities do not necessarily require bacterial lysis, and may be mediated by oxidative stress induced by deformation of the bacterial cell envelope.
- J. Jenkins
- , J. Mantell
- & B. Su
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Article
| Open AccessEndothelialization of arterial vascular grafts by circulating monocytes
Acellular tissue engineered vessels functionalised with VEGF are coated with a layer of endothelial cells after in vivo implantation, but the source of the cells are unknown. Here the authors provide evidence that monocytes expressing VEGF receptors can transdifferentiate into endothelial cells via a macrophage intermediate.
- Randall J. Smith Jr.
- , Bita Nasiri
- & Stelios T. Andreadis
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Article
| Open AccessGenerating high quality libraries for DIA MS with empirically corrected peptide predictions
Data-independent acquisition-mass spectrometry (MS) typically requires many preparatory MS runs to produce experiment-specific spectral libraries. Here, the authors show that empirical correction of in silico predicted spectral libraries enables efficient generation of high-quality experiment-specific libraries.
- Brian C. Searle
- , Kristian E. Swearingen
- & Mathias Wilhelm
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Article
| Open AccessPolychromatic solar energy conversion in pigment-protein chimeras that unite the two kingdoms of (bacterio)chlorophyll-based photosynthesis
The spectra of light used by photosynthetic organisms are determined by their pigmentation colour palettes. Here Liu et al. show that a genetically-encoded chimera of light-harvesting proteins from plants and reaction centres from purple bacteria allows for polychromatic solar energy harvesting.
- Juntai Liu
- , Vincent M. Friebe
- & Michael R. Jones
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Article
| Open AccessTransient non-integrative expression of nuclear reprogramming factors promotes multifaceted amelioration of aging in human cells
Aging involves gradual loss of tissue function, and transcription factor (TF) expression can ameliorate this in progeroid mice. Here the authors show that transient TF expression reverses age-associated epigenetic marks, inflammatory profiles and restores regenerative potential in naturally aged human cells.
- Tapash Jay Sarkar
- , Marco Quarta
- & Vittorio Sebastiano
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Article
| Open AccessUltrasensitive detection of nucleic acids using deformed graphene channel field effect biosensors
Field effect transistors based on graphene hold promise for sensing applications. Here, the authors report a millimeter-sized transistor based on deformed graphene as a biosensor that can detect nucleic acid molecules having detection limit of ~18 molecules of DNA in physiological buffer solution and ~600 molecules in human serum.
- Michael Taeyoung Hwang
- , Mohammad Heiranian
- & Rashid Bashir
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Article
| Open AccessDeveloping a highly efficient hydroxytyrosol whole-cell catalyst by de-bottlenecking rate-limiting steps
Whole-cell catalyst-based hydroxytyrosol production is low. Here, the authors increase the efficiency of its production in E. coli by de-bottlenecking two enzymatic steps catalyzed by monooxygenase and tyramine oxidase using structure-based enzyme redesign or in vivo-directed evolution with the aid of a newly developed biosensor.
- Jun Yao
- , Yang He
- & Shuang-Yan Tang
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Article
| Open AccessAn expanded library of orthogonal split inteins enables modular multi-peptide assemblies
Inteins allow the joining of protein segments through scarless ligation. Here the authors assess 34 inteins to establish a library of 15 mutually orthogonal split inteins for in vivo applications, 10 of which are also active in vitro.
- Filipe Pinto
- , Ella Lucille Thornton
- & Baojun Wang
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Article
| Open AccessA severe leakage of intermediates to shunt products in acarbose biosynthesis
Biosynthetic mechanism for the type 2 diabetes treatment drug acarbose is not fully revealed. Here, the authors show that shunt pathways and inefficient amino-deoxyhexose biosynthesis lead to 1-epi-valienol and valienol accumulation, and minimizing the flux to these shunt products can increase acarbose titer in Actinoplanes species.
- Qinqin Zhao
- , Yuchang Luo
- & Zixin Deng
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Article
| Open AccessAbundance of conserved CRISPR-Cas9 target sites within the highly polymorphic genomes of Anopheles and Aedes mosquitoes
Genetic variation in natural populations could represent gene drive resistant alleles, preventing successful application for population management. Here the authors survey 1280 genomes from three mosquito species and concludes natural variation will not be detrimental to deploying gene drive technology.
- Hanno Schmidt
- , Travis C. Collier
- & Gregory C. Lanzaro
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Article
| Open AccessFiber reinforced GelMA hydrogel to induce the regeneration of corneal stroma
Regeneration of corneal stroma has been a challenge due to its sophisticated structure and the easy transformation of the keratocyte. Here, the authors use a hydrogel reinforced with orthogonally aligned fibres and serum free medium to maintain keratocyte phenotype for the in vivo stromal regeneration.
- Bin Kong
- , Yun Chen
- & Shengli Mi
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