Autologous bone marrow-derived mesenchymal stem cells(BMSCs)have been shown to promote osteogenesis;however,the effects of allogeneic BMSCs(allo-BMSCs)on bone regeneration remain unclear.Therefore,we explored the bone...Autologous bone marrow-derived mesenchymal stem cells(BMSCs)have been shown to promote osteogenesis;however,the effects of allogeneic BMSCs(allo-BMSCs)on bone regeneration remain unclear.Therefore,we explored the bone regeneration promotion effect of allo-BMSCs in 3D-printed autologous bone particle(ABP)scaffolds.First,we concurrently printed scaffolds with polycaprolactone,ABPs,and allo-BMSCs for appropriate support,providing bioactive factors and seed cells to promote osteogenesis.In vitro studies showed that ABP scaffolds promoted allo-BMSC osteogenic differentiation.In vivo studies revealed that the implantation of scaffolds loaded with ABPs and allo-BMSCs into canine skull defects for nine months promoted osteogenesis.Further experiments suggested that only a small portion of implanted allo-BMSCs survived and differentiated into vascular endothelial cells,chondrocytes,and osteocytes.The implanted allo-BMSCs released stromal cell-derived factor 1 through paracrine signaling to recruit native BMSCs into the defect,promoting bone regeneration.This study contributes to our understanding of allo-BMSCs,providing information relevant to their future application.展开更多
In the current settings of osteosarcoma research and drug screening,in vitro three-dimensional(3D)models,which overcome the limitations of traditional models,are favored.In in vitro 3D models,tumor microenvironment si...In the current settings of osteosarcoma research and drug screening,in vitro three-dimensional(3D)models,which overcome the limitations of traditional models,are favored.In in vitro 3D models,tumor microenvironment simulation,particularly of the mechanical microenvironment,is crucial for estimating the biological effects of a tumor.However,current in vitro osteosarcoma model construction is often limited to a single mechanical signal,which fails to simulate the diversity of osteosarcoma mechanical stimuli.In this study,we utilized embedded bioprinting technology and the multiple response properties of calcium ions in soft and hard stiffness systems with osteosarcoma cell biological functions to construct an integrated gradient biomechanical signal-tailored osteosarcoma model(IGBSTOM).We achieved this by printing a fibrinogen bioink containing calcium ions and osteosarcoma tumor spheroids within an extracellular matrix composed of methacryloylated alginate,methacryloylated gelatin,thrombin,and transglutaminase,which is rich in polysaccharides and proteins and exhibits self-healing properties.Our in vitro and in vivo studies showed that the IGBSTOM enhanced tumor stemness,proliferation,and migration,and successfully reproduced the nest-like structure of tumors,providing an in vitro research platform that is more similar to the natural tumor than the existing models.This study proposes a novel IGBSTOM construction and provides a new strategy for the clinical understanding of tumor development,drug screening,and exploration of drug resistance and metastasis mechanisms.展开更多
Degenerative spine pathologies,including intervertebral disc(IVD)degeneration,present a significant healthcare challenge due to their association with chronic pain and disability.This study explores an innovative appr...Degenerative spine pathologies,including intervertebral disc(IVD)degeneration,present a significant healthcare challenge due to their association with chronic pain and disability.This study explores an innovative approach to IVD regeneration utilizing 3D bioprinting technology,specifically visible light-based digital light processing,to fabricate tissue scaffolds that closely mimic the native architecture of the IVD.Utilizing a hybrid bioink composed of gelatin methacrylate(GelMA)and poly(ethylene glycol)diacrylate(PEGDA)at a 10%concentration,we achieved enhanced printing fidelity and mechanical properties suitable for load-bearing applications such as the IVD.Preconditioning rat bone marrow-derived mesenchymal stem cell spheroids with chondrogenic media before incorporating them into the GelMA-PEGDA scaffold further promoted the regenerative capabilities of this system.Our findings demonstrate that this bioprinted scaffold not only supports cell viability and integration but also contributes to the restoration of disc height in a rat caudal disc model without inducing adverse inflammatory responses.The study underscores the potential of combining advanced bioprinting techniques and cell preconditioning strategies to develop effective treatments for IVD degeneration and other musculoskeletal disorders,highlighting the need for further research into the dynamic interplay between cellular migration and the hydrogel matrix.展开更多
The brain exhibits complex physiology characterized by unique features such as a brain-specific extracellular matrix, compartmentalized structure (white and grey matter), and an aligned axonal network. These physiolog...The brain exhibits complex physiology characterized by unique features such as a brain-specific extracellular matrix, compartmentalized structure (white and grey matter), and an aligned axonal network. These physiological characteristics underpin brain function and facilitate signal transduction similar to that in an electrical circuit. Therefore, investigating these features in vitro is crucial for understanding the interactions between neuronal signal transduction processes and the pathology of neurological diseases. Compared to neurons on patterned substrates, three-dimensional (3D) bioprinting-based neural models provide significant advantages in replicating axonal kinetics without physical limitations. This study proposes the development of a 3D bioprinted engineered neural network (BENN) model to replicate the physiological features of the brain, suggesting its application as a tool for studying neurodegenerative diseases. We employed 3D bioprinting to reconstruct the compartmentalized structure of the brain, and controlled the directionality of axonal growth by applying electrical stimuli to the printed neural structure for overcoming spatial constraints. The reconstructed axonal network demonstrated reliability as a neural analog, including the visualization of mature neuronal features and spontaneous calcium reactions. Furthermore, these brain-like neural network models have demonstrated usefulness for studying neurodegeneration by enabling the visualization of degenerative pathophysiology in alcohol-exposed neurons. The BENN facilitates the visualization of region-specific pathological markers in soma or axon populations, including amyloid-beta formation and axonal deformation. Overall, the BENN closely mimics brain physiology, offers insights into the dynamics of axonal networks, and can be applied to studying neurological diseases.展开更多
Optical coherence tomography(OCT)imaging technology has significant advantages in in situ and noninvasive monitoring of biological tissues.However,it still faces the following challenges:including data processing spee...Optical coherence tomography(OCT)imaging technology has significant advantages in in situ and noninvasive monitoring of biological tissues.However,it still faces the following challenges:including data processing speed,image quality,and improvements in three-dimensional(3D)visualization effects.OCT technology,especially functional imaging techniques like optical coherence tomography angiography(OCTA),requires a long acquisition time and a large data size.Despite the substantial increase in the acquisition speed of swept source optical coherence tomography(SS-OCT),it still poses significant challenges for data processing.Additionally,during in situ acquisition,image artifacts resulting from interface reflections or strong reflections from biological tissues and culturing containers present obstacles to data visualization and further analysis.Firstly,a customized frequency domainfilter with anti-banding suppression parameters was designed to suppress artifact noises.Then,this study proposed a graphics processing unit(GPU)-based real-time data processing pipeline for SS-OCT,achieving a measured line-process rate of 800 kHz for 3D fast and high-quality data visualization.Furthermore,a GPU-based realtime data processing for CC-OCTA was integrated to acquire dynamic information.Moreover,a vascular-like network chip was prepared using extrusion-based 3D printing and sacrificial materials,with sacrificial material being printed at the desired vascular network locations and then removed to form the vascular-like network.OCTA imaging technology was used to monitor the progression of sacrificial material removal and vascular-like network formation.Therefore,GPU-based OCT enables real-time processing and visualization with artifact suppression,making it particularly suitable for in situ noninvasive longitudinal monitoring of 3D bioprinting tissue and vascular-like networks in microfluidic chips.展开更多
Treatments for lesions in central nervous system(CNS)are always faced with challenges due to the anatomical and physiological particularity of the CNS despite the fact that several achievements have been made in early...Treatments for lesions in central nervous system(CNS)are always faced with challenges due to the anatomical and physiological particularity of the CNS despite the fact that several achievements have been made in early diagnosis and precision medicine to improve the survival and quality of life of patients with brain tumors in recent years.Understanding the complexity as well as role of the microenvironment of brain tumors may suggest a better revealing of the molecular mechanism of brain tumors and new therapeutic directions,which requires an accurate recapitulation of the complex microenvironment of human brain in vitro.Here,a 3D bioprinted in vitro brain matrix-mimetic microenvironment model with hyaluronic acid(HA)and normal glial cells(HEBs)is developed which simulates both mechanical and biological properties of human brain microenvironment in vivo through the investigation of the formulation of bioinks and optimization of printing process and parameters to study the effects of different concentration of gelatin(GA)within the bioink and different printing structures of the scaffold on the performance of the brain matrix-mimetic microenvironment models.The study provides experimental models for the exploration of the multiple factors in the brain microenvironment and scaffolds for GBM invasion study.展开更多
Due to their special anatomical and physiological features,central nervous system diseases still presented challenges,despite the fact that some advances have been made in early diagnosis and precision medicine.One of...Due to their special anatomical and physiological features,central nervous system diseases still presented challenges,despite the fact that some advances have been made in early diagnosis and precision medicine.One of the complexities in treating tumors is the tumor microenvironment,which includes mesenchymal stem cells(MSCs)that exhibit tumor tropism and can be used for cell therapy.However,whether MSCs promote or suppress gliomas is still unclear,especially in glioma microenvironments.In this study,a coaxial microfiber was designed to mimic the tumor microenvironment and to reveal the effect of MSCs on glioma cells.The fiber shell was composed of MSCs and alginate,and the core was filled with U87 MG(glioblastoma)cells and gelatin methacrylate.This Shell-MSC/Core-U87 MG microenvironment improved the proliferation,survival,invasion,metastasis,and drug resistance of glioma cells,while simultaneously maintaining the stemness of glioma cells.In summary,coaxial extrusion bioprinted Shell-MSC/Core-U87 MG microfiber is an ideal platform for tumor and stromal cell coculture to observe tumor biological behavior in vitro.展开更多
The shortage of skin for grafting continues to be a major problem in the treatment of serious skin injuries.3D bioprinting provides a new way to solve this problem.However,current 3D printed skin is less effective in ...The shortage of skin for grafting continues to be a major problem in the treatment of serious skin injuries.3D bioprinting provides a new way to solve this problem.However,current 3D printed skin is less effective in treatment of large wounds because of severe shrinkage and scarring.In this study,bionically designed bilayer skin was fabricated using an extrusion-based bioprinter and a gelatin/sodium alginate/gelatin methacrylate hydrogel with excellent physical and biological properties.Full-thickness skin wounds were created in the back of nude mice and treated with bioprinted skin or hydrogel.Bioprinted skin accelerated wound healing,reduced wound contraction and scarring,and facilitated wound skin epithelialization compared with the bioprinted hydrogel or untreated wound.The skin from the wound was collected 28 days after grafting for histology and immunofluorescence analysis.The thickness of the dermis and epidermis of the bioprinted skin was similar to that of nude mice.Microvascular formation in the dermis and dense keratinocytes in the epidermis of the bioprinted skin were observed.This study provides a potential treatment strategy for reducing skin contraction and scar in large skin wounds.展开更多
Cholangiocarcinoma(CCA)is characterized by heterogeneous mutations and a refractory nature.Thus,the development of a model for effective drug screening is urgently needed.As the established therapeutic testing models ...Cholangiocarcinoma(CCA)is characterized by heterogeneous mutations and a refractory nature.Thus,the development of a model for effective drug screening is urgently needed.As the established therapeutic testing models for CCA are often ineffective,we fabricated an enabling three-dimensional(3D)-bioprinted CCA-on-a-chip model that to a good extent resembled the multicellular microenvironment and the anatomical microstructure of the hepato-vascular-biliary system to perform high-content antitumor drug screening.Specifically,cholangiocytes,hepatocytes,and vascular endotheliocytes were employed for 3D bioprinting of the models,allowing for a high degree of spatial and tube-like microstructural control.Interestingly,it was possible to observe CCA cells attached to the surfaces of the gelatin methacryloyl(GelMA)hydrogelembedded microchannels and overgrown in a thickening manner,generating bile duct stenosis,which was expected to be analogous to the in vivo configuration.Over 4000 differentially expressed genes were detected in the CCA cells in our 3D coculture model compared to the traditional two-dimensional(2D)monoculture.Further screening revealed that the CCA cells grown in the 3D traditional model were more sensitive to the antitumoral prodrug than those in the 2D monoculture due to drug biotransformation by the neighboring functional hepatocytes.This study provides proof-of-concept validation of our bioprinted CCA-on-a-chip as a promising drug screening model for CCA treatment and paves the way for potential personalized medicine strategies for CCA patients in the future.展开更多
Tunneling wounds create passageways underneath the skin surface with varying sizes and shapes and can have twists and turns,making their treatment extremely difficult.Available wound care solutions only cater to super...Tunneling wounds create passageways underneath the skin surface with varying sizes and shapes and can have twists and turns,making their treatment extremely difficult.Available wound care solutions only cater to superficial wounds,and untreated tunneling wounds pose major health concerns.This study aims to fulfill this challenge by fabricating tunnel wound fillers(TWFs)made of natural polymers that mimic the dermal extracellular matrix.In this study,cellulose microfibers(CMFs)derived from banana stem and fish skin-derived collagen were used to formulate bio-inks with varying CMF contents(25,50,and 75 mg).Tri-layered(CMFs,primary and secondary collagen coatings),drug-eluting(Baneocin),and cell-laden(human mesenchymal stem cells)TWFs were three-dimensional(3D)-printed and extensively characterized.CMFs showed the most suitable rheological properties for 3D printing at 50 mg concentration.The Alamar Blue data showed significantly increased cell proliferation from Day 1 to Day 7,and scratch tests used to evaluate in vitro wound healing revealed that the best coverage of the wound area was achieved using CMFs in combination with collagen and alginate.Finally,the TWF showed promising capability and tunability in terms of wound shape and size upon testing on a chicken tissue model.The results demonstrate the tremendous potential of TWFs in treating deep tunneling wounds with unique advantages,such as patient-specific customization,good wound exudate absorption capability while releasing wound healing drugs,and the inclusion of stem cells for accelerated healing and tissue regeneration.展开更多
It is crucial to maintaining the viability of biofabricated human-sized tissues to ensure their successful survival and function after transplantation.Adenosine is a purine nucleoside that has the function to suppress...It is crucial to maintaining the viability of biofabricated human-sized tissues to ensure their successful survival and function after transplantation.Adenosine is a purine nucleoside that has the function to suppress cellular metabolism and has been previously proposed as a method to prolong cell viability under hypoxia.In this study,we optimized the dose concentration of adenosine for incorporation into bioprinted constructs to preserve long-term cell viability in vitro.Our results showed that muscle cells(C2C12)containing 6,7,or 8 mM adenosine maintained high cell viability for 20 days under hypoxic conditions(0.1%O2),whereas cells without adenosine treatment showed 100%cell death after 11 days.After 20 days under hypoxic conditions,muscle cells treated with adenosine proliferated and differentiated when transferred to normoxic conditions.From these adenosine concentrations,6 mM was picked as the optimized adenosine concentration for further investigations due to its most effective results on improving cell viability.The bioprinted muscle constructs containing adenosine(6 mM)maintained high cell viability for 11 days under hypoxic conditions,while the control constructs without adenosine had no live cells.For in vivo validation,the bioprinted constructs with adenosine implanted under the dorsal subcutaneous space in mice,showed the enhanced formation of muscle tissue with minimal central necrosis and apoptosis,when compared to the constructs without adenosine.These positive in vitro and in vivo results demonstrate that the use of adenosine is an effective approach to preventing the challenge of hypoxia-induced necrosis in bioprinted tissues for clinical translation.展开更多
The growth plate(GP)is a crucial tissue involved in skeleton development via endochondral ossification(EO).The bone organoid is a potential research model capable of simulating the physiological function,spatial struc...The growth plate(GP)is a crucial tissue involved in skeleton development via endochondral ossification(EO).The bone organoid is a potential research model capable of simulating the physiological function,spatial structure,and intercellular communication of native GPs.However,mimicking the EO process remains a key challenge for bone organoid research.To simulate this orderly mineralization process,we designed an in vitro sh Ca_(v)3.3 ATDC5-loaded gelatin methacryloyl(Gel MA)hydrogel model and evaluated its bioprintability for future organoid construction.In this paper,we report the first demonstration that the T-type voltage-dependent calcium channel(T-VDCC)subtype Ca_(v)3.3 is dominantly expressed in chondrocytes and is negatively correlated with the hypertrophic differentiation of chondrocytes during the EO process.Furthermore,Ca_(v)3.3 knockdown chondrocytes loaded with the Gel MA hydrogel successfully captured the EO process and provide a bioink capable of constructing layered and orderly mineralized GP organoids in the future.The results of this study could therefore provide a potential target for regulating the EO process and a novel strategy for simulating it in bone organoids.展开更多
Multicellular three-dimensional(3D)bioprinting technology,a pivotal in vitro approach in tissue engineering and disease modeling,enables the co-culture of multiple cell populations within 3D architectures while preser...Multicellular three-dimensional(3D)bioprinting technology,a pivotal in vitro approach in tissue engineering and disease modeling,enables the co-culture of multiple cell populations within 3D architectures while preserving physiological interactions(1).This strategy facilitates the precise simulation of human histogenesis and pathogenesis through four principal modalities:organoid-based systems that recapitulate tissue self-organization,air-liquid interface(ALI)platforms facilitating epithelial-mesenchymal crosstalk analysis,3D microfluidic devices for spatiotemporal control of biomolecular gradients,and bioprinting techniques achieving micron-level spatial patterning of heterogeneous cells(2).Each modality addresses distinct experimental demands in reconstructing multicellular microenvironments,collectively enhancing the fidelity of drug screening and advancing mechanobiological research.展开更多
Rotator cuff tears are common among physically active individuals and often require surgical intervention owing to their limited self-healing capacity.This study proposes a new bioprinting approach using bone-and tend...Rotator cuff tears are common among physically active individuals and often require surgical intervention owing to their limited self-healing capacity.This study proposes a new bioprinting approach using bone-and tendon tissue-specific bioinks derived from decellularized extracellular matrix,supplemented with hydroxyapatite and TGF-β/poly(vinyl alcohol)to fabricate engineered tendon-to-bone complex tissue.To achieve this goal,a core-shell nozzle system attached to a bioprinter enables the effective and simultaneous fabrication of aligned tendon tissue,a gradient tendon-bone interface(TBI),and a mechanically improved bone region,mimicking the native tendon-to-bone structure.In vitro evaluation demonstrated the well-directed differentiation of human adipose stem cells towards osteogenic and tenogenic lineages in the bone and tendon constructs.In the graded TBI structure,further facilitated fibrocartilage formation and enhanced the integration of tendon-to-bone tissues compared to non-graded structures in vitro.Furthermore,using a rabbit rotator cuff tear model,implantation of the biologically graded constructs significantly promoted the rapid regeneration of full-thickness tendon-to-bone tissue,including the formation of a high-quality TBI in vivo.This bioprinting approach not only improved me-chanical properties and tissue integration but also enhanced angiogenesis and extracellular matrix(ECM)for-mation,demonstrating its potential as a promising platform for the regeneration of tendon-to-bone complex tissues.展开更多
Biomimetic neural substitutes,constructed through the bottom-up assembly of cell-matrix modulus via 3D bioprinting,hold great promise for neural regeneration.However,achieving precise control over the fate of neural s...Biomimetic neural substitutes,constructed through the bottom-up assembly of cell-matrix modulus via 3D bioprinting,hold great promise for neural regeneration.However,achieving precise control over the fate of neural stem cells(NSCs)to ensure biological functionality remains challenging.Cell behaviors are closely linked to cellular dynamics and cell-matrix mechanotransduction within a 3D microenvironment.To address this,a dynamic bioactive bioink is designed to provide adaptable biomechanics and instructive biochemical cues,specifically tailored for the fate commitment of NSCs,through incorporating reversible Schiff-base bonds and bioactive motifs,N-cadherin-mimicking and BDNF-mimicking peptides.We demonstrate that the dynamic properties of 3D bioprinted living fibers alleviate the mechanical confinement on NSCs and significantly enhance their mechanosensing,spreading,migration,and matrix remodeling within the 3D matrix.Additionally,the inclusion of N-cadherin-mimicking and BDNF-mimicking peptides further enhances cells’ability to sense and respond to mechanical and neurotrophic cues provided by the surrounding matrix,which accelerates the selforganization of a functional neural network within the 3D bioprinted construct,leading to significant motor and sensory function recovery in a rat complete spinal cord injury model.This work underscores the critical role of precisely designing cell-instructive bioinks for the advanced functionality of 3D bioprinted living constructs in neural regeneration.展开更多
Cartilage defects are commonly observed in orthopedic clinical studies.Owing to the unique structure of cartilage tissue,current clinical treatments cannot fully address this issue.Cartilage organoids are three-dimens...Cartilage defects are commonly observed in orthopedic clinical studies.Owing to the unique structure of cartilage tissue,current clinical treatments cannot fully address this issue.Cartilage organoids are three-dimensional(3D)active tissue structures constructed in vitro to mimic the structure and function of natural cartilage tissue and can be utilized for disease research and cartilage repair.In this study,we engineered MNPs-BMSCs by introducing magnetic nanoparticles(MNPs)into bone marrow mesenchymal stem cells(BMSCs).Under the influence of the magnetic field induced by the MNPs,MNPs-BMSCs became polarized,significantly enhancingtheir aggregation,migration,andchondrogenic differentiation capabilities.We then used these engineered MNPs-BMSCs as seed cells and applied 3D bioprinting technology to construct an advanced cartilage organoid using a MNPs-BMSC/alginate/gelatin matrix.This structure partially mimics the middle layer of a cartilage.The advanced cartilage organoid demonstrated superior chondrogenic differentiation ability and mechanical properties in vitro.It significantly enhanced tissue repair in cartilage defect areas in vivo,restoring the normal structure of the cartilage layer.Overall,the engineered MNPs-BMSCs/alginate/gelatin advanced cartilage organoids offer a promising approach for studying cartilage tissue in vitro and advancing cartilage repair within the field of tissue engineering.展开更多
ABSTRACT:The repair of large-scale bone defects is still a challenge in clinical orthopedics.Especially,excessive reactive oxygen species(ROS)-induced oxidative stress injury greatly affected bone healing.In this stud...ABSTRACT:The repair of large-scale bone defects is still a challenge in clinical orthopedics.Especially,excessive reactive oxygen species(ROS)-induced oxidative stress injury greatly affected bone healing.In this study,we innovatively developed an antioxidant three-dimensional(3D)-bioprinted MMn_(3)O_(4)@Gel by integrating M-Mn_(3)O_(4) nanozyme into photocrosslinked gelatin methacryloyl(GelMA)for the therapy of bone defects.Results showed that the incorporation of MMn_(3)O_(4) not only enhanced the mechanical properties of the nanocomposite hydrogel with the compressive modulus 141.79%higher than that of pure GelMA,but also maintained excellent 3D printability.In vitro studies confirmed that the 3Dprinted M-Mn_(3)O_(4)@Gel exhibited favorable biocompatibility and cell adhesion.It significantly reduced oxidative stress through efficient ROS scavenging,restored mitochondrial function,and ultimately demonstrated remarkable osteogenic capacity,highlighting the efficacy of control-released nanozymes.More importantly,under near-infrared(NIR)irradiation,MMn_(3)O_(4)@Gel demonstrated further enhanced ROS-scavenging capacity and bone regeneration potential.Mechanistically,MMn_(3)O_(4)@Gel promoted osteogenesis by upregulating heat shock protein 40 kDa(HSP40)and HSP70 expression,effectively mitigating the overactivation of the Nrf2 pathway.This study innovatively combines nanozyme technology with 3D-printed hydrogel materials,offering a novel strategy to address the challenge of oxidative stress in bone regeneration.展开更多
Renal unilateral ischemia-reperfusion injury(UIRI)constitutes a significant global health challenge,with poor recovery leading to chronic kidney disease and subsequent renal fibrosis.Extracellular vesicles(EVs)present...Renal unilateral ischemia-reperfusion injury(UIRI)constitutes a significant global health challenge,with poor recovery leading to chronic kidney disease and subsequent renal fibrosis.Extracellular vesicles(EVs)present substantial potential benefits for renal diseases.However,the limited yield and efficacy of EVs produced through traditional methodologies(2D-EVs)severely restrict their widespread application.Moreover,the efficient and effective strategies for using EVs in UIRI treatment and their mechanisms remain largely unexplored.In this study,we propose an innovative approach by integrating bioprinted mesenchymal stem cell microfiber extracellular vesicles production technology(3D-EVs)with a tail vein injection method,introducing a novel treatment strategy for UIRI.Our comparison of the biological functions of 2D-EVs and 3D-EVs,both in vitro and in vivo,reveals that 3D-EVs significantly outperform 2D-EVs.Specifically,in vitro,3D-EVs demonstrate a superior capacity to enhance the proliferation and migration of NRK-52E cells and mitigate hypoxia/reoxygenation(H/R)-induced injuries by reducing epithelial-mesenchymal transformation,extracellular matrix deposition,and ferroptosis.In vivo,3D-EVs exhibit enhanced therapeutic effects,as evidenced by improved renal function and decreased collagen deposition in UIRI mouse kidneys.We further elucidate the mechanism by which 3D-EVs derived from KLF15 ameliorate UIRI-induced tubular epithelial cells(TECs)ferroptosis through the modulation of SLC7A11 and GPX4 expression.Our findings suggest that bioprinted mesenchymal stem cells microfiberderived EVs significantly ameliorate renal UIRI,opening new avenues for effective and efficient EV-based therapies in UIRI treatment.展开更多
Background:Mammary progenitor cells(MPCs)maintain their reproductive potency through life,and their specific microenvironments exert a deterministic control over these cells.MPCs provides one kind of ideal tools for s...Background:Mammary progenitor cells(MPCs)maintain their reproductive potency through life,and their specific microenvironments exert a deterministic control over these cells.MPCs provides one kind of ideal tools for studying engineered microenvironmental influence because of its accessibility and continually undergoes postnatal developmental changes.The aim of our study is to explore the critical role of the engineered sweat gland(SG)microenvironment in reprogramming MPCs into functional SG cells.Methods:We have utilized a three-dimensional(3D)SG microenvironment composed of gelatin-alginate hydrogels and components from mouse SG extracellular matrix(SG-ECM)proteins to reroute the differentiation of MPCs to study the functions of this microenvironment.MPCs were encapsulated into the artificial SG microenvironment and were printed into a 3D cell-laden construct.The expression of specific markers at the protein and gene levels was detected after cultured 14 days.Results:Compared with the control group,immunofluorescence and gene expression assay demonstrated that MPCs encapsulated in the bioprinted 3D-SG microenvironment could significantly express the functional marker of mouse SG,sodium/potassium channel protein ATP1a1,and tend to express the specific marker of luminal epithelial cells,keratin-8.When the Shh pathway is inhibited,the expression of SG-associated proteins in MPCs under the same induction environment is significantly reduced.Conclusions:Our evidence proved the ability of differentiated mouse MPCs to regenerate SG cells by engineered SG microenvironment in vitro and Shh pathway was found to be correlated with the changes in the differentiation.These results provide insights into regeneration of damaged SG by MPCs and the role of the engineered microenvironment in reprogramming cell fate.展开更多
Large bone defects face a high risk of pathogen exposure due to open wounds,which leads to high infection rates and delayed bone union.To promote successful repair of infectious bone defects,fabrication of a scaffold ...Large bone defects face a high risk of pathogen exposure due to open wounds,which leads to high infection rates and delayed bone union.To promote successful repair of infectious bone defects,fabrication of a scaffold with dual functions of osteo-induction and bacterial inhibition is required.This study describes creation of an engineered progenitor cell line(C3H10T1/2)capable of doxycycline(DOX)-mediated release of bone morphogenetic protein-2(BMP2).Three-dimensional bioprinting technology enabled creation of scaffolds,comprising polycaprolactone/mesoporous bioactive glass/DOX and bioink,containing these engineered cells.In vivo and in vitro experiments confirmed that the scaffold could actively secrete BMP2 to significantly promote osteoblast differentiation and induce ectopic bone formation.Additionally,the scaffold exhibited broad-spectrum antibacterial capacity,thereby ensuring the survival of embedded engineered cells when facing high risk of infection.These findings demonstrated the efficacy of this bioprinted scaffold to release BMP2 in a controlled manner and prevent the occurrence of infection;thus,showing its potential for repairing infectious bone defects.展开更多
基金supported by the Science and Technology Development Fund of the Fourth Military Medical University(No.2016XB051)the Military Medical Promotion Plan of the Fourth Military Medical University(No.2016TSA-005)+2 种基金the Science and Technology Program of Guangzhou(No.201604040002)the Youth Development Project of Air Force Medical University(No.21QNPY072)the Xijing Hospital Booster Program(No.XJZT24CZ10).
文摘Autologous bone marrow-derived mesenchymal stem cells(BMSCs)have been shown to promote osteogenesis;however,the effects of allogeneic BMSCs(allo-BMSCs)on bone regeneration remain unclear.Therefore,we explored the bone regeneration promotion effect of allo-BMSCs in 3D-printed autologous bone particle(ABP)scaffolds.First,we concurrently printed scaffolds with polycaprolactone,ABPs,and allo-BMSCs for appropriate support,providing bioactive factors and seed cells to promote osteogenesis.In vitro studies showed that ABP scaffolds promoted allo-BMSC osteogenic differentiation.In vivo studies revealed that the implantation of scaffolds loaded with ABPs and allo-BMSCs into canine skull defects for nine months promoted osteogenesis.Further experiments suggested that only a small portion of implanted allo-BMSCs survived and differentiated into vascular endothelial cells,chondrocytes,and osteocytes.The implanted allo-BMSCs released stromal cell-derived factor 1 through paracrine signaling to recruit native BMSCs into the defect,promoting bone regeneration.This study contributes to our understanding of allo-BMSCs,providing information relevant to their future application.
基金appreciate financial support from the National Key R&D Program of China(No.2022YFA1104600)2022 Lingang Laboratory“Seeking Outstanding Youth Program”Open Project(No.LGQS-202206-04)+3 种基金Shanghai Ninth People’s Hospital–Shanghai Jiao Tong University School of Medicine–Shanghai University of Science and Technology Cross-funded Collaborative Program(No.JYJC202233)the National Natural Science Foundation of China(No.82372377)Biomaterials and Regenerative Medicine Institute Cooperative Research Project by Shanghai Jiao Tong University School of Medicine(No.2022LHBO8),Shanghai Key Laboratory of Orthopaedic Implants,Department of Orthopaedics by Shanghai Ninth People’s Hospital–Shanghai Jiao Tong University School of Medicine(No.KFKT202206),the Key R&D Program of Jiangsu Province Social Development Project(No.BE2022708)the Project of Shanghai Science and Technology Commission(No.22015820100).
文摘In the current settings of osteosarcoma research and drug screening,in vitro three-dimensional(3D)models,which overcome the limitations of traditional models,are favored.In in vitro 3D models,tumor microenvironment simulation,particularly of the mechanical microenvironment,is crucial for estimating the biological effects of a tumor.However,current in vitro osteosarcoma model construction is often limited to a single mechanical signal,which fails to simulate the diversity of osteosarcoma mechanical stimuli.In this study,we utilized embedded bioprinting technology and the multiple response properties of calcium ions in soft and hard stiffness systems with osteosarcoma cell biological functions to construct an integrated gradient biomechanical signal-tailored osteosarcoma model(IGBSTOM).We achieved this by printing a fibrinogen bioink containing calcium ions and osteosarcoma tumor spheroids within an extracellular matrix composed of methacryloylated alginate,methacryloylated gelatin,thrombin,and transglutaminase,which is rich in polysaccharides and proteins and exhibits self-healing properties.Our in vitro and in vivo studies showed that the IGBSTOM enhanced tumor stemness,proliferation,and migration,and successfully reproduced the nest-like structure of tumors,providing an in vitro research platform that is more similar to the natural tumor than the existing models.This study proposes a novel IGBSTOM construction and provides a new strategy for the clinical understanding of tumor development,drug screening,and exploration of drug resistance and metastasis mechanisms.
基金supported by NIH Grant(T32GM065841),Mayo Foundation for Education and Research.
文摘Degenerative spine pathologies,including intervertebral disc(IVD)degeneration,present a significant healthcare challenge due to their association with chronic pain and disability.This study explores an innovative approach to IVD regeneration utilizing 3D bioprinting technology,specifically visible light-based digital light processing,to fabricate tissue scaffolds that closely mimic the native architecture of the IVD.Utilizing a hybrid bioink composed of gelatin methacrylate(GelMA)and poly(ethylene glycol)diacrylate(PEGDA)at a 10%concentration,we achieved enhanced printing fidelity and mechanical properties suitable for load-bearing applications such as the IVD.Preconditioning rat bone marrow-derived mesenchymal stem cell spheroids with chondrogenic media before incorporating them into the GelMA-PEGDA scaffold further promoted the regenerative capabilities of this system.Our findings demonstrate that this bioprinted scaffold not only supports cell viability and integration but also contributes to the restoration of disc height in a rat caudal disc model without inducing adverse inflammatory responses.The study underscores the potential of combining advanced bioprinting techniques and cell preconditioning strategies to develop effective treatments for IVD degeneration and other musculoskeletal disorders,highlighting the need for further research into the dynamic interplay between cellular migration and the hydrogel matrix.
基金supported by Korean Fund for Regenerative Medicine funded by Ministry of Science and ICT,and Ministry of Health and Welfare(22A0106L1,Republic of Korea)the National Research Foundation of Korea(NRF)grant funded by the Korea government(MSIT)(No.2022M3C1A3081359).
文摘The brain exhibits complex physiology characterized by unique features such as a brain-specific extracellular matrix, compartmentalized structure (white and grey matter), and an aligned axonal network. These physiological characteristics underpin brain function and facilitate signal transduction similar to that in an electrical circuit. Therefore, investigating these features in vitro is crucial for understanding the interactions between neuronal signal transduction processes and the pathology of neurological diseases. Compared to neurons on patterned substrates, three-dimensional (3D) bioprinting-based neural models provide significant advantages in replicating axonal kinetics without physical limitations. This study proposes the development of a 3D bioprinted engineered neural network (BENN) model to replicate the physiological features of the brain, suggesting its application as a tool for studying neurodegenerative diseases. We employed 3D bioprinting to reconstruct the compartmentalized structure of the brain, and controlled the directionality of axonal growth by applying electrical stimuli to the printed neural structure for overcoming spatial constraints. The reconstructed axonal network demonstrated reliability as a neural analog, including the visualization of mature neuronal features and spontaneous calcium reactions. Furthermore, these brain-like neural network models have demonstrated usefulness for studying neurodegeneration by enabling the visualization of degenerative pathophysiology in alcohol-exposed neurons. The BENN facilitates the visualization of region-specific pathological markers in soma or axon populations, including amyloid-beta formation and axonal deformation. Overall, the BENN closely mimics brain physiology, offers insights into the dynamics of axonal networks, and can be applied to studying neurological diseases.
基金supported by the National Key Research and Development Program of China(Nos.2022YFA1104600 and 2022YFA1200208)National Natural Science Foundation of China(No.31927801)Key Research and Development Foundation of Zhejiang Province(No.2022C01123).
文摘Optical coherence tomography(OCT)imaging technology has significant advantages in in situ and noninvasive monitoring of biological tissues.However,it still faces the following challenges:including data processing speed,image quality,and improvements in three-dimensional(3D)visualization effects.OCT technology,especially functional imaging techniques like optical coherence tomography angiography(OCTA),requires a long acquisition time and a large data size.Despite the substantial increase in the acquisition speed of swept source optical coherence tomography(SS-OCT),it still poses significant challenges for data processing.Additionally,during in situ acquisition,image artifacts resulting from interface reflections or strong reflections from biological tissues and culturing containers present obstacles to data visualization and further analysis.Firstly,a customized frequency domainfilter with anti-banding suppression parameters was designed to suppress artifact noises.Then,this study proposed a graphics processing unit(GPU)-based real-time data processing pipeline for SS-OCT,achieving a measured line-process rate of 800 kHz for 3D fast and high-quality data visualization.Furthermore,a GPU-based realtime data processing for CC-OCTA was integrated to acquire dynamic information.Moreover,a vascular-like network chip was prepared using extrusion-based 3D printing and sacrificial materials,with sacrificial material being printed at the desired vascular network locations and then removed to form the vascular-like network.OCTA imaging technology was used to monitor the progression of sacrificial material removal and vascular-like network formation.Therefore,GPU-based OCT enables real-time processing and visualization with artifact suppression,making it particularly suitable for in situ noninvasive longitudinal monitoring of 3D bioprinting tissue and vascular-like networks in microfluidic chips.
基金We would like to thank the support by the National Key Research and Development Program of China(2018YFA0703000)National Natural Science Foundation of China(Grant No.51875518)+1 种基金Key Research and Development Projects of Zhejiang Province(Grant No.2017C01054 and No.2018C03062)the Fundamental Research Funds for the Central Universities(Grant No.2019XZZX003-02,2019FZA4002).
文摘Treatments for lesions in central nervous system(CNS)are always faced with challenges due to the anatomical and physiological particularity of the CNS despite the fact that several achievements have been made in early diagnosis and precision medicine to improve the survival and quality of life of patients with brain tumors in recent years.Understanding the complexity as well as role of the microenvironment of brain tumors may suggest a better revealing of the molecular mechanism of brain tumors and new therapeutic directions,which requires an accurate recapitulation of the complex microenvironment of human brain in vitro.Here,a 3D bioprinted in vitro brain matrix-mimetic microenvironment model with hyaluronic acid(HA)and normal glial cells(HEBs)is developed which simulates both mechanical and biological properties of human brain microenvironment in vivo through the investigation of the formulation of bioinks and optimization of printing process and parameters to study the effects of different concentration of gelatin(GA)within the bioink and different printing structures of the scaffold on the performance of the brain matrix-mimetic microenvironment models.The study provides experimental models for the exploration of the multiple factors in the brain microenvironment and scaffolds for GBM invasion study.
基金Jiangxi Provincial People’s Government and Shangrao East China Institute of Digital Medical Engineering for their support。
文摘Due to their special anatomical and physiological features,central nervous system diseases still presented challenges,despite the fact that some advances have been made in early diagnosis and precision medicine.One of the complexities in treating tumors is the tumor microenvironment,which includes mesenchymal stem cells(MSCs)that exhibit tumor tropism and can be used for cell therapy.However,whether MSCs promote or suppress gliomas is still unclear,especially in glioma microenvironments.In this study,a coaxial microfiber was designed to mimic the tumor microenvironment and to reveal the effect of MSCs on glioma cells.The fiber shell was composed of MSCs and alginate,and the core was filled with U87 MG(glioblastoma)cells and gelatin methacrylate.This Shell-MSC/Core-U87 MG microenvironment improved the proliferation,survival,invasion,metastasis,and drug resistance of glioma cells,while simultaneously maintaining the stemness of glioma cells.In summary,coaxial extrusion bioprinted Shell-MSC/Core-U87 MG microfiber is an ideal platform for tumor and stromal cell coculture to observe tumor biological behavior in vitro.
基金This work was supported by the National Key R&D Program of China(No.2018YFE0207900)the Development projects of Key research of People’s Liberation Army(No.BWS17J036,18-163-13-ZT-003-011-01)the National Natural Science Foundation of China(51835010 and 51375371)。
文摘The shortage of skin for grafting continues to be a major problem in the treatment of serious skin injuries.3D bioprinting provides a new way to solve this problem.However,current 3D printed skin is less effective in treatment of large wounds because of severe shrinkage and scarring.In this study,bionically designed bilayer skin was fabricated using an extrusion-based bioprinter and a gelatin/sodium alginate/gelatin methacrylate hydrogel with excellent physical and biological properties.Full-thickness skin wounds were created in the back of nude mice and treated with bioprinted skin or hydrogel.Bioprinted skin accelerated wound healing,reduced wound contraction and scarring,and facilitated wound skin epithelialization compared with the bioprinted hydrogel or untreated wound.The skin from the wound was collected 28 days after grafting for histology and immunofluorescence analysis.The thickness of the dermis and epidermis of the bioprinted skin was similar to that of nude mice.Microvascular formation in the dermis and dense keratinocytes in the epidermis of the bioprinted skin were observed.This study provides a potential treatment strategy for reducing skin contraction and scar in large skin wounds.
文摘Cholangiocarcinoma(CCA)is characterized by heterogeneous mutations and a refractory nature.Thus,the development of a model for effective drug screening is urgently needed.As the established therapeutic testing models for CCA are often ineffective,we fabricated an enabling three-dimensional(3D)-bioprinted CCA-on-a-chip model that to a good extent resembled the multicellular microenvironment and the anatomical microstructure of the hepato-vascular-biliary system to perform high-content antitumor drug screening.Specifically,cholangiocytes,hepatocytes,and vascular endotheliocytes were employed for 3D bioprinting of the models,allowing for a high degree of spatial and tube-like microstructural control.Interestingly,it was possible to observe CCA cells attached to the surfaces of the gelatin methacryloyl(GelMA)hydrogelembedded microchannels and overgrown in a thickening manner,generating bile duct stenosis,which was expected to be analogous to the in vivo configuration.Over 4000 differentially expressed genes were detected in the CCA cells in our 3D coculture model compared to the traditional two-dimensional(2D)monoculture.Further screening revealed that the CCA cells grown in the 3D traditional model were more sensitive to the antitumoral prodrug than those in the 2D monoculture due to drug biotransformation by the neighboring functional hepatocytes.This study provides proof-of-concept validation of our bioprinted CCA-on-a-chip as a promising drug screening model for CCA treatment and paves the way for potential personalized medicine strategies for CCA patients in the future.
基金supported by the start-up funds from New York University Abu Dhabipartially carried out using the Core Technology Platforms resources at New York University Abu Dhabi。
文摘Tunneling wounds create passageways underneath the skin surface with varying sizes and shapes and can have twists and turns,making their treatment extremely difficult.Available wound care solutions only cater to superficial wounds,and untreated tunneling wounds pose major health concerns.This study aims to fulfill this challenge by fabricating tunnel wound fillers(TWFs)made of natural polymers that mimic the dermal extracellular matrix.In this study,cellulose microfibers(CMFs)derived from banana stem and fish skin-derived collagen were used to formulate bio-inks with varying CMF contents(25,50,and 75 mg).Tri-layered(CMFs,primary and secondary collagen coatings),drug-eluting(Baneocin),and cell-laden(human mesenchymal stem cells)TWFs were three-dimensional(3D)-printed and extensively characterized.CMFs showed the most suitable rheological properties for 3D printing at 50 mg concentration.The Alamar Blue data showed significantly increased cell proliferation from Day 1 to Day 7,and scratch tests used to evaluate in vitro wound healing revealed that the best coverage of the wound area was achieved using CMFs in combination with collagen and alginate.Finally,the TWF showed promising capability and tunability in terms of wound shape and size upon testing on a chicken tissue model.The results demonstrate the tremendous potential of TWFs in treating deep tunneling wounds with unique advantages,such as patient-specific customization,good wound exudate absorption capability while releasing wound healing drugs,and the inclusion of stem cells for accelerated healing and tissue regeneration.
文摘It is crucial to maintaining the viability of biofabricated human-sized tissues to ensure their successful survival and function after transplantation.Adenosine is a purine nucleoside that has the function to suppress cellular metabolism and has been previously proposed as a method to prolong cell viability under hypoxia.In this study,we optimized the dose concentration of adenosine for incorporation into bioprinted constructs to preserve long-term cell viability in vitro.Our results showed that muscle cells(C2C12)containing 6,7,or 8 mM adenosine maintained high cell viability for 20 days under hypoxic conditions(0.1%O2),whereas cells without adenosine treatment showed 100%cell death after 11 days.After 20 days under hypoxic conditions,muscle cells treated with adenosine proliferated and differentiated when transferred to normoxic conditions.From these adenosine concentrations,6 mM was picked as the optimized adenosine concentration for further investigations due to its most effective results on improving cell viability.The bioprinted muscle constructs containing adenosine(6 mM)maintained high cell viability for 11 days under hypoxic conditions,while the control constructs without adenosine had no live cells.For in vivo validation,the bioprinted constructs with adenosine implanted under the dorsal subcutaneous space in mice,showed the enhanced formation of muscle tissue with minimal central necrosis and apoptosis,when compared to the constructs without adenosine.These positive in vitro and in vivo results demonstrate that the use of adenosine is an effective approach to preventing the challenge of hypoxia-induced necrosis in bioprinted tissues for clinical translation.
基金supported by the National Natural Science Foundation of China(No.31800784)the Chongqing Key Laboratory of Precision Medicine in Joint Surgery(No.425Z2138)+2 种基金the Chongqing Excellent Scientist Project(No.425Z2W21)the Chongqing Natural Science Foundation General Project(No.cstc2021jcyjmsxm X0135)the Chongqing Postdoctoral Research Project Special Fund(No.2021XM3033)。
文摘The growth plate(GP)is a crucial tissue involved in skeleton development via endochondral ossification(EO).The bone organoid is a potential research model capable of simulating the physiological function,spatial structure,and intercellular communication of native GPs.However,mimicking the EO process remains a key challenge for bone organoid research.To simulate this orderly mineralization process,we designed an in vitro sh Ca_(v)3.3 ATDC5-loaded gelatin methacryloyl(Gel MA)hydrogel model and evaluated its bioprintability for future organoid construction.In this paper,we report the first demonstration that the T-type voltage-dependent calcium channel(T-VDCC)subtype Ca_(v)3.3 is dominantly expressed in chondrocytes and is negatively correlated with the hypertrophic differentiation of chondrocytes during the EO process.Furthermore,Ca_(v)3.3 knockdown chondrocytes loaded with the Gel MA hydrogel successfully captured the EO process and provide a bioink capable of constructing layered and orderly mineralized GP organoids in the future.The results of this study could therefore provide a potential target for regulating the EO process and a novel strategy for simulating it in bone organoids.
基金supported by the National Natural Science Foundation of China(No.32271470).
文摘Multicellular three-dimensional(3D)bioprinting technology,a pivotal in vitro approach in tissue engineering and disease modeling,enables the co-culture of multiple cell populations within 3D architectures while preserving physiological interactions(1).This strategy facilitates the precise simulation of human histogenesis and pathogenesis through four principal modalities:organoid-based systems that recapitulate tissue self-organization,air-liquid interface(ALI)platforms facilitating epithelial-mesenchymal crosstalk analysis,3D microfluidic devices for spatiotemporal control of biomolecular gradients,and bioprinting techniques achieving micron-level spatial patterning of heterogeneous cells(2).Each modality addresses distinct experimental demands in reconstructing multicellular microenvironments,collectively enhancing the fidelity of drug screening and advancing mechanobiological research.
基金supported by the“Korea National Institute of Health”research project(2022ER130500)grants from the National Research Foundation of Korea(NRF)funded by the Korea government(MSIT)(RS-2024-00336758),(No.NRF2022R1F1A106729811)and(2022R1A2C2091162)+1 种基金supported by the Sung-KyunKwan University and the BK21 FOUR(Graduate School Innova-tion)funded by the Ministry of Education(MOE,Korea)National Research Foundation of Korea(NRF).
文摘Rotator cuff tears are common among physically active individuals and often require surgical intervention owing to their limited self-healing capacity.This study proposes a new bioprinting approach using bone-and tendon tissue-specific bioinks derived from decellularized extracellular matrix,supplemented with hydroxyapatite and TGF-β/poly(vinyl alcohol)to fabricate engineered tendon-to-bone complex tissue.To achieve this goal,a core-shell nozzle system attached to a bioprinter enables the effective and simultaneous fabrication of aligned tendon tissue,a gradient tendon-bone interface(TBI),and a mechanically improved bone region,mimicking the native tendon-to-bone structure.In vitro evaluation demonstrated the well-directed differentiation of human adipose stem cells towards osteogenic and tenogenic lineages in the bone and tendon constructs.In the graded TBI structure,further facilitated fibrocartilage formation and enhanced the integration of tendon-to-bone tissues compared to non-graded structures in vitro.Furthermore,using a rabbit rotator cuff tear model,implantation of the biologically graded constructs significantly promoted the rapid regeneration of full-thickness tendon-to-bone tissue,including the formation of a high-quality TBI in vivo.This bioprinting approach not only improved me-chanical properties and tissue integration but also enhanced angiogenesis and extracellular matrix(ECM)for-mation,demonstrating its potential as a promising platform for the regeneration of tendon-to-bone complex tissues.
基金support from the National Natural Science Foundation of China(Grant No.32271414 and 82301560)support from State Key Laboratory of New Ceramic Materials Tsinghua University(No.KF202409).
文摘Biomimetic neural substitutes,constructed through the bottom-up assembly of cell-matrix modulus via 3D bioprinting,hold great promise for neural regeneration.However,achieving precise control over the fate of neural stem cells(NSCs)to ensure biological functionality remains challenging.Cell behaviors are closely linked to cellular dynamics and cell-matrix mechanotransduction within a 3D microenvironment.To address this,a dynamic bioactive bioink is designed to provide adaptable biomechanics and instructive biochemical cues,specifically tailored for the fate commitment of NSCs,through incorporating reversible Schiff-base bonds and bioactive motifs,N-cadherin-mimicking and BDNF-mimicking peptides.We demonstrate that the dynamic properties of 3D bioprinted living fibers alleviate the mechanical confinement on NSCs and significantly enhance their mechanosensing,spreading,migration,and matrix remodeling within the 3D matrix.Additionally,the inclusion of N-cadherin-mimicking and BDNF-mimicking peptides further enhances cells’ability to sense and respond to mechanical and neurotrophic cues provided by the surrounding matrix,which accelerates the selforganization of a functional neural network within the 3D bioprinted construct,leading to significant motor and sensory function recovery in a rat complete spinal cord injury model.This work underscores the critical role of precisely designing cell-instructive bioinks for the advanced functionality of 3D bioprinted living constructs in neural regeneration.
基金This work was financially supported by Science and Technology Innovation Leading Plan of High-Tech Industry in Hunan Province(No.2020SK2011)The National Natural Science Foundation of China-Youth Science Fund(No.82204538)Macao Special Administrative Region Science and Technology Development Fund(No.0007/2021/AKP).
文摘Cartilage defects are commonly observed in orthopedic clinical studies.Owing to the unique structure of cartilage tissue,current clinical treatments cannot fully address this issue.Cartilage organoids are three-dimensional(3D)active tissue structures constructed in vitro to mimic the structure and function of natural cartilage tissue and can be utilized for disease research and cartilage repair.In this study,we engineered MNPs-BMSCs by introducing magnetic nanoparticles(MNPs)into bone marrow mesenchymal stem cells(BMSCs).Under the influence of the magnetic field induced by the MNPs,MNPs-BMSCs became polarized,significantly enhancingtheir aggregation,migration,andchondrogenic differentiation capabilities.We then used these engineered MNPs-BMSCs as seed cells and applied 3D bioprinting technology to construct an advanced cartilage organoid using a MNPs-BMSC/alginate/gelatin matrix.This structure partially mimics the middle layer of a cartilage.The advanced cartilage organoid demonstrated superior chondrogenic differentiation ability and mechanical properties in vitro.It significantly enhanced tissue repair in cartilage defect areas in vivo,restoring the normal structure of the cartilage layer.Overall,the engineered MNPs-BMSCs/alginate/gelatin advanced cartilage organoids offer a promising approach for studying cartilage tissue in vitro and advancing cartilage repair within the field of tissue engineering.
基金financially supported by the Guangxi Natural Science Foundation(No.2025GXNSFBA069100)the National Natural Science Foundation of China(Nos.52573316 and 82360426)the Basic ability enhancement project for young and middle-aged teachers of universities in Guangxi(No.2024KY0105).
文摘ABSTRACT:The repair of large-scale bone defects is still a challenge in clinical orthopedics.Especially,excessive reactive oxygen species(ROS)-induced oxidative stress injury greatly affected bone healing.In this study,we innovatively developed an antioxidant three-dimensional(3D)-bioprinted MMn_(3)O_(4)@Gel by integrating M-Mn_(3)O_(4) nanozyme into photocrosslinked gelatin methacryloyl(GelMA)for the therapy of bone defects.Results showed that the incorporation of MMn_(3)O_(4) not only enhanced the mechanical properties of the nanocomposite hydrogel with the compressive modulus 141.79%higher than that of pure GelMA,but also maintained excellent 3D printability.In vitro studies confirmed that the 3Dprinted M-Mn_(3)O_(4)@Gel exhibited favorable biocompatibility and cell adhesion.It significantly reduced oxidative stress through efficient ROS scavenging,restored mitochondrial function,and ultimately demonstrated remarkable osteogenic capacity,highlighting the efficacy of control-released nanozymes.More importantly,under near-infrared(NIR)irradiation,MMn_(3)O_(4)@Gel demonstrated further enhanced ROS-scavenging capacity and bone regeneration potential.Mechanistically,MMn_(3)O_(4)@Gel promoted osteogenesis by upregulating heat shock protein 40 kDa(HSP40)and HSP70 expression,effectively mitigating the overactivation of the Nrf2 pathway.This study innovatively combines nanozyme technology with 3D-printed hydrogel materials,offering a novel strategy to address the challenge of oxidative stress in bone regeneration.
基金supported by the Natural Science Foundation of China(Grant No.52075285)the Natural Science Foundation of Guang Dong Province(2019A1515010386,2024A1515010266).
文摘Renal unilateral ischemia-reperfusion injury(UIRI)constitutes a significant global health challenge,with poor recovery leading to chronic kidney disease and subsequent renal fibrosis.Extracellular vesicles(EVs)present substantial potential benefits for renal diseases.However,the limited yield and efficacy of EVs produced through traditional methodologies(2D-EVs)severely restrict their widespread application.Moreover,the efficient and effective strategies for using EVs in UIRI treatment and their mechanisms remain largely unexplored.In this study,we propose an innovative approach by integrating bioprinted mesenchymal stem cell microfiber extracellular vesicles production technology(3D-EVs)with a tail vein injection method,introducing a novel treatment strategy for UIRI.Our comparison of the biological functions of 2D-EVs and 3D-EVs,both in vitro and in vivo,reveals that 3D-EVs significantly outperform 2D-EVs.Specifically,in vitro,3D-EVs demonstrate a superior capacity to enhance the proliferation and migration of NRK-52E cells and mitigate hypoxia/reoxygenation(H/R)-induced injuries by reducing epithelial-mesenchymal transformation,extracellular matrix deposition,and ferroptosis.In vivo,3D-EVs exhibit enhanced therapeutic effects,as evidenced by improved renal function and decreased collagen deposition in UIRI mouse kidneys.We further elucidate the mechanism by which 3D-EVs derived from KLF15 ameliorate UIRI-induced tubular epithelial cells(TECs)ferroptosis through the modulation of SLC7A11 and GPX4 expression.Our findings suggest that bioprinted mesenchymal stem cells microfiberderived EVs significantly ameliorate renal UIRI,opening new avenues for effective and efficient EV-based therapies in UIRI treatment.
基金supported in part by the National Nature Science Foundation of China(81571909,81701906,81830064,81721092)the National Key Research Development Plan(2017YFC1103300)+1 种基金Military Logistics Research Key Project(AWS17J005)Fostering Funds of Chinese PLA General Hospital for National Distinguished Young Scholar Science Fund(2017-JQPY-002).
文摘Background:Mammary progenitor cells(MPCs)maintain their reproductive potency through life,and their specific microenvironments exert a deterministic control over these cells.MPCs provides one kind of ideal tools for studying engineered microenvironmental influence because of its accessibility and continually undergoes postnatal developmental changes.The aim of our study is to explore the critical role of the engineered sweat gland(SG)microenvironment in reprogramming MPCs into functional SG cells.Methods:We have utilized a three-dimensional(3D)SG microenvironment composed of gelatin-alginate hydrogels and components from mouse SG extracellular matrix(SG-ECM)proteins to reroute the differentiation of MPCs to study the functions of this microenvironment.MPCs were encapsulated into the artificial SG microenvironment and were printed into a 3D cell-laden construct.The expression of specific markers at the protein and gene levels was detected after cultured 14 days.Results:Compared with the control group,immunofluorescence and gene expression assay demonstrated that MPCs encapsulated in the bioprinted 3D-SG microenvironment could significantly express the functional marker of mouse SG,sodium/potassium channel protein ATP1a1,and tend to express the specific marker of luminal epithelial cells,keratin-8.When the Shh pathway is inhibited,the expression of SG-associated proteins in MPCs under the same induction environment is significantly reduced.Conclusions:Our evidence proved the ability of differentiated mouse MPCs to regenerate SG cells by engineered SG microenvironment in vitro and Shh pathway was found to be correlated with the changes in the differentiation.These results provide insights into regeneration of damaged SG by MPCs and the role of the engineered microenvironment in reprogramming cell fate.
基金supported by the National Key R&D Program(grant no.2016YFC1102100)a NSFC grant(grant no.81921002)the Shanghai Science and Technology Development Fund(grant no.18DZ2291200 and 18441902700).
文摘Large bone defects face a high risk of pathogen exposure due to open wounds,which leads to high infection rates and delayed bone union.To promote successful repair of infectious bone defects,fabrication of a scaffold with dual functions of osteo-induction and bacterial inhibition is required.This study describes creation of an engineered progenitor cell line(C3H10T1/2)capable of doxycycline(DOX)-mediated release of bone morphogenetic protein-2(BMP2).Three-dimensional bioprinting technology enabled creation of scaffolds,comprising polycaprolactone/mesoporous bioactive glass/DOX and bioink,containing these engineered cells.In vivo and in vitro experiments confirmed that the scaffold could actively secrete BMP2 to significantly promote osteoblast differentiation and induce ectopic bone formation.Additionally,the scaffold exhibited broad-spectrum antibacterial capacity,thereby ensuring the survival of embedded engineered cells when facing high risk of infection.These findings demonstrated the efficacy of this bioprinted scaffold to release BMP2 in a controlled manner and prevent the occurrence of infection;thus,showing its potential for repairing infectious bone defects.
