The combined application of stem cells and three-dimensional bioprinting scaffolds for the repair of spinal cord injury

Spinal cord injury is considered one of the most difficult injuries to repair and has one of the worst prognoses for injuries to the nervous system.Following surgery,the poor regenerative capacity of nerve cells and the generation of new scars can make it very difficult for the impaired nervous system to restore its neural functionality.Traditional treatments can only alleviate secondary injuries but cannot fundamentally repair the spinal cord.Consequently,there is a critical need to develop new treatments to promote functional repair after spinal cord injury.Over recent years,there have been seve ral developments in the use of stem cell therapy for the treatment of spinal cord injury.Alongside significant developments in the field of tissue engineering,three-dimensional bioprinting technology has become a hot research topic due to its ability to accurately print complex structures.This led to the loading of three-dimensional bioprinting scaffolds which provided precise cell localization.These three-dimensional bioprinting scaffolds co uld repair damaged neural circuits and had the potential to repair the damaged spinal cord.In this review,we discuss the mechanisms underlying simple stem cell therapy,the application of different types of stem cells for the treatment of spinal cord injury,and the different manufa cturing methods for three-dimensional bioprinting scaffolds.In particular,we focus on the development of three-dimensional bioprinting scaffolds for the treatment of spinal cord injury.

Innovation leading development:a glimpse into three-dimensional bioprinting in Israel

Three-dimensional(3D)printing has attracted increasing research interest as an emerging manufacturing technology for devel-oping sophisticated and exquisite architecture through hierarchical printing.It has also been employed in various advanced industrial areas.The development of intelligent biomedical engineering has raised the requirements for 3D printing,such as flexible manufacturing processes and technologies,biocompatible constituents,and alternative bioproducts.However,state-of-the-art 3D printing mainly involves inorganics or polymers and generally focuses on traditional industrial fields,thus severely limiting applications demanding biocompatibility and biodegradability.In this regard,peptide architectonics,which are self-assembled by programmed amino acid sequences that can be flexibly functionalized,have shown promising potential as bioinspired inks for 3D printing.Therefore,the combination of 3D printing and peptide self-assembly poten-tially opens up an alternative avenue of 3D bioprinting for diverse advanced applications.Israel,a small but innovative nation,has significantly contributed to 3D bioprinting in terms of scientific studies,marketization,and peptide architectonics,including modulations and applications,and ranks as a leading area in the 3D bioprinting field.This review summarizes the recent progress in 3D bioprinting in Israel,focusing on scientific studies on printable components,soft devices,and tissue engineering.This paper further delves into the manufacture of industrial products,such as artificial meats and bioinspired supramolecular architectures,and the mechanisms,physicochemical properties,and applications of peptide self-assembly.Undoubtedly,Israel contributes significantly to the field of 3D bioprinting and should thus be appropriately recognized.

Three-dimensional bioprinting in ophthalmic care

Three-dimensional(3D)bioprinting is widely used in ophthalmic clinic,including in diagnosis,surgery,prosthetics,medications,drug development and delivery,and medical education.Articles published in 2011–2022 into bioinks,printing technologies,and bioprinting applications in ophthalmology were reviewed and the strengths and limitations of bioprinting in ophthalmology highlighted.The review highlighted the trade-offs of printing technologies and bioinks in respect to,among others,material type cost,throughput,gelation technique,cell density,cell viability,resolution,and printing speed.There is already widespread ophthalmological application of bioprinting outside clinical settings,including in educational modelling,retinal imaging/visualization techniques and drug design/testing.In clinical settings,bioprinting has already found application in pre-operatory planning.Even so,the findings showed that even with its immense promise,actual translation to clinical applications remains distant,but relatively closer for the corneal(except stromal)tissues,epithelium,endothelium,and conjunctiva,than it was for the retina.This review similarly reflected on the critical on the technical,practical,ethical,and cost barrier to rapid progress of bioprinting in ophthalmology,including accessibility to the most sophisticated bioprinting technologies,choice,and suitability of bioinks,tissue viability and storage conditions.The extant research is encouraging,but more work is clearly required for the push towards clinical translation of research.

Bioprinting-Based High-Throughput Fabrication of Three-Dimensional MCF-7 Human Breast Cancer Cellular Spheroids

Cellular spheroids serving as three-dimensional(3D) in vitro tissue models have attracted increasing interest for pathological study and drug-screening applications. Various methods, including microwells in particular, have been developed for engineering cellular spheroids. However, these methods usually suffer from either destructive molding operations or cell loss and non-uniform cell distribution among the wells due to two-step molding and cell seeding. We have developed a facile method that utilizes cellembedded hydrogel arrays as templates for concave well fabrication and in situ MCF-7 cellular spheroid formation on a chip. A custom-built bioprinting system was applied for the fabrication of sacrificial gelatin arrays and sequentially concave wells in a high-throughput, flexible, and controlled manner. The ability to achieve in situ cell seeding for cellular spheroid construction was demonstrated with the advantage of uniform cell seeding and the potential for programmed fabrication of tissue models on chips. The developed method holds great potential for applications in tissue engineering, regenerative medicine, and drug screening.

Bone-targeted lipoplex-loaded three-dimensional bioprinting bilayer scaffold enhanced bone regeneration

Clinical bone-morphogenetic protein 2(BMP2)treatment for bone regeneration,often resulting in complications like soft tissue inflammation and ectopic ossification due to high dosages and non-specific delivery systems,necessitates research into improved biomaterials for better BMP2 stability and retention.To tackle this challenge,we introduced a groundbreaking bone-targeted,lipoplex-loaded,three-dimensional bioprinted bilayer scaffold,termed the polycaprolactone-bioink-nanoparticle(PBN)scaffold,aimed at boosting bone regeneration.We encapsulated BMP2 within the fibroin nanoparticle based lipoplex(Fibroplex)and functionalized it with DSS6 for bone tissue-specific targeting.3D printing technology enables customized,porous PCL scaffolds for bone healing and soft tissue growth,with a two-step bioprinting process creating a cellular lattice structure and a bioink grid using gelatin-alginate hydrogel and DSS6-Fibroplex,shown to support effective nutrient exchange and cell growth at specific pore sizes.The PBN scaffold is predicted through in silico analysis to exhibit biased BMP2 release between bone and soft tissue,a finding validated by in vitro osteogenic differentiation assays.The PBN scaffold was evaluated for critical calvarial defects,focusing on sustained BMP2 delivery,prevention of soft tissue cell infiltration and controlled fiber membrane pore size in vivo.The PBN scaffold demonstrated a more than eight times longer BMP2 release time than that of the collagen sponge,promoting osteogenic differentiation and bone regeneration in a calvarial defect animal.Our findings suggest that the PBN scaffold enhanced the local concentration of BMP2 in bone defects through sustained release and improved the spatial arrangement of bone formation,thereby reducing the risk of heterotopic ossification.

Together but not scrambled:A perspective on chaotic printing/bioprinting

Structures in nature are often multi-material,and their structures have afine bal-ance between segregation and aggregation(mixed,but not scrambled)that provides functionality.Chaotic fabrication,a technology that exploits the ability of chaotic advection to create predictable and reproducible multilayered structures,excels at producing materials where this balance can be achieved andfinely tuned.This method is based on the use of chaotic mixing systems,which can produce constructs with highly organized internal micro-architecture in a simple and cost-effective way.This manuscript provides a perspective on how chaotic printing can be a great enabler in the manufacture of advanced materials,including living tissues.Chaotic printing may overcome many of the critical hurdles that are currently faced in man-ufacturing and biofabrication(e.g.,creating a wide array of interfaces,reaching high resolutions rapidly and at low cost,and producing densely vascularized tissues).The manuscript introduces the technology,explains how the idea originated,presents a timeline that provides a recapitulation of the milestones achieved so far,describes the main characteristics,advantages,limitations,and challenges of the technology,and concludes with future perspectives on the evolution and use of this versatile method.

Droplet 3D cryobioprinting for fabrication of free-standing and volumetric structures

Droplet-based bioprinting has shown remarkable potential in tissue engineering and regenerative medicine.However,it requires bioinks with low viscosities,which makes it challenging to create complex 3D structures and spatially pattern them with different materials.This study introduces a novel approach to bioprinting sophisti-cated volumetric objects by merging droplet-based bioprinting and cryobioprinting techniques.By leveraging the benefits of cryopreservation,we fabricated,for thefirst time,intricate,self-supporting cell-free or cell-laden structures with single or multiple materials in a simple droplet-based bioprinting process that is facilitated by depositing the droplets onto a cryoplate followed by crosslinking during revival.The feasibility of this approach is demonstrated by bioprinting several cell types,with cell viability increasing to 80%–90%after up to 2 or 3 weeks of culture.Furthermore,the applicational capabilities of this approach are showcased by bio-printing an endothelialized breast cancer model.The results indicate that merging droplet and cryogenic bioprinting complements current droplet-based bioprinting techniques and opens new avenues for the fabrication of volumetric objects with enhanced complexity and functionality,presenting exciting potential for biomedical applications.

Jetting-based bioprinting:process,dispense physics,and applications

Jetting-based bioprinting facilitates contactless drop-on-demand deposition of subnanoliter droplets at well-defined positions to control the spatial arrangement of cells,growth factors,drugs,and biomaterials in a highly automated layer-by-layer fabrication approach.Due to its immense versatility,jetting-based bioprinting has been used for various applications,including tissue engineering and regenerative medicine,wound healing,and drug development.A lack of in-depth understanding exists in the processes that occur during jetting-based bioprinting.This review paper will comprehensively discuss the physical considerations for bioinks and printing conditions used in jetting-based bioprinting.We first present an overview of different jetting-based bioprinting techniques such as inkjet bioprinting,laser-induced forward transfer bioprinting,electrohydrodynamic jet bioprinting,acoustic bioprinting and microvalve bioprinting.Next,we provide an in-depth discussion of various considerations for bioink formulation relating to cell deposition,print chamber design,droplet formation and droplet impact.Finally,we highlight recent accomplishments in jetting-based bioprinting.We present the advantages and challenges of each method,discuss considerations relating to cell viability and protein stability,and conclude by providing insights into future directions of jetting-based bioprinting.

Spheroid construction strategies and application in 3D bioprinting

Tissue engineering has been striving toward designing and producing natural and functional human tissues.Cells are the fundamental building blocks of tissues.Compared with traditional two-dimensional cultured cells,cell spheres are threedimensional(3D)structures that can naturally form complex cell–cell and cell–matrix interactions.This structure is close to the natural environment of cells in living organisms.In addition to being used in disease modeling and drug screening,spheroids have significant potential in tissue regeneration.The 3D bioprinting is an advanced biofabrication technique.It accurately deposits bioinks into predesigned 3D shapes to create complex tissue structures.Although 3D bioprinting is efficient,the time required for cells to develop into complex tissue structures can be lengthy.The 3D bioprinting of spheroids significantly reduces the time required for their development into large tissues/organs during later cultivation stages by printing them with high cell density.Combining spheroid fabrication and bioprinting technology should provide a new solution to many problems in regenerative medicine.This paper systematically elaborates and analyzes the spheroid fabrication methods and 3D bioprinting strategies by introducing spheroids as building blocks.Finally,we present the primary challenges faced by spheroid fabrication and 3D bioprinting with future requirements and some recommendations.

Advanced biomedical and electronic dual-function skin patch created through microfluidic-regulated 3D bioprinting

Artificial skin involves multidisciplinary efforts,including materials science,biology,medicine,and tissue engineering.Recent studies have aimed at creating skins that are multifunctional,intelligent,and capable of regenerating tissue.In this work,we present a specialized 3D printing ink composed of polyurethane and bioactive glass(PU-BG)and prepare dual-function skin patch by microfluidic-regulated 3D bioprinting(MRBP)technique.The MRBP endows the skin patch with a highly controlled microstructure and superior strength.Besides,an asymmetric tri-layer is further constructed,which promotes cell attachment and growth through a dual transport mechanism based on hydrogen bonds and gradient structure from hydrophilic to superhydrophilic.More importantly,by combining the features of biomedical skin with electronic skin(e-skin),we achieved a biomedical and electronic dual-function skin patch.In vivo experiments have shown that this skin patch can enhance hemostasis,resist bacterial growth,stimulate the regeneration of blood vessels,and accelerate the healing process.Meanwhile,it also mimics the sensory functions of natural skin to realize signal detection,where the sensitivity reached up to 5.87 kPa1,as well as cyclic stability(over 500 cycles),a wide detection range of 0–150 kPa,high pressure resolution of 0.1%under the pressure of 100 kPa.This work offers a versatile and effective method for creating dual-function skin patches and provide new insights into wound healing and tissue repair,which have significant implications for clinical applications.

3D bioprinting of in vitro porous hepatoma models:establishment,evaluation,and anticancer drug testing

Traditional tumor models do not tend to accurately simulate tumor growth in vitro or enable personalized treatment and are particularly unable to discover more beneficial targeted drugs.To address this,this study describes the use of threedimensional(3D)bioprinting technology to construct a 3D model with human hepatocarcinoma SMMC-7721 cells(3DP-7721)by combining gelatin methacrylate(GelMA)and poly(ethylene oxide)(PEO)as two immiscible aqueous phases to form a bioink and innovatively applying fluorescent carbon quantum dots for long-term tracking of cells.The GelMA(10%,mass fraction)and PEO(1.6%,mass fraction)hydrogel with 3:1 volume ratio offered distinct pore-forming characteristics,satisfactorymechanical properties,and biocompatibility for the creation of the 3DP-7721 model.Immunofluorescence analysis and quantitative real-time fluorescence polymerase chain reaction(PCR)were used to evaluate the biological properties of the model.Compared with the two-dimensional culture cell model(2D-7721)and the 3D mixed culture cell model(3DM-7721),3DP-7721 significantly improved the proliferation of cells and expression of tumor-related proteins and genes.Moreover,we evaluated the differences between the three culture models and the effectiveness of antitumor drugs in the three models and discovered that the efficacy of antitumor drugs varied because of significant differences in resistance proteins and genes between the three models.In addition,the comparison of tumor formation in the three models found that the cells cultured by the 3DP-7721 model had strong tumorigenicity in nude mice.Immunohistochemical evaluation of the levels of biochemical indicators related to the formation of solid tumors showed that the 3DP-7721 model group exhibited pathological characteristics of malignant tumors,the generated solid tumors were similar to actual tumors,and the deterioration was higher.This research therefore acts as a foundation for the application of 3DP-7721 models in drug development research.

Computed tomography three-dimensional reconstruction in the diagnosis of bleeding small intestinal polyps:A case report

BACKGROUND Computed tomography(CT)small bowel three-dimensional(3D)reconstruction is a powerful tool for the diagnosis of small bowel disease and can clearly show the intestinal lumen and wall as well as the outside structure of the wall.The horizontal axis position can show the best adjacent intestinal tube and the lesion between the intestinal tubes,while the coronal position can show the overall view of the small bowel.The ileal end of the localization of the display of excellent,and easy to quantitative measurement of the affected intestinal segments,the sagittal position for the rectum and the pre-sacral lesions show the best,for the discovery of fistulae is also helpful.Sagittal view can show rectal and presacral lesions and is useful for fistula detection.It is suitable for the assessment of inflammatory bowel disease,such as assessment of disease severity and diagnosis and differential diagnosis of the small bowel and mesenteric space-occupying lesions as well as the judgment of small bowel obstruction points.CASE SUMMARY Bleeding caused by small intestinal polyps is often difficult to diagnose in clinical practice.This study reports a 29-year-old male patient who was admitted to the hospital with black stool and abdominal pain for 3 months.Using the combination of CT-3D reconstruction and capsule endoscopy,the condition was diagnosed correctly,and the polyps were removed using single-balloon enteroscopyendoscopic retrograde cholangiopancreatography without postoperative complications.CONCLUSION The role of CT-3D in gastrointestinal diseases was confirmed.CT-3D can assist in the diagnosis and treatment of gastrointestinal diseases in combination with capsule endoscopy and small intestinal microscopy.

Research on Three-Dimensional Simulation of the Internal Arc Gear Skiving

Aiming at the problems that the simulation accuracy which is reduced due to the simplification of the model,a three-dimensional simulation method based on solid modeling is being proposed.By analyzing the motion relationship and positional relationship between the caries knife and the workpiece,the coordinate system of the caries machining was established.With the MATLAB software,the cutting edge model and the blade sweeping surface model of the boring cutter are sequentially established.Boolean operation is performed on the blade swept surface formed by the tooth cutter teeth with time t and the workpiece tooth geometry as well as the undeformed three-dimensional chip geometry model and the instantaneous cogging geometry model are obtained at different times.Through the compare between gear end face simulation tooth profile and the theoretical inner arc tooth profile,we verified the accuracy and rationality of the proposed method.

Three-dimensional bioprinting collagen/silk fibroin scaffold combined with neural stem cells promotes nerve regeneration after spinal cord injury

Many studies have shown that bio-scaffolds have important value for promoting axonal regeneration of injured spinal cord.Indeed,cell transplantation and bio-scaffold implantation are considered to be effective methods for neural regeneration.This study was designed to fabricate a type of three-dimensional collagen/silk fibroin scaffold (3D-CF) with cavities that simulate the anatomy of normal spinal cord.This scaffold allows cell growth in vitro and in vivo.To observe the effects of combined transplantation of neural stem cells (NSCs) and 3D-CF on the repair of spinal cord injury.Forty Sprague-Dawley rats were divided into four groups: sham (only laminectomy was performed),spinal cord injury (transection injury of T10 spinal cord without any transplantation),3D-CF (3D scaffold was transplanted into the local injured cavity),and 3D-CF + NSCs (3D scaffold co-cultured with NSCs was transplanted into the local injured cavity.Neuroelectrophysiology,imaging,hematoxylin-eosin staining,argentaffin staining,immunofluorescence staining,and western blot assay were performed.Apart from the sham group,neurological scores were significantly higher in the 3D-CF + NSCs group compared with other groups.Moreover,latency of the 3D-CF + NSCs group was significantly reduced,while the amplitude was significantly increased in motor evoked potential tests.The results of magnetic resonance imaging and diffusion tensor imaging showed that both spinal cord continuity and the filling of injury cavity were the best in the 3D-CF + NSCs group.Moreover,regenerative axons were abundant and glial scarring was reduced in the 3D-CF + NSCs group compared with other groups.These results confirm that implantation of 3D-CF combined with NSCs can promote the repair of injured spinal cord.This study was approved by the Institutional Animal Care and Use Committee of People’s Armed Police Force Medical Center in 2017 (approval No.2017-0007.2).

A theory for three-dimensional response of micropolar plates

Through combined applications of the transfer-matrix method and asymptotic expansion technique,we formulate a theory to predict the three-dimensional response of micropolar plates.No ad hoc assumptions regarding through-thickness assumptions of the field variables are made,and the governing equations are two-dimensional,with the displacements and microrotations of the mid-plane as the unknowns.Once the deformation of the mid-plane is solved,a three-dimensional micropolar elastic field within the plate is generated,which is exact up to the second order except in the boundary region close to the plate edge.As an illustrative example,the bending of a clamped infinitely long plate caused by a uniformly distributed transverse force is analyzed and discussed in detail.

Oxygen tension modulates cell function in an in vitro three-dimensional glioblastoma tumor model

Hypoxia is a typical feature of the tumor microenvironment,one of the most critical factors affecting cell behavior and tumor progression.However,the lack of tumor models able to precisely emulate natural brain tumor tissue has impeded the study of the effects of hypoxia on the progression and growth of tumor cells.This study reports a three-dimensional(3D)brain tumor model obtained by encapsulating U87MG(U87)cells in a hydrogel containing type I collagen.It also documents the effect of various oxygen concentrations(1%,7%,and 21%)in the culture environment on U87 cell morphology,proliferation,viability,cell cycle,apoptosis rate,and migration.Finally,it compares two-dimensional(2D)and 3D cultures.For comparison purposes,cells cultured in flat culture dishes were used as the control(2D model).Cells cultured in the 3D model proliferated more slowly but had a higher apoptosis rate and proportion of cells in the resting phase(G0 phase)/gap I phase(G1 phase)than those cultured in the 2D model.Besides,the two models yielded significantly different cell morphologies.Finally,hypoxia(e.g.,1%O2)affected cell morphology,slowed cell growth,reduced cell viability,and increased the apoptosis rate in the 3D model.These results indicate that the constructed 3D model is effective for investigating the effects of biological and chemical factors on cell morphology and function,and can be more representative of the tumor microenvironment than 2D culture systems.The developed 3D glioblastoma tumor model is equally applicable to other studies in pharmacology and pathology.

Three-dimensional bioprinting of gelatin methacryloyl (GelMA)

The three-dimensional (3D)bioprinting technology has progressed tremendously over the past decade.By controlling the size, shape,and architecture of the bioprinted constructs,3D bioprinting allows for the fabrication of tissue/organ-like constructs with strong structural-functional similarity with their in vivo counterparts at high fidelity.The bioink,a blend of biomaterials and living cells possessing both high biocompatibility and printability,is a critical component of bioprinting.In particular, gelatin methacryloyl (GelMA)has shown its potential as a viable bioink material due to its suitable biocompatibility and readily tunable physicochemical properties.Current GelMA-based bioinks and relevant bioprinting strategies for GelMA bioprinting are briefly reviewed.

Development of a toroidal soft x-ray imaging system and application for investigating three-dimensional plasma on J-TEXT

A toroidal soft x-ray imaging(T-SXRI)system has been developed to investigate threedimensional(3D)plasma physics on J-TEXT.This T-SXRI system consists of three sets of SXR arrays.Two sets are newly developed and located on the vacuum chamber wall at toroidal positionsφof 126.4°and 272.6°,respectively,while one set was established previously atφ=65.50.Each set of SXR arrays consists of three arrays viewing the plasma poloidally,and hence can be used separately to obtain SXR images via the tomographic method.The sawtooth precursor oscillations are measured by T-SXRI,and the corresponding images of perturbative SXR signals are successfully reconstructed at these three toroidal positions,hence providing measurement of the 3D structure of precursor oscillations.The observed 3D structure is consistent with the helical structure of the m/n=1/1 mode.The experimental observation confirms that the T-SXRI system is able to observe 3D structures in the J-TEXT plasma.

Intraoperative application of three-dimensional printed guides in total hip arthroplasty: A systematic review

BACKGROUND Acetabular component positioning in total hip arthroplasty(THA)is of key importance to ensure satisfactory post-operative outcomes and to minimize the risk of complications.The majority of acetabular components are aligned freehand,without the use of navigation methods.Patient specific instruments(PSI)and three-dimensional(3D)printing of THA placement guides are increasingly used in primary THA to ensure optimal positioning.AIM To summarize the literature on 3D printing in THA and how they improve acetabular component alignment.METHODS PubMed was used to identify and access scientific studies reporting on different 3D printing methods used in THA.Eight studies with 236 hips in 228 patients were included.The studies could be divided into two main categories;3D printed models and 3D printed guides.RESULTS 3D printing in THA helped improve preoperative cup size planning and post-operative Harris hip scores between intervention and control groups(P=0.019,P=0.009).Otherwise,outcome measures were heterogeneous and thus difficult to compare.The overarching consensus between the studies is that the use of 3D guidance tools can assist in improving THA cup positioning and reduce the need for revision THA and the associated costs.CONCLUSION The implementation of 3D printing and PSI for primary THA can significantly improve the positioning accuracy of the acetabular cup component and reduce the number of complications caused by malpositioning.

Computer-assisted three-dimensional individualized extreme liver resection for hepatoblastoma in proximity to the major liver vasculature

BACKGROUND The management of hepatoblastoma(HB)becomes challenging when the tumor remains in close proximity to the major liver vasculature(PMV)even after a full course of neoadjuvant chemotherapy(NAC).In such cases,extreme liver resection can be considered a potential option.AIM To explore whether computer-assisted three-dimensional individualized extreme liver resection is safe and feasible for children with HB who still have PMV after a full course of NAC.METHODS We retrospectively collected data from children with HB who underwent surgical resection at our center from June 2013 to June 2023.We then analyzed the detailed clinical and three-dimensional characteristics of children with HB who still had PMV after a full course of NAC.RESULTS Sixty-seven children diagnosed with HB underwent surgical resection.The age at diagnosis was 21.4±18.8 months,and 40 boys and 27 girls were included.Fifty-nine(88.1%)patients had a single tumor,39(58.2%)of which was located in the right lobe of the liver.A total of 47 patients(70.1%)had PRE-TEXT III or IV.Thirty-nine patients(58.2%)underwent delayed resection.After a full course of NAC,16 patients still had close PMV(within 1 cm in two patients,touching in 11 patients,compressing in four patients,and showing tumor thrombus in three patients).There were 6 patients of tumors in the middle lobe of the liver,and four of those patients exhibited liver anatomy variations.These 16 children underwent extreme liver resection after comprehensive preoperative evaluation.Intraoperative procedures were performed according to the preoperative plan,and the operations were successfully performed.Currently,the 3-year event-free survival of 67 children with HB is 88%.Among the 16 children who underwent extreme liver resection,three experienced recurrence,and one died due to multiple metastases.CONCLUSION Extreme liver resection for HB that is still in close PMV after a full course of NAC is both safe and feasible.This approach not only reduces the necessity for liver transplantation but also results in a favorable prognosis.Individualized three-dimensional surgical planning is beneficial for accurate and complete resection of HB,particularly for assessing vascular involvement,remnant liver volume and anatomical variations.