Effect of degradation of PLGA and PLGA/β-TCP scaffolds on the growth of osteoblasts

Osteoblasts were cultured on porous scaffolds of poly(L-lactide-co-glycolide) (PLGA) and PLGA/β-tricalcium phosphate (β-TCP) to evaluate their cytocompatibility.The proliferation of the cells on both scaffolds was examined before and after in vitro degradation for 4,8 and 12 weeks under static (shaking water bath) and dynamic (cyclic loading) conditions.Results indicate that porous PLGA and PLGA/β-TCP scaffolds have good biocompatibility and can be used as effective templates for guiding the growth of osteoblasts.The degradation of the scaffolds affects the proliferation of osteoblasts and the cell viability decreased with the degradation time.

Study on β-TCP Coated Porous Mg as a Bone Tissue Engineering Scaffold Material

Three-dimensional honeycomb-structured magnesium （Mg） scaffolds with interconnected pores of accurately controlled pore size and porosity were fabricated by laser perforation technique. Biodegradable and bioactiveβ- tricalcium phosphate （β-TCP） coatings were prepared on and the biodegradation mechanism was simply evaluated the porous Mg to further improve its biocompatibility, in vitro. It was found that the mechanical properties of this type of porous Mg significantly depended on its porosity. Elastic modulus and compressive strength similar to human bones could be obtained for the porous Mg with porosity of 42.6%-51%. It was observed that the human osteosarcoma cells （UMR106） were well adhered and proliferated on the surface of the β- TCP coated porous Mg, which indicates that theβ-TCP coated porous Mg is promising to be a bone tissue engineering scaffold material.

In Vitro Characterizations of PLLA/β-TCP Porous Matrix Materials and RMSC-PLLA-β-TCP Composite Scaffolds

To develop a novel degradable poly (L-lactic acid)/β-tricalcium phosphate (PLLA/β-TCP) bioactive materials for bone tissueengineering, β-TCP powder was produced by a new wet process. Porous scaffolds were prepared by three steps, i.e. solventcasting, compression molding and leaching stage. Factors influencing the compressive strength and the degradation behaviorof the porous scaffold, e.g. weight fraction of pore forming agent-sodium chloride (NaCl), weight ratio of PLLA: β-TCP,the particle size of β-TCP and the porosity, were discussed in details. Rat marrow stromal cells (RMSC) were incorporatedinto the composite by tissue engineering approach. Biological and osteogenesis potential of the composite scaffold weredetermined with MTT assay, alkaline phosphatase (ALP) activity and bone osteocalcin (OCN) content evaluation. Resultsshow that PLLA/β-TCP bioactive porous scaffold has good mechanical and pore structure with adjustable compressive strengthneeded for surgery. RMSCs seeding on porous PLLA/β-TCP composite behaves good seeding efficacy, biocompatibility andosteoinductive potential. Osteoprogenitor cells could well penetrate into the material matrix and begin cell proliferation andosteogenic differentiation. Osseous matrix could be formed on the surface of the composite after culturing in vitro. It isexpected that the PLLA/β-TCP porous composites are promising scaffolds for bone tissue engineering in prosthesis surgery.

Preparation and Characterization of β-TCP/CS Scaffolds by Freeze-extraction and Freeze-gelation

The freeze-extraction and freeze-gelation methods were used to prepare highly porous β-TCP/CS scaffolds with different β-TCP/CS ratio.In these methods,the suspending mixture of β-TCP and chitosan was frozen,the frozen mixture was immersed in a non-solvent（0.05mol/L NaOH/ehanol aqueous） bath to allow the exchange between solvent（acetic-acid aqueous） and non-solvent at a temperature lower than the freezing point of the acetic-acid.Then,the β-TCP/CS scaffolds were formed and dried at room temperature.Scanning electron microscopy （SEM）,X-ray diffraction （XRD）,gas chromatography （GC） and omnipotence material testing machine were employed to characterize the β-TCP/CS scaffolds.The results of GC show that the freeze extraction of the β-TCP/chitosan scaffolds was completed when the extraction time is above 24 h.The SEM results show that the β-TCP/CS scaffolds are composed of interconnected pore network.The porosity of the β-TCP/CS scaffolds decrease with the increase of the content of the β-TCP.The β-TCP/CS scaffolds have a highest compressive strength when the chitosan/β-TCP ratio is 30：70.The present work displays that the β-TCP/CS composite scaffolds with appropriate mechanical properties and high porosity can be successfully prepared by the freeze-extraction and freeze-gelation methods.

Insights into Nano-and Micro-Structured Scaffolds for Advanced Electrochemical Energy Storage

Adopting a nano-and micro-structuring approach to fully unleashing the genuine potential of electrode active material benefits in-depth understandings and research progress toward higher energy density electrochemical energy stor-age devices at all technology readiness levels.Due to various challenging issues,especially limited stability,nano-and micro-structured(NMS)electrodes undergo fast electrochemical performance degradation.The emerging NMS scaffold design is a pivotal aspect of many electrodes as it endows them with both robustness and electrochemical performance enhancement,even though it only occupies comple-mentary and facilitating components for the main mechanism.However,extensive efforts are urgently needed toward optimizing the stereoscopic geometrical design of NMS scaffolds to minimize the volume ratio and maximize their functionality to fulfill the ever-increasing dependency and desire for energy power source supplies.This review will aim at highlighting these NMS scaffold design strategies,summariz-ing their corresponding strengths and challenges,and thereby outlining the potential solutions to resolve these challenges,design principles,and key perspectives for future research in this field.Therefore,this review will be one of the earliest reviews from this viewpoint.

Modifi cation of β-TCP/PLGA Scaffold and Its Effect on Bone Regeneration in vivo

In order to look for the best proportion of β-tricalcium phosphate（β-TCP）and poly（lactideco-glycolide）（PLGA）we fabricated porous composites β-TCP/PLGA scaffold using freeze-drying method.Morphologicalcharacterization using scanning electron microscopy showed that the interconnected pore distribution was even and there was no significant difference with the increase of PLGA content.Moreover,the porosity,compressive strength and degradation in vitro were characterized.The fabricated scaffolds with increased PLGA in the composites β-TCP/PLGA scaffolds willget stronger mechanicalproperty and better appearance,furthermore,get suitable environment for cells.According to the evaluation indexes for the tissue engineering scaffold,the group of scaffold（β-TCP/PLGA=6：4）was selected to evaluate the induced celladhesion and proliferative ability of the scaffolds.Then as transplant embed into the bone criticaldefect sites on rats femur.The repairing processes of bone defect sites were characterized by X-ray analysis within 12 weeks.X-ray analysis showed that the bone defect sites alldisplayed the formation of callus obviously,In summary,our data suggest that the scaffold（β-TCP/PLGA=6：4）has a promising clinicalfuture in regeneration of bone criticaldefects.

PLLA/β-TCP支架表面多孔结构的构建及其对细胞黏附的影响

目的通过NaOH溶液的简单处理构建3D打印PLLA/β-TCP骨组织工程支架表面多孔结构,增加支架的粗糙度和亲水性,促进支架表面的细胞黏附。方法通过3D打印熔融沉积成型技术制备PLLA/β-TCP网状支架,并通过NaOH蚀刻的方法进行支架的粗糙化改性,依据支架表面微观形貌、能谱、接触角、力学、细胞黏附等观察NaOH浓度、时间两项反应参数对支架的影响。结果通过熔融沉积成型技术制备的PLLA/β-TCP复合支架存在预先设置的网状结构;经NaOH蚀刻构建了兼有宏观和微观空隙的多孔形态。NaOH浓度、时间中任意一种参数的增加都会导致支架表面微观孔隙的孔径、孔密度增加。NaOH处理参数为0.1 mol/L(9 h)时,可显著减小支架表面水接触角,且对支架的压缩强度无显著影响。体外细胞检测显示,经NaOH蚀刻后的表面多孔复合支架在骨髓间充质干细胞(BMSCs)的黏附增殖上更具优势。结论用NaOH处理3D打印PLLA/β-TCP骨组织工程支架可有效改善支架表面形态,优化该类支架的亲水性及细胞黏附。

Customized scaffolds for large bone defects using 3D‑printed modular blocks from 2D‑medical images

Additive manufacturing(AM)has revolutionized the design and manufacturing of patient-specific,three-dimensional(3D),complex porous structures known as scaffolds for tissue engineering applications.The use of advanced image acquisition techniques,image processing,and computer-aided design methods has enabled the precise design and additive manufacturing of anatomically correct and patient-specific implants and scaffolds.However,these sophisticated techniques can be timeconsuming,labor-intensive,and expensive.Moreover,the necessary imaging and manufacturing equipment may not be readily available when urgent treatment is needed for trauma patients.In this study,a novel design and AM methods are proposed for the development of modular and customizable scaffold blocks that can be adapted to fit the bone defect area of a patient.These modular scaffold blocks can be combined to quickly form any patient-specific scaffold directly from two-dimensional(2D)medical images when the surgeon lacks access to a 3D printer or cannot wait for lengthy 3D imaging,modeling,and 3D printing during surgery.The proposed method begins with developing a bone surface-modeling algorithm that reconstructs a model of the patient’s bone from 2D medical image measurements without the need for expensive 3D medical imaging or segmentation.This algorithm can generate both patient-specific and average bone models.Additionally,a biomimetic continuous path planning method is developed for the additive manufacturing of scaffolds,allowing porous scaffold blocks with the desired biomechanical properties to be manufactured directly from 2D data or images.The algorithms are implemented,and the designed scaffold blocks are 3D printed using an extrusion-based AM process.Guidelines and instructions are also provided to assist surgeons in assembling scaffold blocks for the self-repair of patient-specific large bone defects.

氢氧化钠碱蚀法改善3D打印PCL/β-TCP网状支架表面亲水性及其性能研究

目的为解决聚己内酯(polycaprolactone,PCL)/β-TCP骨组织工程支架表面亲水性不足的问题,通过NaOH蚀刻法改善3D打印PCL/β-TCP支架表面微形貌,进一步影响其亲水性和细胞反应。方法采用3D打印熔融沉积成型(fused deposition modeling,FDM)技术制备PCL/β-TCP网状支架,通过NaOH蚀刻法进行支架表面粗糙化改性,观察NaOH浓度、时间2项反应参数对支架改性前、后在微观形貌、能谱元素、接触角、压缩强度、细胞黏附等性能方面的影响。结果经NaOH蚀刻后成功制备PCL/β-TCP网状支架表面微孔结构。随着NaOH浓度、时间任一种参数的增加均会导致支架表面微孔增大、材料表面接触角减小,但NaOH处理参数为1 mol/L(24 h)或10 mol/L(6 h)的蚀刻支架,其压缩强度与未处理组对比,差异没有统计学意义(P=0.071);蚀刻支架上细胞数目增多,单个细胞铺展面积大,在骨髓间充质干细胞(BMSCs)黏附增殖上更具优势。结论采用NaOH蚀刻法改善3D打印PCL/β-TCP骨组织工程支架亲水性的方法是一种低成本的有效策略,可有效改善支架湿润性和细胞黏附。

室温挤出3D打印HA/β-TCP/DCPA骨支架的复合浆料研究

基于3D打印骨支架的骨组织工程是一种具有应用潜力的骨修复策略。本文将植酸螯合改性的羟基磷灰石(IP6-HA)、β-磷酸三钙(β-TCP)与无水磷酸二氢钙(MCPA)按照不同摩尔比混合的复合粉体作为固相,以不同丙三醇(甘油)含量的水溶液作为液相,通过调整复合粉体中IP6-HA粉末的含量与固化液中甘油的含量,制备了能够应用于室温挤出3D打印骨支架的透钙磷石体系磷酸钙复合浆料。结果表明:复合浆料的水化产物主要为HA、β-TCP和无水磷酸氢钙(DCPA);当固化液中甘油含量为50%(质量分数)时,复合浆料均表现出优异的可打印性能与力学性能,抗压强度最高可达19.8 MPa;残余IP6-HA的存在改变了固化样品表面磷灰石的矿化结构并提升了矿化能力;制备的支架结构完整,孔洞均匀,抗压强度可达5.8 MPa。本研究制备的磷酸钙复合浆料能够通过室温挤出式3D打印技术制备骨支架,在3D打印领域具有良好的应用前景。

Biological scaffold as potential platforms for stem cells:Current development and applications in wound healing

Wound repair is a complex challenge for both clinical practitioners and researchers.Conventional approaches for wound repair have several limitations.Stem cell-based therapy has emerged as a novel strategy to address this issue,exhibiting significant potential for enhancing wound healing rates,improving wound quality,and promoting skin regeneration.However,the use of stem cells in skin regeneration presents several challenges.Recently,stem cells and biomaterials have been identified as crucial components of the wound-healing process.Combination therapy involving the development of biocompatible scaffolds,accompanying cells,multiple biological factors,and structures resembling the natural extracellular matrix(ECM)has gained considerable attention.Biological scaffolds encompass a range of biomaterials that serve as platforms for seeding stem cells,providing them with an environment conducive to growth,similar to that of the ECM.These scaffolds facilitate the delivery and application of stem cells for tissue regeneration and wound healing.This article provides a comprehensive review of the current developments and applications of biological scaffolds for stem cells in wound healing,emphasizing their capacity to facilitate stem cell adhesion,proliferation,differentiation,and paracrine functions.Additionally,we identify the pivotal characteristics of the scaffolds that contribute to enhanced cellular activity.

3D-printed Mg-1Ca/polycaprolactone composite scaffolds with promoted bone regeneration

In bone tissue engineering,polycaprolactone(PCL)is a promising material with good biocompatibility,but its poor degradation rate,mechanical strength,and osteogenic properties limit its application.In this study,we developed an Mg-1Ca/polycaprolactone(Mg-1Ca/PCL)composite scaffolds to overcome these limitations.We used a melt blending method to prepare Mg-1Ca/PCL composites with Mg-1Ca alloy powder mass ratios of 5,10,and 20 wt%.Porous scaffolds with controlled macro-and microstructure were printed using the fused deposition modeling method.We explored the mechanical strength,biocompatibility,osteogenesis performance,and molecular mechanism of the Mg-1Ca/PCL composites.The 5 and 10 wt%Mg-1Ca/PCL composites were found to have good biocompatibility.Moreover,they promoted the mechanical strength,proliferation,adhesion,and osteogenic differentiation of human bone marrow stem cells(hBMSCs)of pure PCL.In vitro degradation experiments revealed that the composite material stably released Mg_(2)+ions for a long period;it formed an apatite layer on the surface of the scaffold that facilitated cell adhesion and growth.Microcomputed tomography and histological analysis showed that both 5 and 10 wt%Mg-1Ca/PCL composite scaffolds promoted bone regeneration bone defects.Our results indicated that the Wnt/β-catenin pathway was involved in the osteogenic effect.Therefore,Mg-1Ca/PCL composite scaffolds are expected to be a promising bone regeneration material for clinical application.Statement of significance:Bone tissue engineering scaffolds have promising applications in the regeneration of critical-sized bone defects.However,there remain many limitations in the materials and manufacturing methods used to fabricate scaffolds.This study shows that the developed Ma-1Ca/PCL composites provides scaffolds with suitable degradation rates and enhanced boneformation capabilities.Furthermore,the fused deposition modeling method allows precise control of the macroscopic morphology and microscopic porosity of the scaffold.The obtained porous scaffolds can significantly promote the regeneration of bone defects.

Constructing a biofunctionalized 3D-printed gelatin/sodium alginate/chitosan tri-polymer complex scaffold with improvised biological andmechanical properties for bone-tissue engineering

Sodium alginate(SA)/chitosan(CH)polyelectrolyte scaffold is a suitable substrate for tissue-engineering application.The present study deals with further improvement in the tensile strength and biological properties of this type of scaffold to make it a potential template for bone-tissue regeneration.We experimented with adding 0%–15%(volume fraction)gelatin(GE),a protein-based biopolymer known to promote cell adhesion,proliferation,and differentiation.The resulting tri-polymer complex was used as bioink to fabricate SA/CH/GEmatrices by three-dimensional(3D)printing.Morphological studies using scanning electron microscopy revealed the microfibrous porous architecture of all the structures,which had a pore size range of 383–419μm.X-ray diffraction and Fourier-transform infrared spectroscopy analyses revealed the amorphous nature of the scaffold and the strong electrostatic interactions among the functional groups of the polymers,thereby forming polyelectrolyte complexes which were found to improve mechanical properties and structural stability.The scaffolds exhibited a desirable degradation rate,controlled swelling,and hydrophilic characteristics which are favorable for bone-tissue engineering.The tensile strength improved from(386±15)to(693±15)kPa due to the increased stiffness of SA/CH scaffolds upon addition of gelatin.The enhanced protein adsorption and in vitro bioactivity(forming an apatite layer)confirmed the ability of the SA/CH/GE scaffold to offer higher cellular adhesion and a bone-like environment to cells during the process of tissue regeneration.In vitro biological evaluation including the MTT assay,confocal microscopy analysis,and alizarin red S assay showed a significant increase in cell attachment,cell viability,and cell proliferation,which further improved biomineralization over the scaffold surface.In addition,SA/CH containing 15%gelatin designated as SA/CH/GE15 showed superior performance to the other fabricated 3D structures,demonstrating its potential for use in bone-tissue engineering.

In vitro investigations on the effects of graphene and graphene oxide on polycaprolactone bone tissue engineering scaffolds

Polycaprolactone(PCL)scaffolds that are produced through additive manufacturing are one of the most researched bone tissue engineering structures in the field.Due to the intrinsic limitations of PCL,carbon nanomaterials are often investigated to reinforce the PCL scaffolds.Despite several studies that have been conducted on carbon nanomaterials,such as graphene(G)and graphene oxide(GO),certain challenges remain in terms of the precise design of the biological and nonbiological properties of the scaffolds.This paper addresses this limitation by investigating both the nonbiological(element composition,surface,degradation,and thermal and mechanical properties)and biological characteristics of carbon nanomaterial-reinforced PCL scaffolds for bone tissue engineering applications.Results showed that the incorporation of G and GO increased surface properties(reduced modulus and wettability),material crystallinity,crystallization temperature,and degradation rate.However,the variations in compressive modulus,strength,surface hardness,and cell metabolic activity strongly depended on the type of reinforcement.Finally,a series of phenomenological models were developed based on experimental results to describe the variations of scaffold’s weight,fiber diameter,porosity,and mechanical properties as functions of degradation time and carbon nanomaterial concentrations.The results presented in this paper enable the design of three-dimensional(3D)bone scaffolds with tuned properties by adjusting the type and concentration of different functional fillers.

Coaxial electrohydrodynamic printing of core–shell microfibrous scaffolds with layer-specific growth factors release for enthesis regeneration

The rotator cuff tear has emerged as a significant global health concern.However,existing therapies fail to fully restore the intricate bone-to-tendon gradients,resulting in compromised biomechanical functionalities of the reconstructed enthesis tissues.Herein,a tri-layered core–shell microfibrous scaffold with layer-specific growth factors(GFs)release is developed using coaxial electrohydrodynamic(EHD)printing for in situ cell recruitment and differentiation to facilitate gradient enthesis tissue repair.Stromal cell-derived factor-1(SDF-1)is loaded in the shell,while basic fibroblast GF,transforming GF-beta,and bone morphogenetic protein-2 are loaded in the core of the EHD-printed microfibrous scaffolds in a layer-specific manner.Correspondingly,the tri-layered microfibrous scaffolds have a core–shell fiber size of(25.7±5.1)μm,with a pore size sequentially increasing from(81.5±4.6)μm to(173.3±6.9)μm,and to(388.9±6.9μm)for the tenogenic,chondrogenic,and osteogenic instructive layers.A rapid release of embedded GFs is observed within the first 2 d,followed by a faster release of SDF-1 and a slightly slower release of differentiation GFs for approximately four weeks.The coaxial EHD-printed microfibrous scaffolds significantly promote stem cell recruitment and direct their differentiation toward tenocyte,chondrocyte,and osteocyte phenotypes in vitro.When implanted in vivo,the tri-layered core–shell microfibrous scaffolds rapidly restored the biomechanical functions and promoted enthesis tissue regeneration with native-like bone-to-tendon gradients.Our findings suggest that the microfibrous scaffolds with layer-specific GFs release may offer a promising clinical solution for enthesis regeneration.

Oxygen vacancy boosting Fenton reaction in bone scaffold towards fighting bacterial infection

Bacterial infection is a major issue after artificial bone transplantation due to the absence of antibacterial function of bone scaffold,which seriously causes the transplant failure and even amputation in severe cases.In this study,oxygen vacancy(OV)defects Fe-doped Ti O2(OV-FeTiO2)nanoparticles were synthesized by nano TiO2and Fe3O4via high-energy ball milling,which was then incorporated into polycaprolactone/polyglycolic acid(PCLGA)biodegradable polymer matrix to construct composite bone scaffold with good antibacterial activities by selective laser sintering.The results indicated that OV defects were introduced into the core/shell-structured OV-FeTiO2nanoparticles through multiple welding and breaking during the high-energy ball milling,which facilitated the adsorption of hydrogen peroxide(H2O2)in the bacterial infection microenvironment at the bone transplant site.The accumulated H2O2could amplify the Fenton reaction efficiency to induce more hydroxyl radicals(·OH),thereby resulting in more bacterial deaths through·OH-mediated oxidative damage.This antibacterial strategy had more effective broad-spectrum antibacterial properties against Gram-negative Escherichia coli(E.coli)and Gram-positive Staphylococcus aureus(S.aureus).In addition,the PCLGA/OV-FeTiO2scaffold possessed mechanical properties that match those of human cancellous bone and good biocompatibility including cell attachment,proliferation and osteogenic differentiation.

Numerical Analysis of Permeability of Functionally Graded Scaffolds

In this work,we numerically study the hydrodynamic permeability of new-generation artificial porous materials used as scaffolds for cell growth in a perfusion bioreactor.We consider two popular solid matrix designs based on triply periodic minimal surfaces,the Schwarz P(primitive)and D(diamond)surfaces,which enable the creation of materials with controlled porosity gradients.The latter property is crucial for regulating the shear stress field in the pores of the scaffold,which makes it possible to control the intensity of cell growth.The permeability of functionally graded materials is studied within the framework of both a microscopic approach based on the Navier-Stokes equation and an averaged description of the liquid filtration through a porous medium based on the equations of the Darcy or Forchheimer models.We calculate the permeability coefficients for both types of solid matrices formed by Schwarz surfaces,study their properties concerning forward and reverse fluid flows,and determine the ranges of Reynolds number for which the description within the Darcy or Forchheimer model is applicable.Finally,we obtain a shear stress field that varies along the sample,demonstrating the ability to tune spatially the rate of tissue growth.

Flame Retardant Material Based on Cellulose Scaffold Mineralized by Calcium Carbonate

Wood-based functional materials have developed rapidly.But the flammability significantly limits its further application.To improve the flame retardancy,the balsa wood was delignified by NaClO2 solution to create a cellulose scaffold,and then alternately immersed in CaCl_(2) ethanol solution and NaHCO3 aqueous solution under vacuum.The high porosity and wettability resulting from delignification benefited the following mineralization process,changing the thermal properties of balsa wood significantly.The organic-inorganic wood composite showed abundant CaCO_(3) spherical particles under scanning electron microscopy.The peak of the heat release rate of delignified balsa-CaCO_(3) was reduced by 33%compared to the native balsa,according to the cone calorimetric characterization.The flame test demonstrated that the mineralized wood was flame retardant and selfextinguish.Additionally,the mineralized wood also displayed lower thermal conductivity.This study developed a feasible way to fabricate a lightweight,fire-retardant,self-extinguishing,and heat-insulating wood composite,providing a promising route for the valuable application of cellulosic biomass.

Advanced strategies for 3D-printed neural scaffolds:materials,structure,and nerve remodeling

Nerve regeneration holds significant potential in the treatment of various skeletal and neurological disorders to restore lost sensory and motor functions.The potential of nerve regeneration in ameliorating neurological diseases and injuries is critical to human health.Three-dimensional(3D)printing offers versatility and precision in the fabrication of neural scaffolds.Complex neural structures such as neural tubes and scaffolds can be fabricated via 3Dprinting.This reviewcomprehensively analyzes the current state of 3D-printed neural scaffolds and explores strategies to enhance their design.It highlights therapeutic strategies and structural design involving neural materials and stem cells.First,nerve regeneration materials and their fabrication techniques are outlined.The applications of conductive materials in neural scaffolds are reviewed,and their potential to facilitate neural signal transmission and regeneration is highlighted.Second,the progress in 3D-printed neural scaffolds applied to the peripheral and central nerves is comprehensively evaluated,and their potential to restore neural function and promote the recovery of different nervous systems is emphasized.In addition,various applications of 3D-printed neural scaffolds in peripheral and neurological diseases,as well as the design strategies of multifunctional biomimetic scaffolds,are discussed.

Enhanced axonal regeneration and functional recovery of the injured sciatic nerve in a rat model by lithium-loaded electrospun nanofibrous scaffolds

Increasing evidence indicates that engineered nerve grafts have great potential for the regeneration of peripheral nerve injuries(PNIs).While most studies have focused only on the topographical features of the grafts,we have considered both the biophysical and biochemical manipulations in our applied nanoscaffold.To achieve this,we fabricated an electrospun nanofibrous scaffold(ENS)containing polylactide nanofibers loaded with lithium(Li)ions,a Wnt/β-catenin signaling activator.In addition,we seeded human adipose-derived mesenchymal stem cells(hADMSCs)onto this engineered scaffold to examine if their differentiation toward Schwann-like cells was induced.We further examined the efficacy of the scaffolds for nerve regeneration in vivo via grafting in a PNI rat model.Our results showed that Li-loaded ENSs gradually released Li within 11 d,at concentrations ranging from 0.02 to(3.64±0.10)mmol/L,and upregulated the expression of Wnt/β-catenin target genes(cyclinD1 and c-Myc)as well as those of Schwann cell markers(growth-associated protein 43(GAP43),S100 calcium binding protein B(S100B),glial fibrillary acidic protein(GFAP),and SRY-box transcription factor 10(SOX10))in differentiated hADMSCs.In the PNI rat model,implantation of Li-loaded ENSs with/without cells improved behavioral features such as sensory and motor functions as well as the electrophysiological characteristics of the injured nerve.This improved function was further validated by histological analysis of sciatic nerves grafted with Li-loaded ENSs,which showed no fibrous connective tissue but enhanced organized myelinated axons.The potential of Li-loaded ENSs in promoting Schwann cell differentiation of hADMSCs and axonal regeneration of injured sciatic nerves suggests their potential for application in peripheral nerve tissue engineering.