The fabrication of hydroxyapatite mineralized hydrogels for bone tissue engineering

Bone is a complex but orderly mineralized tissue with hydroxyapatite(HA)as the inorganic phase and collagen as the organic phase.Inspired by natural bone tissues,HA-mineralized hydrogels have been widely designed and used in bone tissue engineering.HA is majorly utilized for the treatment of bone defects because of its excellent osteoconduction and bone inductivity.Hydrogel is a three-dimensional hydrophilic network structure with similar properties to the extracellular matrix(ECM).The combination of HA and hydrogels produces a new hybrid material that could effectively promote osteointegration and accelerate the healing of bone defects.In this review,the structure and growth of bone and the common strategies used to prepare HA were briefly introduced.Importantly,we discussed the fabrication of HA mineralized hydrogels from simple blending to in situ mineralization.We hope this review can provide a reference for the development of bone repair hydrogels.

Exploring the interconnectivity of biomimetic hierarchical porous Mg scaffolds for bone tissue engineering:Effects of pore size distribution on mechanical properties,degradation behavior and cell migration ability

Interconnectivity is the key characteristic of bone tissue engineering scaffold modulating cell migration,blood vessels invasion and transport of nutrient and waste.However,efforts and understanding of the interconnectivity of porous Mg is limited due to the diverse architectures of pore struts and pore size distribution of Mg scaffold systems.In this work,biomimetic hierarchical porous Mg scaffolds with tailored interconnectivity as well as pore size distribution were prepared by template replication of infiltration casting.Mg scaffold with better interconnectivity showed lower mechanical strength.Enlarging interconnected pores would enhance the interconnectivity of the whole scaffold and reduce the change of ion concentration,pH value and osmolality of the degradation microenvironment due to the lower specific surface area.Nevertheless,the degradation rates of five tested Mg scaffolds were no different because of the same geometry of strut unit.Direct cell culture and evaluation of cell density at both sides of four typical Mg scaffolds indicated that cell migration through hierarchical porous Mg scaffolds could be enhanced by not only bigger interconnected pore size but also larger main pore size.In summary,design of interconnectivity in terms of pore size distribution could regulate mechanical strength,microenvironment in cell culture condition and cell migration potential,and beyond that it shows great potential for personalized therapy which could facilitate the regeneration process.

Reduced graphene oxide-grafted bovine serum albumin/bredigite nanocomposites with high mechanical properties and excellent osteogenic bioactivity for bone tissue engineering

The optimization of the scaffolds to provide a suitable matrix and accelerate the regeneration process is vital for bone tissue engineering.However,poor mechanical and biological characteristics remain the primary challenges that must be addressed.For example,although bredigite(Br)has shown great potential for application in bone tissue engineering,it easily fails in replacement.In the present work,these challenges are addressed by reinforcing the Br matrix with nanosheets of graphene oxide(rGO)that have been reduced by bovine serum albumin(BSA)in order to enhance the mechanical properties and biological behavior.The reduction of graphene oxide by BSA improves the water stability of the nanosheets and provides an electrostatic interaction between theBSA-rGO nanosheets and theBr particles.The high thermal conductivity of theBSA-rGO nanosheets decreases the porosity of the Br by transferring heat to the core of the tablet.Furthermore,the addition of BSA-rGO nanosheets into the Br matrix enhances the adhesion of G-292 cells on the surface of the tablets.These findings suggest that the tablet consisting of BSA-rGO-reinforced Br has encouraging potential for application in bone tissue engineering.

Oxysterols as promising small molecules for bone tissue engineering: Systematic review

BACKGROUND Bone tissue engineering is an area of continued interest within orthopaedic surgery,as it promises to create implantable bone substitute materials that obviate the need for autologous bone graft.Recently,oxysterols–oxygenated derivatives of cholesterol-have been proposed as a novel class of osteoinductive small molecules for bone tissue engineering.Here,we present the first systematic review of the in vivo evidence describing the potential therapeutic utility of oxysterols for bone tissue engineering.AIM To systematically review the available literature examining the effect of oxysterols on in vivo bone formation.METHODS We conducted a systematic review of the literature following PRISMA guidelines.Using the PubMed/MEDLINE,Embase,and Web of Science databases,we queried all publications in the English-language literature investigating the effect of oxysterols on in vivo bone formation.Articles were screened for eligibility using PICOS criteria and assessed for potential bias using an expanded version of the SYRCLE Risk of Bias assessment tool.All full-text articles examining the effect of oxysterols on in vivo bone formation were included.Extracted data included:Animal species,surgical/defect model,description of therapeutic and control treatments,and method for assessing bone growth.Primary outcome was fusion rate for spinal fusion models and percent bone regeneration for critical-sized defect models.Data were tabulated and described by both surgical/defect model and oxysterol employed.Additionally,data from all included studies were aggregated to posit the mechanism by which oxysterols may mediate in vivo bone formation.RESULTS Our search identified 267 unique articles,of which 27 underwent full-text review.Thirteen studies(all preclinical)met our inclusion/exclusion criteria.Of the 13 included studies,5 employed spinal fusion models,2 employed critical-sized alveolar defect models,and 6 employed critical-sized calvarial defect models.Based upon SYRCLE criteria,the included studies were found to possess an overall“unclear risk of bias”;54%of studies reported treatment randomization and 38%reported blinding at any level.Overall,seven unique oxysterols were evaluated:20(S)-hydroxycholesterol,22(R)-hydroxycholesterol,22(S)-hydroxycholesterol,Oxy4/Oxy34,Oxy18,Oxy21/Oxy133,and Oxy49.All had statistically significant in vivo osteoinductive properties,with Oxy4/Oxy34,Oxy21/Oxy133,and Oxy49 showing a dose-dependent effect in some cases.In the eight studies that directly compared oxysterols to rhBMP-2-treated animals,similar rates of bone growth occurred in the two groups.Biochemical investigation of these effects suggests that they may be primarily mediated by direct activation of Smoothened in the Hedgehog signaling pathway.CONCLUSION Present preclinical evidence suggests oxysterols significantly augment in vivo bone formation.However,clinical trials are necessary to determine which have the greatest therapeutic potential for orthopaedic surgery patients.

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.

Morphological Evaluation of PLA/Soybean Oil Epoxidized Acrylate Three-Dimensional Scaffold in Bone Tissue Engineering

Tissue engineering’s main goal is to regenerate or replace tissues or organs that have been destroyed by disease,injury,or congenital disabilities.Tissue engineering now uses artificial supporting structures called scaffolds to restore damaged tissues and organs.These are utilized to attach the right cells and then grow them.Rapid prototyping appears to be the most promising technology due to its high level of precision and control.Bone tissue replacement“scaffolding”is a common theme discussed in this article.The fused deposition technique was used to construct our scaffold,and a polymer called polylactic acids and soybean oil resin were used to construct our samples.The samples were then divided into two groups;the first group was left without immersion in the simulated body fluid and served as a control for comparison.The second group was immersed in the simulated body fluid.The results of the Field Emission Scanning Electron Microscope(FESEM),Energy Dispersive X-ray Spectroscopy(EDX)and X-ray diffraction(XRD)were utilized to interpret the surface attachment to ions,elements,and compounds,giving us a new perspective on scaffold architecture.In this study,an innovative method has been used to print therapeutic scaffold that combines fused deposition three-dimensional printing with ultraviolet curing to create a high-quality biodegradable polymeric scaffold.Finally,the results demonstrate that adding soybean oil resin to the PLA increased ion attachment to the surface while also attracting tricalcium phosphate formation on the surface of the scaffold,which is highly promising in bone tissue replacement.In conclusion,the soybean oil resin,which is new in the field of bone tissue engineering,shows magnificent characteristics and is a good replacement biopolymer that replaces many ceramic and polymeric materials used in this field that have poor morphological characteristics.

Wnt3a-induced ST2 decellularized matrix ornamented PCL scaffold for bone tissue engineering

The limited bioactivity of scaffold materials is an important factor that restricts the development of bone tissue engineering.Wnt3a activates the classicWnt/β-catenin signaling pathway which effects bone growth and development by the accumulation ofβ-catenin in the nucleus.In this study,we fabricated 3D printed PCL scaffold with Wnt3a-induced murine bone marrow-derived stromal cell line ST2 decellularized matrix(Wnt3a-ST2-dCM-PCL)and ST2 decellularized matrix(ST2-dCM-PCL)by freeze-thaw cycle and DNase decellularization treatment which efficiently decellularized>90%DNA while preserved most protein.Compared to ST2-dCM-PCL,Wnt3a-ST2-dCM-PCL significantly enhanced newly-seeded ST2 proliferation,osteogenic differentiation and upregulated osteogenic marker genes alkaline phosphatase(Alp),Runx2,type I collagen(Col 1)and osteocalcin(Ocn)mRNA expression.After 14 days of osteogenic induction,Wnt3a-ST2-dCM-PCL promoted ST2 mineralization.These results demonstrated that Wnt3a-induced ST2 decellularized matrix improve scaffold materials’osteoinductivity and osteoconductivity.

Strontium Substituted Nanohydroxyapatite Incorporated 3D Printing Scaffold for Bone Tissue Engineering

The customized implants which are composed of polycaprolactone( PCL) and strontium substituted nanohydroxyapatite( SrHA) were fabricated successfully by using fused deposition modeling( FDM),which is a simple 3 D printing technology for fabricating personalized products. The physical and chemical properties of composite scaffolds were characterized by transmission electron microscopy( TEM), Fourier transform infrared spectroscopy( FTIR), X-Ray diffraction( XRD) and inductively coupled plasma-atomic emission spectroscopy( ICPAES). The results suggested that strontium element was successfully doped into nanohydroxyapatite and all scaffolds showed the homogeneous network structure. Furthermore, the in vitro biocompatibility of the scaffolds was evaluated by cell counting kit-8( CCK-8) assay. The data indicated that the prepared scaffolds exhibited excellent biocompatibility to bone marrow mesenchymal stem cells( BMSCs). Besides,strontium element can be released from PCL-SrHA scaffolds in a sustained manner. Therefore,the 3 D printing PCL-SrHA scaffolds hold great potential for bone tissue engineering.

Dexamethasone-Loaded PLGA Microspheres Incorporated PLLA/PLGA/PCL Composite Scaffold for Bone Tissue Engineering

The combination of micro-carriers and polymer scaffolds as promising bone grafts have attracted considerable interest in recent decades.The poly(L-lactic acid)/poly(lactic-co-glycolic acid)/polycaprolactone(PLLA/PLGA/PCL)composite scaffold with porous structure was fabricated by thermally induced phase separation(TIPS).Dexamethasone(DEX)was incorporated into PLGA microspheres and then loaded on the PLLA/PLGA/PCL scaffoldtopreparethedesiredcompositescaffold.The physicochemical properties of the prepared composite scaffold were characterized.The morphology of rat bone marrow mesenchymal stem cells(BMSCs)grown on scaffolds was observed using scanning electron microscope(SEM)and fluorescence microscope.The resultsshowedthatthePLLA/PLGA/PCLscaffoldhad interconnected macropores and biomimetic nanofibrous structure.In addition,DEX can be released from scaffold in a sustained manner.More importantly,DEX loaded composite scaffold can effectively support the proliferation of BMSCs as indicated by fluorescence observation and cell proliferation assay.The results suggested that the prepared PLLA/PLGA/PCL composite scaffold incorporating drug-loaded PLGA microspheres could hold great potential for bone tissue engineering applications.

Polymer Nanocomposites Used as Scaffolds for Bone Tissue Regeneration

Scaffolds are three-dimensional biocompatible structures that can mimic the properties of the extracellular matrix (ECM) of a given tissue, like mechanical support and bioactivity, which provides a platform for cellular adherence, proliferation and differentiation. Consequently, scaffolds are frequently used in tissue engineering with the intention of assisting the regeneration of a damaged tissue, and a major application in bone regeneration. An ideal scaffold needs to be biodegradable, biocompatible, and needs to match the biomechanical properties of bone. Polymers are widely used in this field because they fulfil the first two requirements. However, no polymeric material can achieve mechanical properties similar to the bone. For that reason, polymeric nanocomposites, which consist of ceramic/metallic nanoparticles dispersed in a polymer matrix, are being considered for bone scaffold fabrication in order to overcome this problem, since nanoparticles are known to improve composite mechanical strength, and enhance other properties.

Cellulose-based composite scaffolds for bone tissue engineering and localized drug delivery

Natural bone constitutes a complex and organized structure of organic and inorganic components with limited ability to regenerate and restore injured tissues,especially in large bone defects.To improve the reconstruction of the damaged bones,tissue engineering has been introduced as a promising alternative approach to the conventional therapeutic methods including surgical interventions using allograft and autograft implants.Bioengineered composite scaffolds consisting of multifunctional biomaterials in combination with the cells and bioactive therapeutic agents have great promise for bone repair and regeneration.Cellulose and its derivatives are renewable and biodegradable natural polymers that have shown promising potential in bone tissue engineering applications.Cellulose-based scaffolds possess numerous advantages attributed to their excellent properties of non-toxicity,biocompatibility,biodegradability,availability through renewable resources,and the low cost of preparation and processing.Furthermore,cellulose and its derivatives have been extensively used for delivering growth factors and antibiotics directly to the site of the impaired bone tissue to promote tissue repair.This review focuses on the various classifications of cellulose-based composite scaffolds utilized in localized bone drug delivery systems and bone regeneration,including cellulose-organic composites,cellulose-inorganic composites,cellulose-organic/inorganic composites.We will also highlight the physicochemical,mechanical,and biological properties of the different cellulose-based scaffolds for bone tissue engineering applications.

Fabrication and Characterization of Willemite Scaffolds Using Corn Stalk as a Novel Bio Template for Bone Tissue Engineering Applications

In this paper,we used Corn Stalk(CS)as a renewable and economical bio template to fabricate willemite scaffolds with the potential application in skull bone repair.CS was used as a sacrificial template to synthesize the scaffolds.Willemite scaffolds with the chemical formula of Zn2SiO4 and pore size in the range of 3 to 10µm could be successfully synthesized by soaking CS in the willemite solution for 24 h and sintering at 950°C for 5 h.The porosity of the samples was controlled by the soaking time(between 12 and 48 h)in the willemite solution from 5 to 35%,respectively.The properties of these scaffolds showed a good approximation with cranial bone tissue.In addition,cytotoxicity assays(MTT)were performed on Human Bone Marrow Stromal cells(HBMSc)and A172 human glioblastoma cell lines by direct and indirect culture methods to estimate their toxicity for bone and nerve cells,respectively.Alkaline Phosphatase(ALP)activity and DAPI/Phalloidin cell staining were also performed to investigate the efficiency of the scaffolds for bone tissue engineering applications.The results showed that the scaffolds had good biocompatibility with both HBMSC and A172 cells,noticeable improvement on ALP activity,and great apatite formation ability in Simulated Body Fluid(SBF).All the evidence ascertained that willemite scaffolds made by corn stalks could be a useful candidate for bone tissue engineering applications.

MOFs and bone: Application of MOFs in bone tissue engineering and bone diseases

Metal-organic frameworks(MOFs), a class of hybrid materials, consist of organic linkers and bridging metal ions or clusters. Their tunable pore sizes, large surface area, good biocompatibility, structural variability in combination with materials and chemicals, and osteogenic effects provide potential approaches for bone tissue engineering and bone diseases. And there are more and more research on MOFs in the field of osteogenesis in recent years. This review presents an overall summary of the application in the bone tissue engineering and bone diseases of MOFs and their composites, starting with the synthesis of MOFs, which discusses the advantages and disadvantages of different syntheses. Then, the biological functions of MOFs are discussed, which are the basics of MOFs applied in the organism. Importantly,mechanisms and abundant applications of MOFs are detailed in the bone tissue engineering and bone diseases. Finally, some prospects of MOFs are discussed, for instance, exploring whether MOFs can be used to treat other bone diseases.

3D printing method for bone tissue engineering scaffold

3D printing technology is an emerging technology.It constructs solid bodies by stacking materials layer by layer,and can quickly and accurately prepare bone tissue engineering scaffolds with specific shapes and structures to meet the needs of different patients.The field of life sciences has received a great deal of attention.However,different 3D printing technologies and materials have their advantages and disadvantages,and there are limitations in clinical application.In this paper,the technology,materials and clinical applications of 3D printed bone tissue engineering scaffolds are reviewed,and the future development trends and challenges in this field are prospected.

Three-dimensional printing of biomaterials for bonetissue engineering: a review

Processing biomaterials into porous scaffolds for bone tissueengineering is a critical and a key step in defining and controlling their physicochemical,mechanical,and biological properties.Biomaterials such as polymers are commonlyprocessed into porous scaffolds using conventional processing techniques,e.g.,saltleaching.However,these traditional techniques have shown unavoidable limitations andseveral shortcomings.For instance,tissue-engineered porous scaffolds with a complexthree-dimensional(3D)geometric architecture mimicking the complexity of theextracellular matrix of native tissues and with the ability to fit into irregular tissue defectscannot be produced using the conventional processing techniques.3D printing hasrecently emerged as an advanced processing technology that enables the processing ofbiomaterials into 3D porous scaffolds with highly complex architectures and tunableshapes to precisely fit into irregular and complex tissue defects.3D printing providescomputer-based layer-by-layer additive manufacturing processes of highly precise andcomplex 3D structures with well-defined porosity and controlled mechanical propertiesin a highly reproducible manner.Furthermore,3D printing technology provides anaccurate patient-specific tissue defect model and enables the fabrication of a patientspecifictissue-engineered porous scaffold with pre-customized properties.

Recent advances in two-dimensional nanomaterials for bone tissue engineering

Over the last decades,bone tissue engineering has increasingly become a research focus in the field of biomedical engineering,in which biomaterials play an important role because they can provide both biomechanical support and osteogenic microenvironment in the process of bone regeneration.Among these biomaterials,two-dimensional(2D)nanomaterials have recently attracted considerable interest owing to their fantastic physicochemical and biological properties including great biocompatibility,excellent osteogenic capability,large specific surface area,and outstanding drug loading capacity.In this review,we summarize the state-of-the-art advances in 2D nanomaterials for bone tissue engineering.Firstly,we introduce the most explored biomaterials used in bone tissue engineering and their advantages.We then highlight the advances of cutting-edge 2D nanomaterials such as graphene and its derivatives,layered double hydroxides,black phosphorus,transition metal dichalcogenides,montmorillonite,hexagonal boron nitride,graphite phase carbon nitride,and transition metal carbonitrides(MXenes)used in bone tissue engineering.Finally,the current challenges and future prospects of 2D nanomaterials for bone tissue regeneration in process of clinical translation are discussed.

3D Bio-Printed Bone Scaffolds Incorporated with Natural Antibacterial Compounds

3D Bioprinting plays an irreplaceable role in bone tissue engineering. Shellac and curcumin are two natural compounds that are widely used in the food and pharmaceutical sectors. In this study, a new composite scaffold with good biocompatibility and antibacterial ability was manufactured by adding shellac and curcumin into the traditional bone scaffold through low-temperature three-dimensional printing (LT-3DP), and its impact on the osteoimmune microenvironment was evaluated.

Experimental study on advanced bone regeneration by BMP-7 derived-peptide chitosan nanometer hydroxyapatite collagen biomimetic composite

Objective:To investigate the effect of BMP-7 derived-peptide chitosan nanometer hydroxyapatite biomimetic collagen composite on repairing rat critical-sized cranial defects.Methods:The chitosan nanometer hydroxyapatite collagen composite was prepared and the microcosmic appearance of the composite was observed by scanning electron microscope.The BMP-7 derived-peptide was introduced into the composite by vacuum adsorption.The released peptide content from the scaffold was detected using high performance liquid chromatography at different set times.Critical-sized cranial defects were created on both sides of the parietal bone in 24 adult Sprague-Dawley rats.The BMP-7 derived-peptide chitosan nanometer hydroxyapatite biomimetic collagen composites were implanted on the right side as experimental group and the left side was implanted with chitosan nanometer hydroxyapatite biomimetic collagen composites alone as control group.The rats of both groups were killed in batch respectively after 6 and 12 weeks.Macroscopic observation,three-dimensional reconstruction of computed tomography(CT)and histological observation were performed on these samples.Results:The results of scanning electron microscope showed that the surface of the scaffold was porous.The releasing character of BMP-7 derived-peptide belonged to slow release.The result of animal experiment showed that the BMP-7 derived-peptide chitosan nanometer hydroxyapatite biomimetic collagen composite could more effectively promote the repair of cranial bone defects comparing with the chitosan nanometer hydroxyapatite biomimetic collagen composite alone.The difference was statistically significant(p<.05).Conclusions:The BMP-7 derived-peptide chitosan nanometer hydroxyapatite collagen biomimetic composite can effectively promote bone regeneration of bone defects.The composite is a kind of ideal scaffold material for bone tissue engineering.