Scaffolds For Bone Tissue Engineering Research Articles

TPMS Microarchitectures for Vertical Bone Augmentation and Osteoconduction: An In Vivo Study.

Triply periodic minimal surface microarchitectures (TPMS) were developed by mathematicians and evolved in all kingdoms of living organisms. Renowned for their lightweight yet robust attributes, TPMS structures find application in diverse fields, such as the construction of satellites, aircrafts, and electric vehicles. Moreover, these microarchitectures, despite their intricate geometric patterns, demonstrate potential for application as bone substitutes, despite the inherent gothic style of natural bone microarchitecture. Here, we produced three TPMS microarchitectures, D-diamond, G-gyroid, and P-primitive, by 3D printing from hydroxyapatite. We explored their mechanical characterization and, further, implanted them to study their bone augmentation and osteoconduction potential. In terms of strength, the D-diamond and G-gyroid performed significantly better than the P-primitive. In a calvarial defect model and a calvarial bone augmentation model, where osteoconduction is determined as the extent of bony bridging of the defect and bone augmentation as the maximal vertical bone ingrowth, the G-gyroid performed significantly better than the P-primitive. No significant difference in performance was observed between the G-gyroid and D-diamond. Since, in real life, the treatment of bone deficiencies in patients comprises elements of defect bridging and bone augmentation, ceramic scaffolds with D-diamond and G-gyroid microarchitectures appear as the best choice for a TPMS-based scaffold in bone tissue engineering.

Tannic acid and quaternized chitosan mediated puerarin-loaded octacalcium phosphate /sodium alginate scaffold for bone tissue engineering

Current limitations in mechanical performance and foreign body reactions (FBR) often lead to implant failure, restricting the application of bioceramic scaffolds. This study presents a novel 3D-printed scaffold that combines the release of anti-inflammatory drugs with osteogenic stimulation. Initially, the inorganic and organic phases were integrated to ensure the scaffold's mechanical integrity through catechol chemistry and the electrostatic interactions between tannic acid and quaternary ammonium chitosan. Subsequently, layers of polydopamine-encapsulated puerarin-loaded zeolitic imidazolate framework-8 (ZIF-8) were self-assembled onto the stent's surface, creating the drug-loaded scaffold that improved drug release without altering the scaffold's structure. Compared with unloaded scaffolds, the puerarin-loaded scaffold demonstrated excellent osteogenic differentiation properties along with superior anti-inflammatory and osteogenic effects in a range of in vitro and in vivo studies. RNA sequencing clarified the role of the TNF and NF/κB signaling pathways in these effects, further supporting the scaffold's osteogenic potential. This study introduces a novel approach for creating drug-loaded scaffolds, providing a unique method for treating cancellous bone defects.

Customized 3D‐printed heterogeneous porous titanium scaffolds for bone tissue engineering

AbstractBone defect is a common clinical disease. Due to the uncertainty of trauma or infection areas, customized size features are often required for bone substitutes. By inspiration of the natural bone structure, this study designs porous scaffolds with a biomimetic design perspective by using different inner and outer pore units. The outer pore units adopt body‐centered cubic (BCC) structure to simulate the weight‐bearing function of human cortical bone, while inner pore units using I‐Wrapped Package structure, a kind of three periods minimum surface, to obtain a good permeability and simulates the inner layer of cancellous bone. To further regulate the overall modulus of the scaffold within the range of natural bone modulus in the human body, the scaffold was designed to axial gradient structure. Compression experiments were conducted, and the results indicated that when the volume fraction linearly increased from 20% to 50%, the Young's modulus was close to the cortical bone modulus in the human body. In vitro cell experiments further proved that osteoblasts have good cellular activity and spreading morphology on the surface of this scaffold. The customized 3D‐printed heterogeneous porous titanium scaffold has great application potential in bone tissue engineering.

Potential use of a bone tissue engineering scaffold based on electrospun poly (ɛ-caprolactone) - Poly (vinyl alcohol) hybrid nanofibers containing modified cockle shell nanopowder

Today, the construction of scaffolds promoting the differentiation of stem cells is an intelligent innovation that accelerates the differentiation toward the target tissue. The use of calcium and phosphate compounds is capable of elevating the precision and efficiency of the osteogenic differentiation of stem cells. In this research, osteoconductive electrospun poly (ɛ-caprolactone) (PCL) - poly (vinyl alcohol) (PVA) hybrid nanofibrous scaffolds containing modified cockle shell (CS) nanopowder were prepared and investigated. In this regard, the modified CS nanopowder was prepared by grinding and modifying with phosphoric acid, and it was then added to PVA nanofibers at different weight percentages. Based on the SEM images, the optimum content of the modified CS nanopowder was set at 7 wt %, since reaching the threshold of agglomeration restricted this incorporation. In the second step, the PVA-CS7 nanofibrous sample was hybridized with different PCL ratios. Concerning the hydrophilicity and mechanical strength, the sample named PCL50-PVA50-CS7 was ultimately selected as the optimized and suitable candidate scaffold for bone tissue application. The accelerated hydrolytic degradation of the sample was also studied by FTIR and SEM analyses, and the results confirmed that the mineral deposits of CS are available approximately 7 days for mesenchymal stem cells. Moreover, Alizarin red staining illustrated that the presence of CS in the PCL50-PVA50-CS7 hybrid nanofibrous scaffold may potentially lead to an increase in calcium deposits with high precipitates, authenticating the differentiation of stem cells towards osteogenic cells.

Gelatin-containing functionally graded calcium sulfate/bioactive glass bone tissue engineering scaffold

One of the challenges and limitations of bone tissue engineering includes fast degradation rates, reduced bioactivity, donor site morbidity, and unresolved risks of pathogen transmission. In the field of bone tissue engineering, gradient materials are promising for treating bone defects because they can create graded structures and compositions similar to natural bone. By controlling the amount of components in the structure, the degradation rate, bioactivity, and osteogenic capacity can be manipulated. In the current study, a gradient, multilayer, porous nanocomposite of calcium sulfate/glass (Gn CS/BG) with a gelatin coating (Gn CS/BG-Gel) was prepared using rotational casting technique. The SEM results showed an average pore size of approximately 400 μm within the Gn CS/BG-Gel nanocomposite. An elastic modulus (E) of around 240 MPa and an ultimate tensile strength (σ) of approximately 5.5 MPa were achieved for Gn CS/BG-Gel nanocomposite. The degradation analysis in a dynamic tris-buffer environment revealed a degradation rate of 68 % over a 63-day period, with a decrease in degradation rate as the bioactive glass (BG) content increased in each layer, indicating the influence of BG on the degradation process. The bioactivity results in a dynamic simulated body fluid suggested that increased BG content in each layer promoted hydroxyapatite formation, indicating improved bioactive behavior. The evaluation of ion release with ICP-OES, MTT and Acridine Orange assays using human bone marrow-derived mesenchymal stem cells (hBMSCs) confirmed the non-toxic nature of Gn CS/BG-Gel. Assessments of alkaline phosphatase (ALP) activity, calcium content, Alizarin Red staining and cell attachment behavior substantiated the osteogenic potential of Gn CS/BG-Gel. Collectively, the three-dimensional structure along with the Gn CS/BG-Gel composition contributed to the enhanced cellular responses. These findings hold significant promise for the development of biomaterials with potential applications in bone tissue engineering, and they contribute valuable insights to the field.

Cell Instructive Behavior of Composite Scaffolds in a Co-Culture of Human Mesenchymal Stem Cells and Peripheral Blood Mononuclear Cells.

The in vitro evaluation of 3D scaffolds for bone tissue engineering in mono-cultures is a common practice; however, it does not represent the native complex nature of bone tissue. Co-cultures of osteoblasts and osteoclasts, without the addition of stimulating agents for monitoring cellular cross-talk, remains a challenge. In this study, a growth factor-free co-culture of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) and human peripheral blood mononuclear cells (hPBMCs) has been established and used for the evaluation of 3D-printed scaffolds for bone tissue engineering. The scaffolds were produced from PLLA/PCL/PHBV polymeric blends, with two composite materials produced through the addition of 2.5% w/v nanohydroxyapatite (nHA) or strontium-substituted nanohydroxyapatite (Sr-nHA). Cell morphology data showed that hPBMCs remained undifferentiated in co-culture, while no obvious differences were observed in the mono- and co-cultures of hBM-MSCs. A significantly increased alkaline phosphatase (ALP) activity and osteogenic gene expression was observed in co-culture on Sr-nHA-containing scaffolds. Tartrate-resistant acid phosphatase (TRAP) activity and osteoclastogenic gene expression displayed significantly suppressed levels in co-culture on Sr-nHA-containing scaffolds. Interestingly, mono-cultures of hPBMCs on Sr-nHA-containing scaffolds indicated a delay in osteoclasts formation, as evidenced from TRAP activity and gene expression, demonstrating that strontium acts as an osteoclastogenesis inhibitor. This co-culture study presents an effective 3D model to evaluate the regenerative capacity of scaffolds for bone tissue engineering, thus minimizing time-consuming and costly in vivo experiments.

Biodegradable electrospun poly(L-lactide-co-ε-caprolactone)/polyethylene glycol/bioactive glass composite scaffold for bone tissue engineering.

The field of tissue engineering has witnessed significant advancements in recent years, driven by the pursuit of innovative solutions to address the challenges of bone regeneration. In this study, we developed an electrospun composite scaffold for bone tissue engineering. The composite scaffold is made of a blend of poly(L-lactide-co-ε-caprolactone) (PLCL) and polyethylene glycol (PEG), with the incorporation of calcined and lyophilized silicate-chlorinated bioactive glass (BG) particles. Our investigation involved a comprehensive characterization of the scaffold's physical, chemical, and mechanical properties, alongside an evaluation of its biological efficacy employing alveolar bone-derived mesenchymal stem cells. The incorporation of PEG and BG resulted in elevated swelling ratios, consequently enhancing hydrophilicity. Thermal gravimetric analysis confirmed the efficient incorporation of BG, with the scaffolds demonstrating thermal stability up to 250°C. Mechanical testing revealed enhanced tensile strength and Young's modulus in the presence of BG; however, the elongation at break decreased. Cell viability assays demonstrated improved cytocompatibility, especially in the PLCL/PEG+BG group. Alizarin red staining indicated enhanced osteoinductive potential, and fluorescence analysis confirmed increased cell adhesion in the PLCL/PEG+BG group. Our findings suggest that the PLCL/PEG/BG composite scaffold holds promise as an advanced biomaterial for bone tissue engineering.

Design and characterization of 3D printed pore gradient hydrogel scaffold for bone tissue engineering

Macroporous hydrogel scaffolds are widely used in tissue engineering to promote cell growth and proliferation. Aiming to enhance cell seeding efficiency and facilitate the osteodifferentiation of mesenchymal stem cells, this study demonstrates the fabrication of pore gradient biodegradable hydrogel scaffolds inspired by natural bone structure for bone tissue engineering applications. The scaffolds were fabricated via extrusion-based 3D printing, using sequential deposition of three customized Gelatin/Oxidized Alginate - based inks with subsequent cryogenic crosslinking for permanent structure fixation. The resulting constructs were characterized and featured a continuous gradient morphology with pore sizes ranging from 10 to 300 μm. The gradient scaffolds exhibited improved mechanical stability, with a compression resistance of 149 kPa, as opposed to the non-gradient scaffold's 116 kPa at 70 % strain, and a sustained degradation rate with only a 10 % loss of its initial weight within three weeks. Gradient scaffolds demonstrated a doubling of cell seeding efficiency to 47 % with dense and homogeneously distributed cellular layers, as evidenced by confocal and electron microscopy. Furthermore, the gradient scaffolds demonstrated superior osteodifferentiation, with significantly higher ALP and DMP1 production and enhanced extracellular matrix mineralization compared to gradientless macroporous scaffolds. This study provides insights into the design of macroporous scaffolds and emphasizes the advantages of pore gradient over homogeneous gradientless morphologies.

4D-printed dual-responsive bioscaffolds for treating critical-sized irregular bone defects

Currently, treating irregular bone defects with a critical size is still challenging as material implantation in the complex defects with irregular shapes, effective osteogenesis and sufficient vascularization cannot be easily achieved simultaneously via a simple strategy. Herein, a dual-responsive bone tissue engineering scaffold is developed using a 4D printing strategy by intergrating a single type of multifunctional magnetic nanoparticles, Fe3O4@SiO2, with printing inks made of bioceramics and biopolymers. Minimally invasive implantation, fluent navigation of scaffolds as well as the precise boundary matching between scaffolds and the irregular defects are achieved through near-infrared (NIR) irradiation-based temperature responsive shape recovery and static magnetic field (SMF) stimulation. Moreover, improved bone regeneration in critical-sized bone defects is successfully achieved through the activation of PI3K/AKT pathway which significantly promotes osteogenic differentiation and vascularization. Besides, NIR-based photothermal stimulation upregulates the expression of heat shock protein (HSP90) and further promotes the osteogenesis and vascularization. The in vivo study also confirms that our scaffold can precisely fit the bone defect, and induce satisfactory osteogenesis and angiogenesis. This work provides a facile strategy to simultaneously-realize easy scaffold implantation in irregular bone defects and improves osteogenesis and angiogenesis by integrating a single type of multifunctional nanoparticles with scaffold matrices.

Nanocomposite Methacrylated Silk Fibroin-Based Scaffolds for Bone Tissue Engineering.

The treatment of bone defects is a clinical challenge. Bone tissue engineering is gaining interest as an alternative to current treatments, with the development of 3D porous structures (scaffolds) helpful in promoting bone regeneration by ensuring temporary functional support. In this work, methacrylated silk fibroin (SilMA) sponges were investigated as scaffolds for bone tissue engineering by exploiting the combination of physical (induced by NaCl salt during particulate leaching) and chemical crosslinking (induced by UV-light exposure) techniques. A biomimetic approach was adopted to better simulate the extracellular matrix of the bone by introducing either natural (mussel shell-derived) or synthetic-origin hydroxyapatite nanoparticles into the SilMA sponges. The obtained materials were characterized in terms of pore size, water absorption capability and mechanical properties to understand both the effect of the inclusion of the two different types of nanoparticles and the effect of the photocrosslinking. Moreover, the SilMA sponges were tested for their bioactivity and suitability for bone tissue engineering purposes by using osteosarcoma cells, studying their metabolism by an AlamarBlue assay and their morphology by scanning electron microscopy. Results indicate that photocrosslinking helps in obtaining more regular structures with bimodal pore size distributions and in enhancing the stability of the constructs in water. Moreover, the addition of naturally derived hydroxyapatite was observed to be more effective at activating osteosarcoma cell metabolism than synthetic hydroxyapatite, showing a statistically significant difference in the AlamarBlue measurement on day 7 after seeding. The methacrylated silk fibroin/hydroxyapatite nanocomposite sponges developed in this work were found to be promising tools for targeting bone regeneration with a sustainable approach.

Harnessing potential of avian eggshell membrane derived collagen hydrolysate for bone tissue regeneration.

Natural bone grafts are the highly preferred materials for restoring the lost bone, while being constrained of donor availability and risk of disease transmission. As a result, tissue engineering is emerging as an efficacious and competitive technique for bone repair. Bone tissue engineering (TE) scaffolds to support bone regeneration and devoid of aforesaid limitations are being vastly explored and among these the avian eggshell membrane has drawn attention for TE owing to its low immunogenicity, similarity with the extracellular matrix, and easy availability. In this study, the development of bone ingrowth support system from avian eggshell membrane derived collagen hydrolysates (Col-h) is reported. The hydrolysate, cross-linked with glutaraldehyde, was developed into hydrogels with poly-(vinyl alcohol) (PVA) by freeze-thawing and further characterized with ATR-FTIR, XRD, FESEM. The biodegradability, swelling, mechanical, anti-microbial, and biocompatibility evaluation were performed further for the suitability in bone regeneration. The presence of amide I, amide III, and -OH functional groups at 1639cm- 1,1264cm- 1, and 3308cm- 1 respectively and broad peak between 16°-21° (2θ) in XRD data reinstated the composition and form. The maximum ratio of Col-h/PVA that produced well defined hydrogels was 50:50. Though all the hydrogel matrices alluded towards their competitive attributes and applicability towards restorative bone repair, the hydrogel with 40:60 ratios showed better mechanical strength and cell proliferation than its counterparts. The prominent E. coli growth inhibition by the hydrogel matrices was also observed, along with excellent biocompatibility with MG-63 osteoblasts. The findings indicate strongly the promising application of avian eggshell-derived Col-h in supporting bone regeneration.

Biomimetic design and fabrication of thermally induced radial gradient shape memory scaffolds using fused deposition modeling (FDM) for bone tissue engineering

Bone defects pose a significant challenge, often exceeding natural healing capabilities. This study explores the potential of thermally induced radial gradient shape memory (RGSM) scaffolds for minimally invasive bone repair. Inspired by the natural porosity gradient of bone, these scaffolds feature a high-porosity inner zone that mimics cancellous bone and a low-porosity outer zone that resembles cortical bone. When the relationship between porosity and key properties was investigated, it was found that lower-porosity RGSM scaffolds exhibited higher compressive strength but experienced higher residual strain and lower shape recovery ratio compared to their higher-porosity counterparts. Despite this trade-off, the gradient design successfully mimicked the natural bone structure, potentially enhancing osseointegration and bone regeneration. These results demonstrate the feasibility of RGSM scaffolds for bone tissue engineering. This holds promise for advancing minimally invasive surgical techniques and improving the treatment of bone defects.

An in vitro evaluation of the biological activity of biogenic eggshell-derived nanosized bioglass, poly (ε-caprolactone), and zein protein 3D composite scaffolds for bone tissue engineering

An in vitro evaluation of the biological activity of biogenic eggshell-derived nanosized bioglass, poly (ε-caprolactone), and zein protein 3D composite scaffolds for bone tissue engineering

Inhibiting the "isolated island" effect in simulated bone defect repair using a hollow structural scaffold design.

The treatment of bone tissue defects remains a complicated clinical challenge. Recently, the bone tissue engineering (BTE) technology has become an important therapeutic approach for bone defect repair. Researchers have improved the scaffolds, cells, and bioactive factors used in BTE through various existing bone repair material preparation strategies. However, due to insufficient vascularization, inadequate degradation, and fibrous wrapping, most BTE scaffolds impede new bone ingrowth and the reconstruction of grid-like connections in the middle and late stages of bone repair. These non-degradable scaffolds become isolated and disordered like independent "isolated islands", which leads to the failure of osteogenesis. Consequently, we hypothesized that the "island effect" prevents successful bone repair. Accordingly, we proposed a new concept of scaffold modification-osteogenesis requires a bone temporary shelter (also referred to as the empty shell osteogenesis concept). Based on this concept, we consider that designing hollow structural scaffolds is the key to mitigating the "isolated island" effect and enabling optimal bone regeneration and reconstruction.

Optimization of Cobalt-Chromium (Co-Cr) Scaffolds for Bone Tissue Engineering in Endocrine, Metabolic and Immune Disorders.

Approximately 50% of the adult global population is projected to suffer from some form of metabolic disease by 2050, including metabolic syndrome and diabetes mellitus. At the same time, this trend indicates a potential increase in the number of patients who will be in need of implant-supported reconstructions of specific bone regions subjected to inflammatory states. Moreover, physiological conditions associated with dysmetabolic subjects have been suggested to contribute to the severity of bone loss after bone implant insertion. However, there is a perspective evidence strengthening the hypothesis that custom-fabricated bioengineered scaffolds may produce favorable bone healing effects in case of altered endocrine or metabolic conditions. This perspective review aims to share a comprehensive knowledge of the mechanisms implicated in bone resorption and remodelling processes, which have driven researchers to develop metallic implants as the cobalt-chromium (Co-Cr) bioscaffolds, presenting optimized geometries that interact in an effective way with the osteogenetic precursor cells, especially in the cases of perturbed endocrine or metabolic conditions.

Potential Application of Polysaccharide-based Aerogel Scaffolds for Bone Tissue Engineering

Potential Application of Polysaccharide-based Aerogel Scaffolds for Bone Tissue Engineering

Design of Biomimetic Porous Scaffolds for Bone Tissue Engineering

Design of Biomimetic Porous Scaffolds for Bone Tissue Engineering

Effect of hydroxyapatite and pulsed electromagnetic field on osteogenic differentiation of human stem cells seeded onto 3D‐printed polycaprolactone scaffolds for future bone graft transplantation

AbstractBone graft transplantation is a risky procedure mainly due to postoperative complications. Tissue engineering is an alternative to regenerating bone tissue by seeding mesenchymal stem cells into 3D scaffolds by osteoinductive signaling. Poly(ε‐caprolactone) (PCL) is one of the most used biopolymers for fabricating scaffolds for bone tissue engineering. PCL is enriched with hydroxyapatite (HA) to promote the production of the bone‐mineralized matrix. Moreover, the application of pulsed electromagnetic fields (PEMFs) on scaffolds loaded with HA can significantly benefit collagen and HA networks favoring the interaction between organic and mineral phases of bones. In this context, we investigated, for the first time, the effect exerted by a PEMF exposure on the osteogenic differentiation of human adipose stem cells (hASCs) seeded onto 3D printed PCL‐HA scaffolds and cultured in growth and osteogenic medium (OM). After 21 days of culture, the results showed excellent colonization of scaffolds and the deposition of extracellular matrix (ECM) secreted by hASCs. Immunofluorescence staining confirmed the deposition of collagen type I and osteopontin both on scaffolds. Gene expression analysis indicated that both HA crystal and PEMF treatment were able to trigger osteogenic differentiation, as documented by the increase of osteogenic markers. These results suggest that HA, osteogenic factors, and PEMFs converge on the same signaling pathways as there are no additive effects and no change in protein synthesis between scaffolds with HA or treated with PEMFs for cells cultured in OM. The proposed study lays the foundations for exploring new approaches in bone graft transplantation.

Gelatin- zirconium based metal-organic framework (MOF 801) nanocomposite scaffold for bone tissue engineering

Tissue engineering applications in bone defect treatment have shown promising outcomes over the past decade. In this context, taking advantage of bioactive scaffolds mimicking the intrinsic composition of bone has resulted in remarkable effects on bone regeneration. In this study, Zirconium-based metal-organic framework, known as MOF 801, was synthesized and incorporated into the gelatin matrix as an osteoconductive agent to fabricate a nanocomposite bone scaffold by freeze-drying method. Physical and chemical characteristics of MOF 801 and nanocomposite scaffolds were analyzed using FE-SEM, EDS, XRD, ICP and FTIR. Besides, the biological activity of the prepared scaffolds was evaluated by MTT, alizarin red staining, crystal violet staining, and ALP assays. Inclusion of MOF 801 NPs into the scaffolds resulted in enhancement of their compressive strength up to 15 ± 0.05 MPa. The nanocomposite samples containing MOF 801 NPs also exhibited favorable bioactive feature that was confirmed by their apatite-forming ability on their surfaces upon immersion in SBF solution. The Zr ion and fumarate release studies illustrated sustained release profiles from the gelatin matrix, leading to both antioxidant and anti-inflammatory properties. MTT assay results affirmed biocompatibility of scaffolds containing up to 5 % w/w MOF 801, besides the crystal violet assay exhibited proper cell confluency and adhesion. Eventually, Alizarin red and ALP activity assays revealed that an increment of MOF 801 could induce calcium mineralization and Alkaline Phosphatase enzyme production in MG-63 cells confirming the potential of prepared scaffolds for related bone tissue engineering applications.

3D printed Nanohydroxyapatite/Polyamide 66 scaffolds with balanced mechanical property and osteogenic ability for bone repair

The biomaterial requirements for bone tissue repair are extremely strict. It not only requires the biomaterial to have good biocompatibility and biological activity, but also requires sufficient mechanical strength. Polyamide 66 (PA66) and nano-hydroxyapatite (n-HA) have been widely investigated in bone tissue engineering scaffold, however, its composites require further research. In this study, the 3D printing technology was utilized to fabricate personalized n-HA/PA66 bone tissue scaffold. The mechanical properties of n-HA/PA66 composite can be adjusted by configuring different raw material components and applying different porosity. It was found that 50 % porosity n-HA/PA66 composite scaffold showed a uniform porous structure and a compressive strength of 33.9 MPa. The composite scaffold also exhibited excellent biological properties while improving mechanical properties, in vitro cell tests indicated that the 3D printed n-HA/PA66 composite scaffold promoted differentiation of bone marrow mesenchymal stem cells (BMSCs) into osteoblasts. In in vivo animal model proved that the scaffold promoted endogenous bone regeneration in rabbit patellar bone defects, with newly formed bone volume of 20.9 % after two months of implantation. This research proposed a 3D printed polyamide/nano-hydroxyapatite composite with balanced mechanical property and osteogenic ability for customized bone tissue repair, and it has promising application prospects.