Scaffolds For Tissue Engineering Research Articles

Ca-AlN MOFs-loaded chitosan/gelatin scaffolds; a dual-delivery system for bone tissue engineering applications

Creating a scaffold for bone tissue engineering that is bioactive and capable of acting as a local-dual delivery system, releasing bioactive molecules and regulating the bone remodeling process to achieve balanced bone resorption and formation, is a significant challenge. The objective of this research is to create a composite scaffold using chitosan/gelatin (CHS/Gel) and the calcium (Ca)-alendronate (ALN) metal–organic frameworks (MOFs). The scaffold will act as a dual-delivery system, releasing Ca ions and ALN to regulate bone formation. Ca-ALN MOF nanoparticles (NPs) were prepared in mild conditions and studied by FTIR, XRD, FESEM, and TGA. Ca-ALN NPs-loaded CHS/Gel scaffolds were opportunely fabricated through freeze-drying approach. Physicochemical features of the scaffolds after incorporating NPs equated by CHS/Gel scaffold changed, therefore, the attendance of NPs caused a decreasing porosity, decreased swelling, and low rate of degradation. The release profile results showed that the NPs-loaded CHS/Gel scaffolds were able to simultaneously release ALN and Ca ions due to the decomposition of NPs. Additionally, the loading of NPs in the CHS/Gel scaffold led to an increment in alkaline phosphatase (ALP) activity and the quantity of deposited Ca along with osteogenesis gene markers. These findings suggest that the NPs-loaded CHS/Gel scaffold has the potential to enhance the differentiation of human adipose tissue-derived mesenchymal stem cells, making it a promising approach for bone repair.

Sand casting-inspired surface modification of 3D-printed porous polyetheretherketone scaffolds for enhancing osteogenesis

3D-printed polyetheretherketone (PEEK) scaffolds are developed as novel bone substitutes, however, their bioinert surface hinders osteogenesis and modifying PEEK scaffolds without blocking their inter-connected porous structure is challenged. In this study, 45S5 bioactive glasses (BG) were homogeneously coated on PEEK scaffolds inspired by traditional sand-casting. The structure of PEEK scaffolds was preserved under shaping effect of BG fillers. Modulus, coating yield and hydrophilicity of scaffolds after different thermal treatment time were comparatively investigated. Excellent hydroxyapatite-forming ability of BG-coated scaffolds was confirmed by mineralization study. In-vitro assessments, BG-coated scaffolds cultured with MC3T3-E1 cells presented potential as bone implants with excellent cytocompatibility and osteogenic properties. Following similar coating strategy, conductive particles, multi-component particles and template particles were also coated on PEEK surface. The proposed methodology highlights a useful approach towards producing tunable biomedical coatings and microstructure on porous PEEK scaffolds for bone tissue engineering.

Biomimetic mineralization of 3D-printed polyhydroxyalkanoate-based microbial scaffolds for bone tissue engineering

Polyhydroxyalkanoates (PHAs) have gained much attention as a potential alternative to conventional plastic bone scaffolds due to their biocompatibility and biodegradability, among a diverse range of advantageous properties. However, the water resistance of PHA creates an environment that can interfere with cell interactions. In this study, a three-dimensional-printed PHA scaffold was fabricated through fused deposition modeling printing considering the physical properties of PHA. The PHA bone scaffolds were then coated with polydopamine (pDA) and/or hydroxyapatite (HA) in various configurations using a relatively simple and rapid process involving only immersion. The PHA–pDA– HA scaffold showed enhanced cell viability, proliferation, and differentiation, and could thus serve as a versatile platform for bone tissue engineering applications.

A Review on the Electrospinning of Polymer Nanofibers and Its Biomedical Applications

Polymeric nanofibers have emerged as a captivating medium for crafting structures with biomedical applications. Spinning methods have garnered substantial attention in the context of medical applications and neural tissue engineering, ultimately leading to the production of polymer fibers. In comparison with polymer microfibers, polymer nanofibers boasting nanometer-scale diameters offer significantly larger surface areas, facilitating enhanced surface functionalization. Consequently, polymer nanofiber mats are presently undergoing rigorous evaluation for a myriad of applications, including filters, scaffolds for tissue engineering, protective equipment, reinforcement in composite materials, and sensors. This review offers an exhaustive overview of the latest advancements in polymer nanofiber processing and characterization. Additionally, it engages in a discourse regarding research challenges, forthcoming developments in polymer nanofiber production, and diverse polymer types and its applications. Electrospinning has been used to convert a broad range of polymers into nanoparticle nanofibers, and it may be the only approach with significant potential for industrial manufacturing. The basics of these spinning techniques, highlighting the biomedical uses as well as nanostructured fibers for drug delivery, disease modeling, regenerative medicine, tissue engineering, and bio-sensing have been explored.

Framework to optimize fixed-length micro-CT systems for propagation-based phase-contrast imaging.

A laboratory X-ray imaging system with a setup that closely resembles commercial micro-CT systems with a fixed source-to-detector distance of ∼90 cm is investigated for single distance propagation-based phase-contrast imaging and computed tomography (CT). The system had a constant source-to-detector distance, and the sample positions were optimized. Initially, a PTFE wire was imaged, both in 2D and 3D, to characterize fringe contrast and spatial resolution for different X-ray source settings and source-to-sample distances. The results were compared to calculated values based on theoretical models and to simulated (wave-optics based) results, with good agreement being found. The optimization of the imaging system is discussed. CT scans of two biological samples, a tissue-engineered esophageal scaffold and a rat heart, were then acquired at the optimum parameters, demonstrating that significant image quality improvements can be obtained with widely available components placed inside fixed-length cabinets through proper optimization of propagation-based phase-contrast.

Ciliary neurotrophic factor mediated growth of retinal ganglion cell axons on PGS/PCL scaffolds

Ciliary neurotrophic factor (CNTF) promotes survival and/or differentiation of a variety of neuronal cells including retinal ganglion cells (RGCs). Delivery of CNTF requires a suitable medium capable of mediating diffusion and premature release of CNTF within the target tissue. Polymeric tissue-engineered scaffolds have been readily used as substrates for cell transplantation, expansion, and differentiation and, as carriers of cell growth factors. Their functions to CNTF release for RGC proliferation have remained so far unexplored, especially to CNTF affinity to the scaffold and subsequent RGC fate. Electrospun poly(glycerol sebacate)/poly(ϵ-caprolactone) (PGS/PCL) biopolymer scaffolds have recently shown promising results in terms of supporting regeneration of RGC neurites. This work explores covalent immobilization of CNTF on PGS/PCL scaffold and the way immobilised CNTF mediates growth of RGC axons on the scaffold. An ex-vivo three-dimensional model of rodent optic nerve on PGS/PCL revealed that RGC explants cultured in CNTF mediated environment increased their neurite extensions after 20 d of cell culture employing neurite outgrowth measurements. The CNTF secretion on PGS/PCL scaffold was found bio-mimicking natural extracellular matrix of the cell target tissue and, consequently, has shown a potential to improve the overall efficacy of the RGC regeneration process.

Tailoring the Heterogeneous Structure of Macro-Fibers Assembled by Bacterial Cellulose Nanofibrils for Tissue Engineering Scaffolds.

Bacterial cellulose/oxidized bacterial cellulose nanofibrils (BC/oxBCNFs) macro-fibers are developed as a novel scaffold for vascular tissue engineering. Utilizing a low-speed rotary coagulation spinning technique and precise solvent control, macro-fibers with a unique heterogeneous structure with dense surface and porous core are created. Enhanced by a polydopamine (PDA) coating, these macro-fibers offer robust mechanical integrity, high biocompatibility, and excellent cell adhesion. When cultured with endothelial cells (ECs) and smooth muscle cells (SMCs), the macro-fibers support healthy cell proliferation and exhibit a unique spiral SMC alignment, demonstrating their vascular suitability. This innovative strategy opens new avenues for advances in tissue engineering.

"Metal-bone" scaffold for accelerated peri-implant endosseous healing.

Restoring bone defects caused by conditions such as tumors, trauma, or inflammation is a significant clinical challenge. Currently, there is a need for the development of bone tissue engineering scaffolds that meet clinical standards to promote bone regeneration in these defects. In this study, we combined the porous Ti6Al4V scaffold in bone tissue engineering with advanced bone grafting techniques to create a novel "metal-bone" scaffold for enhanced bone regeneration. Utilizing 3D printing technology, we fabricated a porous Ti6Al4V scaffold with an average pore size of 789 ± 22.69μm. The characterization and biocompatibility of the scaffold were validated through in vitro experiments. Subsequently, the scaffold was implanted into the distal femurs of experimental animals, removed after 3months, and transformed into a "metal-bone" scaffold. When this "metal-bone" scaffold was re-implanted into bone defects in the animals, the results demonstrated that, in comparison to a plain porous Ti6Al4V scaffold, the scaffold containing bone tissue achieved accelerated early-stage bone regeneration. The experimental group exhibited more bone tissue generation in the early stages at the defect site, resulting in superior bone integration. In conclusion, the "metal-bone" scaffold, containing bone tissue, proves to be an effective bone-promoting scaffold with promising clinical applications.

Development of a 3D multifunctional collagen scaffold impregnated with peptide LL-37 for vascularised bone tissue regeneration

Bone is a highly dynamic connective tissue that provides structural support, locomotion and acts as a shield for many vital organs from damage. Bone inherits the ability to heal after non-severe injury. In case of severe bone abnormalities due to trauma, infections, genetic disorders and tumors, there is a demand for a scaffold that can enhance bone formation and regenerate the lost bone tissue. In this study, a 3D collagen scaffold (CS) was functionalized and assessed under in vitro and in vivo conditions. For this, a collagen scaffold coated with hydroxyapatite (Ap-CS) was developed and loaded with a peptide LL-37. The physico-chemical characterisation confirmed the hydroxyapatite coating on the outer and inner surfaces of Ap-CS. In vitro studies confirmed that LL-37 loaded Ap-CS promotes osteogenic differentiation of human osteosarcoma cells without showing significant cytotoxicity. The efficacy of the LL-37 loaded Ap-CS for bone regeneration was evaluated at 4 and 12 weeks post-implantation by histopathological and micro-CT analysis in rabbit femur defect model. The implanted LL-37 loaded Ap-CS facilitated the new bone formation at 4 weeks compared with Ap-CS without LL-37. The LL-37 loaded Ap-CS incorporating apatite and peptide LL-37 would be useful as a multifunctional scaffold for bone tissue engineering.

Ultrasound stimulated piezoelectric barium titanate and boron nitride nanotubes in nonconductive poly-ε-caprolactone nanofibrous scaffold for bone tissue engineering

Nanomaterials can provide unique solutions for the problems experienced in tissue engineering by improving a scaffold’s physico-bio-chemical properties. With its piezoelectric property, bone is an active tissue with easy adaptation and remodeling through complicated mechanisms of electromechanical operations. Although poly(ε-caprolactone) (PCL) is an excellent polymer for bone tissue engineering, it is lack of conductivity. In this study, piezoelectric barium titanates (BaTiO3) and boron nitride nanotubes (BNNTs) are used as ultrasound (US) stimulated piezoelectric components in PCL to mimic piezoelectric nature of bone tissue. Electric-responsive Human Osteoblast cells on the scaffolds were stimulated by applying low-frequency US during cell growth. Biocompatibility, cell adhesion, alkaline phosphatase activities and mineralization of osteoblast cells on piezo-composite scaffolds were investigated. BaTiO3 or BNNTs as reinforcement agents improved physical and mechanical properties of PCL scaffolds. In vitro studies show that the use of BaTiO3 or BNNTs as additives in non-conductive scaffolds significantly induces and increases the osteogenic activities even without US stimulation. Although BaTiO3 is one of the best piezoelectric materials, the improvement is more dramatic in the case of BNNTs with the increased mineralization, and excellent chemical and mechanical properties.

Conductive GelMA/alginate/polypyrrole/graphene hydrogel as a potential scaffold for cardiac tissue engineering; Physiochemical, mechanical, and biological evaluations

Cardiac failure can be a life-threatening condition that, if left untreated, can have significant consequences. Functional hydrogel has emerged as a promising platform for cardiac tissue engineering. In the proposed study, gelatin methacrylate (GelMA) and alginate, as a primary matrix to maintain cell viability and proliferation, and polypyrrole and carboxyl-graphene, to improve mechanical and electrical properties, are thoroughly evaluated. Initially, a polymer blend of GelMA/Alginate (1:1) was prepared, and then the addition of 2–5 wt% of polypyrrole was evaluated. Next, the effect of incorporating graphene-carboxyl nanosheets (1, 2, and 3 wt%) within the optimized scaffold with 2 wt% polypyrrole was thoroughly studied. The electrical conductivity of the hydrogels was significantly enhanced from 0.0615 ± 0.007 S/cm in GelMA/alginate to 0.124 ± 0.04 S/cm with the addition of 5 wt% polypyrrole. Also, 3 wt% carboxyl graphene improved the electrical conductivity to 0.27 ± 0.09 S/cm. The compressive strength of carboxyl-graphene-containing hydrogel was in the range of 175 to 520 kPa, and tensile strength was 61 and 133 kPa. The simplicity and low-cost fabrication, tunable mechanical properties, optimal electrical conductivity, blood compatibility, and non-cytotoxicity of GelMA/alginate/polypyrrole/graphene biocomposite hydrogel is a promising construct for cardiac tissue engineering.

The Effect of Spironolactone Loading on the Properties of 3D-Printed Polycaprolactone/Gold Nanoparticles Composite Scaffolds for Myocardial Tissue Engineering

The Effect of Spironolactone Loading on the Properties of 3D-Printed Polycaprolactone/Gold Nanoparticles Composite Scaffolds for Myocardial Tissue Engineering

Hyperspectral Mapping of Human Primary and Stem Cells at Cell-Matrix Interfaces.

Extracellular matrices interface with cells to promote cell growth and tissue development. Given this critical role, matrix mimetics are introduced to enable biomedical materials ranging from tissue engineering scaffolds and tumor models to organoids for drug screening and implant surface coatings. Traditional microscopy methods are used to evaluate such materials in their ability to support exploitable cell responses, which are expressed in changes in cell proliferation rates and morphology. However, the physical imaging methods do not capture the chemistry of cells at cell-matrix interfaces. Herein, we report hyperspectral imaging to map the chemistry of human primary and embryonic stem cells grown on matrix materials, both native and artificial. We provide the statistical analysis of changes in lipid and protein content of the cells obtained from infrared spectral maps to conclude matrix morphologies as a major determinant of biochemical cell responses. The study demonstrates an effective methodology for evaluating bespoke matrix materials directly at cell-matrix interfaces.

Random cellulose acetate nanofibers: a breakthrough for cultivated meat production.

Overcoming the challenge of creating thick, tissue-resembling muscle constructs is paramount in the field of cultivated meat production. This study investigates the remarkable potential of random cellulose acetate nanofibers (CAN) as a transformative scaffold for muscle tissue engineering (MTE), specifically in the context of cultivated meat applications. Through a comparative analysis between random and aligned CAN, utilizing C2C12 and H9c2 myoblasts, we unveil the unparalleled capabilities of random CAN in facilitating muscle differentiation, independent of differentiation media, by exploiting the YAP/TAZ-related mechanotransduction pathway. In addition, we have successfully developed a novel process for stacking cell-loaded CAN sheets, enabling the production of a three-dimensional meat product. C2C12 and H9c2 loaded CAN sheets were stacked (up to four layers) to form a ~300-400 μm thick tissue 2 cm in length, organized in a mesh of uniaxial aligned cells. To further demonstrate the effectiveness of this methodology for cultivated meat purposes, we have generated thick and viable constructs using chicken muscle satellite cells (cSCs) and random CAN. This groundbreaking discovery offers a cost-effective and biomimetic solution for cultivating and differentiating muscle cells, forging a crucial link between tissue engineering and the pursuit of sustainable and affordable cultivated meat production.

Augmented physical, mechanical, and cellular responsiveness of gelatin-aldehyde modified xanthan hydrogel through incorporation of silicon nanoparticles for bone tissue engineering

Bioactive scaffolds fabricated from a combination of organic and inorganic biomaterials are a promising approach for addressing defects in bone tissue engineering. In the present study, a self-crosslinked nanocomposite hydrogel, composed of gelatin/aldehyde-modified xanthan (Gel-AXG) is successfully developed by varying concentrations of porous silicon nanoparticles (PSiNPs). The effect of PSiNPs incorporation on physical, mechanical, and biological performance of the nanocomposite hydrogel is evaluated. Morphological analysis reveals formation of highly porous 3D microstructures with interconnected pores in all nanocomposite hydrogels. Increased content of PSiNPs results in a lower swelling ratio, reduced porosity and pore size, which in turn impeded media penetration and slowed down the degradation process. In addition, remarkable enhancements in dynamic mechanical properties are observed in Gel-AXG-8%Si (compressive strength: 0.6223 MPa at 90 % strain and compressive modulus: 0.054 MPa), along with improved biomineralization ability via hydroxyapatite formation after immersion in simulated body fluid (SBF). This optimized nanocomposite hydrogel provides a sustained release of Si ions at safe dose levels. Furthermore, in-vitro cytocompatibility studies using MG-63 cells exhibited remarkable performance in terms of cell attachment, proliferation, and ALP activity for Gel-AXG-8%Si. These findings suggest that the prepared nanocomposite hydrogel holds promising potential as a scaffold for bone tissue engineering.

The effect of 3D-printed bone tissue scaffolds geometrical designs on bacterial biofilm formation

Bone fractures are recognized as a global health problem. A common strategy to tackle this issue is to employ a tissue engineering scaffold to accelerate tissue healing. However, one of the main challenges that can result in delaying the recovery is the risk of bacterial infections. This study aims to assess the impact of the geometry and the porosity of tissue scaffolds on the Staphylococcus aureus biofilm formation. Three triply periodic minimal surface designs of Schwarz primitive (SP), gyroid (GY), and Schwarz diamond (SD) and re-entrant auxetic (RE) design were examined and compared to a reference design (RD) considering two different porosity levels of 75% and 45%. The amount of biofilm was quantified using crystal violet assay and was visualized using scanning electron microscopy. The SP scaffold, with low porosity, exhibited a significantly less amount of bacterial biofilm formation and was regarded as having the best design among the others, while the SD with low porosity showed the greatest amount of biofilm. The morphological analysis was also in line with the crystal violet assay results. On the other hand, the surface roughness was affected by the complexity, geometrical variations, and limitations of fused filament fabrication three-dimensional printing. For the RD, SP, GY, and SD designs, an increase in surface roughness was demonstrated to increase the production of bacterial biofilms. Without statistical significance, the RE design showed the opposite trend. Contrary to other designs, the increase in pore size of the SP and GY designs was associated with the development of bacterial biofilms. This study suggests that it is possible to minimize the likelihood of bacterial biofilm formation by optimizing the scaffold geometry and its manufacturing.

Piezoelectrically and Topographically Engineered Scaffolds for Accelerating Bone Regeneration.

Bone regeneration remains a critical concern across diverse medical disciplines, because it is a complex process that requires a combinatorial approach involving the integration of mechanical, electrical, and biological stimuli to emulate the native cellular microenvironment. In this context, piezoelectric scaffolds have attracted considerable interest owing to their remarkable ability to generate electric fields in response to dynamic forces. Nonetheless, the application of such scaffolds in bone tissue engineering has been limited by the lack of a scaffold that can simultaneously provide both the intricate electromechanical environment and the biocompatibility of the native bone tissue. Here, we present a pioneering biomimetic scaffold that combines the unique properties of piezoelectric and topographical enhancement with the inherent osteogenic abilities of hydroxyapatite (HAp). Notably, the novelty of this work lies in the incorporation of HAp into polyvinylidene fluoride-co-trifluoro ethylene in a freestanding form, leveraging its natural osteogenic potential within a piezoelectric framework. Through comprehensive in vitro and in vivo investigations, we demonstrate the remarkable potential of these scaffolds to accelerate bone regeneration. Moreover, we demonstrate and propose three pivotal mechanisms─(i) electrical, (ii) topographical, and (iii) paracrine─that collectively contribute to the facilitated bone healing process. Our findings present a synergistically derived biomimetic scaffold design with wide-ranging prospects for bone regeneration as well as various regenerative medicine applications.

Application of metal-organic frameworks-based functional composite scaffolds in tissue engineering.

With the rapid development of materials science and tissue engineering, a variety of biomaterials have been used to construct tissue engineering scaffolds. Due to the performance limitations of single materials, functional composite biomaterials have attracted great attention as tools to improve the effectiveness of biological scaffolds for tissue repair. In recent years, metal-organic frameworks (MOFs) have shown great promise for application in tissue engineering because of their high specific surface area, high porosity, high biocompatibility, appropriate environmental sensitivities and other advantages. This review introduces methods for the construction of MOFs-based functional composite scaffolds and describes the specific functions and mechanisms of MOFs in repairing damaged tissue. The latest MOFs-based functional composites and their applications in different tissues are discussed. Finally, the challenges and future prospects of using MOFs-based composites in tissue engineering are summarized. The aim of this review is to show the great potential of MOFs-based functional composite materials in the field of tissue engineering and to stimulate further innovation in this promising area.

Synergistic Effects of SDS and H2O2 Combinations on Tracheal Scaffold Development: An In Vitro Study Using Goat Trachea.

Currently, a tissue-engineered trachea has been popularly used as a biological graft for tracheal replacement in severe respiratory diseases. In the development of tissue-engineered tracheal scaffolds, in vitro studies play a crucial role in allowing researchers to evaluate the efficacy and safety of scaffold designs and fabrication techniques before progressing to in vivo or clinical trials. This research involved the decellularization of goat trachea using SDS, H2O2, and their combinations. Various quantitative and qualitative assessments were performed, including histological analysis, immunohistochemistry, and biomechanical testing. Hematoxylin and eosin staining evaluated the cellular content, while safranin O-fast green and Masson's trichrome staining assessed glycosaminoglycan content and collagen distribution, respectively. The immunohistochemical analysis focused on detecting MHC-1 antigen presence. Tensile strength measurements were conducted to evaluate the biomechanical properties of the decellularized scaffolds. The results demonstrated that the combination of SDS and H2O2 for goat tracheal decellularization yielded scaffolds with minimal cellular remnants, low toxicity, preserved ECM, and high tensile strength and elasticity. This method holds promise for developing functional tracheal scaffolds to address severe respiratory diseases effectively.

Efficient three-dimensional (3D) human bone differentiation on quercetin-functionalized isotropic nano-architecture chitinous patterns of cockroach wings

Developing cost-effective, biocompatible scaffolds with nano-structured surface that truthfully replicate the physico-(bio)chemical and structural properties of bone tissue's extracellular matrix (ECM) is still challenging. In this regard, surface functionalization of natural scaffolds to enhance capability of mimicking 3D niches of the bone tissue has been suggested as a solution. In the current study, we aimed to investigate the potential of chitin-based cockroach wings (CW) as a natural scaffold for bone tissue engineering. To raise the osteogenic differentiation capacity of such a scaffold, a quercetin coating was also applied (hereafter this scaffold is referred as QCW). Moreover, the QCW scaffold exhibited effective antibacterial properties against gram-positive S. aureus bacteria. With respect to bone regeneration, the QCW scaffold optimally induced the differentiation of adipose-derived human mesenchymal stem cells (AD-hMSCs) into osteoblasts, as validated by mineralization assays, alkaline phosphatase (ALP) activity measurements, expression of pre-osteocyte marker genes, and immunocytochemical staining. Confirmation of the potent biocompatibility and physicochemical characteristics of the QCW scaffold through a series of in vitro and in vivo analysis revealed that surface modification had significant effect on multi-purpose features of obtained scaffold. Altogether, surface modification of QCW made it as an affordable bioinspired scaffold for bone tissue engineering.