Blend Scaffolds Research Articles

Innovative Biocompatible Blend Scaffold of Poly(hydroxybutyrate-co-hydroxyvalerate) and Poly(ε-caprolactone) for Bone Tissue Engineering: In Vitro and In Vivo Evaluation.

This study evaluated the biocompatibility of dense and porous forms of Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV), Poly(ε-caprolactone) (PCL), and their 75/25 blend for bone tissue engineering applications. The biomaterials were characterized morphologically using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR), and the thickness and porosity of the scaffolds were determined. Functional assessments of mesenchymal stem cells (MSCs) included the MTT assay, alkaline phosphatase (ALP) production, and morphological and cytochemical analyses. Moreover, these polymers were implanted into rats to evaluate their in vivo performance. The morphology and FTIR spectra of the scaffolds were consistent with the expected results. Porous polymers were thicker than dense polymers, and porosity was higher than 92% in all samples. The cells exhibited good viability, activity, and growth on the scaffolds. A higher number of cells was observed on dense polymers, likely due to their smaller surface area. ALP production occurred in all samples, but enzyme activity was more intense in PCL samples. The scaffolds did not interfere with the osteogenic capacity of MSCs, and mineralized nodules were present in all samples. Histological analysis revealed new bone formation in all samples, although pure PHBV exhibited lower results compared to the other blends. In vivo results indicated that dense PCL and the dense 75/25 blend were the best materials tested, with PCL tending to improve the performance of PHBV in vivo.

Enzyme-delivery Metal-organic Framework Composite Coatings for Restoration of Hyperglycemia-damaged Osteoblast Differentiation

There is a significant clinical need to develop effective treatments for bone defects in patients with diabetes mellitus (DM), as they are at higher risk of fractures and impaired healing. Guided bone tissue engineering using biocompatible and biodegradable polymers is a promising approach. However, current diabetic bone regenerative therapies often fail due to the accumulation of advanced glycation products, which can affect the integration of traditional tissue engineering scaffolds with native bone. Therefore, novel approaches are needed to improve the efficacy of diabetic bone regeneration. This study presents a proof-of-concept development of a multifunctional polymer composite coating tailored towards restoring diabetes-related damage in osteoblast differentiation. Our composite system involves 3D-printed poly(caprolactone fumarate) (PCLF) and poly(caprolactone) (PCL) blend scaffolds coated with multifunctional chitosan methacrylate (chiMA). The chiMA coating is embedded with a sustained-release formulation of glucose oxidase (GOx) from MIL-127 metal-organic frameworks making the coating a stimuli-responsive biomolecule delivery system. The multifunctional coating is designed for the sustained release of GOx and sodium pyruvate for in vitro glucose modulation and oxidative stress reduction, respectively. We propose that sustained release of GOx from MIL-127 embedded chiMA coatings can modulate the high glucose (HG) cellular milieu towards normal glucose (NG), enhancing osteoblast (OB) differentiation via downstream effects. Our results show successful synthesis of MIL-127, encapsulation of GOx, and fabrication of composite coating on the PCLF/PCL scaffolds with effective enzyme activity measured as a function of lowering glucose concentration in HG media for 144 h to normal levels. In vitro evaluation of OB viability, attachment, proliferation, and differentiation showed an overall decrease in cellular activity in HG conditions, which was restored through the glucose-modulating functionality of the GOx-releasing MIL-127 coatings. Our results also presented preliminary evidence of a statistical correlation between DM-related gene markers and osteogenic markers in vitro that requires further exploration. Although this proof-of-concept study holds promise for advancing precision biomaterials development for diabetic tissue engineering and meeting the unmet clinical need for effective treatments and warrants future in vivo evaluation of the composite coating and molecular biology understanding of correlations between DM and osteogenic markers.

Development of Cerium Oxide-Laden GelMA/PCL Scaffolds for Periodontal Tissue Engineering.

This study investigated gelatin methacryloyl (GelMA) and polycaprolactone (PCL) blend scaffolds incorporating cerium oxide (CeO) nanoparticles at concentrations of 0%, 5%, and 10% w/w via electrospinning for periodontal tissue engineering. The impact of photocrosslinking on these scaffolds was evaluated by comparing crosslinked (C) and non-crosslinked (NC) versions. Methods included Fourier transform infrared spectroscopy (FTIR) for chemical analysis, scanning electron microscopy (SEM) for fiber morphology/diameters, and assessments of swelling capacity, degradation profile, and biomechanical properties. Biological evaluations with alveolar bone-derived mesenchymal stem cells (aBMSCs) and human gingival fibroblasts (HGFs) encompassed tests for cell viability, mineralized nodule deposition (MND), and collagen production (CP). Statistical analysis was performed using Kruskal-Wallis or ANOVA/post-hoc tests (α = 5%). Results indicate that C scaffolds had larger fiber diameters (~250 nm) compared with NC scaffolds (~150 nm). NC scaffolds exhibited higher swelling capacities than C scaffolds, while both types demonstrated significant mass loss (~50%) after 60 days (p < 0.05). C scaffolds containing CeO showed increased Young's modulus and tensile strength than NC scaffolds. Cells cultured on C scaffolds with 10% CeO exhibited significantly higher metabolic activity (>400%, p < 0.05) after 7 days among all groups. Furthermore, CeO-containing scaffolds promoted enhanced MND by aBMSCs (>120%, p < 0.05) and increased CP in 5% CeO scaffolds for both variants (>180%, p < 0.05). These findings underscore the promising biomechanical properties, biodegradability, cytocompatibility, and enhanced tissue regenerative potential of CeO-loaded GelMA/PCL scaffolds for periodontal applications.

Unconventional strategies for liver tissue engineering: plant, paper, silk and nanomaterial-based scaffolds.

The paper highlights how significant characteristics of liver can be modeled in tissue-engineered constructs using unconventional scaffolds. Hepatic lobular organization and metabolic zonation can be mimicked with decellularized plant structures with vasculature resembling a native-hepatic lobule vascular arrangement or silk blend scaffolds meticulously designed for guided cellular arrangement as hepatic patches or metabolic activities. The functionality of hepatocytes can be enhanced and maintained for long periods in naturally fibrous structures paving way for bioartificial liver development. The phase I enzymatic activity in hepatic models can be raised exploiting the microfibrillar structure of paper to allow cellular stacking creating hypoxic conditions to induce in vivo-like xenobiotic metabolism. Lastly, the paper introduces amalgamation of carbon-based nanomaterials into existing scaffolds in liver tissue engineering.

3D Melt-Extrusion Printing of Medium Chain Length Polyhydroxyalkanoates and Their Application as Antibiotic-Free Antibacterial Scaffolds for Bone Regeneration.

In this work, we investigated, for the first time, the possibility of developing scaffolds for bone tissue engineering through three-dimensional (3D) melt-extrusion printing of medium chain length polyhydroxyalkanoate (mcl-PHA) (i.e., poly(3-hydroxyoctanoate-co-hydroxydecanoate-co-hydroxydodecanoate), P(3HO-co-3HD-co-3HDD)). The process parameters were successfully optimized to produce well-defined and reproducible 3D P(3HO-co-3HD-co-3HDD) scaffolds, showing high cell viability (100%) toward both undifferentiated and differentiated MC3T3-E1 cells. To introduce antibacterial features in the developed scaffolds, two strategies were investigated. For the first strategy, P(3HO-co-3HD-co-3HDD) was combined with PHAs containing thioester groups in their side chains (i.e., PHACOS), inherently antibacterial PHAs. The 3D blend scaffolds were able to induce a 70% reduction of Staphylococcus aureus 6538P cells by direct contact testing, confirming their antibacterial properties. Additionally, the scaffolds were able to support the growth of MC3T3-E1 cells, showing the potential for bone regeneration. For the second strategy, composite materials were produced by the combination of P(3HO-co-3HD-co-HDD) with a novel antibacterial hydroxyapatite doped with selenium and strontium ions (Se-Sr-HA). The composite material with 10 wt % Se-Sr-HA as a filler showed high antibacterial activity against both Gram-positive (S. aureus 6538P) and Gram-negative bacteria (Escherichia coli 8739), through a dual mechanism: by direct contact (inducing 80% reduction of both bacterial strains) and through the release of active ions (leading to a 54% bacterial cell count reduction for S. aureus 6538P and 30% for E. coli 8739 after 24 h). Moreover, the composite scaffolds showed high viability of MC3T3-E1 cells through both indirect and direct testing, showing promising results for their application in bone tissue engineering.

Melt electrowriting of poly(ϵ-caprolactone)—poly(ethylene glycol) backbone polymer blend scaffolds with improved hydrophilicity and functionality

Melt electrowriting (MEW) is an additive manufacturing technique that harnesses electro-hydrodynamic phenomena to produce 3D-printed fibres with diameters on the scale of 10s of microns. The ability to print at this small scale provides opportunities to create structures with incredibly fine resolution and highly defined morphology. The current gold standard material for MEW is poly(ϵ-caprolactone) (PCL), a polymer with excellent biocompatibility but lacking in chemical groups that can allow intrinsic additional functionality. To provide this functionality while maintaining PCL’s positive attributes, blending was performed with a Poly(Ethylene Glycol) (PEG)-based Acrylate endcapped Urethane-based Precursor (AUP). AUPs are a group of polymers, built on a backbone of existing polymers, which introduce additional functionality by the addition of one or more acrylate groups that terminate the polymer chain of a backbone polymer. By blending with a 20kDa AUP-PEG in small amounts, it is shown that MEW attributes are preserved, producing high-quality meshes. Blends were produced in various PCL:AUP weight ratios (100:0, 90:10 and 0:100) and processed into both solvent-cast films and MEW meshes that were used to characterise the properties of the blends. It was found that the addition of AUP-PEG to PCL significantly increases the hydrophilicity of structures produced with these polymers, and adds swelling capability compared to the non-swelling PCL. The developed blend (90:10) is shown to be processable using MEW, and the quality of manufactured scaffolds is evaluated against pure PCL scaffolds by performing scanning electron microscopy image analysis, with the quality of the novel MEW blend scaffolds showing comparable quality to that of pure PCL. The presence of the functionalisable AUP material on the surface of the developed scaffolds is also confirmed using fluorescence labelling of the acrylate groups. Biocompatibility of the MEW-processable blend was confirmed through a cell viability study, which found a high degree of cytocompatibility.

Synthesis and Viscoelastic Properties of Polycaprolactone/Polyvinylidene Fluoride/Nanohydroxyapatite Composite Scaffolds

AbstractObtaining a polymer nanocomposite with optimum viscoelastic, thermal, and biocompatibility properties is the main objective when designing nanocomposite systems with potential applications in tissue engineering. For this purpose, a blend of Polycaprolactone (PCL) and Polyvinylidene fluoride (PVDF) in an 85/15 weight ratio, along with a nanocomposite reinforced by nanohydroxyapatite (nHA) particles, is fabricated using a solution casting method in a mold. The impact of nHA content on crystallinity, viscoelastic properties, thermal stability, and the properties–structure relationship of nanocomposites is evaluated using scanning electron microscopy (SEM). Dynamic mechanical thermal (DMTA) analysis is used to determine the William–Landel–Ferry (WLF) constants and the effect of nHA on the nanocomposite's viscoelastic behavior. The PCL/15PVDF/0.5 wt% nHA exhibits the maximum thermal stability (40% residual char value) and 95% increase in storage modulus at 90 °C (rubbery region) in comparison with PCL/15PVDF blend. Water contact angle (WCA) and biocompatibility tests are conducted on the PCL/15PVDF blend and nanocomposite scaffolds to design appropriate nanocomposite systems with potential applications in tissue engineering. The high hydrophilic properties are assigned to PCL/15PVDF/0.5 wt% nHA with a WCA of 67.5°. Finally, in vitro cell culture confirmed 0.5 wt% nHA significantly improves cell adhesion and cytotoxicity with MG‐63 cells.

Biomimicking trilayer scaffolds with controlled estradiol release for uterine tissue regeneration.

Scaffold-based tissue engineering provides an efficient approach for repairing uterine tissue defects and restoring fertility. In the current study, a novel trilayer tissue engineering scaffold with high similarity to the uterine tissue in structure was designed and fabricated via 4D printing, electrospinning and 3D bioprinting for uterine regeneration. Highly stretchable poly(l-lactide-co-trimethylene carbonate) (PLLA-co-TMC, "PTMC" in short)/thermoplastic polyurethane (TPU) polymer blend scaffolds were firstly made via 4D printing. To improve the biocompatibility, porous poly(lactic acid-co-glycolic acid) (PLGA)/gelatin methacryloyl (GelMA) fibers incorporated with polydopamine (PDA) particles were produced on PTMC/TPU scaffolds via electrospinning. Importantly, estradiol (E2) was encapsulated in PDA particles. The bilayer scaffolds thus produced could provide controlled and sustained release of E2. Subsequently, bone marrow derived mesenchymal stem cells (BMSCs) were mixed with gelatin methacryloyl (GelMA)-based inks and the formulated bioinks were used to fabricate a cell-laden hydrogel layer on the bilayer scaffolds via 3D bioprinting, forming ultimately biomimicking trilayer scaffolds for uterine tissue regeneration. The trilayer tissue engineering scaffolds thus formed exhibited a shape morphing ability by transforming from the planar shape to tubular structures when immersed in the culture medium at 37°C. The trilayer tissue engineering scaffolds under development would provide new insights for uterine tissue regeneration.

Snail Mucus-Enhanced Adhesion of Human Chondrocytes on 3D Porous Agarose Scaffolds.

This study reports the preparation of a novel porous 3D scaffold from agarose-snail mucus (AGSMu) for cartilage tissue repair applications. AG is reported for its unique thermal and mechanical properties, biocompatibility, and biodegradability, making it suitable for biomedical applications. Still, it lacks the cell adhesion properties required for tissue engineering applications. SMu is a complex substance identified to contain glycosaminoglycans (GAGs) and other bioactive molecules that promote wound healing and reduce cartilage deterioration and inflammation. Hence, porous 3D blend scaffolds containing AG and SMu were prepared by the freeze-drying method, characterized, and investigated for bioactive effects on human chondrocyte (C28/I2) cells. The scaffolds had a microporous structure with an average pore size of 245 μm. FTIR spectroscopy showed that SMu was successfully incorporated into the scaffolds. The SMu increased the mechanical strength of the composite scaffolds by more than 80% compared to the pristine AG scaffold. The scaffolds were found to be biocompatible with tunable degradation. The human chondrocyte cells attached and proliferated well on the 3D scaffolds in a few days, demonstrating a marked improvement in adhesion due to the presence of SMu. Enhanced cell adhesion and mechanical properties of 3D porous AG scaffolds could make them suitable for articular cartilage repair and regeneration.

PLA blended gelatine-based nanofibrous mats with enhanced hydrophobicity for soft tissue regeneration

Wound healing requires a substantial amount of moisture for faster recovery. Completely hydrophobic or hydrophilic biomaterials are not suitable to be applied for cell growth in wounded areas. The study aimed to prepare a nanofibrous scaffold from the blend of a solution of hydrophobic PLA and a solution of hydrophilic gelatine. The stability of the blend was achieved using a surfactant and an electrospun nanofibrous scaffold was made out of the solution. The optimum composition of gelatine and PLA to make a scaffold of uniform fibre diameter was achieved with the help of conductivity, viscosity and FESEM analysis. The optimum scaffold was characterised by TGA, DSC and XRD analysis. The water contact angle of the optimum sample was observed at 27°. The blended scaffold was found non-toxic to cells and showed a 30% faster healing of wounds in the rat model test compared to the healing rate of the PLA scaffold or the gelatine scaffold alone. The histological assay also supported the blend scaffold as an encouraging material for tissue regeneration.

Multilayered Shape-Morphing Scaffolds with a Hierarchical Structure for Uterine Tissue Regeneration.

Owing to dysfunction of the uterus, millions of couples around the world suffer from infertility. Different from conventional treatments, tissue engineering provides a new and promising approach to deal with difficult problems such as human tissue or organ failure. Adopting scaffold-based tissue engineering, three-dimensional (3D) porous scaffolds in combination with stem cells and appropriate biomolecules may be constructed for uterine tissue regeneration. In this study, a hierarchical tissue engineering scaffold, which mimicked the uterine tissue structure and functions, was designed, and the biomimicking scaffolds were then successfully fabricated using solvent casting, layer-by-layer assembly, and 3D bioprinting techniques. For the multilayered, hierarchical structured scaffolds, poly(l-lactide-co-trimethylene carbonate) (PLLA-co-TMC, "PLATMC" in short) and poly(lactic acid-co-glycolic acid) (PLGA) blends were first used to fabricate the shape-morphing layer of the scaffolds, which was to mimic the function of myometrium in uterine tissue. The PLATMC/PLGA polymer blend scaffolds were highly stretchable. Subsequently, after etching of the PLATMC/PLGA surface and employing estradiol (E2), polydopamine (PDA), and hyaluronic acid (HA), PDA@E2/HA multilayer films were formed on PLATMC/PLGA scaffolds to build an intelligent delivery platform to enable controlled and sustained release of E2. The PDA@E2/HA multilayer films also improved the biological performance of the scaffold. Finally, a layer of bone marrow-derived mesenchymal stem cell (BMSC)-laden hydrogel [which was a blend of gelatin methacryloyl (GelMA) and gelatin (Gel)] was 3D printed on the PDA@E2/HA multilayer films of the scaffold, thereby completing the construction of the hierarchical scaffold. BMSCs in the GelMA/Gel hydrogel layer exhibited excellent cell viability and could spread and be released eventually upon biodegradation of the GelMA/Gel hydrogel. It was shown that the hierarchically structured scaffolds could evolve from the initial flat shape into the tubular structure completely in an aqueous environment at 37 °C, fulfilling the requirement for curved scaffolds for uterine tissue engineering. The biomimicking scaffolds with a hierarchical structure and curved shape, high stretchability, and controlled and sustained E2 release appear to be very promising for uterine tissue regeneration.

Optimization of Electrospun Bilayer Vascular Grafts through Assessment of the Mechanical Properties of Monolayers.

Small-diameter vascular grafts must be obtained with the most appropriate materials and design selection to harmoniously display a variety of features, including adequate tensile strength, compliance, burst strength, biocompatibility, and biodegradability against challenging physiological and hemodynamic conditions. In this study, monolayer vascular grafts with randomly distributed or radially oriented fibers are produced using neat, blended, and copolymer forms of polycaprolactone (PCL) and poly(lactic acid) (PLA) via the electrospinning technique. The blending ratio is varied by increasing 10 in the range of 50-100%. Bilayer graft designs are realized by determining the layers with a random fiber distribution for the inner layer and radial fiber orientation for the outer layer. SEM analysis, wall thickness and fiber diameter measurements, tensile strength, elongation, burst strength, and compliance tests are done for both mono- and bilayer scaffolds. The findings revealed that the scaffolds made of neat PCL show more flexibility than the neat PLA samples, which possess higher tensile strength values than neat PCL scaffolds. Also, in blended samples, the tensile strength values do not show a significant improvement, whereas the elongation values are enhanced in tubular samples, depending on the blending ratio. Also, neat poly(l-lactide-co-caprolactone) (PLCL) samples have both higher elongation and strength values than neat and blended scaffolds, with some exceptions. The blended specimens comprising a combination of PCL and PLA, with blending ratios of 80/20 and 70/30, exhibited the most elevated burst pressures. Conversely, the PLCL scaffolds demonstrated superior compliance levels. These findings suggest that the blending approach and fiber orientation offer enhanced burst strength, while copolymer utilization in PLCL scaffolds without fiber alignment enhances their compliance properties. Thus, it is evident that using a copolymer instead of blending PCL and PLA and combining the PLCL layer with PCL and PLA monolayers in bilayer vascular graft design is promising in terms of mechanical and biological properties.

Modeling and optimization of poly(lactic acid)/poly(ℇ-caprolactone)/Nigella sativa extract nanofibers production for skin wounds healing by artificial neural network and response surface methodology models

Electrospun fibrous scaffolds have great potential for the effective treatment of wounds. Novel blend scaffolds were fabricated from poly(ℇ- caprolactone) (PCL)/poly (lactic acid) (PLA) with Nigella sativa (NS) extract in different concentrations of 10 %, 15 %, 20 %, and 25 % by one nozzle electrospinning. RSM and ANN models were used to determine optimal nanofiber. The results showed that the ANN model had average goodness values of almost 1.992 which was higher than the RSM model with an amount of 1.823. The best sample was determined with the combination of parameters such as PLA/PCL (70:29) concentration, voltage 17 kV, and flow rate 0.2 ml/h in diameter of nanofiber 410 nm by Genetic Algorithm (GA) model with cost value 0.0216 that was lower than cost value (0.0927) of ANN model. The effect of NS extract on nanofibers properties showed that loading high concentrations of NS extract in PLA/PCL polymer solutions caused a decrease in nanofibers diameter, hydrophilicity, and tensile strength. Overall, PLA/PCL/NS 25 % nanofiber was selected as an optimal web with an average diameter of 370 ± 68 nm with a young modulus 5.94 MPa. This scaffold also exhibited the highest antibacterial activity, cell attachment, and cell viability based on the MTT assay.

Konjac glucomannan photocrosslinked hydrogels for in vitro 3D cell culture

Konjac glucomannan (KGM) is an attractive naturally-derived polysaccharide commercially available for use in food industry, but with limited application in tissue engineering (TE) as a standalone polymer. We successfully purified and chemically modified KGM, developing innovative covalently crosslinked, fast-forming and degradable KGM hydrogels with tunable mechanical properties for 3D cell culture. Human dermal fibroblasts embedded in KGM matrices remained viable and metabolically active, supporting its good biocompatibility. We further innovated by grafting a bioactive peptide to KGM backbone applying two different chemical approaches, one of them culminating in the synthesis of a novel pH-responsive KGM derivative. Cells embedded in biofunctionalized KGM hydrogels were able to adhere, spread and establish cell-to-cell interactions, encouraging the formation of a cellular network. This work reports different chemical approaches for targeting the fabrication of bioactive KGM-derived hydrogels for 3D cell culture and compares the biological performance of obtained KGM's covalent and semi-interpenetrating polymer networks (SIPN). These results will pave the way for further exploitation of KGM in TE, where it has been commonly applied as blended scaffold formulations rather than as stable extracellular matrix (ECM)-mimicking materials.

Air-jet spun PHBV/PCL blend tissue engineering scaffolds exhibit improved mechanical properties and cell proliferation

This study aimed to optimize the preparation parameters for air-jet spun blended poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and polycaprolactone (PCL) submicron fibrous scaffolds. Blends of PHBV and PCL were made using different weight ratios of polymers and examined for their mechanical, morphological, and biological properties. The 50/50 ratio PHBV/PCL blend scaffolds showed better overall mechanical properties compared to the scaffolds with higher PHBV content and provided better cell attachment and proliferation compared to PCL scaffolds. The incorporation of PCL improved the degree of crystallinity, thermal stability, and mechanical properties of PHBV/PCL blend scaffolds. In vivo subcutaneous implantation and subsequent histological analysis of the implanted scaffolds indicated that the scaffolds were compatible with animal tissues. The findings show that air-jet spun PHBV/PCL blended scaffolds in a 50/50 ratio can be employed in many tissue engineering applications.

PGS/Gelatin Nanocomposite Electrospun Wound Dressing.

Infectious diabetic wounds can result in severe injuries or even death. Biocompatible wound dressings offer one of the best ways to treat these wounds, but creating a dressing with a suitable hydrophilicity and biodegradation rate can be challenging. To address this issue, we used the electrospinning method to create a wound dressing composed of poly(glycerol sebacate) (PGS) and gelatin (Gel). We dissolved the PGS and Gel in acetic acid (75 v/v%) and added EDC/NHS solution as a crosslinking agent. Our measurements revealed that the scaffolds' fiber diameter ranged from 180.2 to 370.6 nm, and all the scaffolds had porosity percentages above 70%, making them suitable for wound healing applications. Additionally, we observed a significant decrease (p < 0.05) in the contact angle from 110.8° ± 4.3° for PGS to 54.9° ± 2.1° for PGS/Gel scaffolds, indicating an improvement in hydrophilicity of the blend scaffold. Furthermore, our cell viability evaluations demonstrated a significant increase (p < 0.05) in cultured cell growth and proliferation on the scaffolds during the culture time. Our findings suggest that the PGS/Gel scaffold has potential for wound healing applications.

Application of Aloe vera Gel Blended Polymer-Collagen Scaffolds for Bone Tissue Engineering

Application of <i>Aloe vera</i> Gel Blended Polymer-Collagen Scaffolds for Bone Tissue Engineering

Effect of Uniaxial Compression Frequency on Osteogenic Cell Responses in Dynamic 3D Cultures.

The application of mechanical stimulation on bone tissue engineering constructs aims to mimic the native dynamic nature of bone. Although many attempts have been made to evaluate the effect of applied mechanical stimuli on osteogenic differentiation, the conditions that govern this process have not yet been fully explored. In this study, pre-osteoblastic cells were seeded on PLLA/PCL/PHBV (90/5/5 wt.%) polymeric blend scaffolds. The constructs were subjected every day to cyclic uniaxial compression for 40 min at a displacement of 400 μm, using three frequency values, 0.5, 1, and 1.5 Hz, for up to 21 days, and their osteogenic response was compared to that of static cultures. Finite element simulation was performed to validate the scaffold design and the loading direction, and to assure that cells inside the scaffolds would be subjected to significant levels of strain during stimulation. None of the applied loading conditions negatively affected the cell viability. The alkaline phosphatase activity data indicated significantly higher values at all dynamic conditions compared to the static ones at day 7, with the highest response being observed at 0.5 Hz. Collagen and calcium production were significantly increased compared to static controls. These results indicate that all of the examined frequencies substantially promoted the osteogenic capacity.

Characteristic Evaluation of Recombinant MiSp/Poly(lactic-co-glycolic) Acid (PLGA) Nanofiber Scaffolds as Potential Scaffolds for Bone Tissue Engineering.

Biomaterial-based nanofibrous scaffolds are the most effective alternative to bone transplantation therapy. Here, two recombinant minor ampullate spidroins (spider silk proteins), R1SR2 and NR1SR2C, were blended with Poly(lactic-co-glycolic) Acid (PLGA), respectively, to generate nanofiber scaffolds by electrospinning. The N-terminal (N), C-terminal (C), repeating (R1 and R2) and spacer (S) modules were all derived from the minor ampullate spidroins (MiSp). The physical properties and structures of the blended scaffolds were measured by scanning electron microscopy (SEM), water contact angle measurement, Fourier transform infrared spectroscopy (FTIR), Differential scanning calorimetry (DSC), and Tensile mechanical testing. The results showed that blending of MiSp (R1SR2 and NR1SR2C) reduced the diameter of nanofibers, increased the porosity and glass transition temperatures of nanofibrous scaffolds, and effectively improved the hydrophilicity and ultimate strain of scaffolds. It is worth noting that the above changes were more significant in the presence of the N- and C-termini of MiSp. In cell culture assays, human bone mesenchymal stem cells (HBMSCs) grown on NR1SR2C/PLGA (20/80) scaffolds displayed markedly enhanced proliferative and adhesive abilities compared with counterparts grown on pure PLGA scaffolds. Jointly, these findings indicated recombinant MiSp/PLGA, particularly NR1SR2C/PLGA (20/80) blend nanofibrous scaffolds, is promising for bone tissue engineering.

The fabrication and evaluation of silver nanoparticle-based keratin scaffolds.

The biotoxicity caused by focus releasing of Ag, which associated with the Ag loading mode, is a problematic issue that need to be solved for practical utilization of the keratin based wound dressing. In this study, keratin/AgNPs blend scaffolds (Ker/Ag) and keratin scaffolds with AgNPs attached on the scaffold's wall surface (Ag@Ker) were prepared. Structure and physical properties of the scaffolds were tested and investigated. In comparison to the Ag@Ker scaffolds, the Ker/Ag scaffolds with uniform dispersion of AgNPs have larger tensile strength and slower degradation rate. Both kind of scaffolds present excellent antibacterial property with 10μg mL-1 AgNPs addition, while the Ker/Ag displayed a linear Ag releasing ratio in the first 5-7days, which is beneficial for obtaining a continuous antibacterial property and avoiding the biotoxicity caused by focus release of Ag. Correspondingly, cytotoxicity assay further reveals that the continuously slow release of Ag of the Ker/Ag scaffolds accelerated the proliferation of cell. Infectious animal models and histological studies showed that the Ker/Ag scaffolds can effectively inhibit the inflammatory response and accelerate epithelialization. Thus, it can be concluded that the Ker/Ag scaffolds with uniform dispersion of AgNPs are more attractive as wound repair materials.