Potential Bone Tissue Engineering Research Articles

Four-Dimensional Bioprinting: Current Developments and Applications in Bone Tissue Engineering

Four-dimensional (4D) bioprinting, in which the concept of time is integrated with three-dimensional (3D) bioprinting as the fourth dimension, is widely proposed as the next-generation of tissue engineering technology as it presents the possibility of constructing complex, functional structures. 4D bioprinting can be used to fabricate dynamic 3D patterned biological architectures that change their shapes or functions under various stimuli by employing stimuli-responsive materials or novel 3D bioprinting strategies, providing unprecedented potential for bone tissue engineering. The shape memory properties of printed structures cater to the need for personal bone defect repair and biomimetic microenvironments that promote the osteogenic differentiation of stem cells. In this review, we introduce the application of different stimuli-responsive biomaterials in tissue engineering and a series of novel 4D bioprinting strategies based on functional transformation of printed structures through the establishment of biomimetic constructs. Furthermore, we discuss the application of 4D bioprinting in bone tissue engineering, as well as future challenges and perspectives.

Investigation of citric acid-assisted sol-gel synthesis coupled to the self-propagating combustion method for preparing bioactive glass with high structural homogeneity

In this research, the mechanism of an efficient strategy for the synthesis of 58S bioglass with high structural homogeneity by a citric acid assisted sol-gel route was investigated. This is an interesting approach to prepare bioactive glass via the sol-gel method with application potential in bone tissue engineering and also for the development of new biomedical devices. Herein, 58S bioglass was synthesized by two routes: conventional sol-gel method (CSG) and citric acid assisted sol-gel route coupled to the self-propagating combustion method (SPC). The effects of citric acid on the temperature required for 58S vitreous consolidation, long- and short-range ordering were investigated by several analysis techniques. Results suggested that citric acid molecules serve as an effective molecular template formed by molecular network raised from intermolecular forces, especially the hydrogen bonds, resulting from the chemical interactions between the COOH and hydroxyl groups (water, ethanol, POH, SiOH). In this scenario, citric acid controls the phase segregation during the drying and combustion steps of the gel in the SPC method by establishing chemical interactions (hydrogen bonds) with the superficial silanol groups present on the small-sized silica nanoparticles present in the sol governing their growth. Besides these mentioned features, the self-propagating combustion behavior exhibited by the nitrate-citrate in the SPC xerogel during the combustion step allowed the removal of the organic load and the consolidation of the vitreous structure at a temperature considerably lower than the sample obtained by the CSG method. Consequently, the SPC method leads to the formation of a glass structure with high homogeneity for the 58S, whereas the conventional sol-gel method produces a matrix enriched with calcium phosphate crystalline nuclei - glass-ceramic.

Facile fabrication of electrospun regenerated cellulose nanofiber scaffold for potential bone-tissue engineering application

In this study, cellulose acetate (CA) solutions (9–15% w/v) prepared in acetone-water (80:20 & 90:10 v/v) system were subjected to electrospinning for fabricating non-woven nanofibrous CA scaffolds (CAS) with average fiber diameters from 300 to 600 nm. Further, regenerated cellulose scaffold (RCS) was obtained by deacetylation of electrospun CAS in alkaline media for varying time periods to find the ideal time required for complete deacetylation. Following deacetylation, RCS was subjected to varying temperatures (60 °C, 80 °C) to observe the possible positive effect of heat treatment on the improvement of mechanical strength. The RCS was characterized using ATR FTIR, SEM for studying its surface chemistry and morphology along with other physio-chemical characterizations such as micro-tensile strength, swelling property, porosity, degradation rate in acidic conditions. The results were analyzed and co-related with variation of composition in solvent system, deacetylation time and heat treatment temperatures to determine the optimal fabricating conditions for RCS. In vitro studies using MC3T3-E1 osteoblast cells were also conducted on the selected RCS samples to evaluate cell adhesion and cell proliferation using SEM and MTT assay analysis. The primary results indicate positive outcome regarding the viability of RCS as potential biomaterial for bone-tissue engineering.

Injectable Maltodextrin-Based Micelle/Hydrogel Composites for Simvastatin-Controlled Release.

Injectable hydrogels have shown great potential in bone tissue engineering. Simvastatin (SIM), a common hypolipidemic drug, has been suggested as a potential agent to promote bone regeneration. However, due to its hydrophobic nature, the compatibility between SIM and hydrogels is rather poor, thereby greatly affecting the drug release behavior, the mechanical properties, and dimensional stability of the hydrogels. Herein, we presented a novel design to entrap SIM in an injectable maltodextrin-based micelle/hydrogel composite system. Maltodextrin-based micelles were prepared to solubilize and encapsulate SIM. The SIM-loaded aldehyde-modified micelles were anchored to the hydrogel network and served as a cross-linker to realize improved mechanical strength of hydrogel, controlled release, and osteogenic capability of SIM. In all, this study demonstrated a strategy to incorporate drug loaded carriers into hydrogels for drug delivery and tissue engineering applications.

Three-dimensional electrospun nanofibrous scaffolds displaying bone morphogenetic protein-2-derived peptides for the promotion of osteogenic differentiation of stem cells and bone regeneration

Tissue scaffolds with three-dimensional (3D) nanofibrous biomimetic structures have attracted attention in the field of bone regeneration. In recent years, emerging strategies based on electrospinning technologies have facilitated the preparation of 3D nanofibrous scaffolds. Based on these developments, in this study, 3D scaffolds possessing both nanofibrous morphologies and interconnected pores were fabricated for their potential in bone tissue engineering. By combining homogenizing, freeze-drying, and thermal crosslinking techniques, nano-hydroxyapatite/PLLA/gelatin (nHA/PLA/GEL) 3D nanofibrous scaffolds were prepared using pre-fabricated electrospun nanofibers. Then, utilizing a polydopamine (pDA)-assisted coating strategy, bone morphogenetic protein-2 (BMP-2)-derived peptides were further immobilized onto the 3D scaffolds to obtain the resulting nano-hydroxyapatite/PLLA/gelatin-peptide (nHA/PLA/GEL-PEP) 3D nanofibrous scaffolds capable of sustained release. Bone mesenchymal stem cells (BMSCs) were cultured on the 3D nanofibrous scaffolds, then relative cell viability, alkaline phosphatase (ALP) activity, and gene expression assays were performed to study the effects of the scaffolds on cell growth and osteogenic differentiation in vitro. Furthermore, the ability of bone formation in vivo was evaluated using a rat cranial bone defect model. In vitro and in vivo results demonstrated that the 3D nanofibrous scaffolds incorporated with nHA and BMP-2 peptides exhibited favorable biocompatibility and osteoinductivity. Therefore, these nanofibrous scaffolds have excellent potential in bone regenerative medicine.

Improved osteogenic differentiation of human dental pulp stem cells in a layer-by-layer-modified gelatin scaffold.

Dental pulp stem cell is a new type of mesenchymal stem cell that has a potential for tissue regeneration. Gelatin sponges are often used for hemostasis in dental surgery. In this study, we aimed to evaluate the dental pulp stem cells' proliferation and osteogenic differentiation in different layer-by-layer-modified gelatin sponge scaffolds including the G, G + P (gelatin sponge+ poly-l-lysine modification), G + M (gelatin sponge + mineralization modification), and G + M + P (gelatin sponge + mineralization modification + poly-l-lysine modification) groups in vitro and assessed them in vivo. The results showed that dental pulp stem cells had a great potential for osteogenic differentiation. In vitro, the G + M + P group not only enhanced the adhesion and proliferation of dental pulp stem cells but also facilitated their osteogenic differentiation. However, alkaline phosphatase activity was prohibited after modification. In vivo, both dental pulp stem cells and cells from nude mice grew well on the scaffold, and G + M and G + M + P groups could promote the mineralization deposit formation and the expression of osteocalcin in osteogenic differentiation of dental pulp stem cells. In conclusion, the combination of dental pulp stem cells and G + M + P scaffold has a great potential for bone tissue engineering.

In vitro degradation behavior of Mg scaffolds with three-dimensional interconnected porous structures for bone tissue engineering

The degradation behavior in consideration of the pore strut and the interconnectivity of two Mg scaffolds with different three-dimensional interconnected porous structures were evaluated. The interconnectivity of the two scaffolds gradually decreased along with the clogged interconnected pores due to the deposition formation on the pore wall. Mg scaffold with spherical pores and cambered pore strut degraded faster but exhibited better resistance to the deterioration of the interconnectivity compared with Mg scaffold with irregular pores and polygonal pore struts. Direct cell culture of MC3T3-E1 osteoblasts on the two scaffolds indicated a promising potential for bone tissue engineering.

Fabrication and evaluation of carbonate apatite-coated calcium carbonate bone substitutes for bone tissue engineering.

Carbonate apatite-coated calcium carbonate (CO3 Ap/CaCO3 ) was fabricated through a dissolution-precipitation reaction using CaCO3 granules as a precursor to accelerate bone replacement based on superior osteoconductivity of the CO3 Ap shell, along with Ca2+ release from the CaCO3 core and quicker resorption of the CaCO3 core. In the present study, CaCO3 , 10% CO3 Ap/CaCO3 , 30% CO3 Ap/CaCO3 , and CO3 Ap granules were fabricated and examined histologically to evaluate their potential as bone substitutes. Larger contents of CaCO3 in the granules resulted in higher Ca2+ release and promoted cell proliferation of murine preosteoblasts at 6days compared with CO3 Ap. Interestingly, in a rabbit femur defect model, 10% CO3 Ap/CaCO3 induced significantly higher new bone formation and higher material resorption compared with CO3 Ap at 8weeks. Nevertheless, CO3 Ap showed a superior osteoconductive potential compared with 10% CO3 Ap/CaCO3 at 8weeks. All tested granules were most likely resorbed by cell mediation including multinucleated giant cell functions. Therefore, we conclude that CO3 Ap/CaCO3 has a positive potential for bone tissue engineering based on well-controlled calcium release, bone formation, and material resorption.

Novel synthesis and antimicrobial studies of nanoscale titania particles

In the present investigation, a novel strategy of continuous microwave assisted flow synthesis (CMFS) has been adopted in comparison to traditional synthesis procedures (sol-gel and chemical precipitation method) for the quick production of TiO2 nanoparticles with very fine particle properties. The X-ray powder diffraction analysis (XRPD) and transmission electron microscopy (TEM) were two techniques used for analysing the properties related to structure and particle morphology of the resultant samples. It was observed that the particles formed by using continuous flow route were less agglomerated, and particle size (~ 6 nm) was smaller in comparison with others obtained using sol-gel (~ 9 nm) and chemical precipitation method (~ 15 nm). X-ray diffraction impressions established the generation of Anatase phase with preferential [101] dimension. Zeta potential computations were taken to inspect the colloidal stability of nanoparticles. Antimicrobial nature of TiO2 nano-samples was analyzed by using various bacterial and fungal strains. The nanostructured TiO2 particles confirmed outstanding uniformity with respect to chemical and structure. This new ceramic substance with strong antimicrobial activity promised magnificent potential in bone tissue engineering.

Electrospun nanosilicates-based organic/inorganic nanofibers for potential bone tissue engineering.

Although growth factors and drugs (BMP-2, dexamethasone, etc.) have been widely used for bone tissue engineering, they have unignored limits such as adverse effects at high concentrations and easy inactivation in vivo. Accordingly, more osteoinductive supplements without side effects should be considered as alternatives in the design of bone tissue engineering scaffolds. Nanosilicate is a bioactive inorganic nanomaterial consisting of hydrous sodium lithium magnesium silicate, which is recently found to be safe and effective for bone induction. In this study, a range of organic/inorganic nanofibrous scaffolds with varied nanosilicate concentrations (0%, 1%, 5%, and 10% w/w to PCL matrix) were successfully fabricated via electrospinning. The tensile properties of the nanofibers were enhanced at low nanosilicate concentrations, and the incorporation of nanosilicates had no influence on cytocompatibility. Besides, in vitro osteogenesis experiments showed that nanosilicates-doped nanofibers were capable of inducing bone formation better than pure PCL nanofiber samples. More importantly, the results of histological and immunohistochemical assessments further revealed that the nanosilicates-enriched nanofibers had a significant potential of ectopic bone formation in vivo, while the pure PCL samples only induced limited osteogenic cues. All these results indicate that the nanosilicates-based organic/inorganic nanofibers may be potentially efficient for bone tissue engineering.

Physical contact between mesenchymal stem cells and endothelial precursors induces distinct signatures with relevance to the very early phase of regeneration.

Multipotent adult stem cells/precursor cells, especially of the mesenchymal and endothelial lineage, may have great potential for bone tissue engineering. Although their potential is highly recognized, not much is known about the underlying molecular mechanisms that initiate the regeneration process, connect osteogenesis, and angiogenesis and, finally, orchestrate renewal of bone tissue. Our study addressed these questions by generating two in vitro cell culture models to examine the changes in the global gene expression patterns of endothelial precursor cells and mesenchymal stem cells after 24 hours of either humoral (conditioned medium) or direct cell-cell interaction (co-culture). Endothelial precursor cells were isolated from human buffy coat and mesenchymal stem cells from the bone marrow of the femoral head. The comparison of the treated and control cells by microarray analyses revealed in total more than 1500 regulated genes, which were analyzed for their affiliation to angiogenesis and osteogenesis. Expression array analyses at the RNA and protein level revealed data with respect to regulated genes, pathways and targets that may represent a valid basis for further dissection of the systems biology of regeneration processes. It may also be helpful for the reconstitution of the natural composition of a regenerative microenvironment when targeting tissue regeneration both in vitro and in situ.

Hybrid collagen/pNIPAAM hydrogel nanocomposites for tissue engineering application

The present work explores new hybrid bioactive, enzymatically degradable hydrogel nanocomposites. They are composed by stimuli-sensitive semi-interpenetrating collagen/poly(N-isopropyl acrylamide) polymeric matrix, Dellite® 67G and Cloisite® 93A nanoclays, and hydroxyapatite particles. Morphology of the hybrid hydrogel nanocomposites was examined by near infrared chemical imaging, X-ray diffraction, scanning electron microscopy, and transmission electron microscopy techniques. It has been found that the porous hydrogel nanocomposites showed a mix between intercalated and exfoliated structure, with interconnected pores and the inorganic material was evenly distributed in the polymeric matrix because of specific interactions between components. The collagen resistance against enzymatic degradation and the thermal stability were improved due to the protein encapsulation in the synthetic polymer matrix. The hydrogel nanocomposites showed suitable swelling characteristics for tissue engineering. In vitro cytocompatibility and cell viability revealed that the hybrid nanocomposites were non-cytotoxic for rat osteoblasts. These hybrid nanocomposites were designed as potential synthetic bone tissue engineering.

Thermally induced self-agglomeration 3D scaffolds with BMP-2-loaded core-shell fibers for enhanced osteogenic differentiation of rat adipose-derived stem cells.

IntroductionScaffold structure plays a vital role in cell behaviors. Compared with two-dimensional structure, 3D scaffolds can mimic natural extracellular matrix (ECM) and promote cell–cell and cell–matrix interactions. The combination of osteoconductive scaffolds and osteoinductive growth factors is considered to have synergistic effects on bone regeneration.Materials and methodsIn this study, core–shell poly(lactide-co-glycolide) (PLGA)/polycaprolactone (PCL)–BMP-2 (PP–B) fibrous scaffolds were prepared through coaxial electrospinning. Next, we fabricated 3D scaffolds based on PP–B fibers with thermally induced self-agglomeration (TISA) method and compared with conventional PLGA/PCL scaffolds in terms of scaffold morphology and BMP-2 release behaviors. Then, rat adipose-derived stem cells (rADSCs) were seeded on the scaffolds, and the effects on cell proliferation, cell morphology, and osteogenic differentiation of rADSCs were detected.ResultsThe results demonstrated that 3D scaffold incorporated with BMP-2 significantly increased proliferation and osteogenic differentiation of rADSCs, followed by PP–B group.ConclusionOur findings indicate that scaffolds with 3D structure and osteoinductive growth factors have great potential in bone tissue engineering.

Heparin-dopamine functionalized graphene foam for sustained release of bone morphogenetic protein-2.

The recently developed three-dimensional (3D) graphene foam (GrF) is intriguing for potential bone tissue engineering applications because it provides stem cells with a 3D porous substrate for osteogenic differentiation. However, the nature of graphene's structure lacks functional groups, thus making it difficult for further modification such as immobilization or conjugation of growth factors, which are normally required to promote tissue regeneration. To explore the potential of GrF functionalization and sustained release of therapeutic proteins, we fabricated a modified 3D GrF scaffold with bio-inspired heparin-dopamine (Hepa-Dopa) molecules using a highly scalable chemical vapour deposition method. Our data indicated that Hepa-Dopa modification resulted in significantly higher bone morphogenetic protein-2 (BMP2) binding ability and longer release capacity compared with the untreated scaffolds. Importantly, the heparin-functionalized 3D GrF significantly improved the exogenous BMP2-induced osteogenic differentiation. Therefore, our study, for the first time, indicated that the 3D GrF can be biomimetically functionalized with Hepa-Dopa and be used for sustained release of BMP2, thereby inducing osteogenic differentiation and suggesting promising potential as a new multifunctional carrier for therapeutic proteins and stem cells in bone tissue engineering.

Osteogenic Differentiation Modulates the Cytokine, Chemokine, and Growth Factor Profile of ASCs and SHED.

Great efforts have been made to improve bone regeneration techniques owing to a growing variety of sources of stem cells suitable for autologous transplants. Specifically, adipose-derived stem cells (ASCs) and stems cells from human exfoliated deciduous teeth (SHED) hold great potential for bone tissue engineering and cell therapy. After a preliminary characterization of the main biomolecules ASCs and SHED released in their conditioned media, cells were kept both in normal and osteo-inducing conditions. Conventional assays were performed to prove their osteogenic potential such as quantitative real-time polymerase chain reaction (qRT-PCR) (for RUNX-2, collagen type I, osteopontin and osteonectin), alkaline phosphatase activity, osteocalcin production, and von Kossa staining. Conditioned media were tested again after the osteogenic induction and compared to maintaining condition both at base line and after 14 days of culture. The osteogenic condition inhibited the release of all the biomolecules, with the exception, concerning SHED, of growth-regulated alpha protein precursor (GROα), and, to a lesser extent, interleukin (IL)-8. In conclusion, our data support that undifferentiated ASCs and SHED may be preferable to committed ones for general cell therapy approaches, due to their higher paracrine activity. Osteoinduction significantly affects the cytokine, chemokine, and growth factor profile in a differential way, as SHED kept a more pronounced pro-angiogenic signature than ASCs.

Development of mesoporous bioactive glass nanoparticles and its use in bone tissue engineering.

The incidence of bone disorders, from trauma, tissue degeneration due to ageing, pathological conditions to cancer, has been increasing. The pursuit for bone graft substitutes to assist in regenerating large bone defects is ever growing as a result of the shortage in conventional autografts and allografts, in addition to the associated risks of disease transmission. However, the use of alloplastic biomaterials is limited in clinical settings, as further investigations are required to address the properties of synthetic grafts to mimic the native bone tissue and deliver desirable biomolecules to facilitate bone regeneration. This review discusses the fundamental structure and properties of bone with the emphasis on organic and inorganic components that are important for the biomaterial design. The main focus will be on the advancement and usage of bioactive glass (BG) for bone tissue engineering due to its similarity to the natural inorganic constituent of bone. The various BG synthetic processes, modifications of composition, as well as the biomolecule delivery will be discussed in great detail. As the properties of BG are tuneable according to clinical needs, it creates a new paradigm in addition to displaying its superior potential for bone tissue engineering and translational medicine in the field of orthopedic surgery. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2878-2887, 2018.

Aliphatic Polyester Nanofibers Functionalized with Cyclodextrins and Cyclodextrin-Guest Inclusion Complexes.

The fabrication of nanofibers by electrospinning has gained popularity in the past two decades; however, only in this decade, have polymeric nanofibers been functionalized using cyclodextrins (CDs) or their inclusion complexes (ICs). By combining electrospinning of polymers with free CDs, nanofibers can be fabricated that are capable of capturing small molecules, such as wound odors or environmental toxins in water and air. Likewise, combining polymers with cyclodextrin-inclusion complexes (CD-ICs), has shown promise in enhancing or controlling the delivery of small molecule guests, by minor tweaking in the technique utilized in fabricating these nanofibers, for example, by forming core–shell or multilayered structures and conventional electrospinning, for controlled and rapid delivery, respectively. In addition to small molecule delivery, the thermomechanical properties of the polymers can be significantly improved, as our group has shown recently, by adding non-stoichiometric inclusion complexes to the polymeric nanofibers. We recently reported and thoroughly characterized the fabrication of polypseudorotaxane (PpR) nanofibers without a polymeric carrier. These PpR nanofibers show unusual rheological and thermomechanical properties, even when the coverage of those polymer chains is relatively sparse (~3%). A key advantage of these PpR nanofibers is the presence of relatively stable hydroxyl groups on the outer surface of the nanofibers, which can subsequently be taken advantage of for bioconjugation, making them suitable for biomedical applications. Although the number of studies in this area is limited, initial results suggest significant potential for bone tissue engineering, and with additional bioconjugation in other areas of tissue engineering. In addition, the behaviors and uses of aliphatic polyester nanofibers functionalized with CDs and CD-ICs are briefly described and summarized. Based on these observations, we attempt to draw conclusions for each of these combinations, and the relationships that exist between their presence and the functional behaviors of their nanofibers.

Positive effect of wrapping poly caprolactone/polyethylene glycol fibrous films on the mechanical properties of 45S5 bioactive glass scaffolds

Abstract45S5 bioactive glass (BG) scaffolds show great potential in bone tissue engineering due to their superior osteoinductivity and osteoconductivity. However, such scaffolds generally possess poor mechanical properties. Here, inspired by the reinforcing principle of confined concrete elements in civil engineering, poly caprolactone (PCL)/polyethylene glycol (PEG) film‐wrapped 45S5 BG scaffolds were prepared by gluing highly porous foam–replicated BG scaffolds and electrospun PCL/PEG films together with a PCL layer. The results showed that the compressive strength of the PCL/PEG film‐wrapped scaffolds was greatly improved, compared to that of unwrapped BG scaffolds. Moreover, the PCL/PEG film exhibited hydrophilicity, responsible for improved cell activity with respect to the hydrophobic PCL one. Thus, the present work introduces a convenient approach to improve the mechanical properties of highly porous bioceramic scaffolds, and is relevant for future robust scaffold design.

Cell-Sheet-Derived ECM Coatings and Their Effects on BMSCs Responses.

Extracellular matrix (ECM) provides a dynamic and complex environment to determine the fate of stem cells. In this work, light harvested cell sheets were treated with paraformaldehyde or ethanol, which eventually become ECM. Such ECM was then immobilized on titanium substrates via polydopamine chemistry. Their effects on bone marrow mesenchymal stromal cells (BMSCs) behaviors were investigated. It was found that paraformaldehyde-treated ECMcoating (PT-ECM) showed a well-maintained microstructure, whereas that of ethanol-treated (ET-ECM) was completely changed. As a result, different amide structures and distributions of ECM components, such as laminin and collagen I, were exhibited. Alkaline phosphatase activity, osteocalcin secretion, related gene expression, and mineral deposition were evaluated for BMSCs cultured on both ECM coatings. PT-ECM was demonstrated to promote osteogenic differentiation much more efficiently than that of ET-ECM. That is ascribed to the preservation of native ECM milieu of PT-ECM. Such ECM acquirement and immobilization method could establish surfaces being able to direct stem cell responses on various materials. That shows promising potential in bone tissue engineering and other related biomedical applications.

The Investigation of ZnO/Poly(vinylidene fluoride) Nanocomposites with Improved Mechanical, Piezoelectric, and Antimicrobial Properties for Orthopedic Applications.

Studies have shown that piezoelectric materials can be used as bioactively charged surfaces to induce tissue regeneration (like bone reconstruction). β-phase poly(vinylidene fluoride) (PVDF) and zinc oxide (ZnO) are regarded as potential bone tissue engineering materials because of their piezoelectricity and biocompatibility. Additionally, ZnO nanoparticles (NPs) are ideal materials to reduce infection. In this study, PVDF scaffolds doped with ZnO NPs (ZnO/PVDF) were prepared by electrospinning increasing ZnO concentrations and the ratio of the β-phase PVDF resulting in an enhanced the modulus, elongation to failure and load stress. In vitro osteoblast assays were also performed to determine material cytotoxicity and bone reconstruction potential. Results exhibited that the piezo-excited scaffolds had a 30% greater osteoblast density, compared to non-piezoelectric-excited groups after 3 days of culture. To evaluate the scaffold's antimicrobial properties, three common etiologic agents of orthopedic after-surgery infection, Staphylococcus aureus (SA), Methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli), were separately seeded and counted. Results of this in vitro study showed significantly reduced E. coli (45%), SA (48%) and MRSA (37%) density on the 1 mg/ml ZnO/PVDF scaffolds without piezo-excitation (compared to the control). Moreover, compared to the non-piezo-excited scaffolds, results showed for the first time that the density of the same bacteria on the piezo-excited scaffolds reduced by over 23%, 22%, and 33%, respectively. For the first time, the effects of novel ZnO/PVDF scaffolds on osteoblasts and bacteria were investigated here, and significantly decreased bacteria with increased osteoblast density on the piezo-excited scaffolds demonstrated that these scaffolds have a strong potential for numerous antibacterial orthopedic applications.