PVDF Hybrid Nanocomposites with Graphene and Carbon Nanotubes and Their Thermoresistive and Joule Heating Properties.

In recent years, conductive polymer nanocomposites have gained significant attention due to their promising thermoresistive and Joule heating properties across a range of versatile applications, such as heating elements, smart materials, and thermistors. This paper presents an investigation of semi-crystalline polyvinylidene fluoride (PVDF) nanocomposites with 6 wt.% carbon-based nanofillers, namely graphene nanoplatelets (GNPs), multi-walled carbon nanotubes (MWCNTs), and a combination of GNPs and MWCNTs (hybrid). The influence of the mono- and hybrid fillers on the crystalline structure was analyzed by X-ray diffraction (XRD) and differential scanning calorimetry (DSC). It was found that the nanocomposites had increased amorphous fraction compared to the neat PVDF. Furthermore, nanocomposites enhanced the ß phase of the PVDF by up to 12% mainly due to the presence of MWCNTs. The resistive properties of the nanocompositions were weakly affected by the temperature in the analyzed temperature range of 25-100 °C; nevertheless, the hybrid filler composites were proven to be more sensitive than the monofiller ones. The Joule heating effect was observed when 8 and 10 V were applied, and the compositions reached a self-regulating effect at around 100-150 s. In general, the inclusion in PVDF of nanofillers such as GNPs and MWCNTs, and especially their hybrid combinations, may be successfully used for tuning the self-regulated Joule heating properties of the nanocomposites.

Enhancing Multifunctionality: Optimal Properties of Iron-Oxide-Reinforced Polyvinylidene Difluoride Unveiled Through Full Atom Molecular Dynamics Simulations.

Nanocomposites made of magnetite (Fe3O4) nanoparticles (NP)s with different surface chemistry and polyvinyl difluoride (PVDF) polymer were investigated using full atom molecular dynamics (MD) simulation. NPs with hydroxyl (OH), hexanoic, and oleic acid terminations were considered in this study. The effect of each surface chemistry was investigated in terms of the mechanical properties, the distribution of the internal energy around the NP, and the chain polarization gradient from the interface to the bulk. From this investigation, we find that oleic acid termination, although the most popular, is less favorable for interfacial interaction and local polarization. The OH-terminated NP results in the best configuration for the properties investigated. The hexanoic acid-grafted NP presents a good compromise. Hydrogen bonding governs the induced response of the nanocomposites. Although the hexanoic acid grafted NP presents less hydrogen bonding than the OH-terminated case, the conformation of the hexanoic acid acts as a mobility flow inhibitor, leading to a performance comparable to that of the OH-terminated NP composite. This work led to investigating routes to make nanocomposite materials with optimized properties. These results shed light on the multiple combinations offered by nanocomposites that go beyond the conventional effects of size.

Donor variability alters differentiation and mechanical cohesion of tissue-engineered constructs with human endothelial/MSC co-culture.

To move towards clinical applications, tissue engineering (TE) should be validated with human primary cells and offer easy connection to the native vascularisation. Based on a sheet-like bone substitute developed previously, we investigated a mesenchymal stem cells/endothelial cells (MSCs/ECs) coculture to enhance pre-vascularisation. Using MSCs from six independent donors whose differentiation potential was assessed towards two lineages, we focused on donor variability and cell crosstalk regarding bone differentiation. Coculture was performed on calcium phosphate granules in a specific chamber during 1 month. MSCs were seeded first then ECs were added after 2 weeks, with respective monocultures as control groups. Cell viability and organisation (fluorescence, electronic microscopy), differentiation (ALP staining/activity, RT-qPCR) and mechanical cohesion were analysed. Adaptation of the protocol to coculture was validated (high cell viability and proliferation). Activity and differentiation showed strong trends towards synergistic effects between cell types. MSCs reached early mineralisation stage of maturation. The delayed addition of ECs allowed for their attachment on developed MSCs' matrix. The main impact of donor variability could be here the lack of cell proliferation potential with some donors, leading to low differentiation and mechanical cohesion and therefore absence of sheet-like shape successfully obtained with others. We suggest therefore adapting protocols to cell proliferation potentials from one batch of cells to the other in a patient-specific approach.

Monitoring mechanical stimulation for optimal tendon tissue engineering: A mechanical and biological multiscale study.

To understand the effect of mechanical stimulation on cell response, bone marrow stromal cells were cultured on electrospun scaffolds under two distinct mechanical conditions (static and dynamic). Comparison between initial and final mechanical and biological properties of the cell-constructs were conducted over 14 days for both culturing conditions. As a result, mechanically stimulated constructs, in contrast to their static counterparts, showed evident mechanical-induced cell orientation, an effective aligned collagen and tenomodulin extracellular matrix. This orientation provides clues on the importance of mechanical stimulation to induce a tendon-like differentiation. In addition, cell and collagen orientation lead to enhanced storage modulus observed under dynamic stimulation. Altogether mechanical stimulation lead to (a) cell and matrix orientation through the sense of the stretch and (b) a dominant elastic response in the cell-constructs with a minor contribution of the viscosity in the global mechanical behavior. Such a correlation could help in further studies to better understand the effect of mechanical stimulation in tissue engineering.

Multiscale-Engineered Muscle Constructs: PEG Hydrogel Micro-Patterning on an Electrospun PCL Mat Functionalized with Gold Nanoparticles.

The development of new, viable, and functional engineered tissue is a complex and challenging task. Skeletal muscle constructs have specific requirements as cells are sensitive to the stiffness, geometry of the materials, and biological micro-environment. The aim of this study was thus to design and characterize a multi-scale scaffold and to evaluate it regarding the differentiation process of C2C12 skeletal myoblasts. The significance of the work lies in the microfabrication of lines of polyethylene glycol, on poly(ε-caprolactone) nanofiber sheets obtained using the electrospinning process, coated or not with gold nanoparticles to act as a potential substrate for electrical stimulation. The differentiation of C2C12 cells was studied over a period of seven days and quantified through both expression of specific genes, and analysis of the myotubes' alignment and length using confocal microscopy. We demonstrated that our multiscale bio-construct presented tunable mechanical properties and supported the different stages skeletal muscle, as well as improving the parallel orientation of the myotubes with a variation of less than 15°. These scaffolds showed the ability of sustained myogenic differentiation by enhancing the organization of reconstructed skeletal muscle. Moreover, they may be suitable for applications in mechanical and electrical stimulation to mimic the muscle's physiological functions.

The combination of a poly-caprolactone/nano-hydroxyapatite honeycomb scaffold and mesenchymal stem cells promotes bone regeneration in rat calvarial defects.

Bone tissue engineering goes beyond the limitations of conventional methods of treating bone loss, such as autograft-induced morbidity and a lack of integration for large grafts. Novel biomimicry approaches (using three-dimensional [3D] electrospinning and printing techniques) have been designed to offer the most appropriate environment for cells and thus promote bone regeneration. In the present study, we assessed the bone regeneration properties of a composite 3D honeycomb structure from the electrostatic template-assisted deposition process by an alternate deposition of electrospun polycaprolactone (PCL) nanofibers and electrosprayed hydroxyapatite nanoparticles (nHA) on a honeycomb micropatterned substrate. We first confirmed the cytocompatibility of this honeycomb PCL-nHA scaffold in culture with bone marrow-derived mesenchymal stem cells (BM-MSCs). The scaffold was then implanted (alone or with seeded MSCs) for 2 months in a rat critical-sized calvarial defect model. The observation of new bone synthesis in situ (monitored using microcomputed tomography every 2 weeks and a histological assessment upon extraction) demonstrated that the honeycomb PCL-nHA scaffold was osteoconductive. Moreover, the combination of the scaffold with BM-MSCs was associated with significantly greater bone volume and mineralized regeneration during the 2-month experiment. The combination of the biomimetic honeycomb PCL-nHA scaffold with patient mesenchymal stem cells might therefore have great potential for clinical applications and specifically in maxillofacial surgery.

In Vitro Bone Cell Response to Tensile Mechanical Solicitations: Is There an Optimal Protocol?

Bone remodeling is strongly linked to external mechanical signals. Such stimuli are widely used in vitro for bone tissue engineering by applying mechanical solicitations to cell cultures so as to trigger specific cell responses. However, the literature highlights considerable variability in devices and protocols. Here the major biological, mechanical, and technical parameters implemented for in vitro tensile loading applications are reviewed. The objective is to identify which values are used most, and whether there is an optimal protocol to obtain a functional tissue-engineering construct. First, a shift that occurred from fundamental comprehension of bone formation, to its application in rebuilt tissues and clinical fields is shown. Despite the lack of standardized protocols, consensual conditions relevant for in vitro bone development, in particular cell differentiation, could be highlighted. Culture processes are guided by physiological considerations, although out-of-range conditions are sometimes used without implying negative results for the development of rebuilt tissue. Consensus can be found on several parameters, such as strain frequency (1 Hz) or the use of rest periods, but other points have not yet been fully established, especially synergies with other solicitations. It is believed that the present work will be useful to develop new tissue-engineering processes based on stretching.

Biomaterials in Tendon and Skeletal Muscle Tissue Engineering: Current Trends and Challenges.

Tissue engineering is a promising approach to repair tendon and muscle when natural healing fails. Biohybrid constructs obtained after cells’ seeding and culture in dedicated scaffolds have indeed been considered as relevant tools for mimicking native tissue, leading to a better integration in vivo. They can also be employed to perform advanced in vitro studies to model the cell differentiation or regeneration processes. In this review, we report and analyze the different solutions proposed in literature, for the reconstruction of tendon, muscle, and the myotendinous junction. They classically rely on the three pillars of tissue engineering, i.e., cells, biomaterials and environment (both chemical and physical stimuli). We have chosen to present biomimetic or bioinspired strategies based on understanding of the native tissue structure/functions/properties of the tissue of interest. For each tissue, we sorted the relevant publications according to an increasing degree of complexity in the materials’ shape or manufacture. We present their biological and mechanical performances, observed in vitro and in vivo when available. Although there is no consensus for a gold standard technique to reconstruct these musculo-skeletal tissues, the reader can find different ways to progress in the field and to understand the recent history in the choice of materials, from collagen to polymer-based matrices.

Poly(ε-caprolactone)/Hydroxyapatite 3D Honeycomb Scaffolds for a Cellular Microenvironment Adapted to Maxillofacial Bone Reconstruction.

The elaboration of biomimetic materials inspired from the specific structure of native bone is one the main goal of tissue engineering approaches. To offer the most appropriate environment for bone reconstruction, we combined electrospinning and electrospraying to elaborate an innovative scaffold composed of alternating layers of polycaprolactone (PCL) and hydroxyapatite (HA). In our approach, the electrospun PCL was shaped into a honeycomb-like structure with an inner diameter of 160 µm, capable of providing bone cells with a 3D environment while ensuring the material biomechanical strength. After 5 days of culture without any differentiation factor, the murine embryonic cell line demonstrated excellent cell viability on contact with the PCL-HA structures as well as active colonization of the scaffold. The cell differentiation, as tested by RT-qPCR, revealed a 6-fold increase in the expression of the RNA of the Bglap involved in bone mineralization as compared to a classical 2D culture. This differentiation of the cells into osteoblasts was confirmed by alkaline phosphatase staining of the scaffold cultivated with the cell lineage. Later on, organotypic cultures of embryonic bone tissues showed the high capacity of the PCL-HA honeycomb structure to guide the migration of differentiated bone cells throughout the cavities and the ridge of the biomaterial, with a colonization surface twice as big as that of the control. Taken together, our results indicate that PCL-HA honeycomb structures are biomimetic supports that promotes in vitro osteocompatibility, osteoconduction, and osteoinduction and could be suitable for being used for bone reconstruction in complex situations such as the repair of maxillofacial defects.

The Osteogenic and Tenogenic Differentiation Potential of C3H10T1/2 (Mesenchymal Stem Cell Model) Cultured on PCL/PLA Electrospun Scaffolds in the Absence of Specific Differentiation Medium.

The differentiation potential of mesenchymal stem cells (MSC) has been extensively tested on electrospun scaffolds. However, this potential is often assessed with lineage-specific medium, making it difficult to interpret the real contribution of the properties of the scaffold in the cell response. In this study, we analyzed the ability of different polycaprolactone/polylactic acid PCL/PLA electrospun scaffolds (pure or blended compositions, random or aligned fibers, various fiber diameters) to drive MSC towards bone or tendon lineages in the absence of specific differentiation medium. C3H10T1/2 cells (a mesenchymal stem cell model) were cultured on scaffolds for 96 h without differentiation factors. We performed a cross-analysis of the cell-scaffold interactions (spreading, organization, and specific gene expression) with mechanical (elasticity), morphological (porosity, fibers diameter and orientation) and surface (wettability) characterizations of the electrospun fibers. We concluded that (1) osteogenic differentiation can be initiated on pure PCL-based electrospun scaffolds without specific culture conditions; (2) fiber alignment modified cell organization in the short term and (3) PLA added to PCL with an increased fiber diameter encouraged the stem cells towards the tendon lineage without additional tenogenic factors. In summary, the differentiation potential of stem cells on adapted electrospun fibers could be achieved in factor-free medium, making possible future applications in clinically relevant situations.

Towards the development and characterization of an easy handling sheet-like biohybrid bone substitute.

We designed a sheet-like bone substitute capable of adapting to different geometries and becoming a standard tissue-engineered process for bone surgery. Preosteoblastic cells were seeded on to a monolayer of calcium phosphate granules and cultured in a flat parallelepipedic cell culture chamber for 1 month. From the various diameters of the granules examined, the 80-200 µm group exhibited the most homogeneous performances regarding both biological (cell morphology, viability, differentiation, and simple metabolic activity) and mechanical (cohesion and stress-strain behavior) properties. This sheet was easy to handle after extraction from the culture chamber and showed versatile geometry and flexibility, making it easy to use for surgeons, especially for small defects of the maxillofacial area.

Monitoring the viscoelastic properties of skin in liquid environments using quartz crystal microbalance.

Quartz crystal microbalance (QCM) with dissipation can be used to measure the response of the human stratum corneum (SC) attached to the QCM crystal, as it adsorbs or desorbs active ingredients from a liquid medium. The method was demonstrated with the sorption of poly(diallyl dimethyl ammonium chloride), a cationic polymer widely used in formulations for topical and transdermal applications. Using 14-mm diameter SC coupons attached to the QCM crystals with an adhesive, up to five overtones (up to 11th harmonic) were obtained and the response was analyzed using a Voigt model. The adhesive layer could be regarded as a rigid substrate, and the skin with overlaying fluid was modeled as a soft layer underneath a fluid medium. Limited modeling tools that are currently available were used to interpret the observed response in terms of physical parameters such as the changes in thickness, shear modulus, and viscosity. The high sensitivity of the technique demonstrates the possibility of using small samples of human skin for in vitro studies in a variety of topical and transdermal drug delivery applications and in the evaluation of skin care products.