I. Rajzer

Electrospun gelatin/poly(ε-caprolactone) fibrous scaffold modified with calcium phosphate for bone tissue engineering

In this study gelatin (Gel) modified with calcium phosphate nanoparticles (SG5) and polycaprolactone (PCL) were used to prepare a 3D bi-layer scaffold by collecting electrospun PCL and gelatin/SG5 fibers separately in the same collector. The objective of this study was to combine the desired properties of PCL and Gel/SG5 in the same scaffold in order to enhance mineralization, thus improving the ability of the scaffold to bond to the bone tissue. The scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and the wide angle X-ray diffraction (WAXD) measurements confirmed that SG5 nanoparticles were successfully incorporated into the fibrous gelatin matrix. The composite Gel/SG5/PCL scaffold exhibited more enhanced mechanical properties than individual Gel and Gel/SG5 scaffolds. The presence of SG5 nanoparticles accelerated the nucleation and growth of apatite crystals on the surface of the composite Gel/SG5/PCL scaffold in simulated body fluid (SBF). The osteoblast response in vitro to developed electrospun scaffolds (PCL and Gel/SG5/PCL) was investigated by using normal human primary NHOst cell lines. NHOst cell culture studies showed that higher alkaline phosphatase (ALP) activity and better mineralization were obtained in the case of composite materials than in pure PCL scaffolds. The mechanically strong PCL scaffold served as a skeleton, while the Gel/SG5 fibers facilitated cell spreading and mineralization of the scaffold.

Bioactive nanocomposite PLDL/nano-hydroxyapatite electrospun membranes for bone tissue engineering

New nanocomposite membranes with high bioactivity were fabricated using the electrospinning. These nanocomposites combine a degradable polymer poly(l/dl)-lactide and bone cell signaling carbonate nano-hydroxyapatite (n-HAp). Chemical and physical characterization of the membranes using scanning electron microscopy, Fourier transform infrared spectroscopy and the wide angle X-ray diffraction evidenced that nanoparticles were successfully incorporated into the fibers and membrane structure. The incorporation of the n-HAp into the structure increased significantly the mineralization of the membrane in vitro. It has been demonstrated that after a 3-day incubation of composite membrane in the Simulated Body Fluid a continuous compact apatite layer was formed. In vitro experiments demonstrated that the incorporation of n-HAp significantly improved cell attachment, upregulated cells proliferation and stimulated cell differentiation quantified using Alkaline Phosphatase and OsteoImage tests. In conclusion, the results demonstrated that the addition of n-HAp provided chemical cues that were a key factor that regulated osteoblastic differentiation.

In vitro and in vivo studies on biocompatibility of carbon fibres

In the present study we focused on the in vitro and in vivo evaluation of two types of carbon fibres (CFs): hydroxyapatite modified carbon fibres and porous carbon fibres. Porous CFs used as scaffold for tissues regeneration could simultaneously serve as a support for drug delivery or biologically active agents which would stimulate the tissue growth; while addition of nanohydroxyapatite to CFs precursor can modify their biological properties (such as bioactivity) without subsequent surface modifications, making the process cost and time effective. Presented results indicated that fibre modification with HAp promoted formation of apatite on the fibre surface during incubation in simulated body fluid. The materials biocompatibility was determined by culturing human osteoblast-like cells of the line MG 63 in contact with both types of CFs. Both tested materials gave good support to adhesion and growth of bone-derived cells. Materials were implanted into the skeletal rat muscle and a comparative analysis of tissue reaction to the presence of the two types of CFs was done. Activities of marker metabolic enzymes: cytochrome c oxidase (CCO) and acid phosphatase were examined to estimate the effect of implants on the metabolic state of surrounding tissues. Presented results evidence the biocompatibility of porous CFs and activity that stimulates the growth of connective tissues. In case of CFs modified with hydroxyapatite the time of inflammatory reaction was shorter than in case of traditional CFs.

Fabrication of bioactive polycaprolactone/hydroxyapatite scaffolds with final bilayer nano-/micro-fibrous structures for tissue engineering application

In this study, two techniques, namely electrospinning and needle-punching processes, were used to fabricate bioactive polycaprolactone/hydroxyapatite scaffolds with a final bilayer nano-/micro-fibrous porous structure. A hybrid scaffold was fabricated to combine the beneficial properties of nanofibers and microfibers and to create a three-dimensional porous structure (which is usually very difficult to produce using electrospinning technology only). The first part of this work focused on determining the conditions necessary to fabricate nano- and micro-fibrous components of scaffold layers. A characterization of scaffold components, with respect to their morphology, fiber diameter, pore size, wettability, chemical composition and mechanical properties, was performed. Then, the same process parameters were applied to produce a hybrid bilayer scaffold by electrospinning the nanofibers directly onto the micro-fibrous nonwovens obtained in a traditional mechanical needle-punching process. In the second part, the bioactive character of a hybrid nano-/micro-fibrous scaffold in simulated body fluid (SBF) was assessed. Spherical calcium phosphate was precipitated onto the nano-/micro-fibrous scaffold surface proving its bioactivity.

Injectable and fast resorbable calcium phosphate cement for body-setting bone grafts

In this work a calcium phosphate (CPC)/polymer blend was developed with the advantage of being moldable and capable of in situ setting to form calcium deficient hydroxyapatite under physiological conditions in an aqueous environment at body temperature. The CPC paste consists in a mix of R cement, glycerol as a liquid phase carrier and a biodegradable hydrogel such as Polyvinyl alcohol, which acts as a binder. Microstructure and mechanical analysis shows that the CPC blend can be used as an injectable implant for low loaded applications and fast adsorption requirements. The storage for commercial distribution was also evaluated and the properties of the materials obtained do not significantly change during storage at −18°C.

Electrospun polymer scaffolds modified with drugs for tissue engineering

The purpose of this paper was to fabricate nanofibrous scaffolds containing ossein-hydroxyapatite complex (osteogenon) to mimic the native bone extracellular matrix. Polylactide (PLDL) and polycaprolactone (PCL) were used to prepare scaffolds using electrospinning. Unfortunately, both of these biodegradable polymers have poor cell recognition sites leading to poor cell affinity and adhesion, therefore, based on our previous experience, osteogenon-drug was used at the stage of fibers forming by electrospinning. We have compare the physicochemical parameters and mechanical properties of PLDL/osteo and PCL/osteo scaffolds as well as an osteogenon-drug influence on the microstructure of electrospun materials produced for potential application in bone tissue engineering. We have investigated the effect of the microstructure and the chemical composition of electrospun materials on adhesion, proliferation and morphology as well as on the process of differentiation of bone cells. The use of osteogenon improved mineralization, cell adhesion and the rate of cell differentiation.

An ultrasonic through-transmission technique for monitoring the setting of injectable calcium phosphate cement.

An ultrasound through-transmission method to monitor the setting process of injectable calcium phosphate bone cements in body fluids is presented. This method can be used to determine the acoustic properties of the bone cement as it sets, which are linked to its material properties and provide some information about changes occurring within the cement. The development of the methodology of ultrasonic testing and execution of velocity measurements of the longitudinal and transverse waves using the through-transmission method made it possible to determine the material constants of samples during the setting and hardening process of an injectable cement paste in physiological fluids (i.e. the Youngs modulus (E), the Poisson ratio (ν) and the shear modulus (G)), and to determine the degree of anisotropy of wave velocity in the samples. A strong advantage of the proposed method is that it is non-destructive, and the same sample can be used to monitor the whole process of the cement setting. The testing was performed on premixed and injectable calcium phosphate (CPC)/chitosan blend, where glycerol was used as a liquid phase. Comparisons between ultrasonic velocity and empirical tests such as compressive strength, porosity measurement, FTIR, SEM and XRD analysis at different days of immersion in Ringers solutions showed that the ultrasonic velocity can be very useful to provide in situ information about changes occurring within the cement.

Polylactide/polycaprolactone asymmetric membranes for guided bone regeneration

Abstract The aim of this work was to develop bioresorbable, asymmetric membranes for guided bone regeneration (GBR). Two resorbable polymers – polylactide (PLA) and polycaprolactone (PCL) were used in fabrication process. Two different manufacturing methods were applied: electrospinning in the case of PLA and freeze-drying of PCL. Mechanical properties, stability in a water environment and biocompatibility of fabricated membranes were evaluated. Microstructure [scanning electron microscopy (SEM)] of the membranes was assessed in terms of level of porosity, as well as size and shape of the pores. Study showed that combination of electrospinning and freeze-drying methods allows biocompatible PLA/PCL bi-phasic materials of appropriate mechanical properties and diverse microstructure to be produced, that should on the one hand prevent soft tissue growth, and on the other hand be a suitable scaffold for the growth of bone cells.