Polymeric scaffolds prepared via Thermally Induced Phase Separation (TIPS): Tuning of structure and morphology

Scaffolds suitable for tissue engineering applications were prepared by Thermally Induced Phase Separation (TIPS) starting from a ternary solution PLLA/dioxane/water. The experimental protocol consisted of three consecutive steps, a first quench from the homogeneous solution to an appropriate demixing temperature (within the metastable region), a holding stage for a given residence time and a final quench from the demixing temperature to a low temperature (within the unstable region). A large variety of morphologies, in terms of average pore size and interconnection, were obtained upon modifying the demixing time and temperature, owing to the interplay of nucleation and growth processes dur…

Kefiran-based Scaffolds For Biomedical Applications

Kefiran is an exopolysaccharide produced by microorganisms present in kefir grains, with several health promoting properties. A optimized protocol was developed for the separation of kefiran from kefir grains, allowing to reach a yield 4÷5 % without using toxic or expensive chemicals. The capability of kefiran to produce scaffold via Thermally Induced Phase Separation (TIPS) technique was investigated and porous scaffolds structure was obtained. Separated kefiran and scaffolds were analyzed via DSC and different thermal properties between purified kefiran and scaffold were revealed. XRD analysis revealed different structure between kefiran and scaffolds. The porous scaffold structure can be…

Effect of some injection molding processing conditions on weld lines characteristics

The influence of injection and mold temperature as well as holding time on the knit-line characteristics in samples of Nylon 6 obtained by injection molding was analysed. The characteristics of the surface defect seem to be governed by the shrinkage rate and amount in the weld line zone.

Human nasoseptal chondrocytes maintain their differentiated phenotype on PLLA scaffolds produced by thermally induced phase separation and supplemented with bioactive glass 1393

Damage of hyaline cartilage such as nasoseptal cartilage requires proper reconstruction, which remains challenging due to its low intrinsic repair capacity. Implantation of autologous chondrocytes in combination with a biomimetic biomaterial represents a promising strategy to support cartilage repair. Despite so far mostly tested for bone tissue engineering, bioactive glass (BG) could exert stimulatory effects on chondrogenesis. The aim of this work was to produce and characterize composite porous poly(L-lactide) (PLLA)/1393BG scaffolds via thermally induced phase separation (TIPS) technique and assess their effects on chondrogenesis of nasoseptal chondrocytes. The PLLA scaffolds without or…

Evidence of Mechanisms Occurring in Thermally Induced Phase Separation of Polymeric Systems

Thermally induced phase separation is a fabrication technique for porous polymeric structures. By means of easy-to-tune processing parameters, such as system composition and demixing temperature, a vast latitude of average pore dimensions, pore size distributions, and morphologies can be obtained. The relation between demixing temperature and morphology was demonstrated via cloud point curve measurement and foams fabrication with controlled thermal protocols, for the model system poly-l-lactide–dioxane–water. The morphologies obtained at a temperature lower than cloud point showed a closed-pore architecture, suggesting a “nucleation-and-growth” separation mechanism, which produced larger po…

Modeling the interactions between light and crystallizing polymer during fast cooling

In this work, an experimental set-up able to quench thin polymer films whilst recording the sample thermal history as well as the overall and depolarized light intensities of a laser beam emerging from the sample is described. The interactions between the light beam and the crystallizing material have been modeled accounting for absorption and scattering phenomena. The proposed model was found to be able to reproduce the ex- perimentally observed behavior of light intensities and it was validated by comparison with conventional DSC analysis. On the basis of this model, a method to obtain crystallinity evo- lution is proposed and applied to some fast cooling runs. The method was applied to q…

A 3D‑scaffold of PLLA induces the morphological differentiation and migration of primary astrocytes and promotes the production of extracellular vesicles

The present study analyzed the ability of primary rat astrocytes to colonize a porous scaffold, mimicking the reticular structure of the brain parenchyma extracellular matrix, as well as their ability to grow, survive and differentiate on the scaffold. Scaffolds were prepared using poly-L-lactic acid (PLLA) via thermally-induced phase separation. Firstly, the present study studied the effects of scaffold morphology on the growth of astrocytes, evaluating their capability to colonize. Specifically, two different morphologies were tested, which were obtained by changing the polymer concentration in the starting solution. The structures were characterized by scanning electron microscopy, and a…

Computational modeling and experimental characterization of fluid dynamics in micro-CT scanned scaffolds within a multiple-sample airlift perfusion bioreactor

The perfusion of flow during cell culture induces cell proliferation and enhances cellular activity. Perfusion bioreactors offer a controlled dynamic environment for reliable in vitro applications in the tissue engineering field. In this work, to evaluate the effects of the operating parameters of a custom-made bioreactor, numerical simulations were performed to solve the fluid velocity profile inside the bioreactor containing multi-grid support that allows allocating of multiple seeded scaffolds at the same time. The perfusion system exhibited a uniform distribution of liquid velocities within the regions, suitable for cell growth on seeded scaffolds. The effects of the porous microstructu…

Unsuspended mass of solid particles in stirred tanks

On propose une technique originale pour la determination de la fraction de particules solides non suspendues a des vitesses d'agitation inferieures a N js . La technique repose sur le concept des systemes jumeles et est completee par une technique de tracage et de detection pour la phase solide. Les resultats obtenus avec un reservoir chicane agite par une turbine Rushton, montrent que la fraction de solides ne quittant jamais le fond du reservoir peut etre simplement correlee au rapport entre la vitesse d'agitation et la vitesse de suspension complete de Zwietering. On montre qu'a des vitesses de l'agitateur d'environ 80% de la vitesse de suspension complete de Zwietering, pratiquement tou…

Preparation of polymeric foams with a pore size gradient via Thermally Induced Phase Separation (TIPS)

Abstract Foams with a pore size gradient are promising materials for tissue engineering applications where a complex architecture involving morphological variations in space must be mimicked, e.g. in bone tissue repair. In this paper, a technique to obtain a porous scaffold with a pore size gradient is presented. The preparation procedure is based on Thermally Induced Phase Separation (TIPS), by imposing a different thermal history on the two sides of a polymeric solution. In this way, a gradient in thermal history is produced, which will generate a pore size monotonously varying along scaffold thickness. By controlling some parameters easy to manipulate, such as demixing temperature and/or…

A Continuous Pore Size Gradient PLLA Scaffold For Osteochondral Regeneration

Osteochondral (OC) scaffold-based regenerative approaches in the joint are challenging since the scaffold must provide mechanical strength while also mimicking the local cartilage and bone microenvironments. Thermally Induced Phase Separation (TIPS) can produce scaffolds with a wide range of pore size morphologies/distributions. Here, we produced by TIPS a poly-L-lactide (PLL A) scaffold with a continuous pore size gradient along the sample thickness, from ~70μm diameter on one side to ~200 μm diameter on the opposite surface.

Co-Deposition and Characterization of Hydroxyapatite-Chitosan and Hydroxyapatite-Polyvinylacetate Coatings on 304 SS for Biomedical Devices

During the last decades, biomaterials have been deeply studied to perform and improve coatings for biomedical devices. Metallic materials, especially in the orthopedic field, represent the most common material used for different type of devices thanks to their good mechanical properties. Nevertheless, low/medium resistance to corrosion and low osteointegration ability characterizes these materials. To overcome these problems, the use of biocoatings on metals substrate is largely diffused. In fact, biocoatings have a key role to confer biocompatibility properties, to inhibit corrosion and thus improve the lifetime of implanted devices. In this work, the attention was focused on Hydroxyapatit…

PLLA biodegradable scaffolds for angiogenesis via Diffusion Induced Phase Separation (DIPS)

A critical obstacle in tissue engineering is the inability to maintain large masses of living cells upon transfer from the in vitro culture conditions into the host in vivo. Capillaries, and the vascular system, are required to supply essential nutrients, including oxygen, remove waste products and provide a biochemical communication “highway”. For this reason it is mandatory to manufacture an implantable structure where the process of vessel formation – the angiogenesis – can take place. In this work PLLA scaffolds for vascular tissue engineering were produced by dip-coating via Diffusion Induced Phase Separation (DIPS) technique. The scaffolds, with a vessel-like shape, were obtained by p…

A COMPOSITE PLLA SCAFFOLD FOR REGENERATION OF COMPLEX TISSUES

A composite biodegradable scaffold incorporating an integrated network of synthetic blood vessels was designed and prepared, in line with the requirements of a scaffold effectively supporting the regeneration of highly vascularized tissues. In other words, this composite scaffold should allow the regeneration of complex injured tissue (e.g. dermis) and, at the same time, favour the development of a vascular network on its inner, i.e. a 3D polymeric scaffolds embedding synthetic blood vessel-like structures for nutrient supply and metabolite removal. PLLA assures a high degree of biocompatibility and a low level of inflammation response upon implantation, while the embedded tubular vessel-li…

Calcium phosphate/polyvinyl acetate coatings on SS304 via galvanic co-deposition for orthopedic implant applications

Abstract In this work, the galvanic deposition method is used to deposit coatings of brushite/hydroxyapatite/polyvinyl acetate on 304 stainless steel. Coatings are obtained at different temperatures and with different sacrificial anodes, consisting of a mixture of brushite and hydroxyapatite. Samples are aged in a simulated body fluid (SBF), where a complete conversion of brushite into hydroxyapatite with a simultaneous change in morphology and wettability occurred. The corrosion tests show that, compared with bare 304, the coating shifts Ecorr to anodic values and reduces icorr Ecorr, and icorr has different values at different aging times due to chemical interactions at the solid/liquid i…

No-Flow Temperature in Injection Molding Simulation

Most injection molding simulation packages use the no-flow temperature (NFT) as a means of determining whether the polymer flows or is solid. The NFT is not well defined, and a standard method for measuring it does not exist. A sensitivity analysis of the filling stage has been carried out with two different packages [VISI Flow (Vero Software Limited, Gloucestershire, UK) and Moldflow (Autodesk, Inc., San Rafael, CA)] to estimate the influence of the NFT on the main processing parameters. The NFT has a large influence on the thickness of the frozen layer, but it does not appreciably affect the filling pressure. Because the NFT affects the frozen layer, an effect on the estimation of shrinka…

A poly-L-lactic acid/ collagen/glycosaminoglycan matrix for tissue engineering applications

Adhesion of tissue cells to biomaterials is a prerequisite of paramount importance for the effectiveness of a tissue engineering construct (cell and scaffolds). Functionalization of polymeric scaffolds with organic polymers, such as collagen or proteoglycans, is a promising approach in order to improve the cytocompatibility. As a matter of fact, organic polymers, isolated directly from the extracellular matrix, contain a multitude of surface ligand (fibronectin, laminin, vitronectin) and arginine–glycine–aspartic acid-containing peptides that promote cell adhesion. In tissue engineering, the combination of organic and synthetic polymers gives rise to scaffolds characterized simultaneously …

Phenomenological approach to compare the crystallization kinetics of isotactic polypropylene and polyamide-6 under pressure

Reliable experimental data for semicrystalline polymers crystallized under pressure are supplied on the basis of a model experiment in which drastic solidification conditions are applied. The influence of the pressure and cooling rate on some properties, such as the density and microhardness, and on the product morphology, as investigated with wide-angle X-ray scattering (WAXS), is stressed. Results for isotactic polypropylene (iPP) samples display a lower density and a lower microhardness with increasing pressure over a wide range of cooling rates (from 0.01 to 20 °C/s). Polyamide-6 (PA6) samples exhibit the opposite behavior, with the density and microhardness increasing at higher pressur…

CaP-Bioglass composite coating by galvanic deposition

Orthopedic devices are increasingly used in our life to improve the health of patients after bone fractures due to accidents, aging, or sports injuries. Metallic materials (i.e. stainless steel, titanium alloys chromium alloys) are widely employed to fabricate prostheses, screws, and osteosynthesis plates. Although metals could be good mechanical properties like human bone, corrosion phenomena could occur, causing in the worst cases the failure of orthopedic implants. In addition, metal ions released around periprosthetic tissues could arise allergenic and cancerogenic effects. Nowadays, the research was focused on coating science to deal with these issues. In particular, the development of…

Solution-Based Processing for Scaffold Fabrication in Tissue Engineering Applications: A Brief Review

The fabrication of 3D scaffolds is under wide investigation in tissue engineering (TE) because of its incessant development of new advanced technologies and the improvement of traditional processes. Currently, scientific and clinical research focuses on scaffold characterization to restore the function of missing or damaged tissues. A key for suitable scaffold production is the guarantee of an interconnected porous structure that allows the cells to grow as in native tissue. The fabrication techniques should meet the appropriate requirements, including feasible reproducibility and time- and cost-effective assets. This is necessary for easy processability, which is associated with the large …

Current Strategies for Tracheal Replacement: A Review

Airway cancers have been increasing in recent years. Tracheal resection is commonly performed during surgery and is burdened from post-operative complications severely affecting quality of life. Tracheal resection is usually carried out in primary tracheal tumors or other neoplasms of the neck region. Regenerative medicine for tracheal replacement using bio-prosthesis is under current research. In recent years, attempts were made to replace and transplant human cadaver trachea. An effective vascular supply is fundamental for a successful tracheal transplantation. The use of biological scaffolds derived from decellularized tissues has the advantage of a three-dimensional structure based on t…

Poly-left-lactic acid tubular scaffolds via diffusion induced phase separation: Control of morphology

n this work, tubular poly-left-lactic acid scaffolds for vascular tissue engineering applications were produced by an innovative two-step method. The scaffolds were obtained by performing a dip-coating around a nylon fiber, followed by a diffusion induced phase separation process. Morphological analysis revealed that the internal lumen of the as-obtained scaffold is equal to the diameter of the fiber utilized; the internal surface is homogeneous with micropores 1–2 μm large. Moreover, a porous open structure was detected across the thickness of the walls of the scaffold. An accurate analysis of the preparation process revealed that it is possible to tune up the morphology of the scaffold (w…

PLLA scaffolds with controlled architecture as potential microenvironment for in vitro tumor model

Abstract The "microenvironment" where a tumor develops plays a fundamental role in determining its progression, the onset of metastasis and, eventually, its resistance to therapies. Tumor cells can be considered more or less invasive depending both on the nature of the cells and on the site where they are located. Commonly adopted laboratory culture protocols for the investigation of tumor cells take usually place on standard two-dimensional supports. However, such cultures do not allow for reproduction of the biophysical properties of the tumor’s microenvironment, thus causing the cells to lose most of their relevant characteristics. In this work MDA-MB 231 breast cancer cells were cultiva…

In vitro corrosion and biocompatibility of brushite/hydroxyapatite coatings obtained by galvanic deposition on 316LSS

Corrosion behavior and cytotoxicity was reported for mixed brushite (BS)/hydroxyapatite (HA) coatings deposited on 316LSS substrate through a displacement reaction. Corrosion tests, carried out in a simulated body fluid, showed that in comparison with bare 316L, coating shifts Ecorrto anodic values and reduces icorreven if oscillations were observed, which were explained in terms of the chemical interactions at the solid/liquid interface. Cell biocompatibility of the coating was investigated through osteoblastic cell line MC3T3-E1, evidencing the absence of any cytotoxicity Taken together, the results show that galvanic deposition is a simple and cost-effective method for producing bioactiv…

Galvanic Deposition of Hydroxyapatite/Chitosan/Collagen Coatings on 304 Stainless Steel

The galvanic deposition method was used to deposit Hydroxyapatite/Chitosan/Collagen coatings on 304 stainless steel. Galvanic deposition is an alternative and valid way to fabricate bio-coatings with high biocompatibility and good anticorrosion properties. Physical-chemical characterizations were carried out to investigate chemical composition and morphology of the samples. Coatings consist of a mixture of calcium phosphate (Brushite and Hydroxyapatite) with chitosan and collagen. Corrosion tests were performed in the simulated body fluid (SBF) after different aging times. Results show that, in comparison with bare 304 stainless steel, coating shifts corrosion potential to anodic values and…

Polmunary epithelial barrier formation on biodegradable poly-L-lactic-acid (PLLA) membrane

Aims: Investigation of epithelial barrier formation using PLLA membranes for application in bioengineering. Background: The development of functional and biocompatible substitutes for damaged tissue or organs is a major challenge in biomedical engineering. The epithelial barrier plays a central role in tissue homeostasis and immunity preventing damage and contamination of the interstitial tissues. Different in vitro models of the lung and intestinal epithelial barriers have been well characterized, however these tend to use non-biodegradable and/or poorly biocompatible scaffolds. Therefore, there is a need for better supports for epithelial cells for future applications in tissue engineerin…

Measurement of cloud point temperature in polymer solutions.

A temperature-controlled turbidity measurement apparatus for the characterization of polymer solutions has been instrumented and set up. The main features are the coupled temperature-light transmittance measurement and the accurate temperature control, achieved by means of peltier cells. The apparatus allows to measure cloud point temperatures by adopting different cooling protocols: low rate for quasi-equilibrium measurements and high rate for detect kinetic effects. A ternary polymeric solution was adopted as case study system showing that cooling rate affects the measured cloud point temperature.

Physical and biological properties of electrospun poly( d , l ‐lactide)/nanoclay and poly( d , l ‐lactide)/nanosilica nanofibrous scaffold for bone tissue engineering

Electrospun scaffolds exhibiting high physical performances with the ability to support cell attachment and proliferation are attracting more and more scientific interest for tissue engineering applications. The inclusion of inorganic nanoparticles such as nanosilica and nanoclay into electrospun biopolymeric matrices can meet these challenging requirements. The silica and clay incorporation into polymeric nanofibers has been reported to enhance and improve the mechanical properties as well as the osteogenic properties of the scaffolds. In this work, for the first time, the physical and biological properties of polylactic acid (PLA) electrospun mats filled with different concentrations of n…

3D polymeric supports promote the growth and progression of anaplastic thyroid carcinoma.

Abstract Anaplastic thyroid carcinoma (ATC) is a rare and aggressive malignancy that accounts for the majority of deaths from all thyroid cancers. ATC exhibits invasiveness and highly resistance to conventional therapies which include cytotoxic chemotherapy, the combination of BRAF and MEK inhibition and, more recently, immunotherapies, that have shown promising but still limited results. A growing knowledge on ATC tumor biology is needed for developing more effective therapies with significant better survival. Researchers have begun to utilize 3D models to culture cancer cells for in vitro studies. In this work, C643 ATC cell line was cultured on polymeric scaffolds with high-interconnecte…

Phase separation of polymer blends in solution: A case study

Abstract The phase behavior and phase separation features of the quaternary system poly- l -lactide (PLLA)/poly-rac-lactide (PLA)/dioxane/water were investigated. Experiments were performed with fixed total polymer concentration of 6 wt%, by varying the PLLA/PLA weight ratio. Blend weight compositions examined were 100/0, 80/20, 50/50, 20/80 and 0/100, at fixed dioxane/water weight ratio (87/13). Cloud point measurements reported that the demixing temperatures of blends are close to PLLA in the same mixed solvent, in line with the calculated spinodals. As regards to foam preparation, above the PLA cloud point, morphology is similar to pure PLLA foams; conversely, below PLA cloud point, the …

Sterilization of macroscopic poly(l-lactic acid) porous scaffolds withdense carbon dioxide: Investigation of the spatial penetration of thetreatment and of its effect on the properties of the matrix

Abstract In this work the sterilization with dense carbon dioxide of poly( l -lactic acid) (PLLA) porous scaffolds intended for tissue engineering applications was investigated with the main objective of confirming the three-dimensional efficacy of the treatment and of analysing the scaffold properties after CO2 treatment. For this purpose the scaffold was contaminated with a conventional bacterium (Escherichia coli) and with spores (Streptomyces coelicolor), a species more fascinating and difficult to inactivate. Contamination was performed in such a way to soak the whole matrix with bacteria and spores. The effect of pressure and treatment time on the efficacy of the sterilization was eva…

Chitosan-Coating Deposition via Galvanic Coupling

A galvanic method to deposit chitosan coatings on stainless steel substrate is reported. Deposition of suitable coatings is desired to improve biocompatibility and corrosion resistance of metallic medical devices to be implanted in human body. In the present work, a thin hydrogel layer of chitosan was deposited on 304SS by a galvanic displacement reaction, which is advantageous first as it does not require external power supply. 304SS was immersed into an aqueous solution of chitosan/lactic acid and electrochemically coupled with magnesium acting as a sacrificial anode. SEM images showed the formation of a uniform layer of chitosan with a thickness controlled by deposition time. Corrosion t…

Blend scaffolds with polyaspartamide/polyester structure fabricated via TIPS and their RGDC functionalization to promote osteoblast adhesion and proliferation

Target of this work was to prepare a RGDC functionalized hybrid biomaterial via TIPS technique to achieve a more efficient control of osteoblast adhesion and diffusion on the three-dimensional (3D) scaffolds. Starting from a crystalline poly(l-lactic acid) (PLLA) and an amorphous alpha,beta-poly(N-2-hydroxyethyl) (2-aminoethylcarbamate)-d,l-aspartamide-graft-polylactic acid (PHEA-EDA-g-PLA) copolymer, blend scaffolds were characterized by an appropriate porosity and pore interconnection. The PHEA-EDA-PLA interpenetration with PLLA improved hydrolytic susceptibility of hybrid scaffolds. The presence of free amino groups on scaffolds allowed to tether the cyclic RGD peptide (RGDC) via Michael…

Mathematical and numerical modeling of an airlift perfusion bioreactor for tissue engineering applications

The Tissue Engineering (TE) strategy is widely focused on the development of perfusion bioreactors to promote the production of three-dimensional (3D) functional tissues. To optimize tissue production, it is worth investigating the engineering parameters of a bioreactor system for identifying a beneficial range of operation variables. Mathematical and numerical modeling of a perfusion bioreactor is capable to provide relevant insights into the fluid flow and nutrients transport while predicting experimental data and exploring the impact of changing operating parameters, such as fluid velocities. In this work, the hydrodynamic parameters and oxygen transport were investigated using mathemati…

Some Features of Polymeric Membranes for Water Purification via Membrane Distillation

Polymeric membranes are currently adopted in water purification processes, such as reverse osmosis (RO) and membrane distillation (MD). This latter technique is very promising for separation effectiveness and energy savings. A valuable and effective MD unit must be equipped with polymeric membranes that exhibit specific properties, for example, hydrophobicity, a narrow pore size range, a high water penetration pressure, and a large vapor permeability. In this work, we present and examine the main features of membranes for MD processes, with the aim of experimentally evaluating the related performances. Scanning electron microscopy analysis was carried out for a first estimate of the pore si…

Investigation of the Cooling of Hot Walls by Liquid Water Sprays

An experimental study was conducted for the heat transfer from hot walls to liquid water sprays. Four full cone, swirl spray nozzles were used at different upstream pressures, giving mass fluxes impinging on the wall, G, from 8 to 80 kg m(-2) s(-1), mean droplet velocities, U, from 13 to 28 m s(-1) and mean droplet diameters, D, from 0.4 to 2.2 mm. A target consisting of two slabs of beryllium-copper alloy, each 4 x 5 cm in size and 1.1 mm thick, was electrically heated to about 300 degrees C and then rapidly and symmetrically cooled by water sprays issuing from two identical nozzles. The midplane temperature was measured by a fast response, thin-foil thermocouple and the experimental data …

Galvanic deposition of Chitosan-AgNPs as antibacterial coating

Thanks to mechanical properties similar human bones, metallic materials represent the best choice for fabrication of orthopedic implants. Although metals could be widely used in the field of biomedical implants, corrosion phenomena could occur, causing metal ions releasing around periprosthetic tissues leading, in the worst cases, to the development of infections. In these cases, patients need prolonged antibiotic therapies that may cause bacterial resistance. Preventing bacterial colonization of biomedical surfaces is the key to limiting the spread of infections. Antibacterial coatings have become a very active field of research, strongly stimulated by the increasing urgency of identifying…

Effect of Polyhydroxyalkanoate (PHA) Concentration on Polymeric Scaffolds Based on Blends of Poly-L-Lactic Acid (PLLA) and PHA Prepared via Thermally Induced Phase Separation (TIPS)

Hybrid porous scaffolds composed of both natural and synthetic biopolymers have demonstrated significant improvements in the tissue engineering field. This study investigates for the first time the fabrication route and characterization of poly-L-lactic acid scaffolds blended with polyhydroxyalkanoate up to 30 wt%. The hybrid scaffolds were prepared by a thermally induced phase separation method starting from ternary solutions. The microstructure of the hybrid porous structures was analyzed by scanning electron microscopy and related to the blend composition. The porosity and the wettability of the scaffolds were evaluated through gravimetric and water contact angle measurements, respective…

Deposition and characterization of coatings of Hydroxyapatite, Chitosan, and Hydroxyapatite-Chitosan on 316L for biomedical devices

In the last decades, the scientific community has turned on great interest towards the development of increasingly performing biomedical systems. In the orthopedic field, biomedical devices are made up by metallic materials (mainly steel and titanium alloys), which have low/medium resistance to corrosion and a low osteointegration capacity when implanted inside the human body. This can lead to infection or inflammation that can damage the tissues surrounding the implant. The use of biocompatible coatings allows cancelling or mitigating these phenomena. The coating interposing between aggressive environment and biomedical device inhibits corrosion so limiting the metal ions release into the …

Engineering approaches in siRNA delivery.

siRNAs are very potent drug molecules, able to silence genes involved in pathologies development. siRNAs have virtually an unlimited therapeutic potential, particularly for the treatment of inflammatory diseases. However, their use in clinical practice is limited because of their unfavorable properties to interact and not to degrade in physiological environments. In particular they are large macromolecules, negatively charged, which undergo rapid degradation by plasmatic enzymes, are subject to fast renal clearance/hepatic sequestration, and can hardly cross cellular membranes. These aspects seriously impair siRNAs as therapeutics. As in all the other fields of science, siRNAs management ca…

CHARACTERIZATION OF HYDROPHOBIC POLYMERIC MEMBRANES FOR MEMBRANE DISTILLATION PROCESS

Hydrophobic microporous membranes are utilized in membrane distillation (MD) processes, e.g. seawater desalination at moderate temperatures. The vapour permeability of commercial hydrophobic membranes with different pore sizes (0.2-1 micron) was characterized through a simple apparatus designed-on-purpose. A cylindrical vessel had a face closed by the membrane and the other connected to a thin graduate tube. The water level variation in the tube is recorded and related to the vapour flux across the membrane. Measurements were taken in the temperature range 20-80°C. A fan tangential to membrane surface was employed to maintain a constant driving force for vapour transport. Vapour flux did no…

Crystallization kinetics in relation to polymer processing

Phase distribution of quenched samples has been determined by a deconvolution procedure of WAXS spectra in a wide range of cooling rates. The informations collected together with isothermal and DSC results provide a very wide set of data on the crystallization kinetics of polymers relevant which covers conditions encountered in most polymer processing operations. They have been compared with predictions of a non-isothermal crystallization model assuming two independent and parallel crystallization processes competing during solidification.

A High-Throughput Mechanical Activator for Cartilage Engineering Enables Rapid Screening of in vitro Response of Tissue Models to Physiological and Supra-Physiological Loads.

Articular cartilage is crucially influenced by loading during development, health, and disease. However, our knowledge of the mechanical conditions that promote engineered cartilage maturation or tissue repair is still incomplete. Current in vitro models that allow precise control of the local mechanical environment have been dramatically limited by very low throughput, usually just a few specimens per experiment. To overcome this constraint, we have developed a new device for the high throughput compressive loading of tissue constructs: the High Throughput Mechanical Activator for Cartilage Engineering (HiT-MACE), which allows the mechanoactivation of 6 times more samples than current tech…

Behavior of Calcium Phosphate–Chitosan–Collagen Composite Coating on AISI 304 for Orthopedic Applications

Calcium phosphate/chitosan/collagen composite coating on AISI 304 stainless steel was investigated. Coatings were realized by galvanic coupling that occurs without an external power supply because it begins with the coupling between two metals with different standard electrochemical potentials. The process consists of the co-deposition of the three components with the calcium phosphate crystals incorporated into the polymeric composite of chitosan and collagen. Physical-chemical characterizations of the samples were executed to evaluate morphology and chemical composition. Morphological analyses have shown that the surface of the stainless steel is covered by the deposit, which has a very r…

A dynamic air–liquid interface system for in vitro mimicking of the nasal mucosa

The development of an in vitro 3D model for the growth of the nasal mucosa cells can improve the therapy and the study of pathological states for subjects with chronic airway conditions. We have previously characterized a system consisting of a scaffold with an internal channel and a perfusion bioreactor with two independent flows provided by an external and an internal circuit, respectively. In this paper, this system was designed as a model of the nasal cavity, in which cells, grown on the inner surface of the scaffold channel, would be in contact at the same time with both culture medium, supplied by the external circuit, and air, provided with the internal flow. To ensure adequate nutri…

Study on heat transfer coefficients during cooling of PET bottles for food beverages

The heat transfer properties of different cooling systems dealing with Poly-Ethylene-Terephthalate (PET) bottles were investigated. The heat transfer coefficient (Ug) was measured in various fluid dynamic conditions. Cooling media were either air or water. It was shown that heat transfer coefficients are strongly affected by fluid dynamics conditions, and range from 10 W/m2 K to nearly 400 W/m2 K. PET bottle thickness effect on Ug was shown to become relevant under faster fluid dynamics regimes.

Polymeric scaffolds prepared via thermally induced phase separation: Tuning of structure and morphology

Scaffolds suitable for tissue engineering applications like dermal reconstruction were prepared by Thermally Induced Phase Separation (TIPS) starting from a ternary solution PLLA/dioxane/water. The experimental protocol consisted of three consecutive steps, a first quench from the homogeneous solution to an appropriate demixing temperature (within the metastable region), a holding stage for a given residence time, and a final quench from the demixing temperature to a low temperature (within the unstable region). A large variety of morphologies, in terms of average pore size and interconnection, were obtained upon modifying the demixing time and temperature, owing to the interplay of nucleat…

Solidification of syndiotactic polystyrene by a continuous cooling transformation approach

Syndiotactic polystyrene (sPS) was solidified from the melt under drastic conditions according to a continuous cooling transformation methodology developed by the authors, which covered a cooling rate range spanning from approximately 0.03 to 3000 °C/s. The samples produced, structurally homogeneous across both their thickness and surface, were analyzed by macroscopic methods, such as density, wide-angle X-ray diffraction (WAXD), and microhardness (MH) measurements. The density was strictly related to the phase content, as confirmed by WAXD deconvolution. The peculiar behavior encountered (the density first decreasing and then increasing with the cooling rate) was attributed to the singular…

Core-shell PLA/Kef hybrid scaffolds for skin tissue engineering applications prepared by direct kefiran coating on PLA electrospun fibers optimized via air-plasma treatment

Abstract Over the recent years, there is a growing interest in electrospun hybrid scaffolds composed of synthetic and natural polymers that can support cell attachment and proliferation. In this work, the physical and biological properties of polylactic acid (PLA) electrospun mats coated with kefiran (Kef) were evaluated. Gravimetric, spectroscopic (FTIR-ATR) and morphological investigations via scanning electron microscopy confirmed the effective formation of a thin kefiran layer wrapped on the PLA fibers with an easy-tunable thickness. Air plasma pre-treatment carried out on PLA (P-PLA) affected both the morphology and the crystallinity of Kef coating as confirmed by differential scanning…

GENERATION OF PREVASCULARIZED PLLA BIODEGRADABLE SCAFFOLDS BY DIP DRAWING AND DIFFUSION INDUCED PHASE SEPARATION (DIPS).

A critical obstacle in tissue engineering is to develop a massive structure of living cells upon transfer from the in vitro culture conditions into the host in vivo. A vascular network is required to supply essential nutrients, including oxygen, remove metabolic waste products and provide a biochemical communication “highway”. For these reasons to build an implantable structure in which vessel formation (angiogenesis) take place is mandatory. PLLA scaffolds usable in vascular tissue engineering were generated by dip-coating via Diffusion Induced Phase Separation (DIPS) technique. The scaffolds, with a vessel-like shape, were obtained by performing a DIPS process around a nylon fibre whose d…

Preparation and properties of poly(L-lactic acid) scaffolds by thermally induced phase separation from a ternary polymer-solvent system

Poly(L-lactic acid) (PLLA) foams for tissue engineering were prepared via thermally induced phase separation of a ternary system PLLA/dioxane/tetrahydrofuran (THF) followed by double solvent exchange (water and ethyl alcohol) and drying. An extension to solidification from solution of a previously developed method for solidification from the melt was adopted. The technique is based on a continuous cooling transformation (CCT) approach, consisting in recording the thermal history experienced by rapidly cooled samples and then analyzing the resulting sample morphology. Different foams were produced by changing the relative amount of dioxane and THF in the starting solution while the amount of…

Procedimento di preparazione di scaffolds polimerici preangiogenizzati.

Establishment of a pulmonary epithelial barrier on biodegradable poly-L-lactic-acid membranes

Development of biocompatible and functional scaffolds for tissue engineering is a major challenge, especially for development of polarised epithelia that are critical structures in tissue homeostasis. Different in vitro models of the lung epithelial barrier have been characterized using non-degradable polyethylene terephthalate membranes which limits their uses for tissue engineering. Although poly-L-lactic acid (PLLA) membranes are biodegradable, those prepared via conventional Diffusion Induced Phase Separation (DIPS) lack open-porous geometry and show limited permeability compromising their use for epithelial barrier studies. Here we used PLLA membranes prepared via a modification of the…

Injection molding of syndiotactic polystyrene/clay nanocomposites

This study aims at exploring the effect of a commercial organoclay montmorillonite (MMT) on the final properties of syndiotactic polystyrene (sPS) injection-molded samples. To this goal, injection-molded specimens made from neat sPS and commercial MMT modified with various organic compounds were prepared in different molding conditions. Dispersion of clay was attained via melt blending, directly in the injection chamber of the injection-molding machine. The obtained specimens were analyzed by IR spectroscopy, X-ray diffraction, thermogravimetry, and differential thermal analysis, with the aim of elucidating the effect of clay on the microstructures of the samples. Results clearly show that,…

Effect of pressure on the PVT behaviour of iPP as revealed by dilatometric measurements

Isotactic Poly-propylene samples, previously prepared under known conditions of pressure and cooling rate by means of a special apparatus designed and set-up by the authors, were subjected to several isobaric runs at low cooling and heating rate in a confining fluid dilatometer (by GNOMIX). The effect of the previous thermo-mechanical histories and the effect of pressure in the dilatometry on specific volume of the samples was studied. Results show that the initial specific volume depends upon the previous thermo-mechanical histories, which however cancels out after the first heating run. Moreover the reported dilatometric experimental data support the evidence that an increase of pressure …

Optical characterization of phase transitions in pure polymers and blends

To study the optical properties of polymeric samples, an experimental apparatus was designed on purpose and set up. The sample is a thin film enclosed between two glass slides and a PTFE frame, with a very thin thermocouple placed on sample for direct temperature measurement. This sample holder was placed between two aluminum slabs, equipped with a narrow slit for optical measurements and with electrical resistances for temperature control. Sample was enlightened by a laser diode, whereas transmitted light was detected with a photodiode. Measurements were carried out on polyethylene-terephtalate (PET) and two different polyamides, tested as pure polymers and blends. The thermal history impo…

Morphology and thermal properties of foams prepared via thermally induced phase separation based on polylactic acid blends

Blends of poly-l-lactic acid with two different types of polylactic acid with different average molecular weights (50,000 and 175,000 g/mol, respectively) in different proportions (90/10, 80/20 and 70/30) were utilized in order to produce biodegradable and biocompatible scaffolds for soft tissue engineering applications. The scaffolds were produced via thermally induced phase separation starting from ternary systems where dioxane was the solvent and water the non-solvent. Morphology (average pore size and interconnection) was evaluated by scanning electron microscopy. Foams apparent density was also evaluated (porosity ranges from 87% to 92%). Moreover, a differential scanning calorimetry …

Polymer Solidification under Pressure and High Cooling Rates

Abstract Polymer solidification under processing conditions is a complex phenomenon in which the kinetics of flow, high thermal gradients and high pressures determine the product morphology. The study of polymer structure formed under pressure has been mainly made using conventional techniques such as dilatometry and differential scanning calorimetry under isothermal conditions or non isothermal conditions but at cooling rates several orders of magnitude lower than those experienced in industrial processes. A new equipment has been recently developed and improved to study the crystallization of polypropylene when subjected to pressure and cooled rapidly. An experimental apparatus essentiall…

Synthesis, characterization and foaming of PHEA-PLLA, a new graft copolymer for biomedical engineering

Abstract In this study a chemical grafting procedure was set up in order to link high molecular weight poly L-lactic acid (PLLA) chains to the hydrophilic α,β-poly(N-2-hydroxyethyl)-DL-aspartamide (PHEA) backbone. A graft copolymer named PHEA-g-PLLA (or simply PHEA–PLLA) was obtained bearing a degree of derivatization of 1.0 mol.% of PLLA as grafted chain. This new hybrid derivative offers both the opportune crystallinity necessary for the production of scaffolds trough a thermally induced phase separation (TIPS) technique and the proper chemical reactivity to perform further functionalizations with bio-effectors and drugs. PHEA–PLLA porous scaffolds for tissue engineering applications were…

The continuous cooling transformation (CCT) as a flexible tool to investigate polymer crystallization under processing conditions

An experimental route for investigating polymer crystallization over a wide range of cooling rates (from 0.01 to 1000◦C/s) and pressures (from 0.1 to 40 MPa) is illustrated, using a method that recalls the approach adopted in metallurgy for studying structure development in metals. Two types of experimental setup were used, namely an apparatus for fast cooling of thin films (100–200 μm thick) at various cooling rates under atmospheric pressure and a device (based on a on-purpose modified injection molding machine) for quenching massive samples (about 1–2 cm3) under hydrostatic pressure fields. In both cases, ex situ characterization experiments were carried out to probe the resulting struct…

Sistema biocompatibile di supporto per la piantumazione di organismi vegetali in ambiente marino

Il brevetto consiste in un sistema a basso impatto ambientale, realizzato interamente in bioplastica, per il fissaggio sul fondo marino in forma rapida ed efficace di organismi vegetali, allo scopo di garantirne l'attecchimento e la crescita, assecondandone la dinamica naturale di sviluppo.

Isotactic polypropylene solidification under pressure and high cooling rates. A master curve approach

Solidification in industrial processes very often involves flow fields, high thermal gradients and high pressures: the development of a model able to describe the polymer behavior becomes complex. Recently a new equipment has been developed and improved to study the crystallization of polymers when quenched under pressure. An experimental apparatus based on a modified, special injection moulding machine has been employed. Polymer samples can be cooled at a known cooling rate up to 100 °C/s and under a constant pressure up to 40 MPa. Density, Micro Hardness (MH), Wide angle X-ray diffraction (WAXD), and annealing measurements were then used to characterize the obtained sample morphology. Res…

Characterization of PLLA scaffolds for biomedical applications

The porosity and pore size distribution of three-dimensional scaffolds have direct implications on their biomedical applications (tissue engineering, drug delivery, and wound dressing). Accordingly, in this paper, a fast, facile, and conservative method relying on low-field nuclear magnetic resonance (LF-NMR) for the evaluation of mean pore size and pore size distribution of polymeric scaffolds is reported. The applicability of the technique is demonstrated on poly-L-lactic acid scaffolds fabricated using the thermal induced phase separation. Results obtained through LF-NMR are successfully compared to scanning electron microscope and X-ray microcomputed tomography micrographs.

The solidification behavior of a PBT/PET blend over a wide range of cooling rate

In recent years, much attention has been paid to the development of high-performance polyester blends, among which blends of polybutylene terephthalate/polyethylene terephthalate (PBT/PET) are expected to exhibit remarkable properties as far as their crystallization behavior is concerned. Through trial and error, appropriate commercial compositions have been chosen which could not be otherwise explained by a suitable interpretation of the mechanisms determining their solidification behavior. The solidification behavior of a 60/40 w/w PBT/PET blend was studied in a wide range of cooling conditions, according to a continuous cooling transformation (CCT) procedure developed previously, aiming …

Biological evaluation of PLLA membranes, with different pore diameters, to stimulate cell adhesion and growth in vitro

Polymeric membranes prepared via DIPS (Diffusion Induced Phase Separation) are widely studied and utilized as scaffolds for the regeneration of tissue. In this work, poly (L)-lactide membrane are prepared through a DIPS protocol starting from a ternary solution made of polymer, dioxane (solvent) and water (non-solvent). A three-dimensional, porous and mechanically stable membrane is desirable for ingrowth of human bronchial epithelial cells. Polymeric membranes prepared via DIPS (Diffusion Induced Phase Separation) are widely studied and utilized as scaffolds for the regeneration of tissue. In this work, poly (L)-lactide membrane are prepared through a DIPS protocol starting from a ternary …

Double Flow Bioreactor for In Vitro Test of Drug Delivery.

In this work, double-structured polymeric scaffolds were produced, and a double flow bioreactor was designed and set up in order to create a novel system to carry out advanced in vitro drug delivery tests. The scaffolds, consisting of a cylindrical porous matrix, are able to host cells, thus mimicking a three-dimensional tumor mass: moreover, a “pseudo-vascular” structure was embedded into the matrix, with the aim of allowing a flow circulation. The structure that emulates a blood vessel is a porous tubular-shaped scaffold prepared by Diffusion Induced Phase Separation (DIPS), with an internal lumen of 2 mm and a wall thickness of 200 micrometers. The as-prepared vessel was incorporated…

Galvanic deposition and characterization of brushite/hydroxyapatite coatings on 316L stainless steel

In this work, brushite and brushite/hydroxyapatite (BS, CaHPO4·H2O; HA, Ca10(PO4)6(OH)2) coatings were deposited on 316L stainless steel (316LSS) from a solution containing Ca(NO3)2·4H2O and NH4H2PO4 by a displacement reaction based on a galvanic contact, where zinc acts as sacrificial anode. Driving force for the cementation reaction arises from the difference in the electrochemical standard potentials of two different metallic materials (316LSS and Zn) immersed in an electrolyte, so forming a galvanic contact leading to the deposition of BS/HA on nobler metal. We found that temperature and deposition time affect coating features (morphology, structure, and composition). Deposits were char…

A Versatile Technique to Produce Porous Polymeric Scaffolds: The Thermally Induced Phase Separation (TIPS) Method

Among the various scaffold fabrication techniques, thermally induced phase separation (TIPS) is one of the most versatile methods to produce porous polymeric scaffold and it has been largely used for its capability to produce highly porous and interconnected scaffolds. The scaffold architecture can be closely controlled by varying the process parameters, including polymer type and concentration, solvent/non-solvent ratio and thermal history. TIPS technique has been widely employed, also, to produce scaffolds with a hierarchical pore structure and composite polymeric matrix/inorganic filler foams.

PLLA/Fibrin Tubular Scaffold: A New Way for Reliable Endothelial Cell Seeding

In the present work a simple and quick technique for cell seeding into tubular-shaped scaffolds, which allows a homogeneous cell distribution, was tested. The poly-L-lactide (PLLA) scaffolds, prepared via diffusion induced phase separation (DIPS), were filled with fibrin gel in order to obtain a hybrid scaffold for Vascular Tissue Engineering applications. The formation of immobilized fibrin networks on the inner surface of the tubular scaffolds was observed using confocal microscopy and SEM. Morphological analysis of the so-obtained scaffold revealed that the fibrin gel is uniformly distributed on the internal surface of the scaffold, leading to an organized structure. Moreover a penetrati…

Patent number PA2010 A 000023: Sistema biocompatibile di supporto per la piantumazione di organismi vegetali in ambiente marino

Il brevetto consiste in un sistema a basso impatto ambientale, realizzato interamente in bioplastica, per il fissaggio sul fondo marino in forma rapida ed efficace di organismi vegetali, allo scopo di garantirne l'attecchimento e la crescita, assecondandone la dinamica naturale di sviluppo.

PHEA‐PLLA: A New Polymer Blend For Tissue Engineering Applications

One of most important features that a material should have in order to be utilized for tissue engineering applications is its biocompatibility and its chemical surface. These properties are required for a high degree of cell adhesion on the scaffold. Poly‐L‐lactid acid (PLLA) is a biocompatible synthetic polymer approved by the Food and Drug Administration for human clinical applications. It has been largely employed, in the last years, as a constituent of surgical and implantable devices. PHEA is a biocompatible water‐soluble synthetic polymer, with a protein‐like structure, whose use as a drug carrier and as starting material for many other biomedical and pharmaceutical applications has b…

PLLA/PLA scaffolds prepared via Thermally Induced Phase Separation (TIPS): tuning of properties and biodegradability

Foams for tissue engineering applications were prepared via thermally induced phase separation (TIPS). Poly-L-Lactic Acid (PLLA) and blends of PLLA with PLA in different proportions were used (100/0, 90/10, 75/25, 50/50, 0/100 PLLA/PLA wt/wt) starting from ternary systems where dioxane was the solvent and water the non-solvent. Morphology was evaluated by Scanning Electron Microscopy (average pore size and interconnection) and the void fraction was measured by means of Hg porosimetry. Foams apparent density was also evaluated (porosity ranges from 87% to 92%). Biodegradability was estimated in a body mimicking fluid. Results show that structure and morphology (in terms of average pore size …

Biocompatibility evaluation of PLLA scaffolds for vascular tissue engineering

Poly-L-lactic acid (PLLA), a hemicrystalline material, has been extensively studied in applications of engineered tissues, because it is biodegradable, absorbable and it supports cell attachment and growth. The purpose of this study is to evaluate tissue/ material interactions, neovascularization and the biocompatibility of PLLA by optical and scanning electron microscopy in a model of animal implant. PLLA porous disks were implanted into the dorsal subcutis of BALB/C mice for 1, 2, 3, and 8 weeks. The bioptic samples of excised PLLA and the surrounding tissue were evaluated for inflammatory response and tissue ingrowth. The samples were divided in two halves: one was fixed in neutral buffe…

Tailoring PLLA scaffolds for tissue engineering applications: Morphologies for 2D and 3D cell cultures

PLLA scaffold suitable for dermis regeneration were realized by Thermally Induced Phase Separation (TIPS) starting from a ternary solution PLLA/dioxane/water. The reconstruction of a complex tissues as the dermis implies the use of different cellular types (coculture), with different growth behaviour (2D vs. 3D). The scaffolds present an homogeneous porous surface to allow the keratinocytes 2D growth and a porous internal structure for the fibroblasts 3D growth. Our results show that the porosity of the surface can be tuned by changing the chemical nature of the sample holder (aluminium, teflon, polypropylene). A large variety of morphologies, in terms of average pore size and interconnecti…

Functional Activation of Osteoclast Commitment in Chronic Lymphocytic Leukaemia: A Possible Role for RANK/RANKL Pathway

AbstractSkeletal erosion has been found to represent an independent prognostic indicator in patients with advanced stages of chronic lymphocytic leukaemia (CLL). Whether this phenomenon also occurs in early CLL phases and its underlying mechanisms have yet to be fully elucidated. In this study, we prospectively enrolled 36 consecutive treatment-naïve patients to analyse skeletal structure and bone marrow distribution using a computational approach to PET/CT images. This evaluation was combined with the analysis of RANK/RANKL loop activation in the leukemic clone, given recent reports on its role in CLL progression. Bone erosion was particularly evident in long bone shafts, progressively inc…

Polymeric scaffolds based on blends of poly-l-lactic acid (PLLA) with poly-d-l- lactic acid (PLA) prepared via thermally induced phase separation (TIPS): demixing conditions and morphology

Porous scaffolds based on blends of high crystalline Poly-L-lactic acid (PLLA) with low crystalline poly-D-L-lactic acid (PLA) were prepared via Ther- mally Induced Phase Separation (TIPS), with the aim of exploring the possibility to control the degradation behaviour of the PLA-based scaffold, simultaneously pre- serving the morphological characteristics required for tissue engineering applica- tions. Porous foams with different PLLA/PLA weight ratios (from 95/5 to 60/40) were produced and characterised in terms of pore size, porosity, and thermal properties. The scaffolds present an open porosity, with average pore sizes ranging from 30 to 70 lm. Results showed that, when dealing with a P…

Evaluation of vapor mass transfer in various membrane distillation configurations: an experimental study

Vapor mass transfer phenomena in four different membrane distillation (MD) configurations were examined through a self-built laboratory scale experimen- tal apparatus: Air Gap MD, Sweeping Gas MD, Vacuum Sweeping Gas MD and Vacuum MD. Vapor fluxes were measured and compared with those predicted by various models, showing that MD performance under usual processing conditions is severely controlled by the permeate side resistance to mass transfer.

Preparation of Poly(l-lactic acid) Scaffolds by Thermally Induced Phase Separation: Role of Thermal History

Abstract Poly-L-Lactic Acid (PLLA) scaffolds for tissue engineering were prepared via thermally induced phase separation of a ternary system PLLA/dioxane/tetrahydrofurane. An extension to solution of a previously developed method for solidification from the melt was adopted, the technique being based on a Continuous Cooling Transformation (CCT) approach, consisting in recording the thermal history of rapidly cooled samples and analysing the resulting morphology. Different foams were produced by changing the thermal history, the dioxane to THF ratio (50/50, 70/30, 90/10 v/v) and the polymer concentration (2, 2.5, 4 ° wt) in the starting ternary solution. Pore size, porosity, melting and crys…

Demixing time and temperature influence on porosity and interconnection of PLLA scaffolds prepared via TIPS

Scaffolds suitable for tissue engineering applications were prepared by Thermally Induced Phase Separation (TIPS) starting from a ternary solution PLLA/ dioxane/water. The experimental protocol consisted of three consecutive steps, a first quench from the homogeneous solution to an appropriate demixing temperature (within the binodal region), a liquid-liquid demixing stage for a given time and a final quench from the demixing temperature to a low temperature (within the spinodal region). A large variety of morphologies, in terms of average pore size and interconnection were obtained upon modifying the demixing time and temperature, owing to the interplay of nucleation and growth processes d…

PLLA scaffolds produced by thermally induced phase separation (TIPS) allow human chondrocyte growth and extracellular matrix formation dependent on pore size

Damage of hyaline cartilage species such as nasoseptal or joint cartilage requires proper reconstruction, which remains challenging due to the low intrinsic repair capacity of this tissue. Implantation of autologous chondrocytes in combination with a biomimetic biomaterial represents a promising strategy to support cartilage repair. The aim of this work was to assess the viability, attachment, morphology, extracellular matrix (ECM) production of human articular and nasoseptal chondrocytes cultured in vitro in porous poly(L-lactic) (PLLA) scaffolds of two selected pore sizes (100 and 200 μm). The PLLA scaffolds with 100 and 200 μm pore sizes were prepared via ternary thermally induced ph…

Novel dual-flow perfusion bioreactor for in vitro pre-screening of nanoparticles delivery: design, characterization and testing

An advanced dual-flow perfusion bioreactor with a simple and compact design was developed and evaluated as a potential apparatus to reduce the gap between animal testing and drug administration to human subjects in clinical trials. All the experimental tests were carried out using an ad hoc Poly Lactic Acid (PLLA) scaffold synthesized via Thermally Induced Phase Separation (TIPS). The bioreactor shows a tunable radial flow throughout the microporous matrix of the scaffold. The radial perfusion was quantified both with permeability tests and with a mathematical model, applying a combination of Darcy's Theory, Bernoulli's Equation, and Poiseuille's Law. Finally, a diffusion test allowed to in…

Tubular scaffold for vascular tissue engineering application

A critical obstacle in tissue engineering is the inability to maintain large masses of living cells upon transfer from the in vitro culture conditions into the host in vivo. Capillaries, and the vascular system, are required to supply essential nutrients, including oxygen, remove waste products and provide a biochemical communication “highway”. Another goal in this research field is the possibility to tune the biodegradability of the scaffold. After implantation, the scaffold has to be gradually replaced by cells and extra cellular matrix and it is crucial that this replacement takes place with an appropriate dynamics. A premature degradation, in fact, could lead to a collapse of the struct…

PLLA-fibrin scaffolds for Vascular Tissue Engineering

Patent number RM2009A000073: Procedimento di preparazione di scaffold polimerici preangiogenizzati

Classe proposta A61F - Filtri da inserire nei vasi sanguigni; protesi; apparecchi ortopedici e curativi per contraccezione; fomentazione; trattamento o protezione degli occhi e delle orecchie; bendaggi, garze per medicazioni e assorbenti; necessario per il pronto soccorso

On the calculation of free energy of mixing for aqueous polymer solutions with group-contribution models

Abstract Liquid–liquid phase separation processes are currently used as a route to prepare polymeric porous structures for various applications (membranes and scaffolds for tissue engineering). In membrane and foam fabrication technologies, binary and mainly ternary polymer solutions are used. Membrane morphology is strongly affected by phase equilibria of processing solution. In order to achieve a better control of membrane morphology and to explore a wide quantity of solvents, a predictive tool addressing experimentals would be strongly advisable. In this paper, group contribution models were chosen to test the applicability on a PLLA–dioxane–water ternary polymer solution, whose experime…

Poly lactic acid based foams prepared via thermally induced phase separation (TIPS): A method to tune the crystallinity

Blends of Poly-L-Lactic Acid (PLLA) with two Poly-Lactic Acid (PLA) in different proportions (90/10 and 70/30) were utilized in order to produce biodegradable and biocompatible scaffolds for soft tissue engineering applications. The scaffolds were produced via thermally induced phase separation (TIPS) starting from ternary systems where dioxane was the solvent and water the non-solvent. Morphology was evaluated by Scanning Electron Microscopy (average pore size and interconnection). Moreover a DSC analysis was carried out on the as-obtained scaffold in order to obtain information about theirs thermal properties (enthalpy of melt and crystallization). The results showed that is possible to p…

Evaluation of mechanical and morphologic features of PLLA membranes as supports for perfusion cells culture systems

Abstract Porous biodegradable PLLA membranes, which can be used as supports for perfusion cell culture systems were designed, developed and characterized. PLLA membranes were prepared via diffusion induced phase separation (DIPS). A glass slab was coated with a binary PLLA–dioxane solution (8 wt.% PLLA) via dip coating, then pool immersed in two subsequent coagulation baths, and finally dried in a humidity-controlled environment. Surface and mechanical properties were evaluated by measuring pore size, porosity via scanning electron microscopy, storage modulus, loss modulus and loss angle by using a dynamic mechanical analysis (DMA). Cell adhesion assays on different membrane surfaces were a…

PROCEDIMENTO DI PREPARAZIONE DI SCAFFOLD POLIMERICI PREANGIOGENIZZATI

Crystallization kinetics of iPP: Influence of operating conditions and molecular parameters

An analysis of the crystallization kinetics of different grades of isotactic polypropylene (iPP) is here presented. To describe the crystallization kinetics as a function of molecular and operating parameters, the methodological path followed was the preparation of quenched samples of known cooling histories, calorimetric crystallization isotherms tests, differential scanning calorimetry cooling ramps, wide angle X-ray diffraction (WAXD) measurements, and density determination. The WAXD analysis performed on the quenched iPP samples confirmed that during the fast cooling at least a crystalline structure and a mesomorphic one form. The diffractograms were analyzed by a deconvolution procedur…

Una Metodologia Sperimentale per lo Studio della Cristallizzazione dei Polimeri in Condizioni di Processo. L'Influenza della Velocità di Raffreddamento

Preparation, characterization and in vitro test of composites poly-lactic acid/hydroxyapatite scaffolds for bone tissue engineering.

Abstract In this work, the possibility to produce composite Poly-L-lactic acid (PLLA)/Hydroxyapatite (HA) porous scaffolds via Thermally Induced Phase Separation (TIPS) for bone tissue engineering applications was investigated. Several PLLA/HA wt/wt ratios (95/5, 90/10, 70/30, 50/50, 34/66) were tested and the as-obtained scaffolds were characterized via Scanning Electron Microscopy, Wide Angle X-Ray Diffraction, Thermogravimetric analysis, Gas Pycnometry, Differential Scanning Calorimetry and mechanical compression test. Morphological analysis revealed an open structure with interconnected pores and HA particles embedded in the polymer matrix. Finally, cell cultures were carried out into t…

Peltier cells as temperature control elements: Experimental characterization and modeling

Abstract The use of Peltier cells to realize compact and precise temperature controlled devices is under continuous extension in recent years. In order to support the design of temperature control systems, a simplified modeling of heat transfer dynamics for thermoelectric devices is presented. By following a macroscopic approach, the heat flux removed at the cold side of Peltier cell can be expressed as Q ˙ c = γ ( T c − T c eq ) , where γ is a coefficient dependent on the electric current, Tc and T c eq are the actual and steady state cold side temperature, respectively. On the other hand, a microscopic modeling approach was pursued via finite element analysis software packages. To validat…

Tuning of biodegradation rate of PLLA scaffolds via blending with PLA

Blends of Poly-L-Lactic Acid (PLLA) with Poly-Lactic Acid (PLA) in different proportions (95/5, 90/10, 80/20, 70/30 and 60/40) were utilized in order to realize biodegradable and biocompatible scaffolds for soft tissue engineering applications. The scaffolds were produced via thermally induced phase separation (TIPS) starting from ternary systems where dioxane was the solvent and water the non-solvent. Morphology was evaluated by Scanning Electron Microscopy (average pore size and interconnection). Foams’ apparent density was also evaluated (porosity ranges from 87% to 92%). Moreover an in vitro biodegradation test of scaffolds was set-up in order to verify the rate of degradation of the va…

Modulation of physical and biological properties of a composite PLLA and polyaspartamide derivative obtained via thermally induced phase separation (TIPS) technique.

Abstract In the present study, blend of poly l -lactic acid (PLLA) with a graft copolymer based on α,β-poly(N-hydroxyethyl)- dl -aspartamide and PLA named PHEA-PLA, has been used to design porous scaffold by using Thermally Induced Phase Separation (TIPS) technique. Starting from a homogeneous ternary solution of polymers, dioxane and deionised water, PLLA/PHEA-PLA porous foams have been produced by varying the polymers concentration and de-mixing temperature in metastable region. Results have shown that scaffolds prepared with a polymer concentration of 4% and de-mixing temperature of 22.5 °C are the best among those assessed, due to their optimal pore size and interconnection. SEM and DSC…

Galvanic Deposition of Calcium Phosphate/Bioglass Composite Coating on AISI 316L

Calcium phosphate/Bioglass composite coatings on AISI 316L were investigated with regard to their potential role as a beneficial coating for orthopedic implants. These coatings were realized by the galvanic co-deposition of calcium phosphate compounds and Bioglass particles. A different amount of Bioglass 45S5 was used to study its effect on the performance of the composite coatings. The morphology and chemical composition of the coatings were investigated before and after their aging in simulated body fluid. The coatings uniformly covered the AISI 316L substrate and consisted of a brushite and hydroxyapatite mixture. Both phases were detected using X-ray diffraction and Raman spectroscopy.…

Lattice fluid model generalized for specific interactions: An application to ternary polymer solutions

Abstract The phase diagram of the Poly- l -Lactic-Acid (PLLA), dioxane and water system was derived by using the lattice fluid model generalized for specific interactions. A qualitative analysis of Hansen solubility parameters together with empirical observations were adopted to reduce the number of adjustable parameters. The as-selected model parameters were derived by fitting a set of cloud point curves; consequently, the complete phase diagram of the system was computed. The use of specific interactions between species gave out a quantitative agreement with a set of experimental data, whereas changing the polymer polydispersity the agreement is only qualitative. In principle, the present…

Orientation and Crystallinity Measurements in Film Casting Products

Film casting experiments were carried out with iPP under processing conditions causing the crystallization process to occur under orienting flow. Draw ratio and cooling rates were changed by varying mass flow rates and die thickness. The effect of processing conditions on film crystallinity was investigated by combining WAXS and FT-IR transmission methods, while orientation of both phases was measured by IR dichroism (according to Fraser's method) and successfully compared to birefringence measurements on final films. Crystallinity appears to be almost insensitive to draw ratio and cooling rate. Moreover the crystallinity profile turned out to be also constant along the transverse film dire…

Design, build-up and optimization of a fast quenching device for polymeric thin film

In this work an innovative apparatus for the characterization of polymer solidification under very high cooling rates (up to thousand of K/s) is described, according to the continuous cooling transformation approach adopted in metallurgy for studying structure development in metals. The proposed model experiment is addressed to design a method for the characterization of non-isothermal solidification behaviour, encompassing typical cooling conditions of polymer processing. Only temperature history determines the structure formed, as melt solidification takes place in quiescent conditions. With respect to the device previously developed by the authors [1, 3] the present equipment presents th…

No-flow temperature and solidification in injection molding simulation

The no‐flow temperature (NFT) is a parameter representing the rheological solidification temperature of a polymer. A polymer, during injection molding filling stage, can stop its flow because of its high viscosity, although it is not yet fully solidified by means of glass transition or crystallization. The NFT is used in most of injection molding simulation packages: with this simple parameter it is possible to reduce the errors deriving from viscosity extrapolation at relatively low temperatures. The viscosity measurements for polymers are usually carried out at high temperatures, and the viscosity models can fail in prediction at temperatures close to the glass transition or crystallizati…

Indentation test as a tool for monitoring the solidification process during injection molding

An inline method for monitoring the solidification process during the injection molding of semicrystalline polymers is demonstrated. The method has been applied to various poly(ethylene terephthalate) (PET) and poly(buthylene terephthalate) (PBT) samples. The technique is based on a simple device by which an additional ejector pin is pushed onto the injection-molded part with a fixed force at different times during the solidification phase while the mold remains closed. The residual deformation (the so-called indentation depth) due to the applied load is measured offline after ejection. By the performance of indentation at different times during the cooling phase, an indentation depth profi…

Role of thermal history on quiescent cold crystallization of PET

8 pags., 9 figs.

Non-isothermal crystallization kinetics of PET

The crystallization kinetics of poly(ethylene terephthalate) was studied using constant cooling rate, isothermal and quenching experiments. A non-isothermal crystallization kinetics equation based on a single mechanism was used to analyze the data. Different mechanisms of crystallization at low, intermediate, and high cooling rates were hypothesized based on deviation of the experimental data from the single mechanism model.

Heat of crystallization significantly slows down polymer cooling and thus pressure drop increase during mould filling with thermoplastic crystalline polymers. If a correction of thermal diffusivity can account for such a cooling slow down at least as far as the effect on pressure drop is concerned, the use of nonisothermal crystallization kinetics may be avoided in the simulation of mould filling. A procedure to identify such a correction is outlined in this work. Pressure drop values during cavity filling calculated by using a corrected thermal diffusivity in the model proposed by Lord and Williams favourably compare with literature data taken with polypropylene and polyethylene resins. Be…

An experimental methodology to study polymer crystallization under processing conditions. The influence of high cooling rates

Abstract A new experimental route for investigating polymer crystallization under very high cooling rates (up to 2000°C/s) is described. A complete and exhaustive description of the apparatus employed for preparing thin quenched samples (100– 200 μm thick) is reported, the cooling mechanism and the temperature distribution across sample thickness is also analysed, showing that the final structure is determined only by the thermal history imposed by the fast quench apparatus. Details concerning the characterization techniques used to probe the final structure are reported, including density measurements and wide angle X-ray diffraction patterns. Experimental results concerning isotactic poly…

Crystallization of polymer melts under fast cooling. II. High-purity iPP

SYNOPSIS Samples of a high-purity isotactic polypropylene (iPP) were quenched from the melt so as to monitor cooling history. A continuous variation of morphology and crystal structure was obtained with cooling rate. This is discussed in relation to sample thermal history evidencing that cooling history relevant to quenched samples is in the neighborhood of 90°C. In particular the samples are essentially mesomorphic when at this temperature cooling rates larger than 80°C/s were adopted, while below a few tens of °C/s only a­ monocline form is obtained. Densities of quenched samples were compared with predictions of an isokinetic extrapolation of Avrami model of polymer crystallization kinet…

Crystallization Behaviour at High Cooling Rates of Two Polypropylenes

Phase distribution of quenched samples of two isotactic polypropylenes, having different molecular weight distributions, was evaluated by a deconvolution procedure of WAXD spectra. The dependence on cooling rate of the two resins shows the low molecular weights rich polymer is characterized by a faster kinetics with an α-monoclinic to mesomorphic transition taking place at higher cooling rates.

Improvement of osteogenic differentiation of human mesenchymal stem cells on composite poly l-lactic acid/nano-hydroxyapatite scaffolds for bone defect repair.

Tissue engineering offers new approaches to repair bone defects, which cannot be repaired physiologically, developing scaffolds that mimic bone tissue architecture. Furthermore, biomechanical stimulation induced by bioreactor, provides biomechanical cues that regulate a wide range of cellular events especially required for cellular differentiation and function. The improvement of human mesenchymal stem cells (hMSCs) colonization in poly-L-lactic-acid (PLLA)/nano- hydroxyapatite (nHA) composite scaffold was evaluated in terms of cell proliferation (dsDNA content), bone differen- tiation (gene expression and protein synthesis) and ultrastructural analysis by comparing static (s3D) and dynamic…

Analysis of the crystallization behaviour of PBT-rich PBT/PET blends under processing conditions

Among the high‐performance polyesters blends PBT/PET blends are expected to exhibit remarkable properties as far as the crystallization behaviour is concerned. The solidification behaviour of a 60/40 w/w PBT/PET blend was studied in a wide range of cooling conditions, according to a Continuous Cooling Transformation (CCT) procedure developed by the authors, aiming to emulate the typical conditions encountered in polymer processing. A set of several samples characterized by an homogeneous structure was prepared by solidification from the melt through spray cooling, and the resulting structure and properties were evaluated by density, Micro Hardness (MH), Wide Angle X‐ray Diffraction (WAXD) m…

The use of master curves to describe the simultaneous effect of cooling rate and pressure on polymer crystallization

In a previous work a master-curve approach was applied to experimental density data to explain isotactic polypropylene (iPP) behaviour under pressure and high cooling rates. Suitable samples were prepared by solidification from the melt under various cooling rate and pressure conditions with the help of a special apparatus based on a modified injection moulding machine. The approach here reported is more general than the case study previously shown, and is suitable to be applied to several materials and for different measures related to crystalline content. The proposed simple model is able to predict successfully the final polymer properties (density, micro-hardness, crystallinity) by supe…

Analysis of gate freeze-off time in injection molding

Gate solidification time is an important topic in injection molding technology, as it determines cycle time, which itself is an important issue in the economics of the production process. In this work, a study of the effect of both gate and cavity geometries on gate solidification time was conducted, using a commercial polymer, injection molded with constant holding pressure into a rectangular cavity. Three cavity lengths were used, and for each, two cavity thicknesses were adopted. Spe- cial dies containing different gates were assembled in the mold. Gate thickness was found to be the most important factor determining gate sealing time. However, the cavity geometry is also quite important.…

Coagulation bath composition and desiccation environment as tuning parameters to prepare skinless membranes via diffusion induced phase separation

Diffusion Induced Phase Separation (DIPS) is a currently used technique to produce porous membranes for a large variety of applications. A strong limitation is represented by the occurrence of a dense skin, which is formed during the process, highly reducing the membrane permeability. To overcome this issue, two modifications of the standard DIPS protocol were investigated: the use of coagulation baths composed by a solvent/nonsolvent mixture and the desiccation in a controlled environment, by modulating the partial pressure of nonsolvent vapor. An appropriate choice of coagulation bath composition, together with an appropriate desiccation protocol (i.e., the use of a nonsolvent vapor), wil…

Effect of hydroxyapatite concentration and size on morpho-mechanical properties of PLA-based randomly oriented and aligned electrospun nanofibrous mats

The growing demand for nanofibrous biocomposites able to provide peculiar properties requires systematic investigations of processing-structure-property relationships. Understanding the morpho-mechanical properties of an electrospun scaffold as a function of the filler features and mat microstructure can aid in designing these systems. In this work, the reinforcing effect of micrometric and nanometric hydroxyapatite particles in polylactic acid-based electrospun scaffold presenting random and aligned fibers orientation, was evaluated. The particles incorporation was investigated trough Fourier transform infrared spectroscopy in attenuated total reflectance. The morphology of the nanofibers …

Improved experimental characterization of crystallization kinetics

Polymer solidification occurring in many processes, like for instance injection molding, compression molding and extrusion, is a complex phenomenon, strongly influenced by the thermo-mechanical history experienced by the material during processing. From this point of view, characterization of polymer crystallization in the range of processing con- ditions, i.e. including high cooling rate, is of great technological and academic interest. Quiescent, non-isothermal crys- tallization kinetics of two polypropylene resins were investigated using a new method, based on fast cooling of thin samples with air/water sprays and optical detection of the crystallization phenomenon. The range of cooling …

In vitro degradation and bioactivity of composite poly-l-lactic (PLLA)/bioactive glass (BG) scaffolds: comparison of 45S5 and 1393BG compositions

The objective of this study was to compare the effect of two bioglass (BG) compositions 45S5 and 1393 in poly-l-lactic composite scaffolds in terms of morphology, mechanical properties, biodegradation, water uptake and bioactivity. The scaffolds were produced via thermally induced phase separation starting from a ternary polymer solution (polymer/solvent/non-solvent). Furthermore, different BG to polymer ratios have been selected (1, 2.5, 5% wt/wt) to evaluate the effect of the amount of filler on the composite structure. Results show that the addition of 1393BG does not affect the scaffold morphology, whereas the 45S5BG at the highest amount tends to appreciably modify the scaffold archite…

Deposition and characterization of Hydroxyapatite-Chitosan coatings on 304 SS for biomedical devices

During the last years biomaterials have been largely investigated in order to perform and improve biomedical devices. As regards orthopedic field, the most common equipment used (such as implants, bone grafts or screws) are constituted by metallic materials (steel and titanium alloys), characterized by low/medium resistance to corrosion and low osteointegration ability. Furthermore, these factors could produce local inflammations of the tissues surrounding the implants, increasing kinetics of corrosion phenomena. Scientific community has focused the attention on biocoatings interposed between metal and aggressive environment in order to inhibit corrosion. Furthermore, these coatings are abl…

Characterization of PLLA scaffolds for biomedical applications

The porosity and pore size distribution of three-dimensional scaffolds have direct implications on their biomedical applications (tissue engineering, drug delivery, and wound dressing). Accordingly, in this paper, a fast, facile, and conservative method relying on low-field nuclear magnetic resonance (LF-NMR) for the evaluation of mean pore size and pore size distribution of polymeric scaffolds is reported. The applicability of the technique is demonstrated on poly-L-lactic acid scaffolds fabricated using the thermal induced phase separation. Results obtained through LF-NMR are successfully compared to scanning electron microscope and X-ray microcomputed tomography micrographs.

Human nasoseptal chondrocytes maintain their differentiated phenotype on PLLA scaffolds produced by thermally induced phase separation and supplemented with bioactive glass 1393

Damage of hyaline cartilage such as nasoseptal cartilage requires proper reconstruction, which remains challenging due to its low intrinsic repair capacity. Implantation of autologous chondrocytes in combination with a biomimetic biomaterial represents a promising strategy to support cartilage repair. Despite so far mostly tested for bone tissue engineering, bioactive glass (BG) could exert stimulatory effects on chondrogenesis. The aim of this work was to produce and characterize composite porous poly(L-lactide) (PLLA)/1393BG scaffolds via thermally induced phase separation (TIPS) technique and assess their effects on chondrogenesis of nasoseptal chondrocytes. The PLLA scaffolds without or…