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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. Additionally, both analyses revealed that brushite is the primary phase. The presence of Bioglass was verified through energy-dispersive X-ray spectroscopy, which showed the presence of a silicon peak. During aging in simulated body fluid, the coating was subject to a dynamic equilibrium of dissolution/reprecipitation with total conversion in only the hydroxyapatite phase. Corrosion tests performed in simulated body fluid at different aging times revealed that the coatings made with 1 g/L of Bioglass performed best. These samples have a corrosion potential of −0.068V vs. Ag/AgCl and a corrosion current density of 8.87 × 10−7 A/cm2. These values are better than those measured for bare AISI 316L (−0.187 V vs. Ag/AgCl and 2.52 × 10−6 A/cm2, respectively) and remained superior to pure steel for all 21 days of aging. This behavior indicated the good protection of the coating against corrosion phenomena, which was further confirmed by the very low concentration of Ni ions (0.076 ppm) released in the aging solution after 21 days of immersion. Furthermore, the absence of cytotoxicity, verified through cell viability assays with MC3T3-E1 osteoblastic cells, proves the biocompatibility of the coatings.

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 nutrient supply to the cells in the scaffold channel, the radial diffusion of the culture medium through the porous matrix was evaluated first in qualitative and, then, in quantitative terms, demonstrating the capability of the system to control the value and direction of this flux. As a preliminary study, the culture of epithelial cells in the scaffold channel is also discussed in static, maintaining the air-liquid interface condition for up to 3 weeks. Despite minor abnormalities, such as a gap between cell layers and some detachments from the scaffold, the scaffold ensured cell survival and growth during the experimental time.

The intense use of tellurium (Te) in industrial applications, along with the improper disposal of Te-derivatives, is causing their accumulation in the environment, where oxyanion tellurite (TeO32−) is the most soluble, bioavailable, and toxic Te-species. On the other hand, tellurium is a rare metalloid element whose natural supply will end shortly with possible economic and technological effects. Thus, Te-containing waste represents the source from which Te should be recycled and recovered. Among the explored strategies, the microbial TeO32− biotransformation into less toxic Te-species is the most appropriate concerning the circular economy. Actinomycetes are ideal candidates in environmental biotechnology. However, their exploration in TeO32− biotransformation is scarce due to limited knowledge regarding oxyanion microbial processing. Here, this gap was filled by investigating the cell tolerance, adaptation, and response to TeO32− of a Micromonospora strain isolated from a metal(loid)-rich environment. To this aim, an integrated biological, physical-chemical, and statistical approach combining physiological and biochemical assays with confocal or scanning electron (SEM) microscopy and Fourier-transform infrared spectroscopy in attenuated total reflectance mode (ATR-FTIR) was designed. Micromonospora cells exposed to TeO32− under different physiological states revealed a series of striking cell responses, such as cell morphology changes, extracellular polymeric substance production, cell membrane damages and modifications, oxidative stress burst, protein aggregation and phosphorylation, and superoxide dismutase induction. These results highlight this Micromonospora strain as an asset for biotechnological purposes.

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 investigate the efficacy of the radial flow using Polymeric Fluorescent Nanoparticles (FNPs) mimicking drug/colloidal carriers. These tests confirmed the ability of our bioreactor to create a uniform distribution of particles inside porous matrices. All the findings candidate our system as a potential tool for drug pre-screening testing with a cost and time reduction over animal models.

Coatings were grown on the AZ31 Mg alloy by a hard anodizing process in the hot glycerol phosphate-containing electrolyte. Anodizing conditions were optimized, maximizing corrosion resistance estimated by impedance measurements carried out in Hank's solution at 37 °C. A post anodizing annealing treatment (350 °C for 24 h) allowed us to further enhance the corrosion resistance of the coatings mainly containing magnesium phosphate according to energy-dispersive X-ray spectroscopy and Raman analyses. Gravimetric measurements revealed a hydrogen evolution rate within the limits acceptable for application of AZ31 in biomedical devices. In vitro tests demonstrated that the coatings are biocompatible with a preosteoblast cell line.

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 mathematical equations and Computational Fluid Dynamics (CFD) analysis modeling on a novel perfusion bioreactor working as an airlift external loop. Mathematical models and numerical simulation were associated with experimental results at different configurations to evaluate the effect of the reactor parameters on its hydrodynamics. All predictions of gas holdup and liquid circulation velocity from the modeling were in good agreement with the experimental data. A Poly-L-lactic acid (PLLA) scaffold was produced by Thermally Induced Phase Separation (TIPS) and used as 3D support for biological tests in static and dynamic conditions to assess cell viability and proliferation inside the bioreactor. The whole set of fluid-dynamic and biological results showed that our bioreactor environment offers the benefits of an in vitro potential system to produce functional engineered tissues.

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 microstructure of scaffolds on the extracellular matrix deposition also play a crucial role during perfusion cultures. In the present study, a numerical simulation was implemented at the pore level of the scaffold for fluid flow through porous media during perfused culture. Micro-computed tomography was used to obtain the digital 3D image of the complex geometry of a PLLA scaffold, offering a detailed analysis from a volume-based methodology without simplifications of the results as for pore or Darcy's law-models. Predictions about the uniformity of the flow field through the scaffolds-bioreactor system have been assessed by quantifying the cell viability of a perfusion culture while using pre-osteoblastic cells seeded on 24 PLLA scaffolds for up to 6 days.

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 Hydroxyapatite-Chitosan (HA/CS) and Hydroxyapatite-Polyvinylacetate (HA/PVAc) composites, that have been studied as biocoatings for 304 SS based devices. Hydroxyapatite was selected for its osteoconductivity thanks to its chemical structure similar to bones. Furthermore, Chitosan and Polyvinylacetate are largely used yet in medical field (e.g. antibacterial agent or drug deliver) and in this work were used to create a synergic interaction with hydroxyapatite to increase the strength and bioactivity of coating. Biocotings were obtained by galvanic deposition process that does not require an external power supply. It is a spontaneous electrochemical deposition in which materials with different standard electrochemical potential were short-circuited and immersed in an electrolytic solution. Electrons supply for the cathodic reaction in the noblest material comes from oxidation of the less noble material. SEM, EDS, XRD and RAMAN were performed for chemical-physics characterization of biocoatings. Polarization and impedance measurements have been carried out to evaluate corrosion behavior. Besides, in-vitro cytotoxicity assays have been done for the biological features.

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 cultivated within Poly- l -Lactic Acid (PLLA) scaffolds produced via Thermally Induced Phase Separation (TIPS). Starting from a ternary solution (polymer-solvent-nonsolvent) we produced scaffolds with different morphologies, porosities and pore architectures. The influence of porosity and average pore size upon cell adhesion and growth were investigated by using Cell Counting Kit-8 (CCK-8) as cell viability test, a fluorescence assay staining cell with DAPI and Scanning Electron Microscopy (SEM). Our study demonstrates that the average pore size of the polymeric scaffolds influences both the cell adhesion and resulting morphology of the growing breast cancer cells. In particular, the reported data corroborate the evidence that an average pore size ranging from 40 to 50 μm induces tumor cell aggregation and the formation of the irregular tumor masses typically observed in-vivo. In addition, TIPS proved to be a suitable manufacturing technique for finely tuning the scaffolds’ architecture, relevant to developing the most effective microenvironment for an in-vitro tumor cells growth closely mimicking in-vivo conditions.

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 able to decrease the ions release and to improve the lifetime of implanted devices. In this work, hydroxyapatite-chitosan composites were studied as coatings of 304 SS steel substrates. Hydroxyapatite was chosen for its bioactivity thanks to its chemical structure similar to bones that makes suitable for osteoconductive action. Chitosan is a polysaccharide derived from chitin, usually extracted from the shells of crustaceans, largely used in medical field as antibacterial agent and/or for drug deliver. Hydroxyapatite-chitosan composites were realized by an innovative method based on galvanic process that consists in a simple galvanic coupled of metals with different redox potentials, which is advantageous firstly because does not require external power supply.

Teeth extractions are often followed by alveolar bone reabsorption, although an adequate level of bone is required for reliable rehabilitations by dental implants. Leukocyte and platelet-rich fibrin (L-PRF) has been widely applied in regenerative procedures and with antibiotic and antioxidant agents could play an essential role in hard and soft tissue healing. In this work, a nanocomposite (Sponge-C-MTR) consisting of a hyaluronate-based sponge loaded with metronidazole (MTR) and nanostructured lipid carriers containing curcumin (CUR-NLC) was designed to be wrapped in the L-PRF&trade, membrane in the post-extraction sockets and characterized. CUR-NLCs, obtained by homogenization followed by high-frequency sonication of the lipid mixture, showed loading capacity (5% w/w), drug recovery (95% w/w), spherical shape with an average particle size of 112.0 nm, and Zeta potential of &minus, 24 mV. Sponge-C-MTR was obtained by entrapping CUR-NLC in a hydrophilic matrix by a freeze-drying process, and physico-chemical and cytocompatibility properties were evaluated. Moreover, the aptitude of CUR and MTR to the penetrate and/or permeate both L-PRF&trade, and porcine buccal tissue was assessed, highlighting MTR penetration and CUR accumulation promoted by the system. The results positively support the action of nanocomposite in dental tissues regeneration when applied together with the L-PRF&trade

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-interconnected porous matrix. They exhibited distinct viability, proliferation and 3D morphology similar to an in vivo solid tumor mass. We also carried out quantitative real-time PCR experiments for monitoring Cancer Stem Cells enrichment, since they are most probably the cause of tumor resistance, reoccurrence and metastasis. The same tests were performed after cell treatment with the chemotherapic Doxorubicin. An up-regulation of the analyzed stem-cell markers confirmed the high resistance to treatment of these cell line with respect to conventional drugs. In conclusion, 3D scaffolds could be an ideal platform for studying the mechanisms that regulate ACT growth and survival and also improving novel therapeutic approaches for treatment-resistant thyroid cancer.

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 was analyzed through scanning electron microscopy and it was correlated with the viscosity of polymeric solutions studied by rheological measurements. Scaffolds were mechanical characterized with tensile tests in order to find a correlation between the preparation method and the strength of the mats. The influence of hydroxyapatite particles on the crystallinity of the composites was investigated by differential scanning calorimetry. Finally, cell culture assays with pre-osteoblatic cells were conducted on a selected composite scaffold in order to compare the cell proliferation and morphology with that of polylactic acid scaffolds. Based on the results, we can prove that polylactic acid/hydroxyapatite composites can be one of the biomaterials with the greatest potential for bone tissue regeneration.

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 body. Furthermore, the coating must be bio-compatible in order to guarantee the osteointegration of the device. In this work, calcium hydroxylapatite (HA) and chitosan (CS) have been investigated as possible coatings of 316L stainless steel devices. HA has been selected owing to its osteoinductive and osteoconductive features having a composition similar to bones. CS is highly biocompatible and also widely used for controlled drug delivery. HA, CS, and HA/CS composite coatings were fabricated by an innovative method based on galvanic deposition. It is a spontaneous electrochemical-based process where material with different standard electrochemical potential are immersed in an electrolyte and short circuited. The driving force is the electromotive force, between the coupled materials, where the less noble material oxidizes and supplies electrons for the noblest process that leads to the deposition of the material of interest. The process therefore advances without external power supply, without use of specific equipment, and with deposition of both pure and composite coatings. The chemical-physics characterization of the coatings was conducted by SEM, EDS, XRD and RAMAN; the corrosion behaviour was investigated by TAFEL’s curves and impedance measurements; while the biological features were ascertained through in-vitro cytotoxicity assays.

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 range of biomaterials supporting the use of fabrication technologies. This paper presents a review of scaffold fabrication methods starting from polymer solutions that provide highly porous structures under controlled process parameters. In this review, general information of solution-based technologies, including freeze-drying, thermally or diffusion induced phase separation (TIPS or DIPS), and electrospinning, are presented, along with an overview of their technological strategies and applications. Furthermore, the differences in the fabricated constructs in terms of pore size and distribution, porosity, morphology, and mechanical and biological properties, are clarified and critically reviewed. Then, the combination of these techniques for obtaining scaffolds is described, offering the advantages of mimicking the unique architecture of tissues and organs that are intrinsically difficult to design.

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 200Âμm). The PLLA scaffolds with 100 and 200Âμm pore sizes were prepared via ternary thermally induced phase separation (TIPS) technique and analyzed using scanning electron microscopy (SEM). Articular and nasoseptal chondrocytes were seeded on the scaffold and cultures maintained for 7 and 14Âdays. Live/dead staining, (immuno-)histology and gene expression analysis of type II, type I collagen, aggrecan and SOX9 were performed to assess scaffold cytocompatibility and chondrocyte phenotype. The majority of both chondrocyte types survived on both scaffolds for the whole culture period. Hematoxylin-eosin (HE), alcian blue (visualizing glycosaminoglycans) stainings, immunoreactivity and gene expression of ECM proteins and cartilage marker (type II, I collagen, aggrecan, SOX9) of the chondrocyte scaffold constructs indicated that the smaller pore dimensions promoted the differentiation of the chondrocytes compared with the larger pore size. The present work revealed that the scaffold pore size is an important factor influencing chondrocyte differentiation and indicated that the scaffolds with 100Âμm pores serve as a cytocompatible basis for further future modifications.

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 pore size of 20 µm (defined as the average distance between the pore walls) was detected. For comparison, astrocytes were also cultured in the traditional 2D culture system that we have been using since 2003. Then the effects of different substrates, such as collagen I and IV, and fibronectin were analyzed. The results revealed that the PLLA scaffolds, coated with collagen IV, served as very good matrices for astrocytes, which were observed to adhere, grow and colonize the matrix, acquiring their typical morphology. In addition, under these conditions, they secreted extracellular vesicles (EVs) that were compatible in size with exosomes. Their ability to produce exosomes was also suggested by transmission electron microscopy pictures which revealed both EVs and intracellular structures that could be interpreted as multivesicular bodies. The fact that these cells were able to adapt to the PLLA scaffold, together with our previous results, which demonstrated that brain capillary endothelial cells can grow and differentiate on the same scaffold, could support the future use of 3D brain cell co-culture systems.

Despite eye drops generally represent the most convenient, simple and patient-friendly formulations to treat ocular diseases, they suffer from poor retention on the ocular surface and low drug bioavailability leading to the necessity of prolonged and continuous treatment over time. Therefore, ocular insert could represent an innovative way to benefit from ocular topical administration while minimizing all the relevant limitation related to this route of administration. Polymeric non-erodible mucoadhesive ocular inserts should be comfortable and should rapidly adhere on the ocular surface, remain in situ for prolonged period, assure a reproducible and controlled drug release as well as act as transcorneal absorption promoters. In this study, a well-known aliphatic polyester, poly(1,4-butylene succinate) (PBS), was used as starting material to produce hydrophobic microfibrillar scaffolds by means of electrospinning technique. Plasma-assisted chemical surface functionalization of the PBS scaffolds with appropriate biopolymers (inulin, α,β-poly(N-2-hydroxyethyl)-D,L-aspartamide, heparin) was carried out to confer to the final ocular inserts ad hoc properties as wettability, mucoadhesion and cytocompatibility on human corneal epithelial cells, by improving surface hydrophilicity without modifying the bulk properties of the material. The lipophilic drug triamcinolone acetonide was loaded into the obtained ocular insert and release studies were carried out to demonstrate the ability of drug loaded inserts to release the active until 30 days.

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 (d3D) 3D culture conditions at 7 and 21 days. The colonization rate of hMSCs and osteogenic differentiation were amplified by d3D when physical stimulation was provided by a perfusion bioreactor. Increase in dsDNA content (p < 0.0005), up- regulation of RUNX2, ALPL, SPP1 (p < 0.0005) and SOX9 (p < 0.005) gene expression, and more calcium nodule forma- tion (p < 0.0005) were observed in d3D cultures in comparison to s3D ones over time. Dynamic 3D culture, mimicking the mechanical signals of bone environment, improved significantly osteogenic differentiation of hMSCs on PLLA/nHA scaffold, without the addition of growth factors, confirming this composite scaffold suitable for bone regeneration

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 with 1, 2.5, 5% BG1393 were prepared via TIPS technique starting from a ternary solution (polymer/solvent/non-solvent) in a single step. Scaffolds were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) and differential scanning calorimetric analysis (DSC). Human nasoseptal chondrocytes were seeded on the scaffolds with 1 and 2.5% BG for 7 and 14days and cell survival, attachment, morphology and expression of SOX9 and cartilage-specific extracellular cartilage matrix (ECM) components were monitored. The majority of chondrocytes survived on all PLLA scaffolds functionalized with BG for the whole culture period. Also inner parts of the scaffold were colonized by chondrocytes synthesizing an ECM which contained glycosaminoglycans. Type II collagen and aggrecan gene expression increased significantly in 1% BG scaffolds during the culture. Chondrocyte protein expression for cartilage ECM proteins indicated that the chondrocytes maintained their differentiated phenotype in the scaffolds. BG could serve as a cytocompatible basis for future scaffold composites for osteochondral cartilage defect repair. Abbreviations: AB: alcian blue ACAN: gene coding for aggrecan; BG: Bioactive glass; 2D: two-dimensional; 3D: three-dimensional; COL2A1: gene coding for type II collagen; DAPI: 4ʹ,6-diamidino-2-phenylindole; DMEM: Dulbecco’s Modified Eagle’s Medium; DMMB: dimethylmethylene blue; DSC: Differential scanning calorimetric analysis; ECM: extracellular matrix; EDTA: ethylenediaminetetraacetic acid; EtBr: ethidium bromide; FCS: fetal calf serum; FDA: fluorescein diacetate; GAG: glycosaminoglycans; HDPE: high density polyethylene; HE: hematoxylin and eosin staining; HCA: hydoxylapatite; PBE: phosphate buffered EDTA100mM Na2HPO4 and 5mM EDTA, pH8; PBS: phosphate buffered saline; PFA: paraformaldehyde; PG: proteoglycans; PI: propidium iodide; PLLA: Poly-L-Lactic Acid Scaffold; RT: room temperature; SD: standard deviation; SEM: scanning electron microscopy; sGAG: sulfated glycosaminoglycans; SOX9/Sox9: SRY (sex-determining region Y)-box 9 protein; TBS: TRIS buffered saline; TIPS: Thermally Induced Phase Separation; XRD: X-ray diffraction analysis.

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 addition using the maleimide chemistry. Cell culture test carried out on preosteoblastic cells MC3T3-E1 incubated with scaffolds, has evidenced cell adhesion and proliferation. Furthermore, the presence of distributed bone matrix on all scaffolds was evaluated after 70 days compared to PLLA only samples.

Osteochondral defects derived by traumatic injury or aging related disease are often associated with severe joint pain and progressive loss of joint functions for millions of people worldwide and represent a major challenge for the orthopedic community. Tissue engineering offers new therapeutic approach to repair the osteochondral defects, through the production of scaffolds manufactured to mimic their complex architecture, which consists of cartilage and bone layers. Composite scaffolds based on a PLLA polymeric matrix containing hydroxyapatite (HA) as a filler were prepared through a modified thermally induced phase separation (TIPS) protocol. A suspension was prepared by adding sieved HA particles to a ternary poly-L-lactic-acid (PLLA)/dioxane/water solution with a well defined solvent (dioxane) to non-solvent (water) ratio. A demixing protocol based on a well-defined temperature vs. time path was followed in order to achieve a porous structure with an architecture suitable for osteochondral defect repair. This result is obtained by imposing also a spatial gradient of hydroxyapatite on the scaffolds. Scaffolds were characterized via Scannning Electron Microscopy (SEM) and Micro-computed tomography (Micro-CT). Moreover, preliminary cell culture tests in static and dynamic conditions were successfully carried out.

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 the composite scaffolds in order to evaluate the effect of HA on the proliferation and differentiation of osteoblastic cells, showing a higher alkaline phosphatase activity on composite scaffolds compared to neat PLLA ones.

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 architecture interacting with the phase separation process. Bioactivity tests confirmed the formation of a hydroxycarbonateapatite-layer in both types of BGs (detected via scanning electron microscopy, X-ray diffractometry and Fourier Transform Infrared Spectroscopy). Overall, the results showed that 1393BG composition affects the experimental preparation protocol to a minimal extent thus allowing a better control of the scaffold’s morphology compared to 45S5BG.

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 bioactive coatings which enhance corrosion resistance and biocompatibility of the substrate.

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 morphology resulted highly inhomogeneous. This behavior could be related to a change in equilibrium conditions occurring close to PLA cloud point. The equilibrium measurements performed gave out results in line with this hypothesis.

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.

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 penetration of the gel into the porous wall of the scaffold was observed. The in vitro endothelial cell cultures carried out in the scaffolds highlighted a faster cell proliferation inside the hybrid scaffold with respect to simple PLLA scaffold. Results show that the fibrin/PLLA hybrid scaffold may be favourably used for Vascular Tissue Engineering applications.

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 can be advantaged by physical-mathematical descriptions (modeling) in order to clarify the involved phenomena from the preparative step of dosage systems to the description of drug-body interactions, which allows improving the design of delivery systems/processes/therapies. This review analyzes a few mathematical modeling approaches currently adopted to describe the siRNAs delivery, the main procedures in siRNAs vectors’ production processes and siRNAs vectors’ release from hydrogels, and the modeling of pharmacokinetics of siRNAs vectors. Furthermore, the use of physical models to study the siRNAs vectors’ fate in blood stream and in the tissues is presented. The general view depicts a framework maybe not yet usable in therapeutics, but with promising possibilities for forthcoming applications.

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 analysis have been carried out respectively to study scaffold morphology and the influence of PHEA-PLA on PLLA crystallization, while DMF extraction has been carried out in order to quantify PHEA-PLA into the final scaffolds. To evaluate scaffold biodegradability, a hydrolysis study has been performed until 56 days by incubating systems in a media mimicking physiological environment (pH 7.4). Results obtained have highlighted a progressive increase in weight loss with time in PLLA/PHEA-PLA scaffolds, conceivably due to the presence of PHEA-PLA and polymers interpenetration. Viability and adhesion of bovine chondrocytes seeded on the scaffolds have been studied by MTS test and SEM analysis. From results achieved it appears that the presence of PHEA-PLA increases cells affinity, allowing a faster adhesion and proliferation inside the scaffold.

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 evaluated. The E. coli was eradicated from the whole matrix after just 5 min of exposure to supercritical CO2 at 10 MPa and 40 °C. The spores required more severe conditions due to their different and very resistant structures since complete inactivation was obtained after 360 min of exposure at 30 MPa and 40 °C or at 30 MPa and 30 °C after the addition of 200 ppm of H2O2 to near critical CO2. The treatment with dense CO2 did not alter the biocompatibility and the structure of the scaffold as demonstrated by biological culture tests and calorimetric and SEM analyses. Collected data suggest that dense CO2 is a promising alternative to conventional sterilization techniques to sterilize biodegradable PLLA scaffolds.

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 characterized by means of several techniques. The morphology was investigated by scanning electron microscopy, the elemental composition was obtained by X-ray energy dispersive spectroscopy, whilst the structure was identified by Raman spectroscopy and X-ray diffraction. BS was deposited at all investigated temperatures covering the 316LSS surface. At low and moderate temperature, BS coatings were compact, uniform and with good crystalline degree. On BS layers, HA crystals were obtained at 50 °C for all deposition times, while at 25 °C, its presence was revealed only after long deposition time. Electrochemical studies show remarkable improvement in corrosion resistance.

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 (wall thickness, porosity, and average pore dimension), simply by varying some experimental parameters. Preliminary in vitro cell culture tests were carried out inside the scaffold. The results showed that cells are able to grow within the internal surface of the scaffolds and after 3 weeks they begin to form a “primordial” vessel-like structure. Modeling predictions of the dip-coating process display always an underestimate of experimental data (dependence of wall thickness upon extraction rate). In 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 lm 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 (wall thickness, porosity, and average pore dimension), simply by varying some experimental parameters. Preliminary in vitro cell culture tests were carried out inside the scaffold. The results showed that cells are able to grow within the internal surface of the scaffolds and after 3 weeks they begin to form a ‘‘primordial’’ vessel-like structure. Modeling predictions of the dipcoating process display always an underestimate of experimental data (dependence of wall thickness upon extraction rate).

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 into a three-dimensional matrix, prepared by Thermally Induced Phase Separation (TIPS), characterized by a high porosity (about 95%) and pore size adequate to accommodate tumor cells and/or mesenchymal cells. The morphology of the multifunctional scaffolds is easy-tunable in terms of pore size, porosity and thickness and therefore adaptable to various cell or tissue types. At the same time, a double flow bioreactor was designed and built up, in order to be able to carry out biological tests on the scaffold under dynamic conditions. The device allows a separate control of the two flows (one for the tubular scaffold, one for the porous matrix) through the scaffolds. Preliminary characterizations and tests carried out suggest the presented system as a candidate to suitably “in vitro” assess the effects of different drugs on various cell populations.

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), will produce a skinless membrane, and offers the chance to control the morphology of both membrane surfaces. These results underline the importance of post-treatment stage in membrane preparation via phase separation, thus suggesting that membrane washing/drying stage will affect the final morphology. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 42151.

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 buffered formaldehyde for 24 hours, embedded in parraffin blocks, and cut into 8-μm-thick sections. The other half was fixed with 4% glutaraldehyde, dehydrated, gold coated, and was then observed with an scanning electron microscope (FEGESEM QUANTA 200 FEI). The anatomopathological examination of the implant site revealed no significant inflammatory response. The histological analysis revealed a thin fibrous capsule and a minimal inflammatory reaction with polymorphonuclear leukocytes, lymphocytes, monocytes and plasma cells around the implant, which died out during the second week. The presence of neutrophils and lymphocytes was detected until the second week after the implantation and a trama of connective tissue was found on the pores of the scaffold during the first week. The presence of giant cells was detected in the final stage of the inflammatory response and healing, which mainly occurred during the second week after the implantation. In addition, a significant angiogenic activity on the external implant surface was observed during the third week. 8 weeks after the implant, the inflammatory cell response was absolutely minimal, with an intense angiogenic activity occurring within the scaffold. This study shows the following results: PLLA has a high degree of biocompatibility and angiogenic potential; moreover, scaffolds are biocompatible in terms of cell survival and proliferation, as they stimulate a minimal inflammatory response while supporting cell infiltration, the matrix apposition and neo vascularization., Italian Journal of Anatomy and Embryology, Vol 119, No 1 (Supplement) 2014

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 pores at higher holding times. Conversely, the porous structures attained when holding the sample above the cloud point exhibited open pores with dimensions independent of time, denoting a phase separation occurring during sample freezing. Finally, the influence of the cooling rate on final morphology was investigated, showing a clear correlation with microstructure and pore size. © 2014 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2014, 52, 979–983

In vitro cultured endothelial cells forming a continuous monolayer establish stable cell-cell contacts and acquire a "resting" phenotype; on the other hand, when growing in sparse conditions these cells acquire a migratory phenotype and invade the empty area of the culture. Culturing cells in different conditions, we compared expression and clustering of proteolytic enzymes in cells having migratory versus stationary behavior. In order to observe resting and migrating cells in the same microscopic field, a continuous cell monolayer was wounded. Increased expression of proteolytic enzymes was evident in cell membranes of migrating cells especially at sprouting sites and in shed membrane vesicles. Gelatin zymography and western blotting analyses confirmed that in migrating cells, expression of membrane-bound and of vesicle-associated proteolytic enzymes are increased. The enzymes concerned include MMP-2, MMP-9, MT1-MMP, seprase, DPP4 (DiPeptidyl Peptidase 4) and uPA. Shed membrane vesicles were shown to exert degradative activity on ECM components and produce substrates facilitating cell migration. Vesicles shed by migrating cells degraded ECM components at an increased rate; as a result their effect on cell migration was amplified. Inhibiting either Matrix Metallo Proteases (MMPs) or Serine Integral Membrane Peptidases (SIMPs) caused a decrease in the stimulatory effect of vesicles, inhibiting the spontaneous migratory activity of cells; a similar result was also obtained when a monoclonal antibody acting on DPP4 was tested. We conclude that proteolytic enzymes have a synergistic stimulatory effect on cell migration and that their clustering probably facilitates the proteolytic activation cascades needed to produce maximal degradative activity on cell substrates during the angiogenic process.

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 prepare scaffolds of PLLA/PLA via TIPS. Moreover, the PLA seems to influences the TIPS process in terms of demixing temperatures. The data confirm that the morphology and the mechanical properties of the scaffold can be tuned, starting from PLLA blends and using PLA with different molecular weights.

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 analysis was carried out on the as-obtained scaffold, so as to obtain information about their thermal properties (enthalpy of melting and crystallization). The results showed that is possible to prepare scaffolds of poly-l-lactic acid/polylactic acid via thermally induced phase separation with both polylactic acids and to tune their average pore size (from 40 to 70 µm) by changing some experimental parameters (e.g. demixing temperature). Moreover, the average molecular weight of the polylactic acid in the blend seems to influence the thermally induced phase separation process in terms of demixing temperatures, which resulted higher than pure poly-l-lactic acid for the blends containing the high molecular weight polylactic acid, and lower for the blends containing the low molecular weight polylactic acid. Finally, a decrease in the crystallinity of the foams when increasing polylactic acid content in poly-l-lactic acid/polylactic acid blends was observed, as witnessed by a drop in the enthalpy of melting and crystallization. The results confirm that the morphology and the mechanical properties of the scaffold can be tuned up, starting from poly-l-lactic acid and blending it in different proportions with polylactic acid with different molecular weights.

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 been reported in the literature. In this work a copolymer (PHEA‐PLLA) of the aforementioned polymers was synthesized and characterized the possibility to produce porous scaffolds with it was assessed. The results have shown that is possible to prepare scaffolds of PHEA‐PLLA via Thermally Induced Phase Separation (TIPS). The scaffolds as‐obtained present on theirs bulk an open porous structure with interconnected pores whose average pore size was ∼ 20 μm. Moreover a cloud point curve for the system PHEA‐PLLA/Dioxane/water was built in order to find the more appropriate temperatures for the TIPS process.