Publications
28 results found
Ganabady K, Contessi Negrini N, Scherba JC, et al., 2023, High-throughput screening of thiol-ene click chemistries for bone adhesive polymers, ACS Applied Materials and Interfaces, Vol: 15, Pages: 50908-50915, ISSN: 1944-8244
Metal surgical pins and screws are employed in millions of orthopedic surgical procedures every year worldwide, but their usability is limited in the case of complex, comminuted fractures or in surgeries on smaller bones. Therefore, replacing such implants with a bone adhesive material has long been considered an attractive option. However, synthesizing a biocompatible bone adhesive with a high bond strength that is simple to apply presents many challenges. To rapidly identify candidate polymers for a biocompatible bone adhesive, we employed a high-throughput screening strategy to assess human mesenchymal stromal cell (hMSC) adhesion toward a library of polymers synthesized via thiol-ene click chemistry. We chose thiol-ene click chemistry because multifunctional monomers can be rapidly cured via ultraviolet (UV) light while minimizing residual monomer, and it provides a scalable manufacturing process for candidate polymers identified from a high-throughput screen. This screening methodology identified a copolymer (1-S2-FT01) composed of the monomers 1,3,5-triallyl-1,3,5-triazine-2,4,6(1H,3H,5H)-trione (TATATO) and pentaerythritol tetrakis (3-mercaptopropionate) (PETMP), which supported highest hMSC adhesion across a library of 90 polymers. The identified copolymer (1-S2-FT01) exhibited favorable compressive and tensile properties compared to existing commercial bone adhesives and adhered to bone with adhesion strengths similar to commercially available bone glues such as Histoacryl. Furthermore, this cytocompatible polymer supported osteogenic differentiation of hMSCs and could adhere 3D porous polymer scaffolds to the bone tissue, making this polymer an ideal candidate as an alternative bone adhesive with broad utility in orthopedic surgery.
Paradiso A, Volpi M, Rinoldi C, et al., 2023, <i>In vitro</i> functional models for human liver diseases and drug screening: beyond animal testing, BIOMATERIALS SCIENCE, Vol: 11, Pages: 2988-3015, ISSN: 2047-4830
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- Citations: 3
Negrini NC, Franchi A, Danti S, 2023, Biomaterial-Assisted 3D In Vitro Tumor Models: From Organoid towards Cancer Tissue Engineering Approaches, CANCERS, Vol: 15
Contessi Negrini N, Ricci C, Bongiorni F, et al., 2022, An osteosarcoma model by 3D printed polyurethane scaffold and in vitro generated bone extracellular matrix, Cancers, Vol: 14, Pages: 1-20, ISSN: 2072-6694
Osteosarcoma is a primary bone tumor characterized by a dismal prognosis, especially in the case of recurrent disease or metastases. Therefore, tools to understand in-depth osteosarcoma progression and ultimately develop new therapeutics are urgently required. 3D in vitro models can provide an optimal option, as they are highly reproducible, yet sufficiently complex, thus reliable alternatives to 2D in vitro and in vivo models. Here, we describe 3D in vitro osteosarcoma models prepared by printing polyurethane (PU) by fused deposition modeling, further enriched with human mesenchymal stromal cell (hMSC)-secreted biomolecules. We printed scaffolds with different morphologies by changing their design (i.e., the distance between printed filaments and printed patterns) to obtain different pore geometry, size, and distribution. The printed PU scaffolds were stable during in vitro cultures, showed adequate porosity (55–67%) and tunable mechanical properties (Young’s modulus ranging in 0.5–4.0 MPa), and resulted in cytocompatible. We developed the in vitro model by seeding SAOS-2 cells on the optimal PU scaffold (i.e., 0.7 mm inter-filament distance, 60° pattern), by testing different pre-conditioning factors: none, undifferentiated hMSC-secreted, and osteo-differentiated hMSC-secreted extracellular matrix (ECM), which were obtained by cell lysis before SAOS-2 seeding. Scaffolds pre-cultured with osteo-differentiated hMSCs, subsequently lysed, and seeded with SAOS-2 cells showed optimal colonization, thus disclosing a suitable biomimetic microenvironment for osteosarcoma cells, which can be useful both in tumor biology study and, possibly, treatment.
Pitton M, Fiorati A, Buscemi S, et al., 2021, 3D bioprinting of pectin-cellulose nanofibers multicomponent bioinks, Frontiers in Bioengineering and Biotechnology, Vol: 9, Pages: 1-9, ISSN: 2296-4185
Pectin has found extensive interest in biomedical applications, including wound dressing, drug delivery, and cancer targeting. However, the low viscosity of pectin solutions hinders their applications in 3D bioprinting. Here, we developed multicomponent bioinks prepared by combining pectin with TEMPO-oxidized cellulose nanofibers (TOCNFs) to optimize the inks’ printability while ensuring stability of the printed hydrogels and simultaneously print viable cell-laden inks. First, we screened several combinations of pectin (1%, 1.5%, 2%, and 2.5% w/v) and TOCNFs (0%, 0.5%, 1%, and 1.5% w/v) by testing their rheological properties and printability. Addition of TOCNFs allowed increasing the inks’ viscosity while maintaining shear thinning rheological response, and it allowed us to identify the optimal pectin concentration (2.5% w/v). We then selected the optimal TOCNFs concentration (1% w/v) by evaluating the viability of cells embedded in the ink and eventually optimized the writing speed to be used to print accurate 3D grid structures. Bioinks were prepared by embedding L929 fibroblast cells in the ink printed by optimized printing parameters. The printed scaffolds were stable in a physiological-like environment and characterized by an elastic modulus of E = 1.8 ± 0.2 kPa. Cells loaded in the ink and printed were viable (cell viability >80%) and their metabolic activity increased in time during the in vitro culture, showing the potential use of the developed bioinks for biofabrication and tissue engineering applications.
Contessi Negrini N, Sharpe P, Angelova Volponi A, et al., 2021, Tunable crosslinking and adhesion of gelatin hydrogels via bioorthogonal click chemistry, ACS Biomaterials Science and Engineering, Vol: 7, Pages: 4330-4346, ISSN: 2373-9878
Engineering cytocompatible hydrogels with tunable physico-mechanical properties as a biomimetic three-dimensional extracellular matrix (ECM) is fundamental to guide cell response and target tissue regeneration or development of in vitro models. Gelatin represents an optimal choice given its ECM biomimetic properties; however, gelatin cross-linking is required to ensure structural stability at physiological temperature (i.e., T > Tsol–gel gelatin). Here, we use a previously developed cross-linking reaction between tetrazine (Tz)- and norbornene (Nb) modified gelatin derivatives to prepare gelatin hydrogels and we demonstrate the possible tuning of their properties by varying their degree of modification (DOM) and the Tz/Nb ratio (R). The percentage DOM of the gelatin derivatives was tuned between 5 and 15%. Hydrogels prepared with higher DOM cross-linked faster (i.e., 10–20 min) compared to hydrogels prepared with lower DOM (i.e., 60–70 min). A higher DOM and equimolar Tz/Nb ratio R resulted in hydrogels with lower weight variation after immersion in PBS at 37 °C. The mechanical properties of the hydrogels were tuned by varying DOM and R by 1 order of magnitude, achieving elastic modulus E values ranging from 0.5 (low DOM and nonequimolar Tz/Nb ratio) to 5 kPa (high DOM and equimolar Tz/Nb ratio). Human dental pulp stem cells were embedded in the hydrogels and successfully 3D cultured in the hydrogels (percentage viable cells >85%). An increase in metabolic activity and a more elongated cell morphology was detected for cells cultured in hydrogels with lower mechanical properties (E < 1 kPa). Hydrogels prepared with an excess of Tz or Nb were successfully adhered and remained in contact during in vitro cultures, highlighting the potential use of these hydrogels as compartmentalized coculture systems. The successful tuning of the gelatin hydrogel properties here developed by controlling their bioorthogonal cross-linking is promising for t
Contessi Negrini N, Angelova Volponi A, Higgins CA, et al., 2021, Scaffold-based developmental tissue engineering strategies for ectodermal organ regeneration, Materials Today Bio, Vol: 10, Pages: 1-22, ISSN: 2590-0064
Tissue engineering (TE) is a multidisciplinary research field aiming at the regeneration, restoration, or replacement of damaged tissues and organs. Classical TE approaches combine scaffolds, cells and soluble factors to fabricate constructs mimicking the native tissue to be regenerated. However, to date, limited success in clinical translations has been achieved by classical TE approaches, because of the lack of satisfactory biomorphological and biofunctional features of the obtained constructs. Developmental TE has emerged as a novel TE paradigm to obtain tissues and organs with correct biomorphology and biofunctionality by mimicking the morphogenetic processes leading to the tissue/organ generation in the embryo. Ectodermal appendages, for instance, develop in vivo by sequential interactions between epithelium and mesenchyme, in a process known as secondary induction. A fine artificial replication of these complex interactions can potentially lead to the fabrication of the tissues/organs to be regenerated. Successful developmental TE applications have been reported, in vitro and in vivo, for ectodermal appendages such as teeth, hair follicles and glands. Developmental TE strategies require an accurate selection of cell sources, scaffolds and cell culture configurations to allow for the correct replication of the in vivo morphogenetic cues. Herein, we describe and discuss the emergence of this TE paradigm by reviewing the achievements obtained so far in developmental TE 3D scaffolds for teeth, hair follicles, and salivary and lacrimal glands, with particular focus on the selection of biomaterials and cell culture configurations.
Cusanno A, Contessi Negrini N, Villa T, et al., 2021, Post forming analysis and in vitro biological characterization of AZ31B processed by incremental forming and coated with electrospun polycaprolactone, Journal of Manufacturing Science and Engineering, Vol: 143, Pages: 1-11, ISSN: 1087-1357
Main problems related to the adoption of magnesium alloys for temporary orthopedic prostheses manufacturing are (i) the need of an efficient production process and (ii) the high corrosion rate compared with the bone healing time. In this work, the single-point incremental forming (SPIF) process, an effective and flexible solution for manufacturing very small batches even composed by one piece, was investigated. Tests were conducted on AZ31B-H24 sheets and were aimed at understanding the effect of temperature on the mechanical characteristics (microstructure, hardness, and roughness) of the sheet after the above-mentioned forming process and their correlation with both the corrosion rate and the cytocompatibility. In addition, after the forming process, samples processed by SPIF were coated by electrospun polycaprolactone (PCL) to reduce the corrosion rate and to further improve the cytocompatibility. Grain refinement was achieved thanks to the combined effect of temperature and strain rate during forming and finer grain size resulted to improve the magnesium corrosion resistance. In simulated body fluids, the electrospun PCL-coated samples exhibited a slower pH increase compared with uncoated samples. No indirect cytotoxic effects were detected in vitro for MC3T3-E1 cells for both coated and uncoated samples. However, cells colonization was observed only on electrospun PCL-coated samples, suggesting the importance of the polymeric coating in promoting the adhesion and survival of seeded MC3T3-E1 cells on the implant surface.
Michelini L, Probo L, Fare S, et al., 2020, Characterization of gelatin hydrogels derived from different animal sources, MATERIALS LETTERS, Vol: 272, ISSN: 0167-577X
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- Citations: 14
Contessi Negrini N, Toffoletto N, Farè S, et al., 2020, Plant tissues as 3D natural scaffolds for adipose, bone and tendon tissue regeneration, Frontiers in Bioengineering and Biotechnology, Vol: 8, Pages: 1-15, ISSN: 2296-4185
Decellularized tissues are a valid alternative as tissue engineering scaffolds, thanks to the three-dimensional structure that mimics native tissues to be regenerated and the biomimetic microenvironment for cells and tissues growth. Despite decellularized animal tissues have long been used, plant tissue decellularized scaffolds might overcome availability issues, high costs and ethical concerns related to the use of animal sources. The wide range of features covered by different plants offers a unique opportunity for the development of tissue-specific scaffolds, depending on the morphological, physical and mechanical peculiarities of each plant. Herein, three different plant tissues (i.e., apple, carrot, and celery) were decellularized and, according to their peculiar properties (i.e., porosity, mechanical properties), addressed to regeneration of adipose tissue, bone tissue and tendons, respectively. Decellularized apple, carrot and celery maintained their porous structure, with pores ranging from 70 to 420 μm, depending on the plant source, and were stable in PBS at 37°C up to 7 weeks. Different mechanical properties (i.e., Eapple = 4 kPa, Ecarrot = 43 kPa, Ecelery = 590 kPa) were measured and no indirect cytotoxic effects were demonstrated in vitro after plants decellularization. After coating with poly-L-lysine, apples supported 3T3-L1 preadipocytes adhesion, proliferation and adipogenic differentiation; carrots supported MC3T3-E1 pre-osteoblasts adhesion, proliferation and osteogenic differentiation; celery supported L929 cells adhesion, proliferation and guided anisotropic cells orientation. The versatile features of decellularized plant tissues and their potential for the regeneration of different tissues are proved in this work.
Fischetti T, Celikkin N, Contessi Negrini N, et al., 2020, Tripolyphosphate-crosslinked chitosan/gelatin biocomposite ink for 3D printing of uniaxial scaffolds., Front Bioeng Biotechnol, Vol: 8, Pages: 400-400, ISSN: 2296-4185
Chitosan is a natural polymer widely investigated and used due to its antibacterial activity, mucoadhesive, analgesic, and hemostatic properties. Its biocompatibility makes chitosan a favorable candidate for different applications in tissue engineering (TE), such as skin, bone, and cartilage tissue regeneration. Despite promising results obtained with chitosan 3D scaffolds, significant challenges persist in fabricating hydrogel structures with ordered architectures and biological properties to mimic native tissues. In this work, chitosan has been investigated aiming at designing and fabricating uniaxial scaffolds which can be proposed for the regeneration of anisotropic tissues (i.e., skin, skeletal muscle, myocardium) by 3D printing technology. Chitosan was blended with gelatin to form a polyelectrolyte complex in two different ratios, to improve printability and shape retention. After the optimization of the printing process parameters, different crosslinking conditions were investigated, and the 3D printed samples were characterized. Tripolyphosphate (TPP) was used as crosslinker for chitosan-based scaffolds. For the optimization of the printing temperature, the sol-gel temperature of the chitosan-gelatin blend was determined by rheological measurements and extrusion temperature was set to 20°C (i.e., below sol-gel temperature). The shape fidelity and surface morphology of the 3D printed scaffolds after crosslinking was dependent on crosslinking conditions. Interestingly, mechanical properties of the scaffolds were also significantly affected by the crosslinking conditions, nonetheless the stability of the scaffolds was strongly determined by the content of gelatin in the blend. Lastly, in vitro cytocompatibility test was performed to evaluate the interactions between L929 cells and the 3D printed samples. 2% w/v chitosan and 4% w/v gelatin hydrogel scaffolds crosslinked with 10% TPP, 30 min at 4°C following 30 min at 37°C have shown cytocompatible an
Negrini NC, Celikkin N, Tarsini P, et al., 2020, Three-dimensional printing of chemically crosslinked gelatin hydrogels for adipose tissue engineering, BIOFABRICATION, Vol: 12, ISSN: 1758-5082
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- Citations: 62
Contessi Negrini N, Lipreri MV, Tanzi MC, et al., 2020, In vitro cell delivery by gelatin microspheres prepared in water-in-oil emulsion, JOURNAL OF MATERIALS SCIENCE-MATERIALS IN MEDICINE, Vol: 31, ISSN: 0957-4530
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- Citations: 12
Fiorati A, Negrini NC, Baschenis E, et al., 2020, TEMPO-Nanocellulose/Ca2+ hydrogels: ibuprofen drug diffusion and In vitro cytocompatibility, Materials, Vol: 13, Pages: 1-16, ISSN: 1996-1944
Stable hydrogels with tunable rheological properties were prepared by adding Ca2+ ions to aqueous dispersions of 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO)-oxidized and ultra-sonicated cellulose nanofibers (TOUS-CNFs). The gelation occurred by interaction among polyvalent cations and the carboxylic units introduced on TOUS-CNFs during the oxidation process. Both dynamic viscosity values and pseudoplastic rheological behaviour increased by increasing the Ca2+ concentration, confirming the cross-linking action of the bivalent cation. The hydrogels were proved to be suitable controlled release systems by measuring the diffusion coefficient of a drug model (ibuprofen, IB) by high-resolution magic angle spinning (HR-MAS) nuclear magnetic resonance (NMR) spectroscopy. IB was used both as free molecule and as a 1:1 pre-formed complex with β-cyclodextrin (IB/β-CD), showing in this latter case a lower diffusion coefficient. Finally, the cytocompatibility of the TOUS-CNFs/Ca2+ hydrogels was demonstrated in vitro by indirect and direct tests conducted on a L929 murine fibroblast cell line, achieving a percentage number of viable cells after 7 days higher than 70%.
Campiglio CE, Contessi Negrini N, Fare S, et al., 2019, Cross-linking strategies for electrospun gelatin scaffolds, Materials, Vol: 12, ISSN: 1996-1944
Electrospinning is an exceptional technology to fabricate sub-micrometric fiber scaffolds for regenerative medicine applications and to mimic the morphology and the chemistry of the natural extracellular matrix (ECM). Although most synthetic and natural polymers can be electrospun, gelatin frequently represents a material of choice due to the presence of cell-interactive motifs, its wide availability, low cost, easy processability, and biodegradability. However, cross-linking is required to stabilize the structure of the electrospun matrices and avoid gelatin dissolution at body temperature. Different physical and chemical cross-linking protocols have been described to improve electrospun gelatin stability and to preserve the morphological fibrous arrangement of the electrospun gelatin scaffolds. Here, we review the main current strategies. For each method, the cross-linking mechanism and its efficiency, the influence of electrospinning parameters, and the resulting fiber morphology are considered. The main drawbacks as well as the open challenges are also discussed
Milazzo M, Negrini NC, Scialla SA, et al., 2019, Additive Manufacturing Approaches for Hydroxyapatite-Reinforced Composites, ADVANCED FUNCTIONAL MATERIALS, Vol: 29, ISSN: 1616-301X
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- Citations: 95
Negrini NC, Bonnetier M, Giatsidis G, et al., 2019, Tissue-mimicking gelatin scaffolds by alginate sacrificial templates for adipose tissue engineering, ACTA BIOMATERIALIA, Vol: 87, Pages: 61-75, ISSN: 1742-7061
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- Citations: 53
de Melo LP, Negrini NC, Fare S, et al., 2019, Thermomechanical and in vitro biological characterization of injection-molded PLGA craniofacial plates, JOURNAL OF APPLIED BIOMATERIALS & FUNCTIONAL MATERIALS, Vol: 17
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- Citations: 5
Negrini NC, Tarsini P, Tanzi MC, et al., 2019, Chemically crosslinked gelatin hydrogels as scaffolding materials for adipose tissue engineering, JOURNAL OF APPLIED POLYMER SCIENCE, Vol: 136, ISSN: 0021-8995
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- Citations: 26
Tanzi MC, Gantz D, Bertoldi S, et al., 2019, Functionalized polyurethane foams as tissue scaffolds with enhanced cellular response, ISSN: 1526-7547
Statement of Purpose: Recent advancements in Tissue Engineering ask for the development of tailored scaffolds with physicochemical cues able to promote specific and positive responses on cells of the target tissue. In the last 20 years, our research group investigated the design of polyurethane (PU) foams suitable for a use as tissue scaffolds [1-3], acquiring a strong know-how on this line of research. This work was aimed at functionalizing the PU foams with a molecule (i.e., the amino-amide diol PIME, Fig. 1, ref. [4]) predictably able to provide the foams with improved hemo-and cyto-compatibility, resistance to bacterial colonization [5] and, possibly, promote vascularization by a positive interaction with endothelial cells. PIME molecule was developed and successfully used as chain extender in the synthesis of linear polyurethanes [4,5].
Palumbo G, Cusanno A, Garcia Romeu ML, et al., 2019, Single Point Incremental Forming and Electrospinning to produce biodegradable magnesium (AZ31) biomedical prostheses coated with porous PCL, MATERIALS TODAY-PROCEEDINGS, Vol: 7, Pages: 394-401, ISSN: 2214-7853
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- Citations: 11
Gritsch L, Motta FL, Negrini NC, et al., 2018, Crosslinked gelatin hydrogels as carriers for controlled heparin release, MATERIALS LETTERS, Vol: 228, Pages: 375-378, ISSN: 0167-577X
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- Citations: 21
Borzenkov M, Moros M, Tortiglione C, et al., 2018, Fabrication of photothermally active poly(vinyl alcohol) films with gold nanostars for antibacterial applications., Beilstein J Nanotechnol, Vol: 9, Pages: 2040-2048, ISSN: 2190-4286
The unique photothermal properties of non-spherical gold nanoparticles under near-infrared (NIR) irradiation find broad application in nanotechnology and nanomedicine. The combination of their plasmonic features with widely used biocompatible poly(vinyl alcohol) (PVA) films can lead to novel hybrid polymeric materials with tunable photothermal properties and a wide range of applications. In this study, thin PVA films containing highly photothermally efficient gold nanostars (GNSs) were fabricated and their properties were studied. The resulting films displayed good mechanical properties and a pronounced photothermal effect under NIR irradiation. The local photothermal effect triggered by NIR irradiation of the PVA-GNS films is highly efficient at killing bacteria, therefore providing an opportunity to develop new types of protective antibacterial films and coatings.
Contessi Negrini N, Bonetti L, Contili L, et al., 2018, 3D printing of methylcellulose-based hydrogels, Bioprinting, Vol: 10, ISSN: 2405-8866
Methylcellulose-based (MC) hydrogels are optimal substrates to obtain cell sheets for regenerative medicine applications. However, current MC-based hydrogel preparation methods only allow for the obtainment of MC substrates with standardized and simple geometries (i.e., geometry of the container where the hydrogel is produced). Here, we propose the 3D printing of a MC-based hydrogel to obtain substrates with desired and controlled geometries. First, we optimize the printing temperature (i.e., T = 21 °C) of the MC-based hydrogel, so to obtain printed strands reproducing the designed geometry without defects. We investigate the influence of the printing parameters (i.e., needle size, deposition speed and extrusion multiplier) on the printed strands diameters and printing accuracy. A decrease in the sol-gel transition temperature was evidenced, together with an increase in water uptake at 37 °C, for printed MC-based hydrogels compared to not printed samples; the stability at 37 °C and achieved rheological properties were suitable for cell sheet engineering applications. In addition, cell viability higher than 90% was detected after embedding cells in the MC-based hydrogel; moreover, the optimized printing parameters allowed to bioprint C2C12 cells embedded in the MC-based hydrogel with a viability higher than 80%. The printing parameters we optimized could be used to produce MC substrates for cell sheet engineering or cell delivery applications with controlled and complex-shaped geometries.
Cochis A, Bonetti L, Sorrentino R, et al., 2018, 3D printing of thermo-responsive methylcellulose hydrogels for cell-sheet engineering, Materials, Vol: 11, ISSN: 1996-1944
A possible strategy in regenerative medicine is cell-sheet engineering (CSE), i.e., developing smart cell culture surfaces from which to obtain intact cell sheets (CS). The main goal of this study was to develop 3D printing via extrusion-based bioprinting of methylcellulose (MC)-based hydrogels. Hydrogels were prepared by mixing MC powder in saline solutions (Na2SO4 and PBS). MC-based hydrogels were analyzed to investigate the rheological behavior and thus optimize the printing process parameters. Cells were tested in vitro on ring-shaped printed hydrogels; bulk MC hydrogels were used for comparison. In vitro tests used murine embryonic fibroblasts (NIH/3T3) and endothelial murine cells (MS1), and the resulting cell sheets were characterized analyzing cell viability and immunofluorescence. In terms of CS preparation, 3D printing proved to be an optimal approach to obtain ring-shaped CS. Cell orientation was observed for the ring-shaped CS and was confirmed by the degree of circularity of their nuclei: cell nuclei in ring-shaped CS were more elongated than those in sheets detached from bulk hydrogels. The 3D printing process appears adequate for the preparation of cell sheets of different shapes for the regeneration of complex tissues.
Contessi N, Altomare L, Filipponi A, et al., 2017, Thermo-responsive properties of methylcellulose hydrogels for cell sheet engineering, MATERIALS LETTERS, Vol: 207, Pages: 157-160, ISSN: 0167-577X
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- Citations: 46
Angeloni V, Contessi N, De Marco C, et al., 2017, Polyurethane foam scaffold as <i>in vitro</i> model for breast cancer bone metastasis, ACTA BIOMATERIALIA, Vol: 63, Pages: 306-316, ISSN: 1742-7061
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- Citations: 45
Hejazi F, Mirzadeh H, Contessi N, et al., 2017, Novel class of collector in electrospinning device for the fabrication of 3D nanofibrous structure for large defect load-bearing tissue engineering application, JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A, Vol: 105, Pages: 1535-1548, ISSN: 1549-3296
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- Citations: 31
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