Publications (40)111.46 Total impact
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Article: Preliminary Report of In Vitro Reconstruction of a Vascularized Tendonlike Structure: A Novel Application for Adipose-Derived Stem Cells.
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ABSTRACT: INTRODUCTION: A greater supply of tendinous tissue can be obtained through tissue engineering technology with increasing application of adult stem cells. It is well known that adipose-derived stem cells (ADSCs), found in abundance in adipose tissue, have the same differentiating capacity as mesenchymal stem cells yet have the advantage of being easily isolated. In the present study, we combined the great facility of ADSCs to differentiate with the application of an external mechanical stimulus to successfully create an in vitro reconstructed tendonlike structure with a microcapillary network. MATERIALS AND METHODS: Hyalonect meshes (Fidia Advanced Biopolymers, Abano Terme, Padova, Italy) were used as scaffold. Human ADSCs were seeded onto the biomaterials, and the cell/scaffold constructs were cultured under mechanical stress for up to 15 days. Human tenocytes were used in the same conditions as control. Performance was assessed by histology, immunochemistry, ultrastructure, and biomolecular analysis. RESULTS: Adipose-derived stem cells seeded onto Hyalonect adhered and differentiated along the entire surface of the biomaterial and began to infiltrate within its structure. Subsequently, endothelial cells migrated, forming a capillary in the new extracellular matrix. CONCLUSIONS: This technique allowed for the creation of a vascularized tendon equivalent that could easily be detached from the bioreactor, thus facilitating its implant at the lesion site. These results highlight the biologic performance of biodegradable hyaluronic acid-based (HYAFF-11) scaffolds, which were shown to be suitable for deposition of the autologous extracellular matrix critical for ADSCs differentiation.Annals of plastic surgery 02/2013; · 1.29 Impact Factor -
Article: Hyaluronic Acid Induces Activation of the κ-Opioid Receptor.
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ABSTRACT: Nociceptive pain is one of the most common types of pain that originates from an injury involving nociceptors. Approximately 60% of the knee joint innervations are classified as nociceptive. The specific biological mechanism underlying the regulation of nociceptors is relevant for the treatment of symptoms affecting the knee joint. Intra-articular administration of exogenous hyaluronic acid (HA) in patients with osteoarthritis (OA) appears to be particularly effective in reducing pain and improving patient function. We performed an in vitro study conducted in CHO cells that expressed a panel of opioid receptors and in primary rat dorsal root ganglion (DRG) neurons to determine if HA induces the activation of opioid peptide receptors (OPr) using both aequorin and the fluorescent dye Fura-2/AM. Selective agonists and antagonists for each OPr expressed on CHO cells were used to test the efficacy of our in vitro model followed by stimulation with HA. The results showed that HA induces stimulatory effects on the κ receptor (KOP). These effects of HA were also confirmed in rat DRG neurons, which express endogenously the OPr. HA activates the KOP receptor in a concentration dependent manner, with a pEC(50) value of 7.57.PLoS ONE 01/2013; 8(1):e55510. · 4.09 Impact Factor -
Article: Biocompatibility Evaluation of Ionic- and Photo-Crosslinked Methacrylated Gellan Gum Hydrogels: In Vitro and In Vivo Study.
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ABSTRACT: In this study, the stability and biocompatibility of methacrylated gellan gum hydrogels, obtained either by ionic- (iGG-MA) or photo-crosslinking (phGG-MA), were evaluated in vitro and in vivo. Size exclusion chromatography analysis of the methacrylated gellan gum (GG-MA) powder revealed that molecular weight is lower as compared to the non-modified material, i.e., low acyl gellan gum. The water uptake and swelling of iGG-MA and phGG-MA hydrogels were investigated in phosphate-buffered saline solution (pH 7.4). The biocompatibility of the hydrogels was firstly evaluated by producing cell-laden hydrogels. The in vitro cells encapsulation study showed that lung fibroblast cells (L929 cells) and human intervertebral disc (hIVD) cells are viable when cultured within both hydrogels, up to 21 days of culturing. The iGG-MA and phGG-MA hydrogels were also subcutaneously implanted in Lewis rats for 10 and 18 days. Tissue response to the hydrogels implantation was determined by histological analysis (haematoxylin-eosin staining). A thin fibrous capsule was observed around the implanted hydrogels. No necrosis, calcification, and acute inflammatory reaction were observed. The results presented in this study demonstrate that iGG-MA and phGG-MA hydrogels are stable in vitro and in vivo, support L929 and hIVD cells' encapsulation and viability, and were found to be well-tolerated and non-toxic in vivo.Advanced healthcare materials. 11/2012; -
Article: In vitro response of osteoarthritic chondrocytes and fibroblast-like synoviocytes to a 500-730 kDa hyaluronan amide derivative.
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ABSTRACT: The aim of this study was to compare the effects of native hyaluronan (HA) with that of its hexadecylamide derivative (HYADD) on proliferation of fibroblast-like synoviocytes (FLS) and chondrocytes. The production of inflammatory and anti-inflammatory cytokines was also analyzed in FLS cultures. The proliferation of osteoarthritis (OA) chondrocytes was enhanced when cells were treated with 0.5-1.5 mg mL(-1) of HA or HYADD®4-G. This effect was completely suppressed by the anti-CD44 antibody. At 0.5 to 1 mg mL(-1) , HA and HYADD®4-G did not influence the proliferation of normal or pathological FLS; however, at the higher concentration (1.5 mg mL(-1) ), HYADD®4-G did significantly inhibit cell proliferation. As to effects on inflammation, a significant increase in the expression of the IL-10 gene was observed when FLS were pretreated with tumor necrosis factor alpha and then cultured in the presence of 0.5 mg mL(-1) HYADD® 4-G or HA. The effects of HA derivatives on FLS proliferation and production of anti-inflammatory cytokines indicate that they may be of therapeutic benefit in OA. The longer residence time in the joint cavity, the increased viscoelasticity, and the anti-inflammatory potential of HYADD®4-G make it a better candidate than native HA for OA therapy. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2012.Journal of Biomedical Materials Research Part B Applied Biomaterials 08/2012; 100(8):2073-81. · 2.15 Impact Factor -
Chapter: New Developments in Tissue Engineering of Microvascular Prostheses
09/2011; , ISBN: 978-953-307-609-6 -
Article: Temporary coverage of a forehead defect following tumor resection with a hyaluronic acid biological dressing: a case report.
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ABSTRACT: Coverage of large skin defects, especially following tumor resection and in patients who are not good candidates for procedures requiring general anesthesia, may require a staged procedure. The use of dermal substitutes to cover the defect until autologous grafting can be performed has been described. Hyaluronic acid biological dressings (HABD) also have been used for the temporary coverage of partial- to full-thickness posttraumatic or postsurgical wounds. An 82-year-old man with cardiopathy presented with an 8 cm x 4 cm ulcerated squamous cell carcinoma on his forehead. Following surgical removal under local anesthesia, the 12 cm x 8 cm defect was covered with HABD, which was removed immediately before the scheduled surgical closure (21 days later). At that time, well-vascularized granulation tissue completely covered the bone and an autologous skin graft was applied. Punch biopsy results obtained 4 weeks after surgery showed dressing remnants in the dermis and confirmed the graft was totally integrated with the surrounding tissues; there was no evidence of hypertrophy or excessive scarring. In this patient, the staged procedure provided an excellent alternative to a complex surgical procedure. Use of the biological dressing required only a weekly wound and dressing assessment and the cosmetic result was good without evidence of a recurrence at the 2-year follow-up. Studies to ascertain the efficacy and effectiveness of this dressing for the temporary coverage of soft tissue defects are needed.Ostomy/wound management 04/2011; 57(4):56-60. · 1.08 Impact Factor -
Article: Negative pressure therapy potential enhancement with a biointeractive hyaluronan acid scaffold interface: case report.
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ABSTRACT: The authors have indicated no significant interest with commercial supporters.Dermatologic Surgery 03/2011; 37(4):543-6. · 1.80 Impact Factor -
Article: Hyaluronic acid biodegradable material for reconstruction of vascular wall: a preliminary study in rats.
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ABSTRACT: The objective of this preliminary study was to develop a reabsorbable vascular patch that did not require in vitro cell or biochemical preconditioning for vascular wall repair. Patches were composed only of hyaluronic acid (HA). Twenty male Wistar rats weighing 250-350 g were used. The abdominal aorta was exposed and isolated. A rectangular breach (1 mm × 5 mm) was made on vessel wall and arterial defect was repaired with HA made patch. Performance was assessed at 1, 2, 4, 8, and 16 weeks after surgery by histology and immunohistochemistry. Extracellular matrix components were evaluated by molecular biological methods. After 16 weeks, the biomaterial was almost completely degraded and replaced by a neoartery wall composed of endothelial cells, smooth muscle cells, collagen, and elastin fibers organized in layers. In conclusion, HA patches provide a provisional three-dimensional support to interact with cells for the control of their function, guiding the spatially and temporally multicellular processes of artery regeneration.Microsurgery 02/2011; 31(2):138-45. · 1.61 Impact Factor -
Article: ALMS1-deficient fibroblasts over-express extra-cellular matrix components, display cell cycle delay and are resistant to apoptosis.
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ABSTRACT: Alström Syndrome (ALMS) is a rare genetic disorder (483 living cases), characterized by many clinical manifestations, including blindness, obesity, type 2 diabetes and cardiomyopathy. ALMS is caused by mutations in the ALMS1 gene, encoding for a large protein with implicated roles in ciliary function, cellular quiescence and intracellular transport. Patients with ALMS have extensive fibrosis in nearly all tissues resulting in a progressive organ failure which is often the ultimate cause of death. To focus on the role of ALMS1 mutations in the generation and maintenance of this pathological fibrosis, we performed gene expression analysis, ultrastructural characterization and functional assays in 4 dermal fibroblast cultures from ALMS patients. Using a genome-wide gene expression analysis we found alterations in genes belonging to specific categories (cell cycle, extracellular matrix (ECM) and fibrosis, cellular architecture/motility and apoptosis). ALMS fibroblasts display cytoskeleton abnormalities and migration impairment, up-regulate the expression and production of collagens and despite the increase in the cell cycle length are more resistant to apoptosis. Therefore ALMS1-deficient fibroblasts showed a constitutively activated myofibroblast phenotype even if they do not derive from a fibrotic lesion. Our results support a genetic basis for the fibrosis observed in ALMS and show that both an excessive ECM production and a failure to eliminate myofibroblasts are key mechanisms. Furthermore, our findings suggest new roles for ALMS1 in both intra- and extra-cellular events which are essential not only for the normal cellular function but also for cell-cell and ECM-cell interactions.PLoS ONE 01/2011; 6(4):e19081. · 4.09 Impact Factor -
Article: A combining method to enhance the in vitro differentiation of hepatic precursor cells.
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ABSTRACT: The ideal bioartificial liver should be designed to reproduce as nearly as possible in vitro the habitat that hepatic cells find in vivo. In the present work, we investigated the in vitro perfusion condition with a view to improving the hepatic differentiation of pluripotent human liver stem cells (HLSCs) from adult liver. Tissue engineering strategies based on the cocultivation of HLSCs with hepatic stellate cells (ITO) and with several combinations of medium were applied to improve viability and differentiation. A mathematical model estimated the best flow rate for perfused cultures lasting up to 7 days. Morphological and functional assays were performed. Morphological analyses confirmed that a flow of perfusion medium (assured by the bioreactor system) enabled the in vitro organization of the cells into liver clusters even in the deeper levels of the sponge. Our results showed that, when cocultured with ITO using stem cell medium, HLSCs synthesized a large amount of albumin and the MTT test confirmed an improvement in cell proliferation. In conclusion, this study shows that our in vitro cell conditions promote the formation of clusters of HLSCs and enhance the functional differentiation into a mature hepatic population.Tissue Engineering Part C Methods 12/2010; 16(6):1543-51. · 4.64 Impact Factor -
Chapter: Hydrogels and Tissue Engineering
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ABSTRACT: Hydrogels are water-swollen polymeric materials that maintain a distinct three-dimensional structure. They were the first biomaterials designed for use in the human body. Traditional methods of biomaterial synthesis include crosslinking copolymerization, crosslinking of reactive polymer precursors, and crosslinking via polymer-polymer reaction. These methods of hydrogel synthesis were limited in the control of their detailed structure. Other inadequacies of traditional hydrogels have been poor mechanical properties and slow or delayed response times to external stimuli. The huge field of biomaterials research has received a strong revitalization by several novel approaches in hydrogel design. Enhanced biomechanical properties of hydrogel preparation, superporous and comb-type grafted hydrogels with fast response times, and self-assembly are just a few examples of hydrogel biomaterials with a smart future. Potential applications of all the types of hydrogels include: tissue engineering, synthetic extracellular matrix, implantable devices, biosensors, separation systems (valves to control permeability across porous membranes, or materials for affinity separation based on the specific recognition of monomelic strands), materials controlling the activity of enzymes, phospholipid bilayer destabilizing agents, materials controlling reversible cell attachment, nanoreactors with precisely placed reactive groups in three-dimensional space and smart microfluidics. With regard to the applications of hydrogels in recent years, particular attention has been devoted to drug delivery, clinical application as well as to the use of hydrogels as scaffolds for tissue engineering and regenerative medicine. Among the materials used for regenerative applications, hydrogels seem very promising and are receiving increasing attention due to their ability to entrap large amount of water, good biocompatibility and the ability to mimic tissue environment.07/2010: pages 1-8; -
Article: Neural potential of adipose stem cells.
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ABSTRACT: In the last few years, adipose tissue, which has been largely ignored by anatomists and physicians for centuries, has found new brightness thanks to the stem cells contained within. These adipose derived stem cells (ADSC) have the same characteristics of the mesenchymal stem cells (MSC) residing in bone marrow. They have the same cell surface markers and are capable of differentiating into the same cell types, including osteoblasts, chondrocytes, myoblasts, adipocytes, and neuron-like cells. Adipose tissue is ubiquitous and uniquely expandable. Most patients possess excess fat that can be harvested, making adipose tissue the ideal large-scale source for research on clinical applications. In this review focused on the neural potential of adipose-derived stem cells. Current strategies for their isolation, differentiation, and in vitro characterization, as well as their latest in vivo applications for neurological disorders or injury repair, were discussed.Discovery medicine 07/2010; 10(50):37-43. -
Article: Nanoscale particle therapies for wounds and ulcers.
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ABSTRACT: 'Small is beautiful' - this should be the slogan of nanoscientists. Indeed, working with particles less than 100 nm in size, nanotechnology is on the verge of providing a host of new materials and approaches, revolutionizing applied medicine. The obvious potential of nanotechnology has attracted considerable investment from governments and industry hoping to drive its economic development. Several areas of medical care already benefit from the advantages that nanotechnology provides and its application in wound healing will be reviewed in this article.Nanomedicine 06/2010; 5(4):641-56. · 5.05 Impact Factor -
Chapter: Skin Substitutes
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ABSTRACT: The many studies conducted so far reveal that Tissue Engineering of the skin is only at the beginning of its use in human applications. Burns patients were the first targets for such tissue substitutes, then chronic diseases, such as venous ulcers, have followed. The more experience is gained from the surgeon, the more feedback for the basic scientist to improve the product and to broaden clinical indications. Nowadays, progress in cell culture and biomedical material technologies have added two important spare parts: epidermis and dermis to the surgeon's toolbox which can be reconstituted in the laboratory from small biopsies of the same recipient. Other parts will follow in a few years, with the final aim to generate a full transplantable replica of the skin with adnexa and vasculature.03/2010; , ISBN: 978-953-307-079-7 -
Article: Neural potential of a stem cell population in the adipose and cutaneous tissues.
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ABSTRACT: A significant amount of recent interest has been focused on the possibility that adult human stem cells are a realistic therapeutic alternative to embryonic stem cells. Multipotent stem cells that have characteristics reminiscent of embryonic neural crest stem cells have been isolated from several postnatal tissues, including skin, gut, dental pulp and the heart, and are potentially useful for research and therapeutic purposes. However, their neurogenic potential, including their ability to produce electrophysiologically active neurons, is largely unexplored. In the present work, we investigated this issue with regard to skin-derived precursors (SKPs) and adipose-derived stem cells (ADSc) Adult stem cells isolated from skin and from adipose tissue derived from the same adult donor were treated with epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF2). Neurospheres obtained were first expanded and evaluated in term of proliferative ability, and then their neuronal differentiation potential was analysed. Adipose- and skin-derived neurospheres grew in suspension as spheres in the presence of the mitogens FGF2 and EGF. With this protocols, the spheres have been able to proliferate and to originate Schwann and glial-like cells. In summary, we have demonstrated in this work that multipotent adult precursor cell can be isolated and expanded from two accessible adult tissue sources: skin and adipose tissue. The work described in this paper provides the framework for our attempts to use SKPs or ADSc as autologous adult stem cell population for cell replacement and discovery research.Neurological Research 02/2010; 32(1):47-54. · 1.52 Impact Factor -
Article: Hyaluronan-based scaffold for in vivo regeneration of the rat vena cava: Preliminary results in an animal model.
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ABSTRACT: The aim of this study was to develop a prosthetic graft that could perform as a small-diameter vascular conduit for vein regeneration. The difficulty of obtaining significant long-term patency and good wall mechanical strength in vivo has been a significant obstacle in achieving small-diameter vein prostheses. Fifteen Male Wistar rats weighing 250-350 g were used. Tubular structures of hyaluronan (HYAFF-11 tubules, 2 mm diameter, and 1.5 cm length) were implanted in the vena cava of rats as temporary absorbable guides to promote regeneration of veins. Performance was assessed at 30, 60, and 90 days after surgery by histology (hematoxylin-eosin and Weighert solution) and immunohistochemistry (antibodies to von Willebrand factor and to Myosin Light-Chain Kinase). These experiments resulted in two novel findings: (1) sequential regeneration of vascular components led to complete vein wall regeneration 30 days after surgery; (2) the biomaterial used created the ideal environment for the delicate regeneration process during the critical initial phases, yet its biodegradability allowed for complete degradation of the construct 4 months after implantation, at which time, a new vein remained to connect the vein stumps. This work demonstrates the complete vena cava regeneration inside the hyaluronic acid-based prosthesis, opening new perspective of microsurgical applications, like replantation of the upper limb, elongation of vascular pedicle of free flaps, cardiovascular surgery, and pediatric microvascular surgery.Journal of Biomedical Materials Research Part A 10/2009; 93(4):1289-96. · 2.63 Impact Factor -
Article: Isolation method for a stem cell population with neural potential from skin and adipose tissue.
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ABSTRACT: OBJECTIVE: In recent years, research on stem cells has been focused on the development of personalized cell-based therapies. Owing to their homing properties, adult human stem cells are a promising source of autologous cells to be used as therapeutic vehicles. Multiple potential sources for clinically useful stem and progenitor cells have been identified, including autologous and allogenic embryonic, fetal and adult somatic cells from neural, adipose and mesenchymal tissue. In the present report, we describe a simple protocol to obtain an enriched culture of adult stem cells organized in neurospheres from two post-natal tissues: skin and adipose tissue. METHODS: Adult stem cells isolated from skin and adipose tissue derived from the same adult donor were amplified under varying conditions related to the coating of the chamber slide and the presence of serum and/or growth factors, such as with EGF and FGF2. Neurospheres were then expanded and evaluated in terms of proliferation and gene expression. RESULTS: Adipose and skin derived neurospheres were comparable in size, quantity of cells and genes expressed. Cells from both types of tissue grew optimally without slide coating, in the presence of serum and with the combined addition of FGF2 and EGF. DISCUSSION: We describe a method for isolating and improving a population of multipotent adult precursor cells from the two most accessible adult tissue sources: skin and adipose tissue. This autologous adult stem cell population could be used for cell replacement or cell therapies.Neurological Research 09/2009; · 1.52 Impact Factor -
Article: Hyaluronan benzyl ester as a scaffold for tissue engineering.
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ABSTRACT: Tissue engineering is a multidisciplinary field focused on in vitro reconstruction of mammalian tissues. In order to allow a similar three-dimensional organization of in vitro cultured cells, biocompatible scaffolds are needed. This need has provided immense momentum for research on "smart scaffolds" for use in cell culture. One of the most promising materials for tissue engineering and regenerative medicine is a hyaluronan derivative: a benzyl ester of hyaluronan (HYAFF). HYAFF can be processed to obtain several types of devices such as tubes, membranes, non-woven fabrics, gauzes, and sponges. All these scaffolds are highly biocompatible. In the human body they do not elicit any adverse reactions and are resorbed by the host tissues. Human hepatocytes, dermal fibroblasts and keratinocytes, chondrocytes, Schwann cells, bone marrow derived mesenchymal stem cells and adipose tissue derived mesenchymal stem cells have been successfully cultured in these meshes. The same scaffolds, in tube meshes, has been applied for vascular tissue engineering that has emerged as a promising technology for the design of an ideal, responsive, living conduit with properties similar to that of native tissue.International Journal of Molecular Sciences 07/2009; 10(7):2972-85. · 2.60 Impact Factor -
Article: Characteristics of repair tissue in second-look and third-look biopsies from patients treated with engineered cartilage: relationship to symptomatology and time after implantation.
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ABSTRACT: The present study established characteristics of tissue regrowth in patients suffering knee lesions treated with grafts of autologous chondrocytes grown on three-dimensional hyaluronic acid biomaterials. This multicentred study involved a second-look arthroscopy/biopsy, 5 to 33 months post implant (n = 63). Seven patients allowed a third-look biopsy, three of which were performed 18 months post implant. Characteristics of tissues were histologically and histochemically evaluated. The remaining bone stubs were evaluated for cartilage/bone integration. For data analysis, biopsies were further divided into those obtained from postoperative symptomatic patients (n = 41) or from asymptomatic patients (n = 22). The percentage of hyaline regenerated tissues was significantly greater in biopsies obtained after, versus within, 18 months of implantation. Differences were also observed between symptomatic and asymptomatic patients: reparative tissues taken from symptomatic patients 18 months after grafting were mainly fibrocartilage or mixed (hyaline-fibrocartilage) tissue, while tissues taken from asymptomatic patients were hyaline cartilage in 83% of biopsies. In a small group of asymptomatic patients (n = 3), second-look and third-look biopsies taken 18 months after surgery confirmed maturation of the newly formed tissue over time. Cartilage maturation occurred from the inner regions of the graft, in contact with subchondral bone, towards the periphery of the implant. The study indicates that, in asymptomatic patients after chondrocyte implantation, regenerated tissue undergoes a process of maturation that in the majority of cases takes longer than 18 months for completion and leads to hyaline tissue and not fibrous cartilage. Persistence of symptoms might reflect the presence of a nonhyaline cartilage repair tissue.Arthritis research & therapy 12/2008; 10(6):R132. · 4.27 Impact Factor -
Article: Neoarteries grown in vivo using a tissue-engineered hyaluronan-based scaffold.
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ABSTRACT: Vascular tissue engineering has emerged as a promising technology for the design of an ideal, responsive, living conduit with properties similar to that of native tissue. The missing link in tissue-engineered blood vessels is elastin biosynthesis. Several biomaterials are currently used but few support elastin biosynthesis in a 3-D array. In previous studies, we demonstrated that a hyaluronan-based scaffold (HYAFF-11) grafted in the infrarenal rat aorta successfully guided the complete regeneration of a well-functioning small-diameter (2 mm) neoartery. The aim of the present study was to test the ability of HYAFF-11 biodegradable grafts to develop into neovessels of larger size (4 mm) in a porcine model, focusing on extracellular matrix (ECM) remodeling and elastin biosynthesis. HYAFF-11 tubes (diameter 4 mm, length 5 cm) were implanted in an end-to-end fashion in the common carotid artery. Grafts were analyzed for patency with a Duplex scan every 15 days. ECM components were evaluated by histological and molecular biological methods. All the animals survived the observation period without complications. Intimal hyperplasia (initiating at the anastomotic site) and graft thrombosis led to 3 cases of partial or complete occlusion, as demonstrated by histological examination. There were no signs of stenoses or aneurysms in the remaining grafts. After 5 months, the biomaterial was almost completely degraded and replaced by a neoartery segment composed of mature smooth muscle cells, collagen, and elastin fibers organized in layers and was completely covered on the luminal surface by endothelial cells (vWF(+)). Whereas in previous small animal studies, patency rates were not optimal, those obtained in the present study using hyaluronan-based grafts of larger size confirmed the ability of these constructs to guide the development of a well-functioning neoartery, with the remarkable additional attribute of facilitating the formation of organized layers of elastin fibers.The FASEB Journal 05/2008; 22(8):2853-61. · 5.71 Impact Factor
Top co-authors
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Institutions
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2003–2013
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University of Padua
- • Department of Biomedical Sciences - DSB
- • Department of Neurosciences
Padova, Veneto, Italy
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1999–2012
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University-Hospital of Padova
Padova, Veneto, Italy
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1997
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Università degli Studi di Siena
Siena, Tuscany, Italy
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