Viviana De Pasquale

Publications of Viviana De Pasquale

• Contribution of glycosaminoglycans to the microstructural integrity of fibrillar and fiber crimps in tendons and ligaments.

  Authors: Marco Franchi, Viviana De Pasquale, Désirée Martini, Marilisa Quaranta, Maria Macciocca, Alessio Dionisi, Vittoria Ottani

  TheScientificWorldJournal. 01/2010; 10:1932-40.

  The biomechanical roles of both tendons and ligaments are fulfilled by the extracellular matrix of these tissues. In particular, tension is mainly transmitted and resisted by protein (collagen,The biomechanical roles of both tendons and ligaments are fulfilled by the extracellular matrix of these tissues. In particular, tension is mainly transmitted and resisted by protein (collagen, elastin) fibers, whereas compression is opposed by water-soluble glycosaminoglycans (GAGs). GAGs spanning the interfibrillar spaces and interacting with fibrils through the interfibrillar proteoglycans also seem to play a part in transmitting and resisting tensile stresses. Both tendons and ligaments showing similar composition, but different functional roles and collagen array, exhibit periodic undulations of collagen fibers or crimps. Each crimp is composed of many knots of each single fibril or fibrillar crimps. Fibrillar and fiber crimps play a mechanical role in absorbing the initial loading during elongation of both tendons and ligaments, and in recoiling fibrils and fibers when tissues have to return to their original length. This study investigated whether GAGs covalently attached to proteoglycan core proteins directly affect the 3D microstructural integrity of fibrillar crimp regions and fiber crimps in both tendons and ligaments. Achilles tendons and medial collateral ligaments of the knee from eight female Sprague-Dawley rats (90 days old) incubated in a chondroitinase ABC solution to remove GAGs were observed under a scanning electron microscope (SEM). In addition, isolated fibrils of these tissues obtained by mechanical disruption were analyzed by a transmission electron microscope (TEM). Both Achilles tendons and medial collateral ligaments of the rats after chemical or mechanical removal of GAGs still showed crimps and fibrillar crimps comparable to tissues with a normal GAG content. All fibrils in the fibrillar crimp region always twisted leftwards, thus changing their running plane, and then sharply bent, changing their course on a new plane. These data suggest that GAGs do not affect structural integrity or fibrillar crimp functions that seem mainly related to the local fibril leftward twisting and the alternating handedness of collagen from a molecular to a supramolecular level.

• Correlative microscopy of bone in implant osteointegration studies.

  Authors: Alessandra Trirè, Désirée Martini, Ester Orsini, Marco Franchi, Viviana De Pasquale, Beatrice Bacchelli, Mario Raspanti, Alessandro Ruggeri, Vittoria Ottani

  TheScientificWorldJournal. 01/2010; 10:2238-47.

  Routine morphological analyses usually include investigations by light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Each of these techniquesRoutine morphological analyses usually include investigations by light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Each of these techniques provides specific information on tissue morphology and all the obtained results are then combined to give an in-depth morphological overview of the examined sample. The limitations of this traditional comparative microscopy lie in the fact that each technique requires a different experimental sample, so that many specimens are necessary and the combined results come from different samples. The present study describes a technical procedure of correlative microscopy, which allows us to examine the same bone section first by LM and then, after appropriate processing, by SEM or TEM. Thanks to the possibility of analyzing the same undecalcified bone sections both by LM and SEM, the approach described in the present study allows us to make very accurate evaluations of old/new bone morphology at the bone-implant interface.

• Electron microscopy of the remodelling process in hamstring tendon used as ACL graft.

  Authors: Stefano Zaffagnini, Viviana De Pasquale, Leonardo Marchesini Reggiani, Alessandro Russo, Patrizia Agati, Beatrice Bacchelli, Maurilio Marcacci

  Knee surgery, sports traumatology, arthroscopy : official journal of the ESSKA. 09/2009;

  The purpose of the present study was to make a histological analysis of the remodelling process of hamstring tendon graft used as Anterior Cruciate Ligament (ACL). The hamstring graft of eightThe purpose of the present study was to make a histological analysis of the remodelling process of hamstring tendon graft used as Anterior Cruciate Ligament (ACL). The hamstring graft of eight patients was biopsied at different follow-up times from 1 to 10 years. The specimens were analysed with transmission electron microscopy (TEM) at ultrastructural level comparing them with a native ACL and a native hamstring graft. The hamstring graft was found to undergo ultrastructural changes in terms of number and diameter of fibrils with the major changes occurring in the first 2 years. At longer times after surgery (48 and 120 months) no important further changes were evident and the ultrastructure did not vary substantially from 2 to 10 years. In conclusion, the hamstring tendon used as ACL graft undergoes a transformation process but does not match the ultrastructure pattern of a normal ACL up to 10 years.

• Influence of a zirconia sandblasting treated surface on peri-implant bone healing: An experimental study in sheep.

  Authors: Beatrice Bacchelli, Gianluca Giavaresi, Marco Franchi, Désirée Martini, Viviana De Pasquale, Alessandra Trirè, Milena Fini, Roberto Giardino, Alessandro Ruggeri

  Acta biomaterialia. 02/2009;

  A sandblasting process with round zirconia (ZrO(2)) particles might be an alternative surface treatment to enhance the osseointegration of titanium dental implants. Our previous study on sheepA sandblasting process with round zirconia (ZrO(2)) particles might be an alternative surface treatment to enhance the osseointegration of titanium dental implants. Our previous study on sheep compared smooth surface titanium implants (control) with implant surfaces sandblasted with two different granulations of ZrO(2). As the sandblasted surfaces proved superior, the present study further compared the ZrO(2) surface implant with other surface treatments currently employed: machined titanium (control), titanium oxide plasma sprayed (TPS) and alumina sandblasted (Al-SL) at different times after insertion (2, 4 and 12weeks). Twelve sheep were divided into three groups of four animals each and underwent implant insertion in tibia cortical bone under general anaesthesia. The implants with surrounding tissues were subjected to histology, histomorphometry, scanning electron microscopy and microhardness tests. The experimentation indicated that at 2weeks Zr-SL implants had the highest significant bone ingrowth (p<0.05) compared to the other implant surfaces, and a microhardness of newly formed bone inside the threads significantly higher than that of Ti. The present work shows that the ZrO(2) treatment produces better results in peri-implant newly formed bone than Ti and TPS processing, whereas its performance is similar to the Al-SL surface treatment.

• Histological and ultrastructural evaluation of Leeds-Keio ligament 20 years after implant: a case report.

  Authors: Stefano Zaffagnini, Giulio Marcheggiani Muccioli, Vikram Chatrath, Alice Bondi, Viviana De Pasquale, Desiree Martini, Beatrice Bacchelli, Maurilio Marcacci

  Knee surgery, sports traumatology, arthroscopy : official journal of the ESSKA. 10/2008;

  We were capable of undertaking a histological and ultrastructural evaluation of an intact Leeds-Keio ligament implanted 20 years ago to assess the neoligamentization process inside this artificialWe were capable of undertaking a histological and ultrastructural evaluation of an intact Leeds-Keio ligament implanted 20 years ago to assess the neoligamentization process inside this artificial ligament. The histological evaluation disclosed a collagen fibrils orientation very close to the structure of a normal anterior cruciate ligament (ACL) where the collagen fibres are multidirectional [Strocchi et al. in J Anat 180(3):515-519, 1992]. On the other hand we found an unimodal distribution of collagen fibrils in the reconstructed ACL. This suggests that even at long-term follow-up stress exerts a variable influence. The multidirectional arrangement of collagen fibres resembles a normal ACL, but the unimodal distribution of fibrils is quite different from those seen in normal tendon and ligaments which tend to have a bimodal peak [Decker et al. in J Submicrosc Cytol Pathol 23:9-21, 1991; Strocchi et al. in J Anat 180(3):515-519, 1992]. Studies based on biopsy suffer from the potential weakness that the specimen may not have been representative of the entire prosthesis. Further long-term studies, possibly with the entire prosthesis and not only a biopsy, would highlight the behaviour and remodelling of this artificial ligament in greater detail and could be important for the development of future generations of artificial ligaments or tissue engineering ACL reconstruction.

• Effects of freezing on the biomechanical and structural properties of human posterior tibial tendons.

  Authors: Sandro Giannini, Roberto Buda, Francesco Di Caprio, Patrizia Agati, Adriana Bigi, Viviana De Pasquale, Alessandro Ruggeri

  International orthopaedics. 05/2008; 32(2):145-51.

  This work analyzes the effects of storage by fresh-freezing at -80 degrees C on the histological, structural and biomechanical properties of the human posterior tibial tendon (PTT), used for ACLThis work analyzes the effects of storage by fresh-freezing at -80 degrees C on the histological, structural and biomechanical properties of the human posterior tibial tendon (PTT), used for ACL reconstruction. Twenty-two PTTs were harvested from eleven donors. For each donor one tendon was frozen at -80 degrees C and thawed in physiological solution at 37 degrees C, and the other was tested without freezing (control). Transmission electron microscopy (TEM), differential scanning calorimetry (DSC) and biomechanical analysis were performed. We found the following mean changes in frozen-thawed tendons compared to controls: TEM showed an increase in the mean diameter of collagen fibrils and in fibril non-occupation mean ratio, while the mean number of fibrils decreased; DSC showed a decrease in mean denaturation temperature and denaturation enthalpy. Biomechanical analysis showed a decrease in ultimate load and ultimate stress, an increase in stiffness and a decrease in ultimate strain of tendons. In conclusion fresh-freezing brings about significant changes in the biomechanical and structural properties of the human PTT. A high variability exists in the biophysical properties of tendons among individuals and in the effects of storage on tendons. Therefore, when choosing an allograft tendon, particular care is needed to choose a biomechanically suitable graft.

• Different crimp patterns in collagen fibrils relate to the subfibrillar arrangement.

  Authors: Marco Franchi, Mario Raspanti, Carlo Dell'Orbo, Marilisa Quaranta, Viviana De Pasquale, Vittoria Ottani, Alessandro Ruggeri

  Connective tissue research. 02/2008; 49(2):85-91.

  Collagen fibril ultrastructure and course were examined in different connective tissues by PLM, SEM, TEM, and AFM. In tendons, collagen fibrils were large and heterogeneous with a straightCollagen fibril ultrastructure and course were examined in different connective tissues by PLM, SEM, TEM, and AFM. In tendons, collagen fibrils were large and heterogeneous with a straight subfibrillar arrangement. They ran densely packed, parallel, and straight changing their direction only in periodic crimps where fibrils showed a local deformation (fibrillar crimps). Other tissues such as aponeurosis, fascia communis, skin, aortic wall, and tendon and nerve sheaths showed thinner uniform fibrils with a helical subfibrillar arrangement. These fibrils appeared in parallel or helical arrangement following a wavy, undulating course. Ligaments showed large fibrils as in tendon, with fibrillar crimps but less packed. Thinner uniform-sized fibrils also were observed. Fibrillar crimps seem to be related to the subfibrillar arrangement being present only in large fibrils with a straight subfibrillar arrangement. These stiffer fibrils respond mainly to unidirectional tensional forces, whereas the flexible thinner fibrils with helical subfibrils can accommodate extreme curvatures without harm, thus responding to multidirectional loadings.

• Influence of different implant surfaces on peri-implant osteogenesis: histomorphometric analysis in sheep.

  Authors: Marco Franchi, Beatrice Bacchelli, Gianluca Giavaresi, Viviana De Pasquale, Désirée Martini, Milena Fini, Roberto Giardino, Alessandro Ruggeri

  Journal of periodontology. 06/2007; 78(5):879-88.

  BACKGROUND: The present study investigated peri-implant osteogenesis and implant biologic fixation in different zirconia sandblasted endosseous titanium surfaces (SLA-60 and SLA-120) and a turnedBACKGROUND: The present study investigated peri-implant osteogenesis and implant biologic fixation in different zirconia sandblasted endosseous titanium surfaces (SLA-60 and SLA-120) and a turned titanium surface (T) 2 and 4 weeks after surgery. METHODS: Seventy-two implant screws were implanted in tibia of six sheep. Histologic sections of implants (2 and 4 weeks after surgery) were analyzed with light microscopy for histomorphometric analysis of bone-to-implant contact (BIC), bone ingrowth (BI), and bone surface (BS/BV). Histologic blocks were used to perform bone microhardness studies next to the implants. Some implants were also observed with scanning electron microscopy (SEM) and transmission electron microscopy (TEM). RESULTS: In general, the highest values of BIC, BI, BS/BV, and Vickers hardness number (HV) were measured in SLA-60 samples, followed by SLA-120 and T implants. Two weeks after surgery, all the implants appeared biologically fixed by a newly formed woven bone arranged in thin bone trabeculae and filling the gap between implant and host bone. Four weeks after implantation, the thickness of the woven bone trabeculae had increased, especially around the SLA-60 and SLA-120 implants by a gradual deposition of parallel-fiber bone. CONCLUSIONS: Our results suggest that, in the early period of peri-implant healing, the implant surface morphology that seemed to influence the increase of peri-implant osteogenesis, bone turnover, and peri-implant bone maturation was SLA-60. We suggest that this surface, characterized by moderately deep titanium cavities very similar to the osteocyte lacunae, could act as a microscopic scaffold for mesenchymal and/or osteoblast-like cells adhesion.

• Crimp morphology in relaxed and stretched rat Achilles tendon.

  Authors: Marco Franchi, Milena Fini, Marilisa Quaranta, Viviana De Pasquale, Mario Raspanti, Gianluca Giavaresi, Vittoria Ottani, Alessandro Ruggeri

  Journal of anatomy. 02/2007; 210(1):1-7.

  Fibrous extracellular matrix of tendon is considered to be an inextensible anatomical structure consisting of type I collagen fibrils arranged in parallel bundles. Under polarized light microscopyFibrous extracellular matrix of tendon is considered to be an inextensible anatomical structure consisting of type I collagen fibrils arranged in parallel bundles. Under polarized light microscopy the collagen fibre bundles appear crimped with alternating dark and light transverse bands. This study describes the ultrastructure of the collagen fibrils in crimps of both relaxed and in vivo stretched rat Achilles tendon. Under polarized light microscopy crimps of relaxed Achilles tendons appear as isosceles or scalene triangles of different size. Tendon crimps observed via SEM and TEM show the single collagen fibrils that suddenly change their direction containing knots. The fibrils appear partially squeezed in the knots, bent on the same plane like bayonets, or twisted and bent. Moreover some of them lose their D-period, revealing their microfibrillar component. These particular aspects of collagen fibrils inside each tendon crimp have been termed 'fibrillar crimps' and may fulfil the same functional role. When tendon is physiologically stretched in vivo the tendon crimps decrease in number (46.7%) (P<0.01) and appear more flattened with an increase in the crimp top angle (165 degrees in stretched tendons vs. 148 degrees in relaxed tendons, P<0.005). Under SEM and TEM, the 'fibrillar crimps' are still present, never losing their structural identity in straightened collagen fibril bundles of stretched tendons even where tendon crimps are not detectable. These data suggest that the 'fibrillar crimp' may be the true structural component of the tendon crimp acting as a shock absorber during physiological stretching of Achilles tendon.

• Structure relates to elastic recoil and functional role in quadriceps tendon and patellar ligament

  Authors: Marco Franchi, Marilisa Quaranta, Maria Macciocca, Viviana De Pasquale, Vittoria Ottani, Alessandro Ruggeri

  Micron.

  Tendons and ligaments have similar but slightly different structure and composition. Crimps of tendons and ligaments are morphological structures related to the elastic functional properties of theseTendons and ligaments have similar but slightly different structure and composition. Crimps of tendons and ligaments are morphological structures related to the elastic functional properties of these connective tissues. Aim of this study was to investigate the morphological arrangement of collagen fibres, fibrils and crimping pattern of suprapatellar (rectus femoris tendon-RFT and vastus intermedius tendon-VIT) and infrapatellar connective tissues (patellar ligament-PL) to relate their structural aspects to their common function role of leg extension. RFT, VIT and PL were removed from knees of Sprague–Dawley rats and light and electron microscopy (TEM and SEM) performed. Sagittal sections showed that collagen array and crimping pattern were similar in RFT and PL but differed from VIT. Morphometric analysis confirmed that crimp number was about the same in RFT and PL (5.4 ± 1.4 and 6.1 ± 2.8 respectively), but it was almost three times higher in VIT (14.5 ± 4.7). Similarly crimp top angle in RFT and PL (141.5 ± 15.0° and 146.2 ± 12.2° respectively) was significantly higher than in VIT (122.3 ± 14.8°) and the crimp base length was more than twice as wide in RFT (75.5 ± 22.6 μm) and PL (72.3 ± 28.9 μm) than in VIT (36 ± 14.1 μm). The smaller, fewer and most crimped crimps in VIT show that this tendon has a greater elastic recoil and responds to higher forces as among quadriceps muscles the vastus intermedius belly contributes the most during knee extension. By contrast, RFT acting as a “stopper” tendon also plays a ligament role by limiting an excessive flexion of the joint during postural rest position of the knee.