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Orthopaedic Proceedings
Vol. 87-B, Issue SUPP_I | Pages 59 - 59
1 Mar 2005
Specchia N Greco F
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The use of autologous grafts for vertebral arthrodesis is associated with a number of complications that should be properly considered: pain at the harvesting site, increased blood loss, prolonged surgical time, and additional scar. Moreover, in many cases, the amount of autologous bone is insufficient. Novel materials, either natural or synthetic, are therefore needed to be used as bone substitutes in vertebral surgery. For this purpose, a number of synthetic materials have been developed, their characteristics varying considerably in terms of ostoinduction, osteoconduction, biomecanics, and cost.

In particular, clinical and experimental studies have highlighted the potential of demineralized bone matrix (DBM), alone or in combination with autologous grafts, and of collagenic mineralized matrix (Healos).

Aim of this study was the evaluation of the clinical value of these materials in vertebral surgery. We have analyzed a series of 60 patients who underwent vertebral arthrodesis by the addition of either DBM (30 cases) or Healos (30 cases).

Bone substitutes were used both in posterior-lateral arthrodeses (on one side, the other being treated with autologous graft as a control) and in intersomatic arthrodeses in association with titanium or carbon fiber scaffolds.

Patients were followed-up for a minimum 1-year interval, and evaluated with regard to clinical (Oswestry score, SF-36) and radiographic (static and dynamic X-rays, spiral CT, MRI) parameters. The area of arthrodesis was independently analyzed by three independent observers.

Clinical results showed the reliability of both materials as a tool for a stable arthrodesis, since they were found to be able to achieve results comparable to those obtained with autologous grafts in the control arm of the study.


Orthopaedic Proceedings
Vol. 84-B, Issue SUPP_I | Pages - 18
1 Mar 2002
Pagnotta A Specchia N Gigante A Toesca A
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The changes occurring in ligamentum flavum in lumbar spine stenosis are a matter of long–standing controversy. More recently, some studies showed that the posterior spinal structures, including hypertrophied ligamentum flavum, play a major role in the pathogenesis of the lumbar stenosis.

To investigate the pathogenesis of the degenerative changes of the ligamentum flavum occurring in lumbar spine stenosis, yellow ligament cells from patients with lumbar spine stenosis were cultured for the first time and subjected to biochemical, histochemical and immunohistochemical study.

Samples of ligamentum flavum were collected from 4 patients undergoing surgery for lumbar stenosis (mean age 47.2 years). Cell cultures were obtained from each patient and maintained in Dulbecco’s modified essential medium-10% fetal calf serum. Cell characterization was histochemically (Gomori’s and von Kossa staining), immunohistochemically (anti-type I, -type II, -type III and -type X collagen, anti-S100 protein, anti-fibronectin, anti-osteonectin and anti-osteocalcin), biochemically (cAMP activity after human parathyroid hormone stimulation) assessed. Samples collected from 2 age-matched patients who underwent surgery for lumbar fractures were used as controls.

Stenotic ligamentum flavum cells expressed high levels of alkaline phosphatase activity and produced a mineralized matrix rich in type I, type III and type X collagen, fibronectin, osteonectin, and osteocalcin. Stimulation with parathyroid hormone increased intracellular cAMP concentration. These findings indicate that there was significant evidence of osteoblast-like activity in these cells. Staining for type II and type X collagen, and S-100 protein reflected the proliferation of hypertrophic chondrocyte-like cells, confirmed with the co-localization of alkaline phosphatase and collagen type II. Cultures from control patients showed nor hypertrophic chondrocytic nor osteoblastic features. Our data demonstrated the presence of hypertrophic chondrocytes with an osteoblast-like activity in human stenotic ligamentum flavum. The osteoblast-like activity could have a role in the pathophysiology of the heterotopic ossification of ligamentum flavum in lumbar spine stenosis.


Orthopaedic Proceedings
Vol. 84-B, Issue SUPP_I | Pages - 20
1 Mar 2002
Specchia N Pagnotta A Greco F
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The material most widely used in orthopaedics is hydroxyapatite (HA), anyway many differences are still present between synthetic HA and biological HA. The aim of this study was to compare adhesion, proliferation and differentiation of human osteoblast-like cells on hydroxyapatite discs with different porosity and on plastic cultures.

Human osteoblast-like cells were isolated from 4 young patients (mean age 24.5 years old), treated with collagenase and maintained in Dulbecco’s modified essential medium-10% fetal calf serum. Cells were plated on hydroxyapatite discs with 3 different porosities (35%, 35–55% e 55%) and on plastic cultures used as control. The proliferation was determined by the MTT colorimetric method, and alkaline phosphatase (ALP) activity was measured by a spettrophotometric method. Type I collagen and osteonectin production were demonstrated with fluorescence microscopy and osteoblast adhesion was studied by scanning electron microscopic (SEM) analysis. Results were analysed by one-way analysis of variance (ANOVA).

Osteoblast proliferation on HA was three- to six-fold lower then on plastic. At 28 days, 2141 (± 350) cells/well grew on the most porous disks, with highly significant differences from controls. The ALP production was 2–3 fold lower on HA than on plastic. In the most porous disks, the mean ALP activity was of 2.95 (± 0.07) UI/well after 28 days, higher than in the other two groups. The type-I collagen and the osteonectin fluorescence reaction evidenced a cytoplasmic and a matrix labeling on HA at different porosities. SEM analysis showed osteoblasts with a flattened morphology and only few of them were metabolic active.

At 21 and 28 days, proliferation rate and ALP activity on the three HA cultures were significantly different (p< 0.05). A decrease in cell population and increased ALP activity were observed on the most porous material, and high proliferation and poor differentiation rates on the less porous disks.


The Journal of Bone & Joint Surgery British Volume
Vol. 75-B, Issue 1 | Pages 141 - 147
1 Jan 1993
Sanguinetti C Specchia N Gigante A de Palma L Greco F

Four cases are described of solitary spinal neurofibroma, a rare tumour of the spinal cord or nerve roots. Computerised tomography provided an accurate topographical definition of the tumour. Magnetic resonance imaging showed an increased T2-weighted signal and multiple areas of decreased T1- and T2-weighted signals centrally. The MR signals matched the histological examination which showed hyperplastic interfascicular connective tissue, pleomorphic cells, and tightly packed nerve fibres compressed by the surrounding loose connective tissue. Electron microscopy showed three types of cell: Schwann cells, fibroblast-like cells, and mast cells. The histological findings suggests that solitary spinal neurofibroma is a distinct pathological entity which could be diagnosed preoperatively from the MR images.


The Journal of Bone & Joint Surgery British Volume
Vol. 72-B, Issue 3 | Pages 475 - 479
1 May 1990
Sanguinetti C Greco F De Palma L Specchia N Falciglia F

We obtained specimens of growth-plate cartilage from four patients with osteogenesis imperfecta. Light microscopy showed structural changes in the tissue and morphological changes in chondrocytes and matrix, particularly in the hypertrophic zone. There were changes in the process of calcification in the primary mineralisation zone of the cartilage. We also found histochemical changes in the matrix glycosaminoglycans (GAGs) in the zones where physiological mineralisation was disturbed and where the trabeculae were interrupted and poorly mineralised. In addition to the known molecular defects in collagen, changes in GAGs and non-collagenous proteins are important factors in the pathogenesis of the disease.