In vitro effects of concentrated growth factors (CGF) on human SH-SY5Y neuronal cells.

OBJECTIVE: The aim of this study was to test the in vitro differentiation effects of concentrated growth factors (CGF), a platelet rich preparation, using SH-SY5Y cells, derived from human neuroblastoma. MATERIALS AND METHODS: SH-SY5Y cells were cultured in presence of CGF or retinoic acid (RA). After 72 h of treatment, different parameters were investigated: cell proliferation by an automated cell counter; cell viability by thiazolyl blue tetrazolium bromide (MTT) assay; cell differentiation markers, i.e., neuronal nuclear antigen (NeuN), synaptophysin (SYP) and ß3-tubulin, by immunocytochemistry and Western blotting techniques; release of nerve growth factor (NGF) and brain-derived growth factor (BDNF) by enzyme-linked immunosorbent assay (ELISA) and neurite outgrowth by a dedicated image software. RESULTS: In presence of CGF, the cell proliferation rate and viability decreased, as expected for differentiated SH-SY5Y cells. On the contrary, the cellular differentiation markers increased their expression together with the release of growth factors. Moreover, the neurite outgrowth was improved. CONCLUSIONS: The data suggest that CGF treatment positively affects the cell differentiation, regulating the expression of neuronal markers, the release of growth factors and the neurite length. Taken together these results seem to be promising in the development of new approaches for neural regeneration.

Bilateral and multifocal Warthin's tumor of parotid gland: two case reports and review of literature.

Warthin's tumor is the second most common benign neoplasm of the parotid. Most of cases are represented by a single localization, while only a small percentage of patients presents bilateral lesions or unilateral multifocal pattern. Warthin's tumor has an excellent prognosis due to the low rate of recurrence after surgical treatment. Malignant transformation occurs in less than 1% of cases. The aim of this article is to present two unusual cases of Warthin's tumor and an updated review of the latest scientific literature.

Histomorphic-metric evaluation of an implant retrieved from human maxilla after 13 years.

Fixture fracture is the most catastrophic failure of implant components because it usually causes the loss of the implant. Nevertheless, the osseointegrated fractured implants represent a very useful opportunity to study in humans the effects of loading to the peri-implant bone microstructure. The aim of the present study was to evaluate the interplay between microstructure and function of the bone around an implant retrieved from human maxilla after 13 years. There was 1 fractured Dental Implant Line (sand blasted surface from a patient placed in the anterior region of the maxillary bone (2.1) after a bone augmentation procedure, and it was processed for histology. The specimen was analyzed under the scanning electron microscope (SEM), the confocal scanning laser microscope (CSLM) and brightfield light microscope (LM) equipped with circularly polarized light (CPL). The BIC rate of the implant retrieved after 13 years was (mean ±SD) 68.7 ± 3.7. The crestal bone down the implant platform damage appeared to be under modeling process. The transverse collagen fiber orientation (CFO) (mean ±SD) under the lower flank of the threads was 20.4 ± 3.5 x 10(4) pixel while the longitudinal CFO was 19.8 ± 2.8 x 10(4) pixel (P>.05). In the inter-threads region the transverse CFO (mean ±SD) was 15.0 ± 4.0 x 10(4) pixel while the longitudinal CFO was 21.4 ± 3.0 x 10(4) pixel (P>.05). The osteocytes numbers (mean ±SD) was 130 ∓ 34. Under SEM with back scattered electrons (BSE) signal the peri-implant bone appears mainly lamellar and highly mature with several osteons organized in the implant inter-threads areas. The fracture of the implant was most probably correlated to a fatigue of the material mainly associated to a damage of the internal coil. Surprisingly, it was noted a lack of implant site-specific CFO of the bone extracellular matrix facing the threaded dental implant notwithstanding the high level of BIC rate.

Histomorphometric evaluation of an immediately loaded implant retrieved from human mandible after 2 years.

The aim of the present study was to evaluate the interplay between microstructure and function of the bone around an immediately loaded implant retrieved from human maxilla after 23 months due to fracture. A spiral implant of 3.3 mm x 15 mm was placed in a male 53 years old in the anterior region of the mandible bone (4.1) and it was processed for histology. The specimen was analyzed under the confocal scanning laser microscope (CSLM) and brightfield light microscope (LM) equipped with circularly polarized light (CPL). The BIC rate was 76.7 ± 4.9 (mean ±SD). Many cement lines indicates an high remodeling rate of the bone. The transverse collagen fiber orientation (CFO) (mean±SD) under the lower flank of the thread near the tread tip was 55.2 ± 4.8 x 10(4) pixel while the longitudinal CFO was 45.8 ± 2.3 x 10(4) pixel (P<.05). In the inter-threads region the transverse CFO (mean ±SD) was 36.4 ± 2.4 x 10(4) pixel while the longitudinal CFO was 65.6 ± 6.5 x 10(4) pixel (P<.05). The osteocytes numbers (mean ±SD) was 205 ± 45 in the peri-implant bone and 144 ± 53 in the native bone (P=.007). After 2-years of loading the SLA spiral implant was well osseointegrated but still surrounded by woven bone. The osteocytes density was significantly higher in the peri-implant bone than in the native bone. The transverse collagen fibers were significantly associated with the lower flank of the implant threads, while the longitudinal collagen fibers were more represented in the straight surface of the implant. The implant fracture was correlated to crestal bone resorbing and subsequent fatigue yielding.

Calcium sulfate stimulates pulp stem cells towards osteoblasts differentiation.

Calcium sulfate (CaS) is a highly biocompatible material and enhances bone formation in vivo. However, how CaS alters osteoblast activity to promote bone formation is poorly understood. To study how CaS can induce osteoblast differentiation in mesenchymal stem cells, the expression levels of bone related genes and mesenchymal stem cells marker were compared in normal osteoblasts and dental pulp stem cells, using real time Reverse Transcription-Polymerase Chain Reaction. Gene differentially expressed between the two cells type were the trascriptional factor RUNX2, osteopontin (SPP1), COL1A1 (collagen type 1α1) and alkaline phosphatase (ALPL). The obtained results demonstrated that CaS strongly influences the behavior of DPSCs in vitro enhancing proliferation, differentiation and deposition of matrix.

Polylactide-polyglycolide resorbable plates stimulates adipose tissue-derived stem cells towards osteoblasts differentiation.

Polylactide, polyglycolide materials or devices have been utilized routinely during maxillofacial, craniofacial, and orthopaedic reconstructive surgical procedures.(1) These materials combine the benefits of rigid fixation with the advantages of biodegradation, avoiding the need for implant removal and minimizing the risk of other complications.(2) To study how polylactide, polyglycolide acids plates (PLPG plates) can induce osteoblast differentiation and proliferation in mesenchymal stem cells, the expression levels of bone related genes (RUNX2, SP7, ALPL, SPP1, COL1A1, COL3A1 and FOSL1) and mesenchymal stem cells marker (ENG) were measured in adipose derived stem cells (ADSCs) and normal osteoblast (NO) cultivated on PLPG plates after 15 and 30 days of treatment using real time Reverse Transcription-Polymerase Chain Reaction. Significantly differentially expressed genes among ADSCs and NO were SP7, ENG, FOSL1, RUNX, ALPL and SPP1 in the first 15 days of treatment and SP7, ENG FOSL1, COL3A1 COL1A1, SPP1 and ALPL after 30 days. The present study demonstrated that PLPG plates strongly influences the behavior of ADSCs in vitro by enhancing proliferation, differentiation and deposition of matrix.

Overdentures on implants placed in bone augmented with fresh frozen bone.

AIM: In the last decade several studies have been performed to evaluate the clinical outcome of one or two stage loaded implants supporting overdentures. Since fresh frozen bone (FFB) has an ever-increasing number of clinical applications and few reports are available on implants inserted into FFB, we performed a retrospective study on fixtures inserted in FFB and bearing overdentures. METHODS: In the period between December 2003 and December 2006, 17 patients (14 females and 3 males with a median age of about 56 years) were grafted and 60 implants inserted thereafter. A total of 17 overdentures were delivered: 8 in the mandible and 9 in the maxilla. Multiple implant systems were used: 22 Double etched, 7 SLA, 9 Anodic oxidized, and 22 CaPo4 ceramic-blasted. Implant diameter ranged from 3.25 to 4.3 mm and length from 11.5 to 16.0 mm. Implants were inserted to replace 23 incisors, 9 cuspids, 20 premolars and 8 molars. RESULTS: No implants were lost (i.e., survival rate=100%) and no differences were detected among the studied variables. Kaplan Meier algorithm and Cox regression did not reveal any statistical differences among the studied variables also as regards the success rate. CONCLUSION: Implants inserted FFB and bearing overdentures have a high survival rate and success rates, which are comparable to those of implants inserted in non-grafted bone. FFB bone is a reliable material for alveolar ridge augmentation. No difference was detected among removable prostheses supported by two or more implants.

Restoration of locomotion in posttraumatic paraplegics: the multidisciplinary approach and unexpected plasticity of single neurons--facts and fantasy.

In Europe there are about 300,000 paraplegics and in every country approximately 1000 new cases per year. Treatment requires a multidisciplinary approach with scientific cooperation targeted to exchange personal knowledge and expertise. At present a completely disrupted spinal cord cannot heal for recovery of motor and/or sensory functioning, although some promising treatment modalities in laboratory animal experiments have been reported. No interventional stem cell procedure so far has shown evidence to restore impaired functioning in human paraplegics. However, functional electrical stimulation (FES) via an implanted neuroprosthesis (SUAW concept) and central nervous system-peripheral nervous system (CNS-PNS) connection have successfully been used for alternative compensatory strategies for voluntary locomotion. This report is to analyse the authors' experience from two European projects in paraplegic. Factors will be identified that might have caused the one or other pitfall since so far both surgical reconstructive procedures have not been adopted by rehabilitation physicians and/or restorative (neuro-)surgeons despite the promising functional results we have achieved. Unexpected plasticity of single neurons following CNS-PNS by-pass procedures is discussed. Future interventions, for example the present phase 1 prospective multiple centre study on the side effects, effectiveness, and reliability of intrathecal treatment of anti-Nogo-A antibodies, are presented and the Chinese stem cell implantation is critically reviewed.

Experimental neurotisation of the brachial plexus by means of nerve grafting from the corticospinal tract of the proximal spinal cord at the T3-T4 level.

This paper reports experimental work in rats in which the radial nerve was cut at its origin from the brachial plexus and connected by means of a nerve graft to the corticospinal tract of the spinal cord at the T3-T4 level. After surgery, the triceps and extensor muscles of the forearm were able to extend the elbow and the wrist and gave almost normal responses on EMG. This reinnervation was no longer cholinergic, as demonstrated by the Vecuronium test. Because of the drawbacks associated with implantation into the spinal cord, this technique could not be used in all cases of total avulsion of the brachial plexus but it might be useful in those rare cases in which avulsion of all the cervical nerves of the brachial plexus is associated with paraplegia: such cases are rare but are occasionally seen.

Adhesion reduction after knee surgery in a rabbit model by Hyaloglide, a hyaluronan derivative gel.

A randomized controlled experimental trial was performed in a rabbit model of surgical adhesions to investigate the anti-adhesive effects of Hyaloglide, a highly viscous hyaluronan derivative absorbable gel after knee surgery. Twenty New Zealand white rabbits were prepared and randomly divided into two groups of 10 animals each. An intra-articular fibro-adhesive scar was created in the right knee joint of the hind paw of each rabbit using a standardized surgical procedure, and Hyaloglide was administered into the joint cavity of the knee at the end of intervention in the animals belonging to the treatment group. No anti-adhesive treatment was applied in the control group. Additionally, immobilization using a Kirschner wire was applied in order to increase the risk of adhesions. Six weeks after surgery the animals were euthanized and after removal of the immobilization system, adhesions were evaluated both macroscopically and histologically. Results of gross observations using a specific adhesion scoring system showed a significant reduction (p<0.01) of both incidence and severity of adhesions in the hyaluronan-treated group compared to the control group. Histologically, adhesions in the treated group were thinner with less collagenic fibers. In conclusion, Hyaloglide may be considered as a promising absorbable barrier for prevention of post-operative fibrotic adhesions after knee surgery.

Brachial plexus surgery (honorary lecture).

Brachial plexus injuries (B.p.i.) are lesions occurring more and more frequently due to high velocity road and sport traumas. They are severe lesions with disabling sequelae. Surgical procedures and results could greatly be improved in the last 2 decades. Although the anatomy of brachial plexus is well known, less known are the functional maps of the various brachial plexus elements. In this paper treatment modalities for obstetrical, traumatic (adult) and actinic B.p.i. are being described too.

Coupling between "hand" primary sensorimotor cortex and lower limb muscles after ulnar nerve surgical transfer in paraplegia.

Previous neuroimaging evidence revealed an "invasion" of "hand" over "lower limb" primary sensorimotor cortex in paraplegic subjects, with the exception of a rare patient who received a surgical motor reinnervation of hip-thigh muscles by the ulnar nerve. Here, the authors show that a functional reorganization of cortico-muscular and cortico-cortical oscillatory coupling was related to the recovery of the rare patient, as a paradigmatic case of long-term plasticity in human sensorimotor cortex after motor reinnervation of paraplegic muscles. This conclusion was based on electroencephalographic and electromyographic data collected while the patient and normal control subjects performed isometric muscle contraction of the left hand or lower limb. Cortico-muscular and cortico-cortical coupling was estimated by electroencephalographic-electromyographic coherence and directed transfer function of a multivariate autoregressive model.

Restoration of locomotion in paraplegics with aid of autologous bypass grafts for direct neurotisation of muscles by upper motor neurons--the future: surgery of the spinal cord?

OBJECTIVES: Paraplegia means a lifelong sentence of sensory loss, paralysis and dependence. Complete spinal cord lesions cannot heal up to now despite intensive experimental research, remarkable efforts and recent achievements in bio-technology and re-engineering. Traumatic paraplegia due to spinal cord injury (SCI) is a quite frequent condition and related to the socio-economical situation of the population. It is experienced disproportionately by young people. The rise in gunshot wounds is dramatic. SCI has appeared refractory to treatment. PATIENTS AND METHODS: Since 1980 G.A.B. had tried surgical repair of the spinal cord (SC) after experimental bisection in rats, and since 1993 research was done on monkeys (macaca fascicularis) to be closer to human physiology. The sciatic nerve was removed and used as an autologous graft from the lateral bundle of the spinal cord (tractus corticospinalis ventro lateralis) to the three muscles of both legs being known to be most important for locomotion: M. gluteus maximus, M. gluteus medius and M. quadriceps femoris. The first fruitful transplantation in a human being was performed in July 2000. RESULTS: The results in rats were promising and fulfilled the requirements of the American Task Force of the National Institute of Neurological and Communicative Disorders and Stroke of the US. The results in monkeys confirmed the paradigm so that we performed the first operation in a young lady suffering for four months from complete SC lesion T9 after approval by the ethical committee. First voluntary movements of the connected muscles after 17 months. 27 months after op she was able to walk up to 60 steps with the help of a walker and to climb steps in the water. Improvement is still continuing. DISCUSSION: SCI has appeared refractory to any kind of treatment. Compensatory strategies are still experimental in human beings. Autologous nerve grafts from the spinal cord tissue (the lateral spinal bundle) connected to peripheral muscle nerves seem promising in paraplegics. But the physiology is still unclear when the glutamatergic upper motor neuron connected to motor end-plates (cholinergic) does work like in our patient. CONCLUSION: Further studies in primates and paraplegic patients are necessary to clarify the bypass grafting of the SC to muscle groups distal to the complete SCI to restore locomotion.

[Carpal instability with scapho-lunate dissociation treated using the flexor carpi radialis and scaphoid-trapezoid ligament repair: foundations, technique and results of preliminary series]. / Dissociation scapho-lunaire traitée par ligamentoplastie scapho-trapézoïdienne au flexor carpi radialis: fondements, technique et résultat de la série préliminaire.

Carpal instability with scapho-lunate dissociation is still attributed to rupture of the so-called scapho-lunate ligament. Actually, this structure is not a ligament but a loose capsule allowing very different flexion of the scaphoid (92 degrees) and the lunate (20 degrees). As reconstruction of the scapho-lunate "ligament" has often been less than satisfactory we searched for another technique. Sections of the scapho-lunate "ligament" on cadaver specimens never produce scapho-lunate dissociation. This dissociation can not occur because the scaphoid is maintained in the radial facette. It was observed that the only way to produce scapho-lunate dissociation is to section the scapho-trapezo-trapezoid ligament allowing flexion of the scaphoid and dorsal dislocation out of the radial facette. The scapho-trapezo-trapezoid ligament is not well known and is not described in anatomy text books because it is hidden by the flexor carpi radialis tendon. It is confluent with the scaphoid and the trapezoid. This produces a radial (scapho-trapezoid) column which acts like a true external pillar maintaining the height of the carpus and preventing carpal collapse. Finally, dissociation of the proximal pole from the semi-lunate can only occur by posterior displacement. After experimenting the technique on cadaver specimens, we developed a reconstruction method for the palmar scapho-trapezoid ligament using a band of the flexor carpi radialis tendon, applied in 38 patients. The flexor carpi radialis band measured 7 cm and was left attached to the base of the second metacarpal then passed in a tunnel bored anteriorly to posteriorly in the distal pole of the scaphoid. The band was then drawn dorsally while maintaining the scaphoid in place, and sutured to the postero-ulnar border of the radius. The height of the carpus was restored as was scaphoid movement over the lunate. The reduction persisted at mid- and long-term and prevented carpal collapse and joint destruction. Among the 38 operated patients, 33 remained pain free and 5 complained of minor pain under stressful conditions. All were satisfied. Anatomic research and clinical experience has demonstrated that the scapho-trapezoidal ligament is the key to scapho-lunate dissociation and its correction. Currently, this operation is the only way to achieve easy and persistent correction of carpal instability with scapho-lunate dissociation.

[Finkelstein's versus Brunelli's test in De Quervain tenosynovitis]. / Le test de Finkelstein contre le test de Brunelli dans la tenosynovite de De Quervain.

This short paper demonstrates that the Finkelstein's test in De Quervain's tenosynovitis is based on an incorrect assumption. The correct basis for a pathognomic manoeuvre in De Quervain is the provocation of tendons attrition of the first wrist dorsal compartment against their pulley which elicits pain. The Brunelli's test induces this friction and pain by asking the patient to hardly adduct the thumb with the wrist in radial deviation.

Computer added locomotion by implanted electrical stimulation in paraplegic patients (SUAW).

Paraplegia means a live long sentence of sensory loss, paralysis and dependence with approximately 1000 new victims in every European country every year and 11.500 new traumatic SCI cases in the US. respectively. Sixty percent are injured before age 30. More than 90% of SCI victims may survive with nearly normal experience of live. Most patients will recover somewhat from SCI over time but no patient who remained plegic for one year regains voluntary motor function after that time period. Despite remarkable efforts and recent achievements in rehabilitation no treatment can be recommended so far to enhance functional recovery and restoring locomotion in paraplegic humans. FES as a technical compensation has become therefore a challenging treatment to restore muscle function and to prevent atrophy and to improve mobility and quality of life at the same time. In paraplegics FES could be the basis to restore locomotion. One of the advantages of an implanted FES version (neuroprosthesis) is that the FES system, electrodes, and cables remain permanently implanted within the body, so that the patient can stay without cables, the programmer attached to the crutches. The SUAW project, supported under BIOMED II Programme by the European Community was aimed to finalize and to put into practice the results of previous research and development. The novel implant with an ASCI-Chip has 16 channels, 8 on each side, 20 mA for monopolar and 2 mA for bipolar stimulation, only one electrode can be stimulated at a given time. Stimulation of 6 muscle groups of both legs are known to be sufficient for locomotion: M. ileopsoas (erector of the body, hip flexor), M. gluteus maximus (hip extensor), M. gluteus medius (lateral hip stabilisator, knee abductor), Mm. hamstrings (knee flexor) stimulated by epimysial electrodes, Mm. sartorius and rectus femoris (knee extensor) stimulated by neural, bipolar electrodes. Patient's selection criteria were: stable spinal cord lesion between T7 and T11, minimum 1 year after the accident without deformity of the spine, the muscle groups for locomotion responding to external FES with the EXOSTIM programmer with the same programme used later for the neuroprosthesis. Two paraplegic male patients, T8, 38 and 31 years old respectively, were operated on by an international group of surgeons according to the protocol in 09/1999, respectively 7/2000. The postop. course was uneventful. Because the threshold of the primary implant was too low regarding scare tissue around the electrodes, this implant was changed in 01/2000 and worked perfectly. Both patients are happy with the success of the novel treatment modalities.

Direct neurotization of muscles by presynaptic motoneurons.

The spinal cord cannot heal after severance because the central nervous system is "non-permissive" to the advancement of axons that regrow from presynaptic motoneurons. With the aim of overcoming paraplegia, the author has carried out extensive experimental research since 1980, first in rats and subsequently in monkeys, severing the cord and connecting its cephalad stump with the muscular nerve branches by means of peripheral-nerve grafts, and using various surgical protocols. Functional connections were established, ascertained by physical, electrophysiologic, and histologic examinations. In this reported study, it is demonstrated that presynaptic motoneurons are also able to reconstruct the cytoskeleton of peripheral neurons, as well as motor end-plates. The possibility of elongation of the axons of presynaptic motoneurons into the peripheral nerve up to the muscle had not previously even been hypotheized. This possibility, which has now been validated, can open the door to new surgical techniques for spinal-cord lesions. In addition, the author presents preliminary results from a single human case, utilizing the surgical procedures of the preceding animal experiments.

Remodelling of sensorimotor maps in paraplegia: a functional magnetic resonance imaging study after a surgical nerve transfer.

The adult mammalian brain has the capacity of reorganising its neural connections in response to lesions/modifications of the peripheral and central nervous system. We show in vivo, using functional magnetic resonance imaging (fMRI), that in paraplegics the lower-limb sensorimotor cortex is invaded by the arm representation. This functional reshaping appears to be reversible. Indeed, surgical transfer of the ulnar nerve to the ipsilateral quadriceps and hip muscles allowed their contraction in a paraplegic patient. During fMRI, these voluntary movements activated the hip and thigh representation in sensorimotor cortex. We suggest that the functional recovery of the lower-limb functional maps might have been driven by the restored somatosensory inputs from the reactivated periphery. The voluntary movements of the lower-limbs are regained through the 're-awakening' of the corresponding sensorimotor cortex.

Classification criteria and distinction between migraine and tension-type headache in children.

The International Headache Society (IHS) classification system (1988) was developed primarily for headache disorders in adults and its validity for paediatric age is currently under discussion; in 1995 Seshia et al. proposed a revision of the criteria for migraine without aura to make diagnostic criteria more applicable to children. The purposes of the current study were to: (1) compare the IHS classification with the Seshia proposal, (2) compare the children affected by migraine without aura (MO) with the children affected by tension headache (TH) as defined by Seshia, for characteristics which are not included in the classification. The patients are a series of 320 children (mean age 9.9, SD 2.6 years; 144 males, 176 females) with recurrent or chronic headaches referred to a headache clinic in Milan, Italy. Using the Seshia criteria instead of the IHS criteria a higher number of children were included in the MO category: bilateral pain and family history of migraine were the most important factors which allowed a shift of children into this category. However, with the Seshia classification there was no reduction in the number of unclassifiable children. The reason why some children could not be classified was a short duration of attacks; the majority of unclassifiable children were 6 years old or less. No relevant difference was found between children with MO and children with TH for the following variables: occurrence of attacks in the afternoon or evening after school, reduction of attacks during school holidays, full-time schooling, after-school activities on school days, disordered daily life. On the contrary children with MO when compared with those with TH showed a higher number of precipitating factors and for the following factors a significant difference was found: exposure to TV or a computer, sleep deficiency, and strong emotions. Furthermore, children with MO showed a greater severity of attacks.