Decellularization and Delipidation Protocols of Bovine Bone and Pericardium for Bone Grafting and Guided Bone Regeneration Procedures

The combination of bone grafting materials with guided bone regeneration (GBR) membranes seems to provide promising results to restore bone defects in dental clinical practice. In the first part of this work, a novel protocol for decellularization and delipidation of bovine bone, based on multiple steps of thermal shock, washes with detergent and dehydration with alcohol, is described. This protocol is more effective in removal of cellular materials, and shows superior biocompatibility compared to other three methods tested in this study. Furthermore, histological and morphological analyses confirm the maintenance of an intact bone extracellular matrix (ECM). In vitro and in vivo experiments evidence osteoinductive and osteoconductive properties of the produced scaffold, respectively. In the second part of this study, two methods of bovine pericardium decellularization are compared. The osmotic shock-based protocol gives better results in terms of removal of cell components, biocompatibility, maintenance of native ECM structure, and host tissue reaction, in respect to the freeze/thaw method. Overall, the results of this study demonstrate the characterization of a novel protocol for the decellularization of bovine bone to be used as bone graft, and the acquisition of a method to produce a pericardium membrane suitable for GBR applications

I.S.Mu.L.T. Achilles Tendon Ruptures Guidelines

This work provides easily accessible guidelines for the diagnosis, treatment and rehabilitation of Achilles tendon ruptures. These guidelines could be considered as recommendations for good clinical practice developed through a process of systematic review of the literature and expert opinion, to improve the quality of care for the individual patient and rationalize the use of resources. This work is divided into two sessions: 1) questions about hot topics; 2) answers to the questions following Evidence Based Medicine principles. Despite the frequency of the pathology andthe high level of satisfaction achieved in treatment of Achilles tendon ruptures, a global consensus is lacking. In fact, there is not a uniform treatment and rehabilitation protocol used for Achilles tendon ruptures

Clinical and polysomnographic course of childhood narcolepsy with cataplexy.

Our aim was to investigate the natural evolution of cataplexy and polysomnographic features in untreated children with narcolepsy with cataplexy. To this end, clinical, polysomnographic, and cataplexy-video assessments were performed at diagnosis (mean age of 10 ± 3 and disease duration of 1 ± 1 years) and after a median follow-up of 3 years from symptom onset (mean age of 12 ± 4 years) in 21 children with narcolepsy with cataplexy and hypocretin 1 deficiency (tested in 19 subjects). Video assessment was also performed in two control groups matched for age and sex at first evaluation and follow-up and was blindly scored for presence of hypotonic (negative) and active movements. Patients' data at diagnosis and at follow-up were contrasted, compared with controls, and related with age and disease duration. At diagnosis children with narcolepsy with cataplexy showed an increase of sleep time during the 24 h; at follow-up sleep time and nocturnal sleep latency shortened, in the absence of other polysomnographic or clinical (including body mass index) changes. Hypotonic phenomena and selected facial movements decreased over time and, tested against disease duration and age, appeared as age-dependent. At onset, childhood narcolepsy with cataplexy is characterized by an abrupt increase of total sleep over the 24 h, generalized hypotonia and motor overactivity. With time, the picture of cataplexy evolves into classic presentation (i.e., brief muscle weakness episodes triggered by emotions), whereas total sleep time across the 24 h decreases, returning to more age-appropriate levels

Emergency hernia repair in the elderly. multivariate analysis of morbidity and mortality from an Italian registry

Purpose The incidence of inguinal hernia is higher in elderly because of aging-related diseases like prostatism, bronchitis, collagen laxity. A conservative management is common in elderly to reduce surgery-related risks, however watchful waiting can expose to obstruction and strangulation. The aim of the present study was to assess the impact of emergency surgery in a large series of elderly with complicated groin hernia and to identify the independent risk factors for postoperative morbidity and mortality. The predictive performance of prognostic risk scores has been also assessed. Methods This is a prospective observational study carried out between January 2017 and June 2018 in elderly patients who underwent emergency surgery for complicated hernia in 38 Italian hospitals. Pre-operative, surgical and postoperative data were recorded for each patient. ASA score, Charlson's comorbidity index, P-POSSUM and CR-POSSUM were assessed. Results 259 patients were recruited, mean age was 80 years. A direct repair without mesh was performed in 62 (23.9%) patients. Explorative laparotomy was performed in 56 (21.6%) patients and bowel resection was necessary in 44 (17%). Mortality occurred in seven (2.8%) patients. Fifty-five (21.2%) patients developed complications, 12 of whom had a major one. At univariate and multivariate analyses, Charlson's comorbidity index >= 6, altered mental status, and need for laparotomy were associated with major complications and mortality Conclusion Emergency surgery for complicated hernia is burdened by high morbidity and mortality in elderly patients. Preoperative comorbidity played a pivotal role in predicting complications and mortality and therefore Charlson's comorbidity index could be adopted to select patients for elective operation

Perforated peptic ulcer (PPU) treatment: an Italian nationwide propensity score-matched cohort study investigating laparoscopic vs open approach

BackgroundPerforated peptic ulcer (PPU) remain a surgical emergency accounting for 37% of all peptic ulcer-related deaths. Surgery remains the standard of care. The benefits of laparoscopic approach have been well-established even in the elderly. However, because of inconsistent results with specific regard to some technical aspects of such technique surgeons questioned the adoption of laparoscopic approach. This leads to choose the type of approach based on personal experience. The aim of our study was to critically appraise the use of the laparoscopic approach in PPU treatment comparing it with open procedure.MethodsA retrospective study with propensity score matching analysis of patients underwent surgical procedure for PPU was performed. Patients undergoing PPU repair were divided into: Laparoscopic approach (LapA) and Open approach (OpenA) groups and clinical-pathological features of patients in the both groups were compared.ResultsA total of 453 patients underwent PPU simple repair. Among these, a LapA was adopted in 49% (222/453 patients). After propensity score matching, 172 patients were included in each group (the LapA and the OpenA). Analysis demonstrated increased operative times in the OpenA [OpenA: 96.4 +/- 37.2 vs LapA 88.47 +/- 33 min, p = 0.035], with shorter overall length of stay in the LapA group [OpenA 13 +/- 12 vs LapA 10.3 +/- 11.4 days p = 0.038]. There was no statistically significant difference in mortality [OpenA 26 (15.1%) vs LapA 18 (10.5%), p = 0.258]. Focusing on morbidity, the overall rate of 30-day postoperative morbidity was significantly lower in the LapA group [OpenA 67 patients (39.0%) vs LapA 37 patients (21.5%) p = 0.002]. When stratified using the Clavien-Dindo classification, the severity of postoperative complications was statistically different only for C-D 1-2.ConclusionsBased on the present study, we can support that laparoscopic suturing of perforated peptic ulcers, apart from being a safe technique, could provide significant advantages in terms of postoperative complications and hospital stay

Evaluation of appendicitis risk prediction models in adults with suspected appendicitis

Background Appendicitis is the most common general surgical emergency worldwide, but its diagnosis remains challenging. The aim of this study was to determine whether existing risk prediction models can reliably identify patients presenting to hospital in the UK with acute right iliac fossa (RIF) pain who are at low risk of appendicitis. Methods A systematic search was completed to identify all existing appendicitis risk prediction models. Models were validated using UK data from an international prospective cohort study that captured consecutive patients aged 16–45 years presenting to hospital with acute RIF in March to June 2017. The main outcome was best achievable model specificity (proportion of patients who did not have appendicitis correctly classified as low risk) whilst maintaining a failure rate below 5 per cent (proportion of patients identified as low risk who actually had appendicitis). Results Some 5345 patients across 154 UK hospitals were identified, of which two‐thirds (3613 of 5345, 67·6 per cent) were women. Women were more than twice as likely to undergo surgery with removal of a histologically normal appendix (272 of 964, 28·2 per cent) than men (120 of 993, 12·1 per cent) (relative risk 2·33, 95 per cent c.i. 1·92 to 2·84; P < 0·001). Of 15 validated risk prediction models, the Adult Appendicitis Score performed best (cut‐off score 8 or less, specificity 63·1 per cent, failure rate 3·7 per cent). The Appendicitis Inflammatory Response Score performed best for men (cut‐off score 2 or less, specificity 24·7 per cent, failure rate 2·4 per cent). Conclusion Women in the UK had a disproportionate risk of admission without surgical intervention and had high rates of normal appendicectomy. Risk prediction models to support shared decision‐making by identifying adults in the UK at low risk of appendicitis were identified

Progettazione e sviluppo di scaffolds naturali per la rigenerazione ossea

For many years, the regenerative medicine has been using the biometarials like grafts to promote healing of bone tissue. A heterologous graft should not evoke any inflammatory response where it is inserted and its antigenicity is eliminated with a decellularization technique . The aim of this study was to provide a protocol creating a bovine xenograft which easily employed in medicine field for repairing bone defects. For this purpose, the decellularization protocols were applied on 26 rats' femur by means of freezing /disfreezing. The effect of the decellularization in vitro was tested by means of MTT and the light microscope (SEM). The bovine xenograft obtained was inserted surgically in maxillary sinus of 30 sheeps to test the ability of regeneration of biomaterial in vivo. The surgical intervention was carried out bilaterally: one side was experimental side (xenograft) and the other side was control side ( Bio-oss®,Geistlich Pharma AG Wohlhusen, Switzerland). The study subjects were divided in 3 groups .Each group was consisting of 10 animals. They were euthanazied on 15, 30, 60 days from the surgical intervention, respectively. Collected bone samples were decalcified, fixed and colored with hematoxylin-eosin and subsequently, they were evaluated under optical microscope. The MTT tests showed that the duration of freezing at -20°C for 7 days and e -80°C for another 7 days, followed by disfreezing through thermal shock was more efficient for correct decellularization of the bovine extracellular matrix. The RT-PCR results indicate that the bovine xenograft has the optimum osteoinductive competence. In vivo tests showed that the biometarial already present for 15 days, has promoted the capacity of osteoconductive and osteoinductive with absence of inflammatory response. The “gold standard” in medicine manifests the excellent osteoconductive and osteoinductive capacity at 60 days. In conclusion, it can be confirmed that the protocol of decellularization by means of freezing /disfreezing, allows producing an ideal bone replacement for regeneration of bone defects in medicine field.La medicina rigenerativa impiega ormai da molti anni biomateriali in grado di promuovere ed accelerare la guarigione del tessuto osseo in cui vengono innestati. Un innesto osseo eterologo non dovrebbe evocare nessuna risposta infiammatoria nel sito in cui viene inserito. L’antigenicità del tessuto osseo eterologo viene eliminata grazie a tecniche di decellularizzazione ed il protocollo di decellularizzazione impiegato influisce in modo determinante sulle caratteristiche del biomateriale stesso. L'obiettivo di questo studio è stato quello di mettere a punto un protocollo per la creazione di uno xenograft bovino di facile impiego in campo medico per la riparazione di difetti ossei. A questo scopo, sono stati applicati protocolli di decellularizzazione mediante congelamento/scongelamento su 26 femori murini; l'effettiva decellularizzazione in vitro è stata valuta tramite test MTT e mediante microscopia elettronica (SEM). Il protocollo di decellularizzazione più efficace è stato applicato in un secondo momento a tessuto osseo bovino. Successivamente sono state validate in vitro le capacità osteoinduttive dello xenograft bovino mediante l'impiego di cellule staminali. Lo xenograft bovino così ottenuto è stato innestato chirurgicamente nel seno mascellare di 30 pecore per testare in vivo le capacità rigenerative del biomateriale. L'intervento chirurgico è stato eseguito bilateralmente: un lato test (xenograft) e un lato controllo (Bio-oss®,Geistlich Pharma AG Wohlhusen, Switzerland). I soggetti sono stati suddivisi in 3 gruppi da 10 animali ciascuno ed eutanasizzati a 15, 30 e 60 giorni dall’intervento chirurgico. I campioni d’osso prelevati sono stati decalcificati, fissati, colorati con ematossilina ed eosina e successivamente valutati al microscopio ottico. I test MTT hanno dimostrato che i tempi di congelamento a -20°C per 7 giorni e -80°C per altri 7 giorni, seguite da scongelamento tramite shock termico, sono più efficaci per una corretta decellularizzazione della matrice extracellulare bovina. I risultati della RT-PCR indicano che lo xenograft bovino ha ottime capacità osteoinduttive. I test in vivo hanno dimostrato che il biomateriale già a 15 giorni presenta: elevata capacità osteoconduttiva ed osteoinduttiva; assenza di risposta infiammatoria. A 30 e 60 giorni queste caratteristiche non subiscono variazioni significative. Il gold standard in campo medico presenta ottime capacità osteoconduttive ed osteoinduttive a 60 giorni. In conclusione si può affermare che il protocollo di decellularizzazione mediante congelamento/scongelamento da noi sviluppato consente di produrre un sostituto osseo ideale per la rigenerazione di difetti ossei in campo medico; il biomateriale testato promuove la neoangiogenesi e l'osteogenesi a soli 15 giorni dall'innesto

Progettazione e sviluppo di scaffolds naturali per la rigenerazione ossea

For many years, the regenerative medicine has been using the biometarials like grafts to promote healing of bone tissue. A heterologous graft should not evoke any inflammatory response where it is inserted and its antigenicity is eliminated with a decellularization technique . The aim of this study was to provide a protocol creating a bovine xenograft which easily employed in medicine field for repairing bone defects. For this purpose, the decellularization protocols were applied on 26 rats' femur by means of freezing /disfreezing. The effect of the decellularization in vitro was tested by means of MTT and the light microscope (SEM). The bovine xenograft obtained was inserted surgically in maxillary sinus of 30 sheeps to test the ability of regeneration of biomaterial in vivo. The surgical intervention was carried out bilaterally: one side was experimental side (xenograft) and the other side was control side ( Bio-oss®,Geistlich Pharma AG Wohlhusen, Switzerland). The study subjects were divided in 3 groups .Each group was consisting of 10 animals. They were euthanazied on 15, 30, 60 days from the surgical intervention, respectively. Collected bone samples were decalcified, fixed and colored with hematoxylin-eosin and subsequently, they were evaluated under optical microscope. The MTT tests showed that the duration of freezing at -20°C for 7 days and e -80°C for another 7 days, followed by disfreezing through thermal shock was more efficient for correct decellularization of the bovine extracellular matrix. The RT-PCR results indicate that the bovine xenograft has the optimum osteoinductive competence. In vivo tests showed that the biometarial already present for 15 days, has promoted the capacity of osteoconductive and osteoinductive with absence of inflammatory response. The “gold standard” in medicine manifests the excellent osteoconductive and osteoinductive capacity at 60 days. In conclusion, it can be confirmed that the protocol of decellularization by means of freezing /disfreezing, allows producing an ideal bone replacement for regeneration of bone defects in medicine field.La medicina rigenerativa impiega ormai da molti anni biomateriali in grado di promuovere ed accelerare la guarigione del tessuto osseo in cui vengono innestati. Un innesto osseo eterologo non dovrebbe evocare nessuna risposta infiammatoria nel sito in cui viene inserito. L’antigenicità del tessuto osseo eterologo viene eliminata grazie a tecniche di decellularizzazione ed il protocollo di decellularizzazione impiegato influisce in modo determinante sulle caratteristiche del biomateriale stesso. L'obiettivo di questo studio è stato quello di mettere a punto un protocollo per la creazione di uno xenograft bovino di facile impiego in campo medico per la riparazione di difetti ossei. A questo scopo, sono stati applicati protocolli di decellularizzazione mediante congelamento/scongelamento su 26 femori murini; l'effettiva decellularizzazione in vitro è stata valuta tramite test MTT e mediante microscopia elettronica (SEM). Il protocollo di decellularizzazione più efficace è stato applicato in un secondo momento a tessuto osseo bovino. Successivamente sono state validate in vitro le capacità osteoinduttive dello xenograft bovino mediante l'impiego di cellule staminali. Lo xenograft bovino così ottenuto è stato innestato chirurgicamente nel seno mascellare di 30 pecore per testare in vivo le capacità rigenerative del biomateriale. L'intervento chirurgico è stato eseguito bilateralmente: un lato test (xenograft) e un lato controllo (Bio-oss®,Geistlich Pharma AG Wohlhusen, Switzerland). I soggetti sono stati suddivisi in 3 gruppi da 10 animali ciascuno ed eutanasizzati a 15, 30 e 60 giorni dall’intervento chirurgico. I campioni d’osso prelevati sono stati decalcificati, fissati, colorati con ematossilina ed eosina e successivamente valutati al microscopio ottico. I test MTT hanno dimostrato che i tempi di congelamento a -20°C per 7 giorni e -80°C per altri 7 giorni, seguite da scongelamento tramite shock termico, sono più efficaci per una corretta decellularizzazione della matrice extracellulare bovina. I risultati della RT-PCR indicano che lo xenograft bovino ha ottime capacità osteoinduttive. I test in vivo hanno dimostrato che il biomateriale già a 15 giorni presenta: elevata capacità osteoconduttiva ed osteoinduttiva; assenza di risposta infiammatoria. A 30 e 60 giorni queste caratteristiche non subiscono variazioni significative. Il gold standard in campo medico presenta ottime capacità osteoconduttive ed osteoinduttive a 60 giorni. In conclusione si può affermare che il protocollo di decellularizzazione mediante congelamento/scongelamento da noi sviluppato consente di produrre un sostituto osseo ideale per la rigenerazione di difetti ossei in campo medico; il biomateriale testato promuove la neoangiogenesi e l'osteogenesi a soli 15 giorni dall'innesto

Modeling of the magnetic field errors of RFX-mod upgrade

After several years of successful RFX-mod operations, both in RFP and Tokamak configurations, an upgrade of the machine has been conceived. In particular, the vacuum vessel might be removed with the aim of improving passive MHD control and enabling plasma rotation at higher currents. In this paper, a detailed analysis of the new magnetic front-end is presented, with particular emphasis on the computation of the magnetic field errors generated at the poloidal gaps during transient phases of the discharge. At this purpose, a non-linear MHD equilibrium code, MAXFEA, has been used to provide the input data for the 3D electromagnetic analyses in the time domain carried out with the CAFE code. \ua9 2017 Elsevier B.V