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Covid pandemic affected donation activities worldwide, especially for living donation due to the lack of elective surgery. Moreover, the number of heart-beating and non-heart beating donors has recorded a decrease. Fondazione Banca dei Tessuti di Treviso (FBTV) is a non-profit healthcare organisation, located in Veneto Region, tasked with procurement, processing, preserving, validating and distributing human tissue for clinical use. During Covid-19 outbreak, operations in FBTV have never stopped and a great effort was required to maintain a standard trend of activity. The aim of this study was to describe the impact of Sars-CoV-2 on the activity of a multitissue bank in Italy. Moreover, we investigated the presence of the virus in tissues retrieved from two Sars-CoV-2 positive cadaver donors. Our survey demonstrated that the transplantation network of Veneto Region has positively reacted to the pandemic scenario, thanks to the effort of all personnel involved. Statistical analyses underlined that most of the activities of the tissue bank were unaffected during the Sars-CoV-2 pandemic.

Pericardial patches are currently used as reconstructive material in cardiac surgery for surgical treatment of cardiac septal defects. Autologous pericardial patches, either treated with glutaraldehyde or not, can be used as an alternative to synthetic materials or xenograft in congenital septal defects repair. The availability of an allogenic decellularized pericardium could reduce complication during and after surgery and could be a valid alternative. Decellularization of allogenic tissues aims at reducing the immunogenic reaction that might trigger inflammation and tissue calcification over time. The ideal graft for congenital heart disease repair should be biocompatible, mechanically resistant, non-immunogenic, and should have the ability to growth with the patients. The aim of the present study is the evaluation of the efficacy of a new decellularization protocol of homologous pericardium, even after cryopreservation. The technique has proven to be suitable as a tissue bank procedure and highly successful in the removal of cells and nucleic acids content, but also in the preservation of collagen and biomechanical properties of the human pericardium.

The aim of this article is to report the results obtained by the use of HAM in surgical wound healing and the reduction of relapse in patients affected by Medication-related osteonecrosis of the jaw (MRONJ).The study involved patients with the diagnosis of MRONJ, surgically treated between October 2016 and April 2019, in a case–control setting. Enrolled patients were randomly divided into 2 groups. One group will be treated with resective surgery and with the insertion of HAM patch (Group A), while the second group had been treated exclusively with resective surgery (Group B).The patients underwent MRONJ surgical treatment with the placement of amniotic membrane patches at the wound site. Data regarding the long-term complications/functions were evaluated at 3, 6, 12, and 24 months after surgery. Pain measurements were performed before the intervention (T0), 7(T1) and 30(T2) days after surgery. 49 patients were included in the study. 2 patients of GROUP A after 30 days since they were surgically treated showed persistent bone exposure. 5 patients of group B demonstrated a lack of healing of the surgical wound with the persistence of bone exposed to 30 days after surgery. Statistical analysis ruled out any difference in OUTCOME (relapse) between GROUP A and B (p = 0.23). However, the Fisher test highlighted a significant difference between the use of HAM and only surgical treatment in pain at rest (p = 0.032). The use of amniotic membrane implement the patient's quality of life and reduce pain perception. has a learning curve that is fast enough to justify its routine use.

Adipose tissue (AT) is composed of a heterogeneous population which comprises both progenitor and differentiated cells. This heterogeneity allows a variety of roles for the AT, including regenerative functions. In fact, autologous AT is commonly used to repair soft tissue defects, and its cryopreservation could be a useful strategy to reduce the patient discomfort caused by multiple harvesting procedures. Our work aimed to characterize the cryopreserved AT and to validate its storage for up to three years for clinical applications. AT components (stromal vascular fraction-SVF and mature adipocytes) were isolated in fresh and cryopreserved samples using enzymatic digestion, and cell viability was assessed by immunofluorescence (IF) staining. Live, apoptotic and necrotic cells were quantified using cytometry by evaluating phosphatidylserine binding to fluorescent-labeled Annexin V. A multiparametric cytometry was also used to measure adipogenic (CD34+CD90+CD31−CD45−) and endothelial (CD34+CD31+CD45−) precursors and endothelial mature cells (CD34−CD31+CD45−). The maintenance of adipogenic abilities was evaluated using in vitro differentiation of SVF cultures and fluorescent lipid staining. We demonstrated that AT that is cryopreserved for up to three years maintains its differentiation potential and cellular composition. Given our results, a clinical study was started, and two patients had successful transplants without any complications using autologous cryopreserved AT.

Osteoarthritis (OA) is a chronic debilitating disorder causing pain and gradual degeneration of joints. Among various cell therapies, mesenchymal stem cell (MSC) therapy appears to provide encouraging results. Human amniotic suspension allografts (HASA) have anti-inflammatory and chondroregenerative potential and represent a promising treatment strategy. The purpose of the present study was to prospectively assess the safety, clinical effectiveness, and feasibility of intra-articular injections of human amniotic suspension allograft (HASA) in unilateral knee OA in order to assess the improvement of symptoms and delay the necessity for invasive surgical procedures. A total of 25 symptomatic patients, affected by knee OA were treated with 3 mL of HASA. Clinical evaluations before the treatment and after 3, 6, and 12 months were performed through International Knee Documentation Committee (IKDC) score and Visual Analogue Scale (VAS) scores. Adverse events were recorded. No severe complications were noted during the treatment and the follow-up period. A statistically significant improvement from basal evaluation to the 3-, 6-, and 12-month follow-up visits was observed. The present pilot study indicates that a single intra-articular injection of HASA seems safe and able to provide positive clinical outcomes, potentially offering a new minimally invasive therapeutic option for patients with knee OA.

OBJECTIVES The aim of this study was to evaluate early- and long-term outcomes of cryopreserved aortic homograft (CAH) implantation for aortic valve replacement (AVR) or aortic root replacement (ARR) in patients with or without complex infective endocarditis. METHODS All adult patients undergoing AVR or ARR with CAH at our institution between January 1993 and July 2021 were included in the study. RESULTS One hundred four patients, 75 males and 29 females, aged 59 ± 17 years, underwent AVR or ARR with CAH for infective endocarditis (n = 94, 90%) or aortic valve disease (n = 10, 10%). There were 33 (35%) native valve endocarditis and 61 (65%) prosthetic valve endocarditis, which were complicated by annular abscess in 77 (82%) patients, mitral valve endocarditis in 13 (14%) and tricuspid valve endocarditis in 13 (14%). The mean cardiopulmonary bypass time was 214 ± 80 min and the mean aortic cross-clamping time was 164 ± 56 min. There were 12 (12%) hospital deaths and 7 (7%) postoperative low cardiac output syndrome requiring extracorporeal membrane oxygenation in 4 patients and intra-aortic balloon pump in 3. Thirty-nine (42%) patients died during the follow-up (94% complete). The mean survival time was 13.9 ± 1.2 years. Twenty-five patients (26%) underwent late reoperation for aortic homograft degeneration (n = 17, 18%), homograft endocarditis (n = 6, 7%), homograft dehiscence (n = 1, 1%) and mitral valve regurgitation (n = 1, 1%). The mean survival free from reintervention was 15.7 ± 1.2 years. CONCLUSIONS AVR or ARR with a CAH for complex endocarditis is associated with satisfactory hospital survival, considering the critical patient presentation at surgery, and excellent survival free from recurrent infection. Need for reoperation late after surgery is similar to other biological prostheses.

STUDY QUESTIONCan risks associated with novelties in assisted reproduction technologies (ARTs) be assessed in a systematic and structured way?SUMMARY ANSWERAn ART-specific risk assessment tool has been developed to assess the risks associated with the development of novelties in ART (EuroGTP II-ART).WHAT IS KNOWN ALREADYHow to implement new technologies in ART is well-described in the literature. The successive steps should include testing in animal models, executing pre-clinical studies using supernumerary gametes or embryos, prospective clinical trials and finally, short- and long-term follow-up studies on the health of the offspring. A framework categorizing treatments from experimental through innovative to established according to the extent of the studies conducted has been devised. However, a systematic and standardized methodology to facilitate risk evaluation before innovations are performed in a clinical setting is lacking.STUDY DESIGN, SIZE, DURATIONThe EuroGTP II-ART risk assessment tool was developed on the basis of a generic risk assessment algorithm developed for tissue and cell therapies and products (TCTPs) in the context of the project ‘Good Practices for demonstrating safety and quality through recipient follow-up European Good Tissue and cells Practices II (EuroGTP II)’. For this purpose, a series of four meetings was held in which eight ART experts participated. In addition, several tests and simulations were undertaken to fine-tune the final tool.PARTICIPANTS/MATERIALS, SETTING, METHODSThe three steps comprising the EuroGTP II methodology were evaluated against its usefulness and applicability in ART. Ways to improve and adapt the methodology into ART risk assessment were agreed and implemented.MAIN RESULTS AND THE ROLE OF CHANCEAssessment of the novelty (Step 1), consisting of seven questions, is the same as for other TCTPs. Practical examples were included for better understanding. Identification of potential risks and consequences (Step 2), consisting of a series of risks and risk consequences to consider during risk assessment, was adapted from the generic methodology, adding more potential risks for processes involving gonadic tissues. The algorithm to score risks was also adapted, giving a specific range of highest possible risk scores. A list of strategies for risk reduction and definition of extended studies required to ensure effectiveness and safety (Step 3) was also produced by the ART experts, based on generic EuroGTP II methodology. Several explanations and examples were provided for each of the steps for better understanding within this field.LIMITATIONS, REASONS FOR CAUTIONA multidisciplinary team is needed to perform risk assessment, to interpret results and to determine risk mitigation strategies and/or next steps required to ensure the safety in the clinical use of novelties.WIDER IMPLICATIONS OF THE FINDINGSThis is a dynamic tool whose value goes beyond assessment of risk before implementing a novel ART in clinical practice, to re-evaluate risks based on information collected during the process.STUDY FUNDING / COMPETING INTEREST(S)This study was called EUROGTP II and was funded by the European Commission (Grant agreement number 709567). The authors declare no competing interests concerning the results of this study.

Recently, in the field of abdominal wall repair surgery, some minimally invasive procedures introduced the use of staplers to provide a retromuscular prosthetic repair. However, to the knowledge of the authors, there are little data in the literature about the outcomes of stapled sutures adoption for midline reconstruction. This study aims to investigate the biomechanics of stapled sutures, simple (stapled), or oversewn (hybrid), in comparison with handsewn suture. From the results obtained, we tried to draw indications for their use in a clinical context.Human cadaver fascia lata specimens, sutured (handsewn, stapled, or hybrid) or not, underwent tensile tests. The data on strength (maximal stress), ultimate strain (deformability), Young's modulus (rigidity), and dissipated specific energy (ability to absorb mechanical energy up to the breaking point) were recorded for each type of specimens and analyzed.Stapled and hybrid suture showed a significantly higher strength (handsewn 0.83 MPa, stapled 2.10 MPa, hybrid 2.68 MPa) and a trend toward a lower ultimate strain as compared to manual sutures (handsewn 344%, stapled 249%, hybrid 280%). Stapled and hybrid sutures had fourfold higher Young's modulus as compared to handsewn sutures (handsewn 1.779 MPa, stapled 7.374 MPa, hybrid 6.964 MPa). Handsewn and hybrid sutures showed significantly higher dissipated specific energy (handsewn 0.99 mJ-mmStapled sutures can resist high loads, but are less deformable and rigid than handsewn suture. This suggests a safer employment in case of small defects or diastasis ( W1 in accord to EHS classification), where the presumed tissutal displacement is minimal. Oversewing a stapled suture improves its efficiency, becoming crucial in case of larger defects ( W1 in accord to EHS classification) where the expected tissutal displacement is maximal. Hybrid sutures seem to be a good compromise.

Mucogingival surgery has the objective to obtain coverage of the recession, with a favorable long-term prognosis, but also to minimize postoperative problems and surgery-related risks. Human amniotic membrane is increasingly employed for periodontal tissue repair in order to promote epithelialization and to reduce pain and scar tissue formation. A 38-year-old female patient reports dental hypersensitivity resulting from gingival recession affecting tooth 4.1. A mucogingival surgical procedure was proposed and a partial-thickness flap of epithelial-connective tissue was harvested from the palate and grafted on to the receiving site. The amniotic membrane was positioned at the donor site to reduce postoperative morbidity and to encourage rapid palatal healing. One week after the application of HAM (human amniotic membrane), the wound was healed and 1 month after the donor site was completely re-epithelialized. The present case report suggests that in the surgical treatment of gingival recession with palatal epithelial-connective tissue graft, HAM promotes rapid epithelialization of the palatal donor site wound with a reduction in morbidity.

Purpose The purpose of this study was to evaluate the clinical course of bone reconstruction of the alveolar crest using homologous fresh-frozen bone harvested from deceased donors. Methods A retrospective survey was based on the Castelfranco Veneto Hospital database, in which 3264 clinical records with a primary or secondary diagnosis of alveolar atrophy were collected over a 10-year period. A random sample of 483 patients with at least 5 years’ follow-up was included in the survey. Patients were contacted by telephone and administered a questionnaire with specific questions to build a significant sample. Results Of the patients, 449 (93% of the sample) had an uneventful follow-up after surgery and 93.2% received at least one implant, with a mean of 3.4 implants per patient. At the time of the survey, 93% of the patients were wearing a dental prosthesis, 86.9% had not lost any implants, and 6.7% had lost at least one implant, while 6.4% still had implants but presented some clinical problems. Finally, patients were asked to provide an index score (1–10 points) on the therapy as a whole, i.e., bone graft, implants, and prostheses. A score of insufficient (up to 5 points) was given by 5.3% of patients, of sufficient (6 to 7 points) by 6.1%, and of good/very good (over 7) by 88.6%. Conclusions Homologous bone for alveolar crest reconstruction can be a valid alternative to autologous grafting if specific tissue limitations are considered when planning therapy. Creeping substitution is partial and slower than in autologous grafts, especially in cases where cortical bone is thick or volume graft is very large. The quality of soft tissue coverage and mucosa lining is also important, possibly due to slower tissue revascularization, so future implants should predictably be positioned primarily within the original host bone.

BACKGROUND Allograft venous thrombosis is a severe complication after kidney transplantation (KT). Early diagnosis and prompt treatment are crucial in preserving the survival of the allograft. In this study, we aimed to describe an emergent strategy for the management of acute allograft venous thrombosis. CASE PRESENTATION A 4-year-old girl, weighing 13.5 kg, was diagnosed with bilateral congenital renal hypodysplasia, urogenital sinus and anorectal malformation. The patient was referred to our department for living-donor KT. Her mother was eligible as a donor, presenting a body weight ratio of 1:4.5. Thrombosis of the inferior vena cava (ICV) was also identified, without any predisposing factor for thrombophilia. KT was performed by an extraperitoneal approach without complications. Venous anastomosis required a human vascular graft sutured to the ICV, and renal artery was anastomosed to the aorta. On postoperative day (POD) 8, acute abdominal pain and hematuria led to the diagnosis of an allograft venous thrombosis. An emergent laparotomy was required to explant the allograft, followed by bench surgery. The allograft was irrigated with thrombolytic agents and lactated Ringer's solution and then after removing the venous vascular graft, it was reimplanted through vascular anastomosis with the ICV and aorta. The recovery of perfusion and function was good with diuresis since day 4 after re-surgery. At 2-year follow-up, the child presented normal allograft function with an estimated GFR of 65 ml/min/1.73 m2 . CONCLUSION According to our experience, explantation of the kidney allograft, followed by irrigation with thrombolytics in bench surgery, and reimplantation resulted in unexpected optimal outcomes in the case of allograft venous thrombosis.

Junctional epidermolysis bullosa (JEB) is a clinically and genetically heterogeneous skin fragility disorder frequently caused by mutations in genes encoding the epithelial laminin isoform, laminin-332. JEB patients also present mucosal involvement, including painful corneal lesions. Recurrent corneal abrasions may lead to corneal opacities and visual impairment. Current treatments are merely supportive. We report a novel JEB phenotype distinguished by the complete resolution of skin fragility in infancy and persistent ocular involvement with unremitting and painful corneal abrasions. Biallelic LAMB3 mutations c.3052-5C&gt, G and c.3492_3493delCG were identified as the molecular basis for this phenotype, with one mutation being a hypomorphic splice variant that allows residual wild-type laminin-332 production. The reduced laminin-332 level was associated with impaired keratinocyte adhesion. Then, we also investigated the therapeutic power of a human amniotic membrane (AM) eyedrop preparation for corneal lesions. AM were isolated from placenta donors, according to a procedure preserving the AM biological characteristics as a tissue, and confirmed to contain laminin-332. We found that AM eyedrop preparation could restore keratinocyte adhesion in an in vitro assay. Of note, AM eyedrop administration to the patient resulted in long-lasting remission of her ocular manifestations. Our findings suggest that AM eyedrops could represent an effective, non-invasive, simple-to-handle treatment for corneal lesions in patients with JEB and possibly other EB forms.

PurposeHuman amniotic membrane (HAM) has biological properties which are important for the experimental and clinical applications in managing patients of various medical specialties such as oral and maxilla-facial surgery.The aim was to evaluate the use of HAM as improvement in surgical wound healing and reduction of relapse in patient affected by medication-related osteonecrosis of the jaw (MRONJ). Materials and MethodsA prospective cohort of patients with the diagnosis of MRONJ at the unit of Maxillofacial Surgery of the Ca Foncello Hospital in Treviso, Italy was selected for the study from October 2016 and May 2019. The patients underwent surgical treatment with the placement of amniotic membrane patches. Staging of the lesions was performed according to classification of MRONJ set by the Italian Society of oral medicine with Italian Society of maxillofacial surgery. A multivariate logistic regression was conducted to evaluate the effects of surgical therapy on disease resolution.Results27 patients were included in this study (a total of 34 surgical sites) with a mean age of 68.48 years. The sample involved 6 male and 21 female. 16 patients were assuming Zoledronic acid. Antiresorptive/Antiangiogenic agents were being taken by 16 oncological patients (57%). The median follow-up period was 14,39 months.Only 2 patients after 30 days since they were surgically treated showed persistent bone exposure. Both patients were retreated with success. ConclusionsThe use of the amniotic membrane has a learning curve that is fast enough to justify its routine use, in order to improve the predictability of the interventions for the management of MRONJ. In particular, implement the patient's quality of life and reduce the risk of relapse. The work stands as a pilot study in order to further enhance the results obtained, also introducing a control group.

A gold-standard technique has yet to be found for the treatment of temporomandibular joint ankylosis (TMJa), particularly in patients with recurring ankylosis. A 58-year-old male patient, with a history of multiple TMJ surgeries and severe limitation of mouth opening (maximum interincisal distance [MID] was 10 mm). Computerised tomography (CT) imaging highlighted a bilateral type IV ankylosis. The surgical guides were manufactured using a 3D printing method after obtaining a proper design of the osteotomy lines. The positioning of the fossa and condyle components of the custom TMJ prosthesis was digitally performed. Osteotomies were carried out using surgical guides and TMJ prostheses were placed as per the virtual planning. A human amniotic mambrana is inserted between the two prosthetic components to avoid ranchylosis. The post-operative CT showed the correct positioning of the condylar prosthesis. MID after 10 days was 37 mm. Total joint reconstruction surgery using 3D virtual surgical planning may be an effective surgical option for achieving a precise surgical outcome and making use of a single-stage approach in cases of TMJa and the use of the amniotic membrane, thanks to its healing properties and reduction of pain perception, seems to improve the quality of the immediate post-operative period.

Introduction A dura mater substitute in decompressive craniectomies must protect the brain while providing a dissection plane between the cortex and myocutaneous layer. The human amniotic membrane (AM) has anti-inflammatory, wound healing, and differentiation properties. We tested AM properties as a dural substitute by comparing the outcomes to biological ones. Methods We prospectively collected data on 25 patients who randomly underwent decompressive craniectomy with lyophilized AM patches and 25 in which biological substitutes were utilized between 2015 and 2019. The AM was laid with the epithelial side facing the brain because of the anti-adhesive proprieties, while the chorion facing the myocutaneous flap. We collected data on demographics, neurological status, comorbidities, and surgical outcomes. Additionally, we created a score – dura mimicking score– and reviewed postoperative imaging and pathological specimens. Results The majority (96%) of AM grafts were integrated into native dura. Thirteen patients scored as excellent and 11 good on our “dura mimicking score”, showing tissue integration ability but no cerebral cortex adhesion. The histopathological analysis showed that AM had thick plates of dense fibrous tissue with small reactive vessels, reactive fibroblasts, and lymphocytes infiltrate. The AM group's first outcomes were not different from the biological substitute patients but higher integration rate to the dura and less adhesion to the myocutaneous flap in AM patients. Conclusions We documented the anti-adhesive, protective, and integrative properties of AM dural substitute patches in patients who underwent decompressive craniectomies, comparing the intraoperative differences and postoperative outcomes to biological dural substitutes.

Background Facial palsy treatment comprises static and dynamic techniques. Among dynamic techniques, local temporalis transposition represents a reliable solution to achieve facial reanimation. The present study describes a modification of the temporalis tendon transfer using a cryopreserved fascia allograft. Case presentation Between March 2015 and September 2018, seven patients with facial palsy underwent facial reanimation with temporalis tendon transfer and fascia lata allograft. Patients with long-term palsy were considered, and both physical and social functions were evaluated. The mean follow-up time was 21.5 months. No immediate complications were observed. Patients reported improvement in facial symmetry both in static and dynamic. Improvement was noticed also in articulation, eating, drinking, and saliva control. The Facial Disability Index revealed an improvement both in physical function subscale and in the social/well-being function subscale. Conclusions This modified orthodromic technique allows to reduce the operative time and the risk of complications connected to the use of autologous tissues. The use of the cryopreserved fascia allografts from cadaveric donors seems to provide promising and long-standing results in the treatment of facial palsy.

Background: Implantation of the amniotic membrane and their derivatives can have a beneficial effect on tissue repair and regeneration. We report for the first time the implant of an amniotic membrane in a patient affected by cryptoglandular anal fistula. Methods: A patch of human amniotic membrane was implanted in a female patient affected by an anterior transphincteric fistula. Following an accurate curettage of the anal fistula, the cryopreserved amniotic membrane was thawed and then washed in the operating room; one side of the membrane was transfixed with a resorbable suture thus creating an implantable fusiform patch. The membrane was subsequently implanted into the fistula tract from the external to the internal opening. The inner and outer parts of the membrane were then sutured to the internal and external fistula openings. Results: No intraoperative or postoperative complications occurred. The patient was discharged one day after the procedure after an uneventful hospitalization. At the 1-week, 1- and 3-month follow-up visits no pain (VAS 0) was referred by the patient and no inflammation was evident at the level of the previous external fistula opening. Conclusions: The implant of human amniotic membrane in a patient affected by cryptoglandular anal fistula was safely and easily performed. Moreover, future studies to assess the efficacy in the long-term follow-up are needed.

Although careful donor selection reduces tissue contamination, close microbiological control of harvested allografts remains a key task of tissue banks. To guarantee the safety of human tissues for allograft transplantation, a decontamination regimen must be adopted which, as recommended by European guidelines, is active against the majority of microorganisms isolated in tissues. Antibiotic decontamination methods differ from one tissue bank to another in terms of antimicrobial agents, temperature and length of exposure. After identifying the most effective antibiotics against the bacterial strains most commonly isolated in allografts, Treviso Tissue Bank Foundation demonstrated the efficacy of an antibiotic cocktail for tissue decontamination containing Gentamicin, Vancomycin and Meropenem. The aim of this study was to analyse the degradation kinetics of the three antibiotics according to preparation method and use. The results show that only Meropenem is unstable at + 4 °C, while Gentamicin and Vancomycin are valid for over 10 days. We thus established to add Meropenem before the start of the tissue decontamination phase.

Cryopreserved allogeneic conduits are the elective biocompatible choice among currently available substitutes for surgical replacement in end-stage valvulopathy. However, degeneration occurs in 15 years in adults or faster in children, due to recipient's immunological reactions to donor's antigens. Here, human aortic valves were decellularized by TRICOL, based on Triton X-100 and sodium cholate, and submitted to standard cryopreservation (TRICOL-human aortic valves (hAVs)). Tissue samples were analyzed to study the effects of the combined procedure on original valve architecture and donor's cell removal. Residual amounts of nucleic acids, pathological microorganisms, and detergents were also investigated. TRICOL-hAVs proved to be efficaciously decellularized with removal of donor's cell components and preservation of valve scaffolding. Trivial traces of detergents, no cytotoxicity, and abrogated bioburden were documented. TRICOL-hAVs may represent off-the-shelf alternatives for both aortic and pulmonary valve replacements in pediatric and grown-up with congenital heart disease patients.

Mario Martelloni,1 Giulia Montagner,2 Diletta Trojan,2 Roberta Abate3 1Private Dental Practice, Roma 00179, Italy; 2Treviso Tissue Bank Foundation Onlus, Treviso, 31100, Italy; 3Private Dental Practice, Roma 00132, ItalyCorrespondence: Diletta TrojanFondazione Banca dei Tessuti di Treviso Onlus, Piazzale Ospedale, 1, Treviso 31100, ItalyTel +39 0422 322951Fax +39 0422 322849Email dtrojan@fbtv-treviso.orgAbstract: Mucogingival surgery has the objective to obtain coverage of the recession, with a favorable long-term prognosis, but also to minimize postoperative problems and surgery-related risks. Human amniotic membrane is increasingly employed for periodontal tissue repair in order to promote epithelialization and to reduce pain and scar tissue formation. A 38-year-old female patient reports dental hypersensitivity resulting from gingival recession affecting tooth 4.1. A mucogingival surgical procedure was proposed and a partial-thickness flap of epithelial-connective tissue was harvested from the palate and grafted on to the receiving site. The amniotic membrane was positioned at the donor site to reduce postoperative morbidity and to encourage rapid palatal healing. One week after the application of HAM (human amniotic membrane), the wound was healed and 1 month after the donor site was completely re-epithelialized. The present case report suggests that in the surgical treatment of gingival recession with palatal epithelial-connective tissue graft, HAM promotes rapid epithelialization of the palatal donor site wound with a reduction in morbidity.Keywords: gingival recession, amniotic membrane, palatal epithelial-connective tissue, human tissue

The amniotic membrane (AM) has an increasing role as a scaffold for tissue repair due to its unique biological properties. The implantation of AM on pancreatic anastomosis after pancreaticoduodenectomy might improve the anastomotic healing and strengthen its structure, however has never been used in pancreatic surgery. We present the first application of AM after a pancreaticoduodenectomy was performed for a malignant tumor of the pancreatic head. After completing the pancreatic anastomosis, the AM was placed around the pancreatic anastomosis and fixed to it with single stiches. The AM, due to its physical characteristics, could be easily manipulated and adapted to the pancreatic anastomosis. This interesting and unique case shows that covering a pancreatic anastomosis with the AM is safe and technically feasible. The AM has no adverse effects while it may eventually provide a beneficial impact over the anastomotic healing.

Background The aim of this study was to evaluate radiological and histological characteristics of fresh-frozen homologous bone as grafting material for maxillary sinus floor augmentation. Radiological, histological and clinical evaluations were made. Methods Twenty-three patients with a 2 mm to 6 mm alveolar ridge height in the posterior maxilla have been enrolled. Unilateral or bilateral sinus floor augmentations were performed with fresh frozen morcelized homologous bone. Together with implant placement, 7 months after surgery, a bone core was harvested for histological analysis. Radiological measurements were obtained by superimposition of CT scans carried out at the surgery time and six months later. A total of 93 implants were positioned. Results A mean (±SD) increase in mineralized tissue height of 10.74±2.82 mm was noticed by comparing the CT scans. Histological analysis revealed the presence of newly formed bone in the grafted sites. The follow up period after the prosthetic load ranged from 8 to 31 months. One implant failure occurred. Conclusions Fresh frozen homologous bone seems to have a good healing pattern and to be a successful and steady grafting material for the treatment of maxillary ridge atrophy. It might be considered a valid alternative to autologous bone in sinus floor augmentation procedures.

Background. Temporomandibular joint (TMJ) arthritis is a degenerative pathology that may cause pain and dysfunction. Nonsurgical therapy is the traditional treatment of TMJ diseases but if ineffective, TMJ surgery can be performed and may include arthroplasty with interposition of autograft. The encouraging results reported with the use of human amniotic membrane (HAM) in different surgical fields have highlighted its potential, but approaches providing the positioning of HAM within the intra-articular space of arthritic TMJs have never been investigated. Case Presentation. A 48-year-old woman was presented with limited mouth opening and pain with palpation at the left joint. A severe TMJ degeneration was diagnosed, and a surgical treatment was necessary. In the present case report, the authors describe the application of a cryopreserved HAM patch within the joint space as a disc-replacing film during major surgeries for discectomy and arthroplasty. Three months after the intervention, the patient reported an overall improvement in chewing efficiency as well as the absence of pain. Conclusions. According to the regenerative effects of HAM, the design of trials on the topic should be encouraged for its possible inclusion within the field of TMJ disease practice.

The aim of this work was to compare the effects on human amniotic membrane of freeze-drying and γ-irradiation at doses of 10, 20 and 30 kGy, with freezing. For this purpose, nine cytokines (interleukin 10, platelet-derived growth factor-AA, platelet-derived growth factor-BB, basic fibroblast growth factor, epidermal growth factor, transforming growth factor beta 1, and tissue inhibitors of metalloproteinase-1, -2, and -4) were titrated in 5 different preparations for each of 3 amniotic membranes included in the study. In addition, the extracellular matrix structure of each sample was assessed by transmission electron microscopy. While freeze-drying did not seem to affect the biological structure or cytokine content of the different amniotic membrane samples, γ-irradiation led to a significant decrease in the tissue inhibitors of metalloproteinase-4, basic fibroblast growth factor and epidermal growth factor, and induced structural damage to the epithelium, basement membrane and lamina densa. The higher the irradiation dose the more severe the damage to the amniotic membrane structure. In conclusion, the Authors recommend processing amniotic membrane under sterile conditions to guarantee safety at every step rather than final sterilization with γ-irradiation, thereby avoiding alteration to the biological characteristics of the amniotic membrane.

Microbiological contamination of retrieved tissues has become an issue of key importance and is a critical aspect of allograft safety, especially in the case of multi-tissue donations, which frequently become contaminated during retrieval and handling. We analysed contamination in 11,129 tissues with a longitudinal contamination profile for each individual tissue. Specifically, 10,035 musculoskeletal tissues and 1094 cardiovascular tissues were retrieved from a total of 763 multi-tissue donors, of whom 105 heart-beating organ donors and 658 deceased tissue donors. Of the 1955 tissues found to be contaminated after the first decontamination step, 1401 tissues (72%) were contaminated by the same species as the one(s) isolated at retrieval (Time1) and 554 (28%) by different species. Among the 113 tissues testing positive after the 2nd decontamination (Time3), 36 tissues (32%) were contaminated by the same species detected at Timel while the contaminating species differed from Time1 in 77 tissues (68%). The higher the number of contaminating species per tissue the higher the percentage of tissues in which contamination changed over time compared to Time1. The analysis revealed a 28% incidence of new species in tissues already testing positive after retrieval and of 3.5% of tissues becoming positive after admission to the tissue bank. Of these, coagulase-negative Staphylococcus accounted for over 70% of new contaminations.

The use of bisphosphonates has led to a new disease, bisphosphonate-related osteonecrosis of the jaw (BRONJ). There is currently no effective treatment for this disease; the surgical approach is controversial. The widespread use of human amniotic membrane (HAM) in surgery and the results obtained have highlighted its many potential properties, including antimicrobial, anti-inflammatory, antifibrotic and antiapoptotic, as well as its capacity for encouraging epithelialisation and cell differentiation. These properties are believed to encourage the recovery of patients with BRONJ, facilitating the wound healing process after surgical debridement of the bone. We report our experience with the use of HAM: two patients treated with patches of HAM. The follow-up to date, which includes x-rays and clinical assessments, demonstrates good levels of epithelialisation and absence of infections and pain. To conclude, the use of HAM in patients with BRONJ seems to be a promising therapeutic alternative to current conventional treatments.

Microbiological contamination of retrieved tissues is a critical aspect of allograft safety and tissue banks must continuously implement decontamination procedures to minimize tissue contamination. In this study we compared the decontamination efficacy of a new antibiotic cocktail (cocktail B: BASE medium with Gentamicin, Meropenem and Vancomycin) with the cocktail previously adopted at Treviso Tissue Bank Foundation (FBTV) (cocktail A: RPMI medium with Ceftazidime, Lincomycin, Polymyxin B and Vancomycin). Two decontamination steps were carried out, the first immediately after retrieval, the second after processing. The contamination rate was calculated before processing (Time 1) and cryopreservation (Time 2) for total tissues, musculoskeletal tissues and cardiovascular tissues, and the bacterial species involved were analyzed. Cocktail A was used to decontaminate 3548 tissues, of which 266 were cardiovascular and 3282 musculoskeletal tissues. For cocktail A, total tissue contamination was 18.6% at Time 1 and 0.9% at Time 2, with 15.7% contaminated musculoskeletal tissues at Time 1 and 0.4% at Time 2, respectively, while cardiovascular tissues were 50% contaminated at Time 1 and 6.4% at Time 2. Cocktail B was used to decontaminate 3634 tissues of which 318 were cardiovascular and 3316 musculoskeletal tissues. For cocktail B, total tissue contamination was 8.6% at Time 1 and 0.2% at Time 2, with 7.6% contaminated musculoskeletal tissues at Time 1 and 0.03% at Time 2, respectively. Contamination of cardiovascular tissues was 20.4% at Time 1 and 1.9% at Time 2. Intergroup and intragroup contamination rates decreased statistically significantly (p

Approaches providing the positioning of human amniotic membrane (HAM) within the intra-articular space of arthritic TMJs have never been investigated. This contrasts with the increasing amount of evidence suggesting the potential positive effects of HAM on a number of surgical conditions, even included the interpositional arthroplasty for TMJ ankylosis. Thus, the possible usefulness of HAM to restore joint functions in severely damaged TMJs could be hypothesized. Based on these premises, the clinical research question "Is human amniotic membrane positioning effective to reduce symptoms and restore jaw function in patients with severe inflammatory-degenerative disorders of the temporomandibular joint?" has been addressed by performing a systematic review of the literature. Out of potential 11988 and 8883 citations in the PubMed and Scopus databases, respectively, only five were of possible interest for inclusion in the review, but none of them addressed specifically the clinical research question. Thus, the hypothetical background for usefulness was discussed. The benefits of HAM positioning in TMJs with severe inflammatory-degenerative disorders could be related with its anti-inflammatory and anti-microbial and analgesic properties as well as its low immunogenicity. Studies in which HAM is positioned within the joint space of patients with severe TMJ degeneration, either as a disc-replacing film during major surgeries for discectomy and arthroplasty or as an injectable solution that can be needle-inserted after an arthrocentesis procedure, should be designed to test the hypothesis.

Microbiological contamination of retrieved tissues has become a very important topic and a critical aspect in the safety of allografts. We have analysed contamination in 11,129 tissues with a longitudinal contamination profile for each individual tissue. More specifically, 10,035 musculoskeletal tissues and 1,094 cardiovascular tissues were retrieved from a total of 763 multi-tissue donors, of whom 105 were heart-beating donors as well as organ donors, while the remaining 658 were non-heart beating donors and tissue donors only. All tissues were decontaminated twice, the first time immediately after retrieval and the second time after processing. Each tissue was submitted to microbiological culture three times, i.e., upon retrieval (Time 1), after the first decontamination (Time 2) and after the second decontamination (Time 3). The contamination rate for musculoskeletal tissues was 52%, 16.2% and 0.5% at Time 1, 2 and 3, respectively. The contamination rate for cardiovascular tissues was 84%, 42% and 6%. More than one strain was simultaneously present in 10.8% of musculoskeletal tissues and 44.6% of cardiovascular tissues. Out of 8,560 non-heart-beating donor musculoskeletal tissues, 4,689 (54.8%), 1,383 (16.2%) and 42 (0.5%) were contaminated at Time 1, Time 2 and Time 3, respectively. Out of 1,475 heart-beating donor musculoskeletal tissues, 522 (35.4%) 113 (7.7%) and 2 (0.1%) tissues were found to be contaminated at Time 1, 2 and 3, respectively. Out of 984 non-heart beating donor cardiovascular tissues, 869 (88.3%), 449 (45.6%) and 69 (7%) proved positive at Time 1, 2 and 3 respectively, while 50 (45.5%) and 10 (9.1%) heart-beating donor cardiovascular tissues were contaminated at Time 1 and 2. No tissue was contaminated at Time 3. Based on our methods, the two-step decontamination approach is mandatory in order to drastically reduce the number of tissues found to be positive at the end of the process.

Links between cancer and stem cells have been proposed for many years. As the cancer stem cell (CSC) theory became widely studied, new methods were developed to culture and expand cancer cells with conserved determinants of “stemness”. These cells show increased ability to grow in suspension as spheres in serum-free medium supplemented with growth factors and chemicals. The physiological relevance of this phenomenon in established cancer cell lines remains unclear. Cell lines have traditionally been used to explore tumor biology and serve as preclinical models for the screening of potential therapeutic agents. Here, we grew cell-forming spheres (CFS) from 25 established colorectal cancer cell lines. The molecular and cellular characteristics of CFS were compared to the bulk of tumor cells. CFS could be isolated from 72 % of the cell lines. Both CFS and their parental CRC cell lines were highly tumorigenic. Compared to their parental cells, they showed similar expression of putative CSC markers. The ability of CRC cells to grow as CFS was greatly enhanced by prior treatment with 5-fluorouracil. At the molecular level, CFS and parental CRC cells showed identical gene mutations and very similar genomic profiles, although microarray analysis revealed changes in CFS gene expression that were independent of DNA copy-number. We identified a CFS gene expression signature common to CFS from all CRC cell lines, which was predictive of disease relapse in CRC patients. In conclusion, CFS models derived from CRC cell lines possess interesting phenotypic features that may have clinical relevance for drug resistance and disease relapse.

Bacterial contamination of tissues retrieved from cadaveric donors is a common feature worldwide, and every tissue bank, albeit using different methods, conducts decontamination to guarantee safe tissues suitable for clinical use. The effectiveness of the methods used to eradicate pathogens differs. In order to reduce the tissue bioburden at retrieval, we have introduced a new method involving rinsing tissues in a sodium hypochlorite solution. To test its effectiveness we analyzed two comparable groups of tissues: Group A: 1881 tissues, all rinsed with isotonic saline solution after retrieval, and Group B: 1968 tissues immersed in an isotonic saline solution containing sodium hypochlorite (final concentration 0.1 %) for different lengths of time and subsequently rinsed with isotonic saline. The rinsing solution of each tissue was then sampled for microbiological cultures in both groups. The resultant overall contamination rate was 40.5 % for Group A and 6.7 % for Group B, with an 82.8 % difference in the reduction of contamination between the two groups. This was especially the case for commensal skin bacteria in musculoskeletal tissue, which accounted for over half the overall contamination. Our data highlighted that decontamination with sodium hypochlorite was helpful in reducing the bacterial bioburden in tissues retrieved from cadaveric donors.

The use of amniotic membrane (AM) is a widespread clinical practice for eye surgeries and the treatment of an increasing number of ocular surface pathologies. Here we describe the AM collection methods and donor selection criteria adopted by our tissue bank to distribute 5349 amniotic membrane patches over the last 12 years for the treatment of several ocular pathologies. Specific quality control measures are described and the long term results attained using the reported procedure are presented. A case of AM utilized to treat severe ocular ulceration is also described as an example of AM transplantation. Collective data for the total amniotic membrane patches deployed to treat various ocular diseases are discussed and success rates for AM transplantations are reported. An extensive follow-up is illustrated. The results suggest that the procedures and protocols used by the Treviso Tissue Bank Foundation and Veneto Eye Bank Foundation for collection, preservation, distribution and follow-up are of an optimal standard. Accordingly, the authors conclude that the safety and efficiency of the proposed procedure for the therapeutic use of AM to treat various ocular pathologies are reproducible, with additional evidence favoring the use of AM as an alternative to conventional medical treatment for certain ocular conditions.