–
For decades, dermal tissue grafts have been used in various regenerative, reconstructive, and augmentative procedures across the body. To eliminate antigenicity and immunogenic response while still preserving the individual components and collective structural integrity of the extracellular matrix (ECM), dermis can be decellularized. Acellular dermal matrix (ADM) products like such are produced to accurately serve diverse clinical purposes. The aim of the present study is to evaluate the efficacy of a novel decellularization protocol of the human dermis, which eliminates residual human genetic material without compromising the biomechanical integrity and collagenous content of the tissue. Moreover, a freeze-drying protocol was validated. The results showed that though our decellularization protocol, human dermis can be decellularized obtaining a biocompatible matrix. The procedure is completely realized in GMP aseptic condition, avoiding tissue terminal sterilization.
https://link.springer.com/article/10.1007/s10561-024-10131-6
20 Settembre 2023
Musculoskeletal allografts represent an important practice in orthopedic surgeries and the demand for them has been growing. For this reason, in order to reduce clinical risk and to more efficiently manage the increase of allograft usage and also to optimize timing of the surgeries, the thawing and washing processes with aseptic technique were centralized in the department of Hospital Pharmacy. This study describe the design and execution of an adapted Media Fill Test (MFT) to demonstrate aseptic thawing and washing of allografts. For this specific and innovative setting, to better simulate the actual processing steps, a surrogate system was developed to simulate the tendon allograft. The aseptic technique of four operators was assessed and an initial performance validation and the first revalidation were described. All MFT were completed successfully, with no observation of turbidity. The readapted MFT shown in this study can provide insight into this innovative and growing field to other health professionals who want to implement this service.
https://pubmed.ncbi.nlm.nih.gov/37728671/
Antonella Galeone, Jacopo Gardellini, Diletta Trojan, Venanzio Di Nicola, Renato Di Gaetano, Giuseppe Faggian, Giovanni Battista Luciani
The objective of this study was to evaluate early and long-term outcomes of patients with aortic prosthetic valve endocarditis (a-PVE) treated with a prosthetic aortic valve (PAV), prosthetic valved conduit (PVC), or cryopreserved aortic homograft (CAH). A total of 144 patients, 115 male and 29 female, aged 67 ± 12 years, underwent surgery for a-PVE at our institution between 1994 and 2021. Median time from the original cardiac surgery was 1.9 [0.6-5.6] years, and 47 (33%) patients developed an early a-PVE. Of these patients, 73 (51%) underwent aortic valve replacement (AVR) with a biological or mechanical PAV, 12 (8%) underwent aortic root replacement (ARR) with a biological or mechanical PVC, and 59 (42%) underwent AVR or ARR with a CAH. Patients treated with a CAH had significantly more circumferential annular abscess multiple valve involvement, longer CPB and aortic cross-clamping times, and needed more postoperative pacemaker implantation than patients treated with a PAV. No difference was observed in survival, reoperation rates, or recurrence of IE between patients treated with a PAV, a PVC, or a CAH. CAHs are technically more demanding and more often used in patients who have extensive annular abscess and multiple valve involvement. However, the use of CAH is safe in patients with complex a-PVE, and it shows excellent early and long-term outcomes.
https://pubmed.ncbi.nlm.nih.gov/37623351/
Andrea Ossato, Valeria Mezzadrelli, Giulia Montagner, Diletta Trojan, Giuseppe Giovagnoni, Michele Giannini, Carlotta Trabucchi, Chiara Angelini, Francesca Realdon, Lorenza Cipriano, Nicola Realdon, Teresa Zuppini, Roberto Tessari
The transplantation of human tissues is a greatly expanding field of medicine with unquestionable benefits that raise questions about safety, quality and ethics. Since 1 October 2019, the Fondazione Banca dei Tessuti del Veneto (FBTV) stopped sending thawed and ready to be transplanted cadaveric human tissues to hospitals. A retrospective analysis of the period 2016-2019 found a significant number of unused tissues. For this reason, the hospital pharmacy has developed a new centralised service characterised by thawing and washing human tissues for orthopaedic allografts. This study aims to analyse the hospital cost and benefit derived from this new service.
https://pubmed.ncbi.nlm.nih.gov/37316166/
Elena Stocco, Silvia Barbon, Marco Mammana, Diletta Trojan, Alice Bianchin, Francesca Favaretto, Martina Contran, Giovanni Zambello, Andrea Vogliardi, Marta Confalonieri, Silvia Todros, Piero G Pavan, Filippo Romanato, Maria Teresa Conconi, Veronica Macchi, Raffaele De Caro, Federico Rea, Andrea Porzionato
Tracheal reconstruction represents a challenge when primary anastomosis is not feasible. Within this scenario, the study aim was to develop a new pig-derived decellularized trachea (DecellT) to be compared with the cryopreserved counterpart (CryoT) for a close predictive analysis. Tracheal segments underwent decellularization by a physical + enzymatic + chemical method (12 cycles); in parallel, cryopreserved samples were also prepared. Once decellularized (histology/DNA quantification), the two groups were characterized for Alpha-Gal epitopes/structural proteins (immunohistochemistry/histology/biochemical assays/second harmonic generation microscopy)/ultrastructure (Scanning Electron Microscopy (SEM))/mechanical behaviour. Cytotoxicity absence was assessed in vitro (extract-test assay/direct seeding, HM1SV40 cell line) while biocompatibility was verified in BALB/c mice, followed by histological/immunohistochemical analyses and SEM (14 days). Decellularization effectively removed Alpha-Gal epitopes; cartilage histoarchitecture was retained in both groups, showing chondrocytes only in the CryoT. Cryopreservation maintained few respiratory epithelium sparse cilia, not detectable in DecellT. Focusing on ECM, preserved structural/ultrastructural organization and collagen content were observed in the cartilage of both; conversely, the GAGs were significantly reduced in DecellT, as confirmed by mechanical study results. No cytotoxicity was highlighted by CryoT/DecellT in vitro, as they were also corroborated by a biocompatibility assay. Despite some limitations (cells presence/GAGs reduction), CryoT/DecellT are both appealing options, which warrant further investigation in comparative in vivo studies
https://pubmed.ncbi.nlm.nih.gov/36980229/
Francesca Favaretto, Chiara Compagnin, Elisa Cogliati, Giulia Montagner, Francesco Dell’Antonia, Giorgio Berna, Roberto Vettor, Gabriella Milan, Diletta Trojan
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.
https://pubmed.ncbi.nlm.nih.gov/37175896/
Giulia Montagner, Antonia Barbazza, Andrea Tancredi Lugas, Mara Terzini, Gianpaolo Serino, Cristina Bignardi, Matilde Cacciatore, Vladimiro L. Vida, Massimo A. Padalino, Diletta Trojan
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 infammation and tissue calcifcation 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 efcacy 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.
https://link.springer.com/article/10.1007/s10561-023-10072-6
Montagner G, De Vettor R, Favaretto F, Vici D, Del Vecchio C, Franchin E, Trojan D, Feltrin G.
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.
https://pubmed.ncbi.nlm.nih.gov/35235097/
2019
Implantation of amniotic membrane over pancreatic anastomosis after pancreaticoduodenectomy: report of the first case.
9 maggio 2019
J Surg Case Rep. 2019 May 9;2019(5):rjz097. doi: 10.1093/jscr/rjz097
Frigerio I, Bannone E, Trojan D, Montagner G, Bergamaschi G, Butturini G
Abstract
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.
https://www.ncbi.nlm.nih.gov/pubmed/?term=PMID%3A+31086645
Human Amniotic Membrane Positioning in the Surgical Treatment of Temporomandibular Joint Degenerative Disorder.
3 marzo 2019
Case Rep Surg. 2019 Mar 3;2019:6037191.
Guarda-Nardini L, Trojan D, Montagner G, Cogliati E, Bendini M, Manfredini D.
Abstract
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.
https://www.ncbi.nlm.nih.gov/pubmed/?term=PMID%3A+30944749
2018
Contamination profile in allografts retrieved from multitissue donors: longitudinal analysis.
21 novembre 2018
Paolin A, Montagner G, Petit P, Trojan D.
Cell Tissue Bank. 2018 Dec;19(4):809-817
Abstract
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.
Stability analysis of the antibiotic cocktail used by Treviso Tissue Bank Foundation for tissues decontamination.
11 settembre 2018
Montagner G, Trojan D, Cogliati E, Manea F, Vantini A, Paolin A.
Cell Tissue Bank. 2018 Dec;19(4):721-726.
Abstract
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.
Evaluation of allograft decontamination with two different antibiotic cocktails at the Treviso Tissue Bank Foundation.
2 agosto 2018
Paolin A, Spagnol L, Battistella G, Trojan D.
PLoS One. 2018 Aug 2;13(8):e0201792.
Abstract
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<0.05). Our results have shown that cocktail B was more effective than cocktail A in killing bacteria in both cardiovascular and musculoskeletal tissues during the two decontamination cycles.
https://www.ncbi.nlm.nih.gov/pubmed/?term=PMID%3A+30071084
Homologous cryopreserved amniotic membrane in the repair of myelomeningocele: preliminary experience.
1 giugno 2018
Marton E, Giordan E, Gioffrè G, Canova G, Paolin A, Mazzucco MG, Longatti P.
Acta Neurochir (Wien). 2018 Aug;160(8):1625-1631.
Abstract
OBJECTIVE: Surgical management of spinal dysraphism often requires the use of dural substitutes. Amniotic membrane (AM) has drawn the interest of clinicians for its valuable concentration of cytokines and factors capable of promoting wound healing, re-epithelialization, inhibiting fibrosis and regulating angiogenesis. These beneficial qualities could make AM an interesting dural substitute for spina bifida repair. In this study, we describe the use of banked homologous AM as a dural substitute for the repair of spinal dysraphism in newborns. Our purpose is to test the mechanical characteristics, as well as the safety and effectiveness of AM in preventing postoperative complications and re-tethering.
METHODS: The AM patch was carefully detached from the chorion of donors undergoing caesarean section, rinsed in saline solution, and cryopreserved in liquid nitrogen. Five newborns were treated using AM: three affected by open spinal dysraphism and two by spina bifida occulta. The AM patch was used as a dural substitute with two different positions and purposes: the amnion-side down covering the placode to prevent adhesions or placed extradurally facing the dura to avoid scarring and facilitating the sliding of the dural sac itself under the extradural tissue layers.
RESULTS: No adverse events occurred, and the surgical wounds healed without complications. MRI scans taken at 3 and 6 months after surgery showed a satisfying de-tethering of the spinal cord with no obvious evidence of new adherence formation.
CONCLUSIONS: We present a multimodal interposition technique using AM as a reconstructive and anti-adhesive tissue for the treatment of open myelomeningocele (MMC) and lipomeningocele (LMC) treatment. In our experience, AM proved its efficacy in restoring the dural sac integrity without complications. We support the use of AM as a promising dural substitute, speculating on how the use of AM could potentially change reconstructive strategies for spinal dysraphism.
Use of amniotic membrane in the treatment of patients with BRONJ: two case reports.
20 aprile 2018
Ragazzo M, Trojan D, Spagnol L, Paolin A, Guarda Nardini L.
J Surg Case Rep. 2018 Apr 20;2018(4):rjy073.
Abstract
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.
https://www.ncbi.nlm.nih.gov/pubmed/?term=PMID%3A+29692884
2017
Analysis of potential factors affecting allografts contamination at retrieval.
17 ottobre 2017
Cell Tissue Bank. 2017 Dec;18(4):539-545.
Paolin A, Romualdi C, Romagnoli L, Trojan D.
Abstract
The microbiological contamination of retrieved tissues has become a very important topic and it is a critical aspect in the safety of allografts, especially from multi-tissue donors whose tissues are frequently contaminated as a consequence of retrieval. We analysed a total of 10,107 tissues, 8178 musculoskeletal and 1929 cardiovascular tissues, retrieved from 978 multi-tissue donors. Of these, 159 heart-beating donors (HBD) were also organ donors, while the remaining 819 non-heart-beating donors (NHBD) were tissue donors only. A multivariate logistic model was used to determine the factors affecting contamination risk during retrieval. In the model, the dependent variable was the presence/absence of contamination while the covariates included were: gender, type of donor, age of donor, cause of death, previous skin donation, cadaver time, number of people attending the retrieval, number of tissues retrieved. Moreover, a second log-linear model was used to determine the number of strains isolated per tissue. Tissue contamination was statistically correlated with gender, type of donor, cadaver time, number of people attending the retrieval and season. In conclusion, to minimize the risk of bacterial contamination, aseptic techniques should be used at retrieval, with the number of retrieval team members kept to a minimum. In addition, cadaver time should be as short as possible and the donor should be refrigerated within a few hours after death.
Management of temporomandibular joint degenerative disorders with human amniotic membrane: Hypothesis of action.
29 maggio 2017
Guarda-Nardini L, Trojan D, Paolin A, Manfredini D.
Med Hypotheses. 2017 Jul;104:68-71.
Abstract
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.
Decellularized Cryopreserved Allografts as Off-the-Shelf Allogeneic Alternative for Heart Valve Replacement: In Vitro Assessment Before Clinical Translation.
9 marzo 2017
Iop L, Paolin A, Aguiari P, Trojan D, Cogliati E, Gerosa G.
J Cardiovasc Transl Res. 2017 Apr;10(2):93-103.
Abstract
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.
Evaluation of allograft contamination and decontamination at the Treviso Tissue Bank Foundation: A retrospective study of 11,129 tissues.
7 marzo 2017
Paolin A, Trojan D, Petit P, Coato P, Rigoli R.
PLoS One. 2017 Mar 7;12(3):e0173154.
Abstract
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.
https://www.ncbi.nlm.nih.gov/pubmed/?term=PMID%3A+28267776
2016
Analysis of the effectiveness of Sodium Hypochlorite decontamination of cadaveric human tissues at retrieval.
18 ottobre 2016
Cell Tissue Bank. 2016 Dec;17(4):611-618.
Paolin A, Trojan D, Carniato A, Tasca F, Massarin E, Tugnoli A, Cogliati E.
Abstract
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.
https://www.ncbi.nlm.nih.gov/pubmed/?term=PMID%3A+27757727
Evaluation of new antibiotic cocktails against contaminating bacteria found in allograft tissues.
7 settembre 2016
Serafini A, Riello E, Trojan D, Cogliati E, Palù G, Manganelli R, Paolin A.
Cell Tissue Bank. 2016 Dec;17(4):619-628.
Abstract
Contamination of retrieved tissues is a major problem for allograft safety. Consequently, tissue banks have implemented decontamination protocols to eliminate microorganisms from tissues. Despite the widespread adoption of these protocols, few comprehensive studies validating such methods have been published. In this manuscript we compare the bactericidal activity of different antibiotic cocktails at different temperatures against a panel of bacterial species frequently isolated in allograft tissues collected at the Treviso Tissue Bank Foundation, a reference organization of the Veneto Region in Italy that was instituted to select, recover, process, store and distribute human tissues. We were able to identify at least two different formulations capable of killing most of the bacteria during prolonged incubation at 4 °C.
https://www.ncbi.nlm.nih.gov/pubmed/?term=PMID%3A+27604467
Cytokine expression and ultrastructural alterations in fresh-frozen, freeze-dried and γ-irradiated human amniotic membranes.
12 aprile 2016
Paolin A, Trojan D, Leonardi A, Mellone S, Volpe A, Orlandi A, Cogliati E.
Cell Tissue Bank. 2016 Sep;17(3):399-406.
Abstract
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.
https://www.ncbi.nlm.nih.gov/pubmed/?term=PMID%3A+27072557
2015
Amniotic membranes in ophthalmology: long term data on transplantation outcomes.
11 luglio 2015
Paolin A, Cogliati E, Trojan D, Griffoni C, Grassetto A, Elbadawy HM, Ponzin D.
Cell Tissue Bank. 2016 Mar;17(1):51-8.
Abstract
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.
https://www.ncbi.nlm.nih.gov/pubmed/?term=PMID%3A+26162811
Cardiac mesenchymal stromal cells are a source of adipocytes in arrhythmogenic cardiomyopathy.
20 novembre 2015
Sommariva E, Brambilla S, Carbucicchio C, Gambini E, Meraviglia V, Dello Russo A, Farina FM, Casella M, Catto V, Pontone G, Chiesa M, Stadiotti I, Cogliati E, Paolin A, Ouali Alami N, Preziuso C, d’Amati G, Colombo GI, Rossini A, Capogrossi MC, Tondo C, Pompilio G.
Eur Heart J. 2016 Jun 14;37(23):1835-46
Abstract
AIM:Arrhythmogenic cardiomyopathy (ACM) is a genetic disorder mainly due to mutations in desmosomal genes, characterized by progressive fibro-adipose replacement of the myocardium, arrhythmias, and sudden death. It is still unclear which cell type is responsible for fibro-adipose substitution and which molecular mechanisms lead to this structural change. Cardiac mesenchymal stromal cells (C-MSC) are the most abundant cells in the heart, with propensity to differentiate into several cell types, including adipocytes, and their role in ACM is unknown. The aim of the present study was to investigate whether C-MSC contributed to excess adipocytes in patients with ACM.
METHODS AND RESULTS: We found that, in ACM patients’ explanted heart sections, cells actively differentiating into adipocytes are of mesenchymal origin. Therefore, we isolated C-MSC from endomyocardial biopsies of ACM and from not affected by arrhythmogenic cardiomyopathy (NON-ACM) (control) patients. We found that both ACM and control C-MSC express desmosomal genes, with ACM C-MSC showing lower expression of plakophilin (PKP2) protein vs.
CONTROLS: Arrhythmogenic cardiomyopathy C-MSC cultured in adipogenic medium accumulated more lipid droplets than controls. Accordingly, the expression of adipogenic genes was higher in ACM vs. NON-ACM C-MSC, while expression of cell cycle and anti-adipogenic genes was lower. Both lipid accumulation and transcription reprogramming were dependent on PKP2 deficiency.
CONCLUSIONS: Cardiac mesenchymal stromal cells contribute to the adipogenic substitution observed in ACM patients’ hearts. Moreover, C-MSC from ACM patients recapitulate the features of ACM adipogenesis, representing a novel, scalable, patient-specific in vitro tool for future mechanistic studies.
https://www.ncbi.nlm.nih.gov/pubmed/?term=PMID%3A+26590176
2013
Nerve regeneration across cryopreserved allografts from cadaveric donos: a novel approach for peripheral nerve reconstruction
26 luglio 2013
Giovanna Squintani, Bruno Bonetti, Adolfo Paolin, Daniela Vici, Elisa Cogliati, Bruno Murer, Giorgio Stevanato
J Neurosurg. 2013 Oct;119(4):907-13.
Abstract
OBJECT. The use of allografts from cadaveric donors has attracted renewed interest in recent years, and pretreatment with cryopreservation adn immunosuppression methods has been investigated to maximize axonal regrowth and minimize allograft rejection. The authors wanted to assess the outcome of treatments of brachial plexus stretch injuries with cryopreserved allografts from cadaveric donors in nonimmunosuppressed patients.
METHODS. Ten patients with brachial plexus lesions were submitted to electromyography (EMG) testing 1 and 3 months after a traumatic event and 1 week before surgery to localize and identify the type of lesion. Intraoperative EMG recordings were performed for intraoperative monitoring to select the bet surgical strategy, and postoperative EMG was used to follow up patients and determine surgical outcomes. If nerve action potentials (NAPs) were present intraoperatively, neurolysis was performed, whereas muscular/nerve neurotization was performed if NAPs were absent. Cryopreserved allografts obtained from selected cadaveric donors and provided by the tissue bank of Treviso were used for nerve reconstruction in patients who were not treated with immunosuppressive drugs.
RESULTS. The surgical strategy was selected according to the type and site of the nerve lesion and on the basis of IOM results: 14 cryopreserved allografts were used for 7 muscular neurotizations and for 7 nerve neurotizations, and 5 neurolysis procedures were performed. All of the patients had regained motor function at the 1- and 2-year follow-ups.
CONCLUSIONS. Some variables may affect functional recovery after allograft surgery, and the outcome of peripheral nerve reconstruction is more favorable when patients are carefully evaluated and selected for the surgery. The authors demonstrated that using cryopreserved allografts from cadaveric donors is a valid surgical strategy to restore function of the damaged nerve without the need for any immunosuppressive treatments. This approach offers new perspectives on procedures for extensive reconstruction of brachial and lumbosacral plexures.
2010
Fresh-frozen bone: case series of a new grafting material for sinus lift and immediate implants.
Agosto 2010
Viscioni A, Dalla Rosa J, Paolin A, Franco M.
J Ir Dent Assoc. 2010 Aug-Sep;56(4):186-91.
Abstract
STATEMENT OF THE PROBLEM: Although autologous bone is considered to be the gold standard grafting material, it needs to be harvested from patients, a process that can be off-putting and can lead to donor site morbidity. For this reason, homologous fresh-frozen bone (FFB) was used in the current study as an alternative graft material.
PURPOSE OF THE STUDY: The aim of this study was to evaluate the effectiveness of FFB as a grafting material in complex maxillary sinus lift with immediate implant insertion.
METHODS: FFB was obtained from the Veneto Tissue Bank and preserved at -80 degrees C. Twenty-one patients were surgically treated with FFB block grafts in 26 maxillary sinus rehabilitations, with 47 immediate implant insertions, with a reopening phase after six months. All patients underwent orthopanoramic X-rays and CT scans before, immediately after and four months (X-ray only) post surgery. Bone biopsies were performed in order to evaluate the volume and density of the bone grafts, which all showed optimal adherence without complications. RESULTS: Four months post surgery, 64% of grafts showed no evidence of bone resorption or resizing. In all other cases resorption was slight. All implants were clinically osseointegrated, with only one implant failure during the provisional prosthetic loading stage (97.8% success rate). Histological studies confirmed these results, showing the presence of new bone and sparse osteoclastic activity four months post implantation, with 80% mature bone material observed after 12 months.
CONCLUSIONS: Use of FFB permits effective bone-adding surgery and immediate implant insertion under local anaesthesia, decreasing both chair time and patient discomfort.
Effectiveness of fresh frozen and cryopreserved homologue iliac crest grafts used in sinus lifting: a comparative study.
6 luglio 2010
Viscioni A, Franco M, Paolin A, Cogliati E, Callegari M, Zollino I, Sollazzo V, Carinci F.
Cell Tissue Bank. 2011 Nov;12(4):263-71.
Abstract
In the last decade, several investigators have reported that autologous and homologous fresh frozen bones (FFB) are effective materials to restore alveolar ridges previous to insert dental implants. Recently we have used cryopreserved homologue grafts (CFFB). Here we reported a retrospective comparative study between implants inserted in FFB and CFFB evaluate their clinical outcome. Patients were treated with a split mouth scheme for bone grafting with FFB and CFFB and spiral family implants (SPI) were inserted in the same surgical time. Several variables (patient, grafts, anatomic site, implant, prosthetic restoration) were investigated. Implant’ failure and peri-implant bone resorption were considered as predictor of clinical outcome. A total of 84 SFIs were inserted in 12 patients. Implants were inserted to replace 8 incisors, 4 cuspids, 31 premolars and 41 molars. The mean follow-up was 14 months. Three out of 84 implants was lost (i.e. survival rate SVR = 96.4%) and no differences were detected among the studied variables. Similar result was obtained by analyzing the crestal bone resorption around implant’ neck (i.e. success rate). FFB and CFFB have high and comparable survival and success rate. Implants inserted with one step surgical procedure in native (i.e. not grafted) bone, FFB and CFFB have similar clinical outcome.
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