BeST-Graft viewer, a new system to improve the bone allograft-recipient matching process.

INTRODUCTION: Tissue establishments are responsible for processing, testing, preserving, storing, and distributing allografts from donors to be transplanted into recipients. In some situations, a matching process is required to determine the allograft that best fits the recipient. Allograft morphology is a key consideration for the matching process. The manual procedures applied to obtain these parameters make the process error-prone. MATERIAL AND METHODS: A new system to manage bone allograft-recipient matching for tissue establishments is proposed. The system requires bone allografts to be digitalized and the resulting images to be stored in a DICOM file. The system provides functionalities to: (i) manage DICOM files (registered in the PACs) from both allografts and recipients; (ii) reconstruct 3D models from DICOM images; (iii) explore 3D models using 2D, 3D, and multiplanar reconstructions; (iv) take allograft and recipient measurements; and (v) visualize and interact with recipient and allograft data simultaneously. The system has been installed in the Barcelona Tissue Bank (Banc de Sang i Teixits), which has digitalized the bone allografts to test the system. RESULTS: A use case with a femur is presented to test all the viewer functionalities. In addition, the recipient-allograft workflow is evaluated to show the steps of the procedure where the viewer can be used. CONCLUSIONS: The bone allograft-recipient matching procedure can be optimized using software tools with functionalities to visualize, interact, and take measurements.

Quality by Design: Development of Safe and Efficacious Full-Thickness Acellular Dermal Matrix Based on EuroGTPII Methodologies.

Background: The activities of tissue establishments are constantly and rapidly evolving. The development of a new type of allograft, full-thickness acellular dermal matrix, with high mechanical properties to be used in tendon repair surgeries and abdominal wall reconstruction, has determined the need for quality by design process in order to assess evidence of quality, safety and efficacy. The EuroGTPII methodologies were specifically tailored to perform the risk assessment, identify and suggest tests in order to mitigate the potential risk consequences of a novel tissue preparation implementation. Methods: The new allograft and associated preparation processes were assessed using the EuroGTP methodologies and characterized to properly evaluate the novelty (Step 1), identify and quantify the potential risks and risk consequences (Step 2), and define the extent of pre-clinical and clinical assessments required to mitigate the risks identified in the assessment (Step 3). Results: Four risk consequences associated with the preparation process were identified: (i) implant failure related with tissue procurement and the reagents used during the decellularization protocol; (ii) unwanted immunogenicity related with the processing; (iii) disease transmission linked with the processing, reagents used, reduction in the reliability of microbiology testing and the storage conditions; and (iv) toxicity related to the reagents used and handling of the tissue during clinical application. The outcome of the risk assessment was a low level of risk. Nevertheless, it determined the need for a series of risk mitigation strategies proposed to reduce each individual risk and to provide additional evidence of the safety and efficacy of full-thickness acellular dermal matrix grafts. Conclusion: EuroGTPII methodologies allow us to identify the risks and ensure the correct definition of pre-clinical assessments required to address and mitigate the potential risk consequences, before proceeding with clinical use of the new allografts in patients.

Decellularized Graft for Repairing Severe Peripheral Nerve Injuries in Sheep.

BACKGROUND AND OBJECTIVES: Peripheral nerve injuries resulting in a nerve defect require surgical repair. The gold standard of autograft (AG) has several limitations, and therefore, new alternatives must be developed. The main objective of this study was to assess nerve regeneration through a long gap nerve injury (50 mm) in the peroneal nerve of sheep with a decellularized nerve allograft (DCA). METHODS: A 5-cm long nerve gap was made in the peroneal nerve of sheep and repaired using an AG or using a DCA. Functional tests were performed once a month and electrophysiology and echography evaluations at 6.5 and 9 months postsurgery. Nerve grafts were harvested at 9 months for immunohistochemical and morphological analyses. RESULTS: The decellularization protocol completely eliminated the cells while preserving the extracellular matrix of the nerve. No significant differences were observed in functional tests of locomotion and pain response. Reinnervation of the tibialis anterior muscles occurred in all animals, with some delay in the DCA group compared with the AG group. Histology showed a preserved fascicular structure in both AG and DCA; however, the number of axons distal to the nerve graft was higher in AG than in DCA. CONCLUSION: The decellularized graft assayed supported effective axonal regeneration when used to repair a 5-cm long gap in the sheep. As expected, a delay in functional recovery was observed compared with the AG because of the lack of Schwann cells.

Are Hamstring Grafts a Predisposing Factor to Infection in R-ACL Surgery? A Comparative In Vitro Study.

BACKGROUND: The objective of the present study was to evaluate the formation of biofilms in bone patellar tendon bone grafts (BPTB grafts), and to compare it to the formation of biofilm formation in quadrupled hamstring anterior cruciate ligament grafts (4×Ht graft). METHODS: A descriptive in vitro study was conducted. One 4×Ht graft and one BPTB graft were prepared. They were then contaminated with a strain of S. epidermidis. Later, a quantitative analysis was conducted by means of microcalorimetry and sonication with plating. Additionally, a qualitative analysis was conducted by means of electron microscopy. RESULTS: No significant differences were found between the bacterial growth profiles of the 4×Ht graft and the BPTB graft in microcalorimetry and colony counting. In the samples analyzed with electron microscopy, no specific biofilm growth pattern was identified upon comparing the BPTB graft to the 4×Ht graft. CONCLUSIONS: There were no significant differences found at either the quantitative or qualitative level when comparing bacterial growth in the BPTB graft to that in the 4×Ht graft. Therefore, the presence of sutures in the 4×Ht graft cannot be established as a predisposing factor for increased biofilm growth in this in vitro study.

Early Postoperative CT Scan Provides Prognostic Data on Clinical Outcomes of Fresh Osteochondral Transplantation of the Knee.

BACKGROUND: There is a lack of information regarding the ability of imaging studies to predict clinical outcomes after fresh osteochondral allograft (FOCA) transplantation of the knee. PURPOSE: To determine the value of computed tomography (CT) scans to predict the clinical outcome of FOCA transplantation using the assessment computed tomography osteochondral allograft (ACTOCA) score. STUDY DESIGN: Cohort study; Level of evidence, 3. METHODS: We prospectively collected data from all consecutive patients who underwent FOCA transplantation for osteochondral knee lesions at one institution between August 2017 and August 2019. All patients were followed up for a minimum of 2 years. CT scans performed 6 months after surgery were evaluated by a musculoskeletal radiologist using the ACTOCA scoring system. The radiologist was blinded to the patient's medical history. Clinical outcomes were assessed preoperatively and at 12 and 30 months postoperatively using the International Knee Documentation Committee (IKDC) score, the Kujala score, the Tegner activity scale, and the Western Ontario Meniscal Evaluation Tool (WOMET) score. RESULTS: A total of 38 cases were included. The ACTOCA score at 6 months after surgery showed a statistically significant correlation with clinical results at 12 and 30 months. The correlation was better at 30 months, showing a high negative correlation with the IKDC score (-0.663) and a moderate negative correlation with the Kujala, WOMET, and Tegner scores (-0.593; -0.547, and -0.593, respectively) (P < .001). CONCLUSION: A statistically significant correlation between the mean ACTOCA score on CT scans at 6 months and the clinical results measured by the IKDC, Kujala, WOMET, and Tegner scores at 30 months confirmed the predictive value of the ACTOCA score for use in clinical practice.

A Comparative Study Using Fluorescent Confocal Microscopy and Flow Cytometry to Evaluate Chondrocyte Viability in Human Osteochondral Allografts.

The preservation conditions of fresh osteochondral allografts for clinical applications are critical due their objective: to transplant mature hyaline cartilage containing viable chondrocytes, maintaining their metabolic activity and also preserving the structural and functional characteristics of the extracellular matrix. The aim of the present study was to compare fluorescence confocal microscopy and flow cytometry techniques to evaluate the viability of the chondrocytes present in the osteochondral tissue, in order to determine their effectiveness and thus ensure reproducibility and robustness of the analysis. To this end, osteochondral allografts from human cadaveric donors were preserved at 4 °C for 3 weeks in a preservation medium supplemented with antibiotic and antifungal agents. Cell viability of chondrocytes was determined by monitoring the cartilage for 3 weeks of preservation by confocal fluorescence microscopy and flow cytometry, obtaining cell viabilities of 83.7 ± 2.6% and 55.8 ± 7.8% for week three, respectively. The confocal fluorescence microscopy approach is more advantageous and accurate, as it correlates better with actual cell viability values for monitoring osteochondral graft preservation, detecting only the cells that died a natural death associated with the preservation method.

A novel decellularized nerve graft for repairing peripheral nerve long gap injury in the rat.

Decellularized nerve allografts are an alternative to autograft for repairing severe nerve injuries, since they have higher availability and do not induce rejection. In this study, we have assessed the regenerative potential of a novel decellularization protocol for human and rat nerves for repairing nerve resections, compared to the gold standard autograft. A 15-mm gap in the sciatic nerve was repaired with decellularized rat allograft (DC-RA), decellularized human xenograft (DC-HX), or fresh autograft (AG). Electrophysiology tests were performed monthly to evaluate muscle reinnervation, whereas histological and immunohistochemical analyses of the grafts were evaluated at 4 months. A short-term study was also performed to compare the differences between the two decellularized grafts (DC-RA and DC-HX) in early phases of regeneration. The decellularization process eliminated cellularity while preserving the ECM and endoneurial tubules of both rat and human nerves. Higher amount of reinnervation was observed in the AG group compared to the DC-RA group, while only half of the animals of the DC-HX showed distal muscle reinnervation. The number of regenerating myelinated axons in the mid-graft was similar between AG and DC-RA and lower in DC-HX graft, but significantly lower in both DC grafts distally. At short term, fibroblasts repopulated the DC-RA graft, supporting regenerated axons, whereas an important fibrotic reaction was observed around DC-HX grafts. In conclusion, the decellularized allograft sustained regeneration through a long gap in the rat although at a slower rate compared to the ideal autograft, whereas regeneration was limited or even failed when using a decellularized xenograft.

Impact of Early Referral to Palliative Care in Patients with Interstitial Lung Disease.

Background: Interstitial lung diseases (ILDs) have a major impact on survival and quality of life but only a small percentage of patients are referred for palliative care (PC). Objective: To assess the impact of early PC referral on hospital admissions, emergency department visits, and place of death in the last year of life. Design: This is a single-center retrospective observational study. Setting/Subjects: Subjects were patients with ILDs who attended the respiratory department of Hospital Santa Creu i Sant Pau (Barcelona, Spain) between 2011 and 2019. Results: Of the 51 included patients, 45% received early PC referral. Logistic regression indicated that early PC referral was independently associated with a lower risk of hospital admissions in the last year of life (OR = 0.16; 95% CI 0.03-0.75; p = 0.02) and a lower risk of dying in hospital (OR = 0.11; 95% CI 0.02-0.5; p = 0.009). Conclusion: Early PC referral reduces the need for hospitalization and enables domiciliary death.

Computerized tomography scan evaluation after fresh osteochondral allograft transplantation of the knee correlates with clinical outcomes.

PURPOSE: To determine the correlation between the assessment computed tomography osteochondral allograft (ACTOCA) scoring system and clinical outcomes scores. The hypothesis was that the ACTOCA score would show sufficient correlation to support its use in clinical practice. METHODS: We prospectively collected data from all consecutive patients who underwent cartilage restitution with fresh osteochondral allograft (FOCA) transplantation for osteochondral lesions of the knee and had a minimum follow-up of two years. CT scans were performed at three, six and 24 months post-operatively. A musculoskeletal radiologist blinded to the patients' medical history evaluated the scans using the ACTOCA scoring system. Clinical outcomes collected preoperatively and at three, six and 24 months postoperatively were evaluated using the International Knee Documentation Committee (IKDC), Kujala, the Western Ontario Meniscal Evaluation Tool (WOMET), and the Tegner Activity Scale. RESULTS: The mean total ACTOCA score showed a statistically significant correlation with the clinical outcome. The correlation was optimal at 24 months. We found a high negative correlation with the IKDC, Kujala and Tegner (- 0.737; - 0.757, and - 0.781 respectively), and a moderate negative correlation with WOMET (- 0.566) (p < 0.001). IKDC, Kujala, WOMET, and Tegner scores showed a significant continuous improvement in all scores (p < 0.001). CONCLUSION: The mean total ACTOCA score showed a linear correlation with clinical results in IKDC, Kujala, WOMET, and Tegner scores, being the highest at 24 months post-surgery. This finding supports the use of ACTOCA to standardize CT scan reports following fresh osteochondral allograft transplantation in the knee.

"Off-the-Shelf" Nerve Matrix Preservation.

Background: Decellularized human nerves overcome the limitations of the current treatments for large peripheral nerve injuries. However, the use of decellularized nerves requires an "off-the-shelf" availability for useful and actual clinical application. In this study, we addressed the preservation of the native and decellularized human nerve matrix in an integrative approach for tissue scaffold production. Materials and Methods: For native nerve matrix preservation analysis, we used histological examination and immunofluorescence to examine the structure, biomechanical assays to evaluate the tensile strength and Young's modulus, and analyzed the extracellular matrix (ECM) composition using enzyme-linked immunosorbent assay (ELISA) and biochemical assays for laminin, collagen and sulfated glycosaminoglycans (sGAG). After decellularization, nuclear remnants and DNA content were evaluated using 4',6-diamidino-2-phenylindole (DAPI) staining and the picogreen quantification assay, as well as immunofluorescence or ELISA for cell rests (S100 protein and myelin staining) evaluation. Decellularized cryopreserved scaffolds were assayed for biomechanics, ECM composition, and structural maintenance. Cytotoxicity assays were performed to evaluate the biocompatibility of the nerve matrix extracts after cryopreservation. Results: We compared different strategies for native nerve storage and found that preservation up to 7 days at 4°C in Roswell Park Memorial Institute medium maintained biomechanical properties, such as Young's modulus and tensile strength, along with the structure and ECM composition, regarding laminin, collagen, and sGAG. After a successful decellularization process, that eliminated cell remnants, nerve scaffolds were frozen in an "in house" formulated cryoprotectant, using an automatic controlled rate freezer. Nerve structure, ECM composition, and biomechanical properties were maintained before and after the freezing process in comparison with native nerves. The extracts of the nerve scaffolds after thawing were not cytotoxic and the freezing process sustained good viability in 3T3 cells (graphical abstract). Conclusion: Since our approach facilitates transport, storage, and provide a ready-to-use alternative, it could be used in a clinical application for the treatment of long-gap peripheral nerve injuries in regenerative medicine.

4†New strategies in the barcelona eye bank to minimize the impact of the COVID-19 pandemic.

Since the start of the pandemic, the tissue donation in Catalonia (Spain) has decreased drastically. At the beginning of the lockdown (from March to May 2020) there was a drop of around 70% in donation of corneas and of approximately 90% in donation of placentas. Despite the fast updating of standard operating procedures, we had big difficulties in different points. For instance, in the availability of the transplant coordinator for the donor detection and evaluation, in obtaining the necessary PPE (personal protective equipment), or in the resources available in the quality control laboratories for screening. This, added to the collapse that hospitals suffered due to the large number of patients hospitalized each day, made donation levels slowly rebound.In order to provide solutions to all patients, we tried to adapt quickly to these emerging changes.In the case of corneas, we found a scenario that we had never had before. Although the cornea transplant plummeted at the beginning of the confinement (decreased by 60% compared to 2019), we run out of corneas -even for emergency situations- at the end of March.This situation led us to develop a new type of therapeutic solution in our Eye Bank. The cryopreserved cornea for tectonic purposes is a tissue that is kept frozen at -196°C and can be preserved for up to 5 years. Therefore, it is a tissue that allows us to respond to possible emergencies in subsequent similar situations.Regarding amniotic membrane for ocular care indications, the strategy was completely different. For this kind of tissue, we carried out an adaptation of our processing with two different purposes. On the one hand, to make sure that we could inactivate the SARS-CoV-2 virus, if it was there. On the other hand, to increase the donation of placentas. For this, changes in the transport medium and in the antibiotic cocktail were performed. In addition, an irradiation step was added to the final product.Little by little, it seems that the donations of corneas and placentas have been recovering. However, it is necessary to think about future contingency strategies in case a stop in donation is repeated.

A new computed tomography scoring system to assess osteochondral allograft transplantation for the knee: inter-observer and intra-observer agreement.

AIM OF THE STUDY: To describe a new semiquantitative computed tomography (CT) scoring system for multi-feature analysis of cartilage defect repair by osteochondral allografts for the knee and to assess its intra-observer and inter-observer variability. METHOD: A semiquantitative assessment CT osteochondral allograft (ACTOCA) scoring system was designed based on fresh osteochondral allograft transplantations for the knee. The system includes five CT features: density relative to host bone, integration at the host-graft junction, surface percentage with a discernible cleft at the host-graft junction, cystic changes, and intra-articular fragments. Inter-observer variability was calculated by three observers blinded to the patient's medical history and treatment. Intra-observer variability was also determined. RESULTS: Inter-observer agreement was moderate to substantial for all CT score components and intra-observer agreement was moderate to almost perfect for all CT score components (κ > 0.5, p < 0.05). CONCLUSION: The ACTOCA score is a reliable tool to evaluate integration of osteochondral allograft transplantations. It provides an accurate evaluation of bone changes and may help to standardize CT scan reports following osteochondral allograft transplantation for the knee.

Art Therapy in a Palliative Care Unit: Symptom Relief and Perceived Helpfulness in Patients and Their Relatives.

CONTEXT: Creative arts therapies aim to expand conventional palliative care interventions by making clinical care more holistic. OBJECTIVES: The objective of the present study was to evaluate the benefits of an art therapy intervention in a tertiary hospital palliative care unit, directly in adult cancer inpatients and indirectly in their relatives. METHODS: We evaluated the intensity of pain, anxiety, depression, and well-being using the Edmonton Symptom Assessment Scale before and after the first, third, and fifth art therapy sessions. After the third and fifth sessions, perceived helpfulness was assessed via a questionnaire developed by the palliative care team, combining open-ended questions and a checklist. We categorized the narrative data into three predetermined types: generally helpful (some positive experience), helpfulness related to a dyadic relationship (patient-art therapist), and helpfulness related to a triadic relationship (patient-image-art therapist). RESULTS: We observed a significant reduction in anxiety, depression, and pain as well as a significant increase in well-being at each of the time points evaluated. Ninety-eight percent of the patients considered the art therapy helpful, which could be categorized as generally helpful in 54.8%, related to a triadic relationship in 32.9%, and to a dyadic relationship in 12.3%. Relatives gave similar opinions regarding the effects on patients and, in addition, reported an indirect helpful effect for themselves. The most frequently selected experiences from the checklist were feeling calm, being entertained, and expressing and communicating emotions. CONCLUSION: This art therapy intervention was beneficial in reducing symptom intensity. Almost all the participants directly or indirectly involved in the creative art process considered it helpful. They reported a wide variety of sensory, emotional, cognitive, and spiritual experiences.

Extrinsic modulation of integrin α6 and progenitor cell behavior in mesenchymal stem cells.

Mesenchymal stem cells (MSC) are heterogeneous cells of complex nature that show different potentials while different culture conditions can modify their functionalities through interactions with the microenviroment. Here, we found that bone marrow (BM) MSC from different donor sources and passages that expressed higher levels of α6 integrin subunit (ITGA6), showed higher clonogenicity, migration and differentiation potential. ITGA6 showed important roles improving these potentials and regulating proliferation through protein kinase B (AKT) pathway and cell cycle inhibitor proteins p53 and p21. Moreover, ITGA6 downregulation impaired migration. Cell confluence regulated ITGA6, increasing its expression in low density cultures and decreasing in high density cultures. Besides, ITGA6- cells expressed ITGA6 when seeded at low densities. We found higher ITGA6 expression on fibronectin substrates at lower confluency. Fibronectin increased proliferation, clonogenicity, activation of AKT, decreased cell cycle inhibitor proteins and augmented growth factors expression. Spheres-derived MSC showed higher ITGA6 expression and enhanced potentials for migration, clonogenicity and proliferation. In conclusion, though there is an intrinsic regulation of ITGA6 expression, associated to the progenitor potential of BM-MSC, this expression is regulated by culture conditions and is translated in changes in cell behavior and proliferation. This knowledge could be used to enhance the potential of BM-MSC for clinical application.

Which type of bone releases the most vancomycin? Comparison of spongious bone, cortical powder and cortico-spongious bone.

Bone infections can be challenging to treat and can lead to several surgeries and relapses. When a graft is needed, cavitary bone loss can be grafted with cancellous or cortical bone. Both can be used for grafting. However, the antibiotic releasing capacity of these grafts has not been compared. Which type of bone is best at releasing the most antibiotic has not been well established. The aim of this study was to determine which type of bone is best for antibiotic release when the bone is suffused with antibiotics by the surgeon. The hypothesis is that there would be a difference between the type of bone tested due to different release capacities of cortical and cancellous bone. This was an experimental study. Cortical spongy bone in chips, Spongy bone in chips and demineralized cortical bone powder were compared. For each type of bone, 5 samples were tested. Processed and decontaminated grafts were freeze-dried to be kept at room temperature. The primary endpoint was the amount of vancomycin released by the graft as it affects the concentration of antibiotic around the graft in clinical practice. The procedure for the study consisted of full graft immersion in a vancomycin solution. Then, the liquid was removed with aspiration. In order to measure the quantity of antibiotic released, the bone was put into distilled water in agitation in a heated rocker at 37 °C. After 30 min of soaking, 1 mL of the liquid was removed. The same extraction process was also carried out after 60 min soaking, 2 h, 3 h, 24 h, and 48 h. No differences were found between each type of bone relative to the concentration of vancomycin released at each time of the assessment. There was a significant difference in the weight of the bone with a higher weight for the cortical powder (1.793 g) versus cortical spongy bone and spongy bone (1.154 g and 1.013 g) with a p value < 0.0001. A significant difference was seen in the weight of the bone with vancomycin after the aspiration of the liquid with 3.026 g for cortical powder, 2.140 g and 2.049 g for the cortical spongy bone and the spongy bone with a p value < 0.0001. In daily clinical practice, one can use cancellous bone, cortico-cancellous bone or cortical powder in order to add vancomycin to a bone graft. Our results show the release kinetics of the soaked allografts. With a maximum of 14 mg/mL in the first minutes and a rapid decrease it shows a pattern comparable to antibiotic loaded bone cement. The method used appears favourable for prophylactic use, protecting the graft against contamination at implantation, but is not sufficient for treating chronic bone infection. LEVEL OF EVIDENCE: V.

Fresh Osteochondral Resurfacing of the Patellofemoral Joint.

Large osteochondral lesions of the knee in young patients continue to be a challenge for orthopaedic surgeons and the focus of continual research. This is particularly true if the injury is a consequence of a dysplastic trochlea and involves both articular surfaces of the biomechanically complex patellofemoral joint. To obtain a healthy and congruent patellofemoral joint, the use of a bipolar fresh osteochondral allograft transplantation of the patella and trochlea is one of the few options to biologically treat these injuries. This would achieve a replacement of the entire articular surface of the patellofemoral joint with a high number of viable chondrocytes and respect the unique structural characteristics of the cartilage. The aim of this study was to obtain symptomatic and functional improvements while delaying the timing of prosthetic surgery. We present a reproducible although demanding surgical technique to perform a bipolar fresh osteochondral allograft transplantation of the patella and trochlea.

Fresh Osteochondral Patellar Allograft Resurfacing.

The treatment of articular cartilage lesions in young patients is certainly a complex matter and subject of continuous research, particularly for those located at the patellofemoral joint, given its peculiar biomechanical characteristics. Osteochondral grafts can be of relatively small size when the defect is focal and in an area that allows good stability and consequently the graft's integration. In case of large or multifocal lesions, it is possible to consider an osteochondral transplantation of the entire articular surface of the patella. We present a simple and reproducible technique to perform a patellar fresh osteochondral allograft resurfacing attempting to reduce the symptoms and delay a prosthetic implant in young patients with advanced patellar chondral injuries.

Fresh Osteochondral and Meniscus Allografting for Post-traumatic Tibial Plateau Defects.

Large post-traumatic osteochondral defects of the proximal tibia in young active patients can be challenging because total or partial arthroplasties are to be avoided. The use of a fresh osteochondral allograft including its meniscus is one of the few options to biologically treat these injuries. Although the use of a fresh allograft is not easily accessible in some places and carries considerable logistical limitations, it is an alternative that provides viable chondrocytes to the defect. The inclusion of the meniscus in the osteochondral graft improves the results but also makes the technique even more demanding. We present a thorough description of this allograft transplantation to make it as reproducible as possible.