Cell and Tissue Banking最新文献

Human Tissue Banks (BTH) must validate the storage of collected/processed tissues ensuring physical integrity, sterility, and microbial protection for up to 5 years. (1) Is it safe to use bone tissue for transplants collected by a BTH after 5 years of storage? (2) Do the packaging of stored tissues present physical integrity, sterility, and microbial protection after 5 years? (3) What are the morphological results of bone tissues after 5 years?. 20 femoral heads were used with a storage time of between 9 and 10 years at -80 °C. From each femoral head, the following were carried out: microbiological tests for aerobic and anaerobic bacteria and fungi, collecting 3 fragments of bone tissue, 3 Stuart swabs from the inner surface of the packaging, and a sample of 0.9% SF for Bioburden examination; tissue histology; the quantitative and qualitative mechanical resistance test and pyrogenicity/cytotoxicity test were used on the packaging, pre and post storage. No bone tissue samples showed pathogenicity. Histological findings showed morphologically preserved osteocytes with points of bone degeneration and necrotic adipose tissue. No packaging showed contamination, cytotoxicity, or pyrogenicity. The mechanical properties of these packages demonstrated uniformity in thickness, high tension, and relative stiffness even after storage (p = 0.001). It is concluded that the packaging used in the study presented physical integrity, sterility, and microbial protection for bone tissues after 9 and 10 years of storage. A possible increase in the shelf life of fabrics is contemplated to up to 10 years. Such results expand future research directions to continuously improve the quality of products and services offered by BTH.

Prosthetic valves derived from bovine pericardium (BP) are crucial for heart valve replacement, yet current crosslinking methods with glutaraldehyde can lead to immune responses and calcification. This study evaluated the effects of reducing the glutaraldehyde crosslinking time from 10 to 5 days in bovine pericardial patches for use as heart valve substitutes. In addition to examining the physical properties of the BP, the study analyzed the biocompatibility, tissue structure, and calcification of the pericardial tissue. BPs were processed using two protocols based on the fixation time with glutaraldehyde: BP10d (10 days) and BP5d (5 days). All samples were treated with glutamic acid to neutralize residual aldehyde groups from the glutaraldehyde. Subsequently, the resulting material was assessed for mechanical and thermal properties and histologically using light and scanning electron microscopy. Post-implantation histological evaluation and calcium content determination were conducted after 7, 14, 30, 60 and 120 days. The calcification was a rare occurrence. However, some samples from the BP10d group displayed positive Von Kossa staining, indicating mineral deposition. Chemical analysis using ICP-OES revealed low calcium concentrations in the explants of both groups, with higher concentrations observed in the BP10d group during the later analysis periods. Mechanical and thermal stability assessments showed no significant differences between experimental groups. Histological examination revealed more collagen and elastic fibers deformation, and inflammation in the BP10d group compared to the BP5d group. The revised manufacturing protocol, with a 5-day fixation time, showed promising anti-calcifying activity, biocompatibility, and tissue preservation.

Sinus lift surgery is essential after pneumatization caused by loss of posterior teeth. Leukocyte-Platelet-Rich Fibrin (L-PRF) accelerates bone healing by releasing growth factors that promote angiogenesis, cell differentiation, and inflammatory modulation. To evaluate the efficacy of L-PRF in bone healing and repair in sinus lift surgeries, in addition to investigating its role in angiogenesis and inflammatory modulation. The systematic review protocol included definition of the research question, search strategy, inclusion and exclusion criteria, study types, effect measures, screening methods, and data analysis. The search resulted in 860 studies. After removal of duplicates, 704 articles remained, of which 11 met the inclusion criteria. After careful evaluation, 4 studies were considered highly relevant and included in the systematic review. Evidence indicates that the combination of L-PRF with bone grafts, such as DBBM, can accelerate bone formation and allow early implant placement, supported by increased expression of protein markers essential for osteogenesis. The addition of L-PRF to DBBM demonstrated significant benefits in promoting a more favorable bone environment, reducing the time required for osseointegration.

The body has evolved three types of cartilage: hyaline, elastic, and fibrocartilage. Modern tissue engineering techniques can harvest different types of chondrocytes, expand them in vitro, and use them to repair various cartilage defects. However, the modulatory effect of different cartilaginous niches on the type of regenerated cartilage after the implantation of chondrocytes from different origins remains unknown. In this study, three typical types of cartilage-auricular (elastic cartilage), articular (hyaline cartilage), and meniscus (fibrocartilage)-were investigated. Chondrocytes derived from these cartilages were mixed with Pluronic gel and implanted into three different cartilaginous niches for one month using a goat model. Our results demonstrated that in the articular cartilage environment, regenerated cartilage from auricular chondrocytes lost elastin expression, and cartilage from meniscus chondrocytes lacked a fibrous structure, showing reduced type I collagen and increased type II collagen expression, all resembling a hyaline cartilage-like structure. In the auricular cartilage environment, regenerated cartilage from articular chondrocytes did not express elastin, maintaining a hyaline cartilage-like structure, while fibrocartilage chondrocytes failed to form regenerated cartilage. In the fibrocartilage environment, regenerated cartilage from auricular and meniscus chondrocytes did not exhibit a fibrous structure, with weak type I collagen expression and positive type II collagen expression. Regenerated cartilage from auricular chondrocytes did not express elastin and did not transform into fibrocartilage. This study provides valuable insights into how different cartilaginous niches influence the characteristics of regenerated cartilage, offering potential implications for improving cartilage repair strategies in tissue engineering.

The study explores the effects of the COVID-19 pandemic on the Musculoskeletal Tissue Bank (MSTB) in Milan, with a particular focus on tissue harvesting and its subsequent use in surgical procedures. A retrospective descriptive epidemiological analysis compared data from the pre-pandemic period (2018-2019) with that of the pandemic period (2020-2022), revealing a 24.8% reduction in tissue retrievals during the pandemic. Although there was a decrease in the number of eligible donors not collected (from 93 to 67, from 36.05 to 34.54%), this reduction was not statistically significant. The decline in tissue retrievals was due to decreased non-COVID-related pathologies, a lower number of potential donors from reduced accidents and increase in COVID-positive deaths. However, the MSTB successfully met tissue demands throughout this period. Notably, the reduction in retrievals at the MSTB was lower than national averages (- 24.8 vs. - 47.5%). Logistic regression analysis showed no significant organizational issues in donor collection. Despite the challenges, the MSTB remained resilient and adaptable, continuing its essential services. This underscores the broader impact of the pandemic on healthcare systems and emphasizes the importance of a flexible healthcare infrastructure during public health emergencies.

A prospective clinical study was conducted to evaluate the change in horizontal alveolar ridge width following horizontal ridge augmentation using Freeze-Dried Bone Allograft (FDBA) block graft and Amnion Chorion Membrane (ACM) for the management of atrophic ridges. The study included a total of 10 subjects of either sex, aged between 18 and 60 years, presenting with deficient residual alveolar ridge width. All subjects were treated with horizontal ridge augmentation using an indigenously prepared FDBA block graft and ACM. The ridge width at the alveolar crest and 5 mm apical to the crest were recorded at baseline, immediately after augmentation, and after 6 months, followed by dental implant placement. Also, this study focused on the clinical efficacy and any adverse effects of the indigenously prepared FDBA block graft and ACM. The indigenously prepared FDBA block graft and ACM exhibit regenerative properties that significantly enhance horizontal alveolar ridge width in all cases of deficient ridges, thereby greatly facilitating dental implant placement. The mean difference after 6 months at the crest was 2.21 ± 0.89 mm (p < 0.05) and at 5 mm apical to the crest was 2.8 ± 0.91 mm (p = 0.001)This significant increase in ridge width achieved through indigenously prepared FDBA and ACM serves as an excellent alternative to commercially sourced or imported allograft materials, making it a cost-efficient choice.

The purpose of this study was to compare two methods for harvesting human dental pulp stem cells (hDPSCs) and investigate their impact on the DPSC viability and differentiation capacity. Healthy premolar teeth were collected from children aged 9 - 17 years old undergoing orthodontic treatment with two or four premolar extractions. The included premolars were randomly allocated to the apical group (removal of the dental pulp through the apical foramen after tooth extraction) or the coronal group (isolating the pulp tissue through the crown prior to tooth extraction). A total of 148 healthy premolar teeth from 46 patients were extracted, including 74 pulp tissues for the apical group and 74 for the coronal group. Due to bacterial and fungal infections, 37 (18 apical and 19 coronal) DPSC lines were obtained. Flow cytometric analysis and MTT assays showed no statistical differences in cell viability between the two methods. Odontogenic differentiation, assessed through immunocytochemical staining and Alizarin Red staining, also showed no visual or statistical differences between the apical and coronal approaches. In conclusion, the biological characteristics of hDPSCs harvested through the crown were equal to apically prelevated hDPSCs.

Cryobags play a critical role in freezing, storing, and transporting cellular therapy products but are prone to fractures, which can disrupt patient outcomes and facility workflows. This study evaluated the incidence, risk factors, and impact of cryobag fractures in a large inventory of cellular therapy products in Brazil. A retrospective cohort study included 4514 cryobags from 2262 peripheral blood stem cell collections processed between 2015 and 2024 at a single center supporting nine transplant facilities. Cryobags were frozen at - 80 °C and stored in nitrogen tanks. Fractures and leaks were identified through routine visual inspections. Among the cryobags, 15 (0.3%) fractured, with 12 detected at processing facility and 3 after release. The fracture rate was 0.37 per 100 bag-years, with a cumulative incidence of 1% at 3.62 years. Of these, 8 were discarded, and 7 were salvaged and infused into six patients. Two salvaged cryobags underwent bedside recovery, while five were recovered aseptically in the processing facility. Positive bacterial cultures were commonly found in salvaged products. In multivariate analysis, a higher total nucleated cell count per cryobag remained an independent risk factor for fracture (OR = 1.005; 95% CI: 1.0002-1.0099; p = 0.046). Following implementation of quality improvement initiatives based on root cause analysis, no further fractures were observed. These findings highlight the importance of monitoring cell concentration and adjusting cryopreservation protocols to mitigate risks. Adding overwraps may provide additional protection for cryobags at higher risk, reducing the likelihood of microbial contamination and improving the safety and reliability of cellular therapy products.

The purpose of this study was to investigate the role of Calcium-Sensing Receptor in the chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and to provide a new target for cartilage defect repair. BMSCs were cultured in vitro, and cultured in the complete culture medium with gradient concentration of calcium sensitive receptor inhibitor and activator, and the optimum dose was selected by CCK-8 experiment. The experiment was divided into four groups. After 7, 14 and 21 days of intervention, the intracellular calcium concentration was detected by laser confocal microscope, the differentiation of cartilage was detected by toluidine blue staining, and the expression of cartilage marker proteins (Col- II, Agg and Sox9) was detected by immunocytochemical staining and Western Blot. The CCK-8 assay results showed that the optimal concentrations of Gd and NPS were 300 μM and 10 μM, respectively. After 7, 14, and 21 days of culture, intracellular calcium fluorescence decreased, with notably higher cartilage differentiation in the NPS inhibitor group. Col-II, Agg and Sox9 chondrocyte marker proteins increased with culture time in all groups, with significantly higher levels in the inhibitor group compared to others, followed by the cartilage induction solution group, and then the activator group. Inhibition of calcium sensitive receptors can promote chondrogenic differentiation of rat BMSCs by regulating Sox9, affecting Col- II and Agg.

The transmissibility rate of the SARS-CoV-2 virus, which causes COVID-19, in organ and tissue donation is not known. Considering all issues related to the risk of SARS-CoV-2 transmission and the risk of tissue shortages due to donor exclusion, this in vitro study evaluated the detection of SARS-CoV-2 RNA in bone tissue models after bone processing for tissue bank. Bone processing has activity against SARS-CoV-2 and not RNA was detected in the end of the process. The use of bone tissue can be considered in case of shortage.