Cell and Tissue Banking最新文献

In tissue engineering, natural and synthetic nanofibers that can regenerate body damage have been successfully used in the repair of many lesion types, including peripheral neural lesions, in recent years. So, we developed three different nanofibers that we think can regenerate peripheral nerve damage. Three different nanofibers based on biodegradable poly-ε-caprolactone (PCL); Pure PCL (PCL) nanofiber, 70% PCL and 30% bioactive glass (PCL/BG) hybrid nanofiber, and 0.1% vitamin B12 added (PCL/BG)-B12 hybrid nanofiber were produced by electrospinning. Sol-gel method was used in the synthesis of biomaterials containing bioactive glass. The nanofibers were characterized by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), and fourier transform infrared spectroscopy (FT-IR). Cell viability assays were performed with healthy L929 fibroblast cells and PC12 cells to evaluate the biocompatibility of nanofibers. Neuronal differentiation of PC12 cells were stimulated by nerve growth factor (NGF). To assess the differentiation levels of PC12 cells, the length of neurites and number of outgrowing neurites per cell was evaluated morphologically, and NGF production levels of the cells were determined by ELISA. The results suggest that these biocompatible nanofibers stimulated PC12 cell survival and neuronal differentiation. Among these scaffolds, PCL/BG-B12 nanofibers strikingly triggered NGF production of PC12 cells as a hallmark of neuroregeneration. Thus, the nanofibers are capable of neuroprotective properties due to their safe, supporting proliferation, and NGF-releasing capacity. Additionally, it could be suggested that the PCL/BG nanofiber and vitamin B12 have the potential to be used in further studies for neurodegenerative diseases.

Tissue engineering (TE) combines cells, biomaterials, and bioactive molecules to create functional tissue constructs aimed at restoring tissue function and improving patient outcomes. The human amniotic membrane (HAM) is a widely studied biological scaffold for various biomedical applications. Decellularization of HAM (dHAM) is necessary to reduce graft rejection but depletes stem cells and growth factors, potentially limiting regenerative potential. This study investigates the recellularization of dHAM with adipose-derived mesenchymal stem cells (AdMSCs) to enhance its bioactivity using a novel 3D seeding technique. Decellularized HAM (dHAM) was recellularized with AdMSCs employing a novel 3D seeding method to achieve uniform cell distribution within the scaffold. The viability, differentiation potential, and morphology of AdMSCs were assessed in both 2D and 3D culture systems. Flow cytometry was used to evaluate the differentiation capacity of AdMSCs into osteogenic, chondrogenic, and adipogenic lineages. Field emission scanning electron microscopy (FESEM) was utilized to analyze cell morphology and penetration depth within the scaffold. AdMSC viability was comparable between 2 and 3D cultures, indicating that dHAM scaffolds effectively support cell survival regardless of the culture technique. The composition and properties of dHAM preserved cell functions in both culture systems. Flow cytometry confirmed the multilineage differentiation potential of AdMSCs. FESEM imaging revealed AdMSCs with extending filopodia on the scaffold surface and cell penetration up to 17.68 µm into the dHAM matrix. The successful 3D recellularization of dHAM with AdMSCs demonstrates its potential as a biological scaffold for stem cell delivery. This approach holds promise for tissue repair and wound healing applications, enhancing the regenerative efficacy of dHAM-based constructs.

The gap between the clinical demand for transplantable organs/tissues and the limited availability of deceased donors motivates the search for alternative donor types. This growing need has prompted exploration into non-traditional donor populations, including those who have died due to circulatory causes, elderly individuals, and donors previously excluded from consideration, such as those with extensive burns. Burned deceased donors are often perceived as unsuitable due to the detrimental effects of burn injury on multiple organ systems, including the phenomenon known as "burn disease." This condition, characterized by systemic inflammatory responses, metabolic derangements, and sepsis risk, can potentially compromise organ viability.However, proper assessment and management of these donors, including rigorous evaluation of organ function, the extent of burn injury, and potential infection, can enable successful multi-organ and multi-tissue donation. Several reports suggest that, under appropriate clinical and logistical circumstances, organs such as the kidneys, liver, heart, and lungs, as well as various tissues, can be procured from burned donors.Nonetheless, expanding research in this area is essential to establish the guidelines for safe utilization of burn deceased donor population.

The current research was conducted to assess wound healing activity and antibacterial properties of carboxymethyl cellulose (CMC) hydrogels loaded with silver nanoparticles (AgNPs) against excisional wounds (15 × 15 mm²) infected with Pseudomonas aeruginosa and Staphylococcus aureus in a rat model.CMC/AgNPs hydrogels were synthesized using varying concentrations of AgNPs and subsequently lyophilized. A comprehensive range of in vitro tests were conducted, including nanoparticle characterization, scanning electron microscopy (SEM) morphology study, water uptake (WUE) study, blood uptake capacity study (BUC), weight loss study (WLA), pH, hemolysis percentage (HP), blood coagulation index (BCI), antibacterial activity (minimum inhibitory concentration [MIC] and minimum bactericidal concentration [MBC]), and cell viability through the MTT assay. In vivo wound healing studies were conducted using infected excisional wound models in rats. SEM confirmed a porous structure with a mean pore size ranging from 68 to 152 μm. The hydrogels exhibited dosage-dependent swelling and sustained physiological pH (7.4-7.6) for a period of time. The 125 μg/mL AgNPs formulation showed a BUC of 97.68% in 22 h. Hemocompatibility assay showed minimal hemolysis and acceptable coagulation indices for all concentrations of AgNPs. MIC and MBC against both strains of bacteria were found to be 250 μg/mL and 500 μg/mL, respectively. CMC/AgNPs hydrogel with the concentration of 250 μg/mL showed the optimal cell viability and the optimal in vivo wound healing result. The findings indicate that AgNPs-loaded CMC hydrogels possess favorable physicochemical, biocompatible, and antimicrobial properties, suggesting their potential as a wound dressing for managing infected wounds and supporting the wound healing process.

The present study aimed to evaluate the effectiveness of super-activated platelet lysate (sPL), as well as sPL in combination with bone marrow mesenchymal stem cells (BMSCs) in treating bone defects in rats. Ninety rats with induced radial defects were divided into three groups: sPL/BMSCs, sPL, and control. The healing process was evaluated using X-rays, Lane Sandhu scores, HE staining, RT-qPCR, immunohistochemistry, and ELISA. Both sPL and sPL/BMSCs facilitate bone repair. Compared to the other groups, the sPL/BMSCs group showed higher OCN, OSX, and RUNX2 gene expression until 10 weeks after treatment ALP levels in the sPL group increased until week 6 and remained elevated until week 10, with the sPL/BMSCs group showing significantly higher peak ALP levels. IGF-1, TGF-β, and VEGF levels in both treatment groups were higher than in the control group. Our study demonstrated that both sPL and sPL/BMSC transplants at the bone defect site can promote bone healing and modulate bone metabolism-related factors. However, the effect of sPL/BMSCs was superior to sPL alone.

Bone grafts and its substitutes are commonly used in periodontics to enhance bone formation and periodontal regeneration. Allografts, derived from human donors, support new bone growth and are biocompatible, with reduced disease transmission risk due to rigorous screening. Combining bone grafts with growth factors can further improve treatment outcomes. Platelet Rich Fibrin (PRF) releases growth factors that enhance regeneration. This study evaluates the effectiveness of combining processed fresh frozen bone allograft (FFBA) with PRF mambrane for treating periodontal bone defects. The FFBA was prepared from bone chips collected from systemically healthy patients in an aseptic environment during orthopedic surgeries, which was then processed and tested for microbiological and cytotoxicity concerns using Human Osteosarcoma (MG-63) and Normal Mouse Fibroblast (L929) cell lines. PRF was prepared by centrifuging blood collected from patients at 2300 rpm for 12 min. A case study comprising of ten patients with chronic periodontitis was conducted, Clinical and radiographic measurements, including plaque index (PI), gingival index (GI), probing depth (PD), and bone defect fill percentage using radiographs were recorded at baseline and nine months. Descriptive statistics (Mean ± SD), a 5% significance level, and 90% confidence intervals were used. Paired t-tests showed significant improvements in GI (p < 0.001), PI (p < 0.001), PPD (p < 0.001), and Bone defect fill (p < 0.001). The study concluded that using processed fresh frozen bone allograft with PRF positively affects the treatment of periodontal osseous defects.

To summarize the evidence examining the outcomes of Descemet membrane endothelial keratoplasty (DMEK) using eye bank pre-stripped versus surgeon prepared grafts. Systematic review and meta-analysis. This study was conducted following the preferred reporting items for systematic reviews and meta-analyses consensus statement (PROSPERO ID: CRD42023457120). Searches of medline (Ovid), EMBASE, EMCARE, the Cochrane register of controlled trials, and grey literature were performed until April 2025. All comparative studies comparing DMEK outcomes after eye bank prepared (pre-stripped only) with surgeon-prepared tissue were included. Two independent reviewers completed data extraction and performed quality assessments. The primary outcomes were the rebubbling rate and corrected distance visual acuity (CDVA). Results were summarized using a random-effects meta-analysis. Five studies totalling 750 eyes receiving DMEK grafts were included. Four were retrospective observational studies and one was a non-randomized prospective study. There were no significant differences between eye bank pre-stripped and surgeon-prepared graft outcomes with regards to rebubbling rate (odds ratio, 1.11; 95% confidence interval [CI] 0.65 to 1.90) and postoperative logMAR CDVA (mean difference - 0.11, 95% CI - 0.23 to 0.01). No statistically significant difference was noted in postoperative corneal thickness, endothelial cell loss or density, and postoperative complications.Eye bank pre-stripped grafts and surgeon-prepared grafts yield comparable outcomes in terms of CDVA and rebubbling rates following DMEK.However, there is limited evidence, as only five studies were included in this analysis.

To evaluate long‑term outcomes of corneal patch grafting (CPG) and to determine prognostic factors for anatomical and functional success. This retrospective study included 35 eyes from 35 patients who underwent CPG between April 2016 and September 2022 at Adana City Training and Research Hospital. Collected data included age, sex, preoperative and postoperative best-corrected visual acuity (BCVA), graft localization and size, anterior segment findings, graft survival, secondary surgical procedures, and rates of anatomical and functional success. Anatomical success was achieved in 82.9% (29/35), and functional success in 45.7% (16/35). The most common reason for CPG was non-infectious, non-traumatic perforation related to immune disorders (37.1%). Anatomical success was not significantly associated with the primary etiology (p = 0.73), whereas functional success was significantly higher in patients with traumatic corneal perforations (p = 0.029). A statistically significant improvement was observed in postoperative BCVA for the study group (p < 0.001). Functional success was significantly associated with younger age and peripheral graft location (p = 0.005 and p < 0.001, respectively). Smaller graft diameter was significantly correlated with both higher anatomical and functional success (p = 0.031 and p = 0.007, respectively), while no significant association was found with horizontal graft diameter (p = 0.27 and p = 0.068, respectively). Corneal patch grafting is a highly effective technique for restoring anatomical integrity in most patients and provides acceptable functional outcomes in selected cases. Younger age, peripheral graft location, and smaller graft diameter appear to be favorable prognostic factors for functional recovery.

In the United States, the use of Food & Drug Administration (FDA)-licensed, approved, or cleared tests is required for infectious disease screening and determining the eligibility of deceased donors for all Human Cells, Tissues, and Cellular and Tissue-Based Products (HCT/Ps). With the discontinuation of two manual enzyme-linked immunoassay (EIA) tests, automated Chemiluminescent Microparticle Immunoassay (CMIA) technology was introduced as the primary alternative. This study compares serologic reactivity rates between manual EIA and automated CMIA methods. Serology testing was performed on blood specimens from deceased tissue and cornea donors using either manual EIA assays or automated CMIA platforms. A retrospective analysis of over 160,000 donors was conducted to evaluate assay-specific reactive rates. Concordance between serology and Nucleic Acid Testing (NAT) results was also assessed. Reactive rates for HBsAg, HIV-1/2 antibodies, and HTLV-I/II antibodies increased following the transition to CMIA-based testing compared to manual EIA methods. However, these increases were not associated with a corresponding rise in NAT reactive results, indicating a potential increase in false-positive or non-viremic results. Automated testing offers improved quality control and reduces variability associated with manual techniques. However, the shift to CMIA assays resulted in a higher number of reactive serology results, leading to increased donor ineligibility despite negative NAT results. The data presented here highlights the potential impact on donor deferral rates for tissue and cornea donors due to assay platform changes.

Although many preclinical and clinical studies are ongoing on amniotic membrane extract (AME), an amniotic membrane-derived product developed to support ocular surface healing, the effect of AME on the basic cellular functions and properties of human corneal epithelial cells (hCECs) has not been clearly defined. In this study, we aimed to evaluate the effect of AME supplementation to the culture media, on basic cellular functions of hCECs and on expression of specific cell markers of hCECs, as well as to determine its effectiveness in an experimental in vitro wound model. hCECs were seeded with the constant cell density in 6, 24 and 48 well plates. The next day, the media was refreshed with 0 mg/ml, 0.75 mg/ml, 1.5 mg/ml and 3 mg/ml final concentration of AME supplemented complete growth medium. Cellular morphology, viability, metabolic activity, proliferation assessments and immunocytochemistry were conducted on the time points at day 3, 6 and 9. Then, in vitro wound healing assay was performed on hCECs under the effect of AME. AME did not affect cellular morphology, viability, metabolic activity and proliferation. AME supplementation induced CK3 and CK12 expression of hCECs significantly (p < 0.05) higher than without AME group. In vitro wound healing assay revealed that while control and AME-treated cultures both exhibited healing, AME-treated mechanical abrasions closed at a greater rate compared to control. Our study shows that AME promotes in vitro wound healing and cell characteristics in terms of CK3 and CK12 protein expressions, while preserving basic cellular functions of corneal epithelial cells.