Cell and Tissue Banking最新文献

Regeneration of articular cartilage disorders is one of the critical challenges in musculoskeletal medicine. Tissue engineering could represent a therapeutic option to support cartilage regeneration. Natural and biological materials are appropriate for fabricating tissue engineering scaffolds because of their similarity to natural tissues. The properties of amniotic membranes, including low immunogenicity, anti-inflammatory role, cell loading capability, expression of various growth factors, and chondroprotective effect, make them an interesting option for cartilage regeneration. This review studied the structure of articular cartilage and potential applications of the human amniotic membrane (AM) for articular cartilage regeneration. In addition, processing and decellularization methods of AM and the most common forms of amniotic membrane used in cartilage regeneration, including sheet, injectable form, and 3D forms, were studied. This review highlights the benefits of amniotic membrane applications in cartilage regeneration and clinical trial studies.

Platelet lysate is a derivative of platelet-rich plasma that is used as supplement for in vitro cell culture media. A variety of protocols for its preparation have been described. However, its potential use in the clinical setting has been poorly studied. In the present work, the effect of several protocols on cell proliferation has been comparatively analyzed. Additionally, the effect of exposing bone and vascular tissues to different concentrations of platelet lysate has also been analyzed. Human fibroblast-like cells were used to test preparation protocols. Thawed skull and artery fragments were incubated with platelet lysate and seeded as explants in culture plates. Cell growth was evaluated quantitatively in the first assay (cell count) and qualitatively in the second (presence of growing cell colonies). The presence of leukocytes in the raw material to obtain the platelet lysate was correlated with higher cell proliferation. In all cultures from arteries and 71.4% of those from bones, the presence of viable cells was detected. No statistically significant differences that correlated with the percentage of platelet lysate used during the post-thaw incubation were observed. The main findings of this study revealed that: there is a contribution of bioactive substances for cell growth by lymphocytes, incubation with platelet lysate had no significant activating effect on cells in thawed tissues, arteries stored in liquid nitrogen retained cell viability for long periods (over 5 years), and cell viability in bones stored at - 80 °C decreased after 3 months.

Cardiovascular allografts are essential for patients with severe cardiovascular diseases. Yet, microbial contamination of the grafts poses a life-threatening risk to recipients. Tissue banks utilize various decontamination methods during cardiovascular tissue processing, often involving antibiotic solutions. This study compares the efficacy of an in-house prepared antibiotic cocktail (tissue bank cocktail) and a commercially available solution (BASE.128) in decontaminating cardiovascular tissues. For this study, the efficacy of the two antibiotic cocktails was compared through quantitative comparisons against challenge microorganisms, and retrospective analysis of routine sterility tests. Both solutions demonstrated comparable decontamination efficiency against challenge strains, achieving significant reductions in bacterial load. However, retrospective sterility tests revealed that while both antibiotic solutions were highly effective in decontaminating cardiovascular allografts, the use of BASE.128 followed a tenfold increase in contamination rates compared to the tissue bank cocktail, primarily due to a slow-growing non-tuberculous mycobacteria strain. These findings highlight the importance of tailored decontamination protocols that consider prevalent microbial contaminants while preserving tissue quality.

Peripheral nerve injuries often require surgical intervention when end-to-end coaptation is not feasible, with autologous nerve grafts being the current gold standard. However, limitations such as donor-site defects drive the search for alternative methods. This study explores the efficacy of acellular nerve allografts obtained through a feasible protocol as a potential off-the-shelf substitute for autografting in a 14-mm rat sciatic nerve defect. Thirty-two female Wistar rats were divided into four groups: autograft, lyophilized acellular allograft, fresh acellular allograft and silicone tube. Functional assessments and histological examinations were performed at 14 and 20 weeks post-surgery, respectively. Results showed comparable axonal regeneration between acellular nerve allografts and autografts. Histomorphometric analysis revealed no significant differences in axonal characteristics between groups. Muscle histomorphometry indicated superior recovery in animals treated with fresh acellular allografts, who exhibited the least muscle atrophy and larger muscle fiber diameter compared to lyophilized processed allografts and autografts. Functional assessments revealed no significant intergroup differences. Processed acellular allografts promote axonal regeneration similar to autografts in a 14-mm rat sciatic nerve defect. Fresh acellular allografts achieve better muscle reinnervation in the medial gastrocnemius muscle. However, axonal regeneration does not consistently correlate with functional or histomorphological outcomes of the hind leg muscle. The successful decellularization protocol and lack of immune rejection pave the way for adapting it to human nerve grafts. These could revolutionize clinical practice in our country, becoming an example of leveraging existing resources and replacing collagen conduits and autografts for treating certain injuries.

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.