Search

Tooth extractions can result in alveolar bone dimensional changes, necessitating additional bone grafting for implant placement. Alveolar ridge preservation (ARP) aims to counteract postextraction changes. This case series evaluates the bone regenerative properties of a freeze-dried bone allograft (FDBA) and the clinical outcomes of implants in grafted extraction sites. A total of 33 patients were enrolled, undergoing single/multiple tooth extractions followed by ARP. Biopsy samples were harvested during implant placement for histologic and histomorphometric analysis. Clinical outcomes included marginal bone loss and pink esthetic score (PES). Twenty-five patients completed the study. FDBA-augmented sockets exhibited new bone formation adjacent to graft particles. Implants (n = 25) showed 100% survival and success rates at 1 and 2 years. PES improved significantly over time (P < .001), while marginal bone loss did not significantly differ at 1 and 2 years (P = .096). Specimens showed trabecular bone, residual FDBA particles, and marrow spaces. High magnification revealed immature bone and woven bone bridges around graft particles. No inflammatory cells were observed. This case series provides valuable insights into ARP performed with FDBA (as implants were placed after 3 months of healing without any additional bone augmentation), the histologic outcomes were favorable, and implants were successful after a 2-year follow-up period. [ABSTRACT FROM AUTHOR]

By employing brute-force molecular dynamics, umbrella sampling, and seeding simulations, we investigate homogeneous nucleation during melting and freezing of hard spheres. We provide insights into these opposing phase transitions from the standpoint of classical nucleation theory. We observe that melting has both a lower driving force and a lower interfacial tension than freezing. The lower driving force arises from the vicinity of a spinodal instability in the solid and from a strain energy. The lower interfacial tension implies that the Tolman lengths associated with melting and freezing have opposite signs, a phenomenon that we interpret with Turnbull's rule. Despite these asymmetries, the nucleation rates for freezing and melting are found to be comparable. [ABSTRACT FROM AUTHOR]

Objective: The predictability and long-term success of periodontal regeneration begins with oral hygiene education, disease management, and an individually tailored periodontal maintenance protocol. The treatment outcomes could be enhanced when biologics and bone grafts are combined. The aim of this report was to describe the outcome of two complex infrabony defects in the same patient treated with recombinant human platelet-derived growth factor-BB (rhPDGF-BB) and freeze-dried bone allograft (FDBA) over 10 years. Case presentation: Two complex infrabony defects were treated following guided tissue regeneration principles and procedures. Fullthickness flaps were raised to allow visualization of the defects. The areas were debrided, and exposed root surfaces were planed. FDBA and rhPDGF-BB were combined to fill both defects. A collagen membrane was used over the bone graft in one case. The flaps were reapproximated to achieve primary closure. The patient was seen for regular periodontal maintenance visits and clinical and radiographic follow-ups over 10 years. Throughout the examination periods, the probing depths improved without bleeding on probing, and there was radiographic evidence of bone regeneration. Conclusion: The growth factorinfused bone graft was successfully utilized for periodontal regeneration in complex bony defects. [ABSTRACT FROM AUTHOR]

Extracellular vesicles (EVs) are widely investigated for their implications in cell-cell signaling, immune modulation, disease pathogenesis, cancer, regenerative medicine, and as a potential drug delivery vector. However, maintaining integrity and bioactivity of EVs between Good Manufacturing Practice separation/filtration and end-user application remains a consistent bottleneck towards commercialization. Milk-derived extracellular vesicles (mEVs), separated from bovine milk, could provide a relatively low-cost, scalable platform for large-scale mEV production; however, the reliance on cold supply chain for storage remains a logistical and financial burden for biologics that are unstable at room temperature. Herein, we aim to characterize and engineer a freeze-dried, mEV formulation that can be stored at room temperature without sacrificing structure/bioactivity and can be reconstituted before delivery. In addition to undertaking established mEV assays of structure and function on our preparations, we introduce a novel, efficient, high throughput assay of mEV bioactivity based on Electric Cell Substrate Impedance Sensing (ECIS) in Human dermal fibroblast monolayers. By adding appropriate excipients, such as trehalose and tryptophan, we describe a protective formulation that preserves mEV bioactivity during long-term, room temperature storage. Our identification of the efficacy of tryptophan as a novel additive to mEV lyophilization solutions could represent a significant advancement in stabilizing small extracellular vesicles outside of cold storage conditions. [ABSTRACT FROM AUTHOR]

Permanent preservation of genetic resources may be indispensable for the future of humanity. This requires liquid nitrogen, as is the case for preserving animal sperm. However, this technique is expensive and poses a risk of irrecoverable sample loss on non-replenishment of liquid nitrogen in case of natural disasters. In this study, we demonstrate that lyophilization may be used as a reliable method for long-term preservation of mouse sperm at room temperature. Sperm from four mouse strains were freeze-dried and stored in a non-temperature controlled room for 5–6 years. Although the ability of the stored sperm to activate oocytes had diminished slightly, healthy offspring were obtained by artificially activating the oocytes after sperm injection. Moreover, the birth rate did not decrease even after ≤ 6 years of storage. Furthermore, owing to its low cost, safety, and ease of storage at any location, we believe that this method could be a major mode of preserving mammalian genetic resources in the future. [ABSTRACT FROM AUTHOR]

The aim of our study was to upscale the production of Lysobacter enzymogenes strain LEC from shake flasks to a laboratory bioreactor. The obtained fermentation broth was to be downstream-processed by lyophilisation and fluidised bed-drying, followed by testing of the dried preparations against plant pathogens. Different batches of broth from shake flasks and from individual fermenter runs were characterised and compared. Number of viable cells, in vitro efficacy (ED90 value) against the apple scab pathogen Ventura inaequalis, as well as efficacy against Phytophthora infestans on potato plants were all reproducible and not impaired by upscaling the production volume from shake flask to the bioreactor. Fermentation broth was processed by lyophilisation and fluidised bed-drying. In bioassays, the activity against downy mildew (Pseudoperonospora cubensis) on cucumber were comparable for the unformulated fermentation broth and the fluidised bed-dried product, while the lyophilised product was significantly more effective. Strain LEC showed reproducible pathogen suppressive activity, independent from production scale and batch. Production upscaling and formulation by drying are possible without loss of efficacy. [ABSTRACT FROM AUTHOR]

Antibodies and antibody mimics are extensively used in the pharmaceutical industry, where stringent safety standards are required. Implementing heat sterilization during or after the manufacturing process could help prevent contamination by viruses and bacteria. However, conventional antibodies and antibody mimics are not suitable for heat sterilization because they irreversibly denature at high temperatures. In this study, we focused on the refolding property of the cold shock protein from the hyperthermophile Thermotoga maritima (TmCSP), which denatures at elevated temperatures but regains its native structure upon re‐cooling. We designed and constructed a mutant library of TmCSP in which amino acid residues in its three surface loops were diversified. From the library, mutant TmCSPs that bind to each of eight target proteins were selected by phage and yeast surface display methods. We confirmed that the secondary structure and binding affinity of all the selected mutants were restored after heat treatment followed by cooling. Additionally, freeze‐drying did not impair their binding affinity. The crystal structure of a mutant TmCSP in complex with its target, the esterase from Alicyclobacillus acidocaldarius, revealed specific interactions between them. These results clearly demonstrate the feasibility of creating heat‐sterilizable antibody mimics using TmCSP as a scaffold. [ABSTRACT FROM AUTHOR]

AbstractDimethylcurcumin (DMC) is a derivative of curcumin. Studies have shown that DMC can enhance the degradation of androgen receptors (AR), thereby inhibiting AR-related diseases. However, the extremely poor water solubility and fast metabolism in vivo of DMC greatly limit its clinical applications. In the present study, liposomes loaded with DMC were prepared by an improved membrane hydration method. The formulation was modified to obtain DMC-liposomes with particle size between 110 and 150 nm, moderate encapsulation efficiency, and good freeze-drying stability. In vitro release studies showed that DMC-liposomes displayed sustained drug release. Cellular uptake studies showed that cellular uptake of DMC-liposomes was enhanced compared to free DMC. Compared with free DMC, DMC-liposomes were more toxic to 22Rv1 and HepG2 cells in vitro. These results suggest that DMC-liposomes are a promising DMC formulation for anticancer DMC delivery. [ABSTRACT FROM AUTHOR]

For over a century African swine fever (ASF) has been causing outbreaks leading to devastating losses for the swine industry. The current pandemic of ASF has shown no signs of stopping and continues to spread causing outbreaks in additional countries. Currently control relies mostly on culling infected farms, and strict biosecurity procedures. Recently a vaccine, ASFV-G-ΔI177L was approved for use in Vietnam. In this study we evaluate the long-term stability of lyophilized ASFV-G-ΔI177L. Understanding the stability of different formulations of vaccines is information necessary for deployment of vaccines to ASF outbreak areas, particularly those that do not have a reliable well established cold chain to ensure conservation of vaccine quality. In this report, we determined that ASFV-G-ΔI177L, when lyophilized under specific conditions, is stable for up to one year at 4 °C, with similar vaccine titers after storage. Next-generation sequencing analysis also determined that lyophilization and long-term storage under these conditions had no effect on the genome of ASFV as the genome remained genetically identical to the original non-lyophilized form. [ABSTRACT FROM AUTHOR]

AbstractAgaricus bisporus, the most widely produced edible mushroom globally, faces a challenge with its short shelf life, complicating post-harvest distribution. This study aimed to produce high-quality, calcium-fortified dried Paris mushrooms to enhance their value and extend their shelf life. The effects of calcium vacuum impregnation, convective drying at various temperatures, and freeze-drying with vacuum freezing at the initial stage were evaluated regarding process kinetics and quality parameters. Vacuum impregnation efficiently fortified the mushrooms with calcium, increasing their concentration by 59 to 76 times in 10 min. While convective drying at 60 °C accelerated the dehydration process compared to 40 °C and 50 °C, it hurt the mushrooms’ color, antioxidant activity, rehydration capacity, and texture. In contrast, samples subjected to vacuum freezing exhibited higher freeze-drying rates than those frozen by contact and demonstrated superior quality across all evaluated parameters, particularly when compared to samples dried using the convective method. These fortified mushrooms can meet FAO/WHO daily calcium intake recommendations, offering nutritional benefits and enhanced post-harvest preservation. [ABSTRACT FROM AUTHOR]

Fermentation is of great importance to the food industry as it extends the shelf life of food while imparting desirable nutritional and sensory properties. However, the unpredictable fermentation period, inconsistency of the final product quality, and contamination with spoilage microorganisms of spontaneously fermented food remain a challenge. The aim of this study was to explore the storage stability of Lactobacillus plantarum and Streptococcus thermophilus prepared by freeze‐drying using coconut powder (CCN) (25%) as a carrier medium in comparison to skim milk powder (SMP). Freeze‐dried lactic acid bacteria (LAB) were evaluated for cell viability, vitality, moisture content, water activity (aw), acidification kinetics, cell morphology, and glass transition temperature (Tg). In general, LAB stored at 4°C showed better survival after 112 days than LAB stored at room temperature (RT). With cell viability, the reduction in S. thermophilus and L. plantarum counts was not significantly (p < 0.05) affected by the type of carrier media (CCN or SMP) after 112 days at 4°C. SMP supported better cell vitality (approx. 2.0) at 4°C than CCN after 112 days for L. plantarum. The moisture content and aw were within acceptable levels throughout storage on all samples. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) depicted a normal cell structure at 4°C for S. thermophilus and L. plantarum in SMP and CCN, while negative changes were observed at RT. The Tg on all samples was above 75°C, and this suggested long‐term stability of the LAB strains. Overall, our findings show that CCN shows similar protection of L. plantarum and S. thermophilus strains to that of SMP during storage at 4°C. Further work is needed to confirm the fermentation performance of CCN‐preserved strains. [ABSTRACT FROM AUTHOR]

The enhancement of cellulose degradation is important for improving the quality of corn-stalk silage. However, the rapid drop in pH caused by the propagation of lactic acid bacteria (LAB) can influence the degradation of cellulose by cellulose-degrading microorganisms (CDMs) during the mixed fermentation process of ensilage. In this study, a CDM (Bacillus methylotrophic 2–4, BM 2–4) was isolated, and its lyophilization condition was studied. Moreover, a slow-release strategy was developed to delay the release of LAB (Lactobacillus plantarum S-1, LP S-1) by embedding technology to provide time for BM 2–4 to degrade cellulose during the corn-stalk ensilage process. The results showed that BM 2–4 had a higher survival rate (89.53%) under the following conditions: cell collection (5,600 r/min in 4.4°C for 9.5-min centrifugation) and lyophilization using cryoprotectants [skim milk (10.4%), peptone (10.4%), and glucose (5.2%)] at −50°C with a vacuum pressure of <10 Pa. Based on the results of the previous study on embedded LP S-1, simultaneous inoculation of embedded LP S-1 and lyophilized BM2-4 at a 2:1 ratio, with an inoculum size of 6% and moisture content of 85%, significantly degraded CF by 3.8% and increased CP by 3.7% (p < 0.05). This treatment did not significantly influence the final pH of corn-stalk silage (p > 0.05) after 7 days of fermentation. [ABSTRACT FROM AUTHOR]

Electrostatic spray drying (ESD) of a milk protein matrix comprising whey protein isolate (WPI), skim milk powder (SMP), and lactose was compared to conventional spray drying (CSD) and freeze-drying (FD). ESD and CSD were used to produce powders at low (0.12–0.14), medium (0.16–0.17), and high (0.31–0.36) levels of water activity (aw), while FD powders targeted low aw (0.12). Maillard reaction indicators were studied after drying and during storage for up to 28 days at 20, 40, or 60 °C by measuring free -NH2 groups, as an indicator of available lysine, and 5-hydroxymethylfurfural (HMF). After drying, levels of residual free -NH2 groups were ~15% higher in ESD and FD powders than in their CSD counterparts. CSD powders also had ~14% higher HMF concentrations compared to their ESD and FD counterparts. Storage led to reductions in free -NH2 groups and increases in HMF content in all powders, the extent of which increased with increasing storage temperature. Reductions in free -NH2 groups followed first-order reaction kinetics at 20 and 40 °C but second-order reaction kinetics at 60 °C. Lactose crystallization was detected in high-aw CSD powders after 14 d at 40 °C and in both CSD and ESD powders after 7 d at 60 °C. Overall, we found that ESD is a gentle drying technology which enables production of powders with lower Maillard reaction markers. [ABSTRACT FROM AUTHOR]