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Aims. To assess the bone dimensional changes after extraction and alveolar ridge preservation (ARP) using primary coverage (closed flap technique, CFT) or healing by secondary intention (open flap technique, OFT). Materials and Methods. Ten patients (split mouth design) were planned for extraction and ARP. All sites received ARP with freeze-dried bone allograft (FDBA) and nonresorbable membrane after extraction. Clinical standardized measurements were used to assess the dimensional alterations of the alveolar ridge. Results. All patients completed the study, and a total of 20 sites were randomized to CFT or OFT group. Center height (mean difference of 8.1 mm, SD =1.9 CFT, and 7.5 mm, SD= 1.8 OFT) and buccal height (mean difference of 0.8 mm, SD =1.0 CFT, and 0.3 mm, SD= 1.1 OFT) were significantly different within the same group. However, there was no statistically significant difference between groups. In the OFT group, the keratinized tissue width was higher and the pain VAS scores at 24 hours were lower compared with the CFT (p = 0.004 and p = 0.006, respectively). Conclusions. Leaving the flap open did not have any effects on the dimensional changes of bone height or width. However, there was a wider band of keratinized tissue and less pain with the CFT compared with the OFT. The study protocol was registered at ClinicalTrials.gov, Identifier NCT03136913.

Postextraction ridge resorption compromises the aesthetics and function of the subsequent implant therapy or prosthetic rehabilitation. Several techniques are employed to reduce ridge resorption, ranging from basic socket fillers such as blood clot preservation to more advanced techniques using barrier membranes and bone grafts such as autografts, allografts, xenografts, and synthetic materials. The aim of this study is to evaluate and compare the effectiveness of alveolar socket preservation by using demineralized freeze-dried bone allograft (DFDBA) with titanium platelet-rich fibrin (T-PRF) and autologous dentin graft (ADG) with T-PRF. This assessment will be performed through comprehensive clinical, radiographic, and histomorphometric analyses. The primary objective of this study is to histologically evaluate the new bone formation in the extraction sockets preserved using either DFDBA with T-PRF or ADG with T-PRF. The secondary objective is to evaluate the dimensional changes, including ridge width and height at these healing points, as measured by clinical and radiographic methods. We will evaluate and compare the clinical, radiographic, and histomorphometric outcomes of using DFDBA with T-PRF versus ADG with T-PRF for maintaining socket integrity after tooth extraction. We will also ascertain which grafting technique facilitates the production of new bone during the healing phase and preserves alveolar ridge dimensions. This randomized controlled trial will involve 16 patients aged 22-60 years requiring tooth extractions and subsequent implant placement. Participants will be randomly assigned to one of the 2 groups: (1) socket preservation using DFDBA in combination with T-PRF or (2) socket preservation using ADG with T-PRF. At baseline, all the clinical variables will be assessed using UNC-15 probe and cone beam computed tomography radiographs. Extraction will be done atraumatically with minimal flap reflection by using periotomes. Postextraction sockets will be preserved using DFDBA+T-PRF or ADG+T-PRF. At 4 months, clinical and radiographic evaluations will be done, and the implant will be placed using a 2-stage protocol. Histomorphometric analysis will be performed after receiving bone samples during implant placement. At 3 months after implant placement, the second-stage surgery will be done. Participant enrollment commenced in March 2024, and the study is scheduled to conclude postassessments and analyses by the end of 2025. The results of this study are anticipated to be accessible in late 2025. This study is not funded, and the results are expected to be published by 2026. This study represents valuable insights into the clinical effectiveness of 2 biologically driven socket preservation techniques. We hypothesize that the use of ADG combined with T-PRF will show similar or more effective outcomes in alveolar socket preservation demonstrated by enhanced bone formation and better maintenance of socket dimensions compared to DFDBA combined with T-PRF without increased morbidity. Clinical Trials Registry - India CTRI/2024/05/068192; https://ctri.nic.in/Clinicaltrials/pmaindet2.php?EncHid=MTI1NzIz&Enc=&userName=. DERR1-10.2196/70649.

In this review, recent advances in the methods of pre-treatment of plant material for the extraction of secondary metabolites with high biological activity are presented. The correct preparation of the material for extraction is as important as the selection of the extraction method. This step should prevent the degradation of bioactive compounds as well as the development of fungi and bacteria. Currently, the methods of preparation are expected to modify the particles of the plant material in such a way that will contribute to the release of bioactive compounds loosely bonded to cell wall polymers. This review presents a wide range of methods of preparing plant material, including drying, freeze-drying, convection drying, microwave vacuum drying, enzymatic processes, and fermentation. The influence of the particular methods on the structure of plant material particles, the level of preserved bioactive compounds, and the possibility of their release during the extraction were highlighted. The plant material pre-treatment techniques used were discussed with respect to the amount of compounds released during extraction as well their application in various industries interested in products with a high content of biologically active compounds, such as the pharmaceutical, cosmetics, and food industries.

The success of mRNA vaccines against SARS-CoV-2 has prompted interest in mRNA-based pharmaceuticals due to their rapid production, adaptability, and safety. Despite these advantages, the inherent instability of mRNA and its rapid degradation in vivo underscores the need for an encapsulation system for the administration and delivery of RNA-based therapeutics. Lipid nanoparticles (LNPs) have proven the most robust and safest option for in vivo applications. However, the mid- to long-term storage of mRNA-LNPs still requires sub-zero temperatures along the entire chain of supply, highlighting the need to develop alternatives to improve mRNA vaccine stability under non-freezing conditions to facilitate logistics and distribution. Lyophilization presents itself as an effective alternative to prolong the shelf life of mRNA vaccines under refrigeration conditions, although a complex optimization of the process parameters is needed to maintain the integrity of the mRNA-LNPs. Recent studies have demonstrated the feasibility of freeze-drying LNPs, showing that lyophilized mRNA-LNPs retain activity and stability. However, long-term functional data remain limited. Herein, we focus on obtaining an optimized lyophilizable mRNA-LNP formulation through the careful selection of an optimal buffer and cryoprotectant and by tuning freeze-drying parameters. The results demonstrate that our optimized lyophilization process maintains LNP characteristics and functionality for over a year at refrigerated temperatures, offering a viable solution to the logistical hurdles of mRNA vaccine distribution.

Lyophilization is the “gold standard” for drying plant extracts, which is important in preserving their quality and extending their shelf-life. Compared to other methods of drying plant extracts, lyophilization is costlier due to equipment, material and operational expenses. An alternative method is post-extraction oven-drying, but the effects of this process on extract quality are unknown. In this study, crude extracts from Arthrocnemum macrostachyum shoots were compared using three post-extraction drying methods (lyophilization and oven drying at 40 and 60 °C) and two extraction solvents (water and aqueous 50% ethanol). Untargeted metabolomics coupled with chemometrics analysis revealed that post extraction oven-drying resulted in the loss of up to 27% of molecular features when compared to lyophilization in water extracts only. In contrast, only 3% of molecular features were lost in aqueous 50% ethanol extracts when subjected to oven drying. That is to say, ethanol used as a solvent has a stabilizing effect on metabolites and enhances their resistance to thermal transformation in the oven. Collectively, oven-drying of extracts was as effective as lyophilization in preserving metabolites in extracts only when 50% ethanol was used as a solvent. The results presented in this paper demonstrate the value of selecting solvent-appropriate post-extraction drying methods.

The availability, biocompatibility, non-toxicity, and ease of chemical modification make cellulose a promising natural polymer for the production of biomedical materials. Cryogelation is a relatively new and straightforward technique for producing porous light and super-macroporous cellulose materials. The production stages include dissolution of cellulose in an appropriate solvent, regeneration (coagulation) from the solution, removal of the excessive solvent, and then freezing. Subsequent freeze-drying preserves the micro- and nanostructures of the material formed during the regeneration and freezing steps. Various factors can affect the structure and properties of cellulose cryogels, including the cellulose origin, the dissolution parameters, the solvent type, and the temperature and rate of freezing, as well as the inclusion of different fillers. Adjustment of these parameters can change the morphology and properties of cellulose cryogels to impart the desired characteristics. This review discusses the structure of cellulose and its properties as a biomaterial, the strategies for cellulose dissolution, and the factors affecting the structure and properties of the formed cryogels. We focus on the advantages of the freeze-drying process, highlighting recent studies on the production and application of cellulose cryogels in biomedicine and the main cryogel quality characteristics. Finally, conclusions and prospects are presented regarding the application of cellulose cryogels in wound healing, in the regeneration of various tissues (e.g., damaged cartilage, bone tissue, and nerves), and in controlled-release drug delivery.

Phage therapy is one of the promising alternatives to combat the increasing problem of antibiotic resistance. Lyophilization is used for the preparation of pharmaceutical products to improve their stability in long-term storage. The aim of this study was to improve the stability of lyophilized bacteriophages using different excipients. Three lytic bacteriophages Escherichia phage ECP311, Klebsiella phage KPP235 and Enterobacter phage ELP140 were subjected to lyophilization using six different excipients: glucose, sucrose, gelatin, mannitol, polyethylene glycol and sorbitol. The lyophilized phages were stored at 4 °C and 37 °C and rehydrated using biological saline to test their viability at 5 months interval up to 20 months. The results showed that the use of sucrose, gelatin and their combination was beneficial in maintaining the viability of phages post-lyophilization. When lyophilized phages were stored at 4 °C, their viability was maintained up to 20 months, but at 37 °C there was a reduction in activity after 10 months. This is one of the few studies to report the lyophilization of phage cocktails to have viability for up to 10 months. Our study identified promising lyophilization excipients to effectively lyophilize bacteriophages for pharmaceutical applications and long-term storage.

Fecal microbiota transplantation (FMT) is increasingly being used for treatment of recurrent Clostridium difficile infection (R-CDI) that cannot be cured with antibiotics alone. In addition, FMT is being investigated for a variety of indications where restoration or restructuring of the gut microbial community is hypothesized to be beneficial. We sought to develop a stable, freeze-dried encapsulated preparation of standardized fecal microbiota that can be used for FMT with ease and convenience in clinical practice and research. We systematically developed a lyophilization protocol that preserved the viability of bacteria across the taxonomic spectrum found in fecal microbiota and yielded physicochemical properties that enabled consistent encapsulation. We also treated a cohort of R-CDI patients with a range of doses of encapsulated microbiota and analyzed the associated changes in the fecal microbiome of the recipients. The optimized lyophilized preparation satisfied all our preset goals for physicochemical properties, encapsulation ease, stability at different temperatures, and microbiota viability in vitro and in vivo (germ-free mice). The capsule treatment was administered to 49 patients. Overall, 43/49 (88%) of patients achieved a clinical success, defined as no recurrence of CDI over 2 months. Analysis of the fecal microbiome demonstrated near normalization of the fecal microbial community by 1 month following FMT treatment. The simplest protocol using the lowest dose (2.1-2.5 × 10 bacteria in 2-3 capsules) without any colon purgative performed equally well in terms of clinical outcomes and microbiota engraftment. A single administration of encapsulated, freeze-dried fecal microbiota from a healthy donor was highly successful in treating antibiotic-refractory R-CDI syndrome.

Demineralized freeze-dried bone allograft (DFDBA) has been used extensively in periodontal therapy. Questions have been raised however, about the osteogenic potential of the variety of grafts available. In India the cost factor is another important consideration. The aim of this study therefore was to evaluate the clinical efficiency of the low priced, indigenously prepared DFDBA obtained from the Tata Memorial Hospital (TMH) Tissue Bank, in periodontal regeneration in infrabony periodontal defects, as compared to DFDBA obtained from the Pacific Coast Tissue Bank (DEMBONE). The latter was used as the control. 16 patients with bilaterally similar periodontal infrabony defects were selected, and randomly allotted to the test and control groups. At baseline, using standardized protocol, recession, probing depths (PD), and clinical attachment levels (CAL) were measured, following which periodontal surgery was carried out, with placement of the respective graft materials. Patients were recalled after 6 months for re-assessment. Statistically significant improvement was obtained for PD reduction and CAL gain for both groups ( p  < 0.05). However, no significant difference was observed between the test and control groups. It was therefore concluded that both the materials from different tissue banks are equally effective clinically, with the test material being additionally cost effective.

Freeze-drying (FD) is the most extensive drying technique in pharmaceutical and biopharmaceutical industries. It relies on three main steps: freezing, primary, and secondary drying, where the sample is frozen and then dried by ice sublimation. FD possesses several features, mainly its suitability for heat-sensitive materials and its ability to produce dry products with improved physicochemical characteristics. Although FD is a gentle drying process, it can cause numerous stresses that induce chemical and physical instabilities. Herein, the addition of suitable excipients along with optimizing the process parameters is critical in attaining lyophilizates with high-quality attributes. Besides, the freeze-drying method has been explored as a unique route to produce porous materials with different applications. This work aims to dismantle the basics of freeze-drying and its role in developing porous materials, mainly amorphous and co-amorphous solid-dispersions, orodispersible tablets and films, as well as porous dressings and 3D scaffolds for effective wound healing and tissue engineering, respectively. The challenges and limitations of lyophilization have also been addressed. Highlights Freeze drying is a propitious drying method, especially for thermolabile materials. The freeze-drying technique can form products with high-porous features. The successful lyophilization cycle depends on understanding both the formulation variables and the process parameters. Recent studies have shown that lyophilization under controllable conditions can develop porous materials with different applications and high-quality attributes. Graphical Abstract