Abstract In this review, we systematically categorize diverse organoid engineering strategies—including cellular programming, material engineering, and platform- or system-level innovations—according to their impact on reproducibility and scalability, and highlight representative applications and emerging directions. By reframing organoid generation as a manufacturing process, these technological advances pave the way toward standardized and high-fidelity organoid production for both fundamental research and translational applications.

Seminal cryopreservation is a widely used technique in assisted reproduction. The technique is used for humans and animal samples to preserve cellular genetic material for long periods of time in low temperatures. However, this procedure can cause cellular damage by increasing the reactive oxygen species (ROS), which may negatively affect DNA and acrosome integrity, sperm motility and mitochondria function. Therefore, research are necessary to prevent such damage. Recent studies have investigated supplementation of cryopreservation media with nanoproducts, mainly with antioxidant potential, aiming to reduce ROS and improve sample quality after thawing. This literature review analyzed 13 studies that used nanotechnology for this purpose. The articles were arranged on a table with their information and then some frequent topics were discussed.

Cryogenic focused ion beam (Cryo-FIB) milling has become a standard step in the cryogenic electron tomography (Cryo-ET) workflow and is required to thin cells to electron-semitransparency. However, this destructive process removes the vast majority of the cellular material and raises a critical question: what thin section should be preserved for Cryo-ET analysis? Using a tri-coincident cryogenic FIB-SEM-LM system, we identify an interferometric optical response that can be used for targeting lamella production to fluorescently labeled structures with accuracy beyond the diffraction limit. Here we demonstrate this approach using synthetic samples of fluorescent beads embedded in micron-scale droplets of amorphous ice. We then apply the approach to capture virions inside host cells. Successful targeting is confirmed by Cryo-ET revealing clusters of virions in intracellular vesicles. The method does not require any fluorescent fiducials or axial registration and can be performed on any fluorescently labeled structure that is visible in widefield fluorescence microscopy.

Trace DNA can be deposited onto a wide range of surface types which can include substrates with a coating of oil: examples being firearm components, power tools, and kitchen utensils. We report on the impacts of oil on DNA recovery and downstream DNA processing. Thumbprints were made on 60 separate glass slides, then stained with Diamond Dye and fluorescent cellular material counted as an estimation of the cells deposited. Aliquots of five different mineral oils, chosen due to their common uses and variation in viscosity, were spread across the entire deposited thumbprint. The thumbprints coated with one of the five oils were left at room temperature for either 1 day or 7 days. A swab was used to collect cellular material which was then processed manually through a DNA extraction process, quantified and STR alleles amplified using the VeriFiler™ Plus STR kit. It was observed that all five oils impacted the initial DNA recovery process by saturating the swab. The presence of oils also impacted the DNA extraction process by interacting with the magnetic resin. Oils with higher viscosity accentuated these observed effects during the DNA extraction process, as the presence of a white precipitate was seen to be carried over into the final eluate. A total of 56% of the 1-day samples and 72% of the 7-day samples resulted in DNA profiles comprised of 12 or more alleles. The results of this study highlighted the possible impacts of collecting samples with oils present on the surface.

Abstract Introduction and aims Many classic three-dimensional (3D) skin models contain animal-derived materials e.g. bovine/rat collagen I. In this pioneering project, animal-derived materials were replaced with animal-component-free materials to evaluate the cellular interactions so that an animal-component-free full-thickness 3D skin model can be created. The first step to creating an animal-component-free 3D skin model is to change the culture medium of the cells. There are defined animal-component-free media available; however, they can be expensive. This project evaluates the most beneficial cost-effective media. The aim of this project was to evaluate whether animal-component-free media affects human primary skin cells compared with cells grown in animal-derived media in a two-dimensional cell culture. Methods Normal human dermal fibroblasts (NHDFs) and normal human epidermal keratinocytes (NHEKs) were grown in animal-component-free media and animal-derived media. The cells were treated with both commercial defined media or in-house animal-component-free-media containing various different supplements including growth hormones and cytokines and left to grow for 7 days. On days 3, 5 and 7, the morphology, cell growth and viability were analysed. The morphology was imaged using a Leica DMi8 microscope. From this, average cell size was calculated using Image J. The cells were counted with trypan blue in an automated cell counter for cell growth and viability analysis. Expression studies and an enzyme-linked immunosorbent assay were done with cellular material. An Anova test was performed for statistical significance. Results The results show that as the duration of culture increased, the cells grown in animal-component-free media had an increased growth rate (in-house media and commercial defined media) compared with animal-derived media. There was a change in the morphology of primary skin cells grown in different animal-component-free media. Conclusions This project analyses the effects of animal-component-free media compared with animal-derived media on NHEKs and NHDFs, examining multiple factors including cellular growth and gene expression.

DNA alloys: The enduring story of touch DNA on metals.

A NIR-responsive upconversion implant for wireless photodynamic therapy of tumors.

Auricular repair: symphony between scaffolds and stem cells.

Biopsy is a fundamental interventional procedure that enables tissue sampling from suspicious lesions for diagnostic purposes. Image-guided biopsies are minimally invasive approaches that increase diagnostic accuracy. Targeted tissue sampling of lesions can be performed with high success rates under the guidance of methods such as ultrasonography, computed tomography, fluoroscopy, or magnetic resonance imaging. Under the guidance of these imaging modalities, safe and controlled sampling with a low risk of complications is possible, especially in lesions involving salivary glands, cortical bone perforation, or soft tissue components. Image guidance offers advantages such as determining the entry path, avoiding surrounding neurovascular structures, assessing lesion vascularity, and providing real-time guidance during the procedure.In dentistry, fine-needle aspiration biopsy and Tru-Cut biopsy are the most commonly used biopsy methods. Fine-needle aspiration biopsy is a minimally invasive method with a low risk of complications, often preferred for fluid-containing or superficial lesions. Obtaining cellular material allows for cytological evaluation, but procedure repetition may be necessary in case of inadequate sampling. Tru-Cut biopsy, on the other hand, allows for obtaining a tissue core with thicker needles and preserves the histological structure and tissue architecture. It has higher diagnostic accuracy in solid or bone lesions, although the risk of complications is somewhat increased compared to fine-needle aspiration biopsy.This review discusses the techniques, procedural steps, advantages and disadvantages, complications, and techniques of fine-needle aspiration biopsy and Tru-Cut biopsy performed under image guidance in dentistry, in light of the literature.

Abstract. Purpose. To study the character and peculiarities of cytomorphological changes of cellular sediment material of fluid collections (FC) as criterion for objective assessment of the clinical course severity in acute complicated pancreatitis (ACP). Materials and methods. The results of cytological researches of peripancreatic sediments of FC obtained during interventional ultrasonography in 51 (88%) and videolaparoscopy in 7 (12%) observations in 58 patients on ACP were analysed. Results. It was found that in intraabdominal FC (n=37) in 10 (27%) cases the dominating of proliferative mesothelium with its polymorphism was observed. In 15 cases (41%) degenerative-dystrophic changes of cytoplasm whippings with the appearance of large “ring-similar” cells were manifested. In 12 studies (32%), atypically altered mesothelium cells with anisocytosis and anisonucleosis, as well as cytomorphological changes similar to the neoplastic process were detected. In retroperitoneal localization of FC (n=21) more significant changes were observed in granulocyte-nuclei cells, in particular, in 9 cases (43%), irreversible violations of their structure were identified as phenomena of cariopicnosis, carioresix and cariolysis. Was established that depth of structural changes of mesotheliocyte cells and neutrophil granulocytes depended on degree of ACP severity. Conclusions. Estimation of cytomorphological changes of the FC cell components can serve as objective criterion of the ACP severity.

Tissue regeneration is essential for wound healing, organ function restoration, and overall patient recovery. Its success significantly impacts medical procedures in fields like internal medicine and orthopedics, enhancing patient quality of life. Recent advances in regenerative medicine, particularly the combination of advanced drug delivery systems (DDS) and bioengineering, have enabled customized methods to improve tissue regeneration outcomes. However, conventional tissue engineering techniques have drawbacks, often using static scaffolds that lack the dynamic properties of real tissues, leading to subpar healing outcomes. The use of 3D printing and other advanced scaffolding techniques allows for the creation of bio functional scaffolds that deliver bioactive molecules at precise locations and times. The optimal integration of biological systems with enhanced material properties for personalized treatment options remains unclear. There is a need for more research into the complex interactions between cellular biology, drug delivery, and material technology to improve tissue regeneration. Despite progress in developing bioactive scaffolds and localized drug delivery methods, the interactions among different scaffold materials, bioactive agents, and cellular behaviors within the regenerative ecosystem are not fully understood. While there is extensive research on 3D-printed scaffolds in tissue engineering, there is a lack of studies integrating bio printing with in vivo biological reactions in real time. Limited research on the dynamic integration of patient-specific parameters in regeneration methods highlights the need for customized approaches that consider individual physiological differences and the complex biological environment at injury sites. Additionally, challenges arise when translating laboratory results into effective therapeutic applications, underscoring the necessity for interdisciplinary collaboration and innovative design approaches that align advanced material properties with biological needs.

This study investigates the usefulness of extemporaneous cytological examination performed during imaging-guided biopsies in dogs and cats. In veterinary diagnostics, imaging techniques such as ultrasound and computed tomography are essential for identifying lesions, but they cannot distinguish between inflammatory and neoplastic processes. Biopsies are therefore required, and this study explores whether immediate cytological evaluation of tissue cores, rolled onto slides and examined under a microscope, can help determine sample adequacy and reduce the number of biopsies needed. Seventy-nine animals with soft tissue or bone masses underwent ultrasound or CT-guided biopsies, followed by extemporaneous cytology and histopathological analysis. Cellular material representative of the lesion was identified in 81.1% of cases, allowing clinicians to conclude the procedure without further sampling. The diagnostic accuracy of cytology compared to histology was 68.3%, with particularly high accuracy for lipomas, melanomas, and mast cell tumors. When excluding non-diagnostic samples due to poor preparation or lesion characteristics, the accuracy reaches 92.1%. These findings suggest that extemporaneous cytology is a useful tool for verifying sample adequacy in real time, potentially reducing procedural risks and improving diagnostic efficiency. While histology remains the gold standard, this method offers immediate feedback and may enhance clinical decision-making in veterinary interventional diagnostics.

Membrane curvature: a key regulator of membrane protein structure and function.

Abstract Since the introduction of wood cellular material, or Dendrolight®, the process of creating dimensionally stable wood panels (Berger et al ., 2016) has evolved towards the creation of a similar, but significantly different new material - Lightweight Stabilised Blockboard (LSB). It was patented (Roziņš, M. Vašuks & P. Vašuks, 2014a) with the invented profiled solid wood boards (Roziņš, M. Vašuks & P. Vašuks, 2014b). The LSB design has introduced a smaller groove or kerf height, which hypothetically allows for a reduction in energy consumption during its manufacturing process. Considering that LSB is a new material, most suitable parameters of cutting regime for the LSB mechanical machining have not been developed. To develop machining processes for LSB, it is necessary to determine the optimal cutting tool, machine parameters, expected feed speed and cutting power. The study analyses the parameters of cutting regime that affect the cutting power, and performs theoretical calculations that reveal the expected and most suitable cutting process indicators in the industrial production of LSB, as well as compares the machining of LSB and Dendrolight®. A new evaluation parameter has been introduced that characterises the ratio between the cutting power and the permissible feed speed. It was concluded that grooving LSB lamellae requires significantly less cutting power, and that higher feed speeds are possible compared with the grooving of analogous Dendrolight® lamellae. However, the cutting power required for the LSB grooving is still quite energy-intensive, which potentially limits the possible width of LSB lamellae and the number of simultaneously operating cutting tools.

Crosstalk between lipid droplets and autophagy in cancer: A nexus for therapeutic targeting.

Cryo-imaging in cellular biology provides the means to visualize the cellular interior at close-to-native conditions. A cornerstone in the field has been cryo-correlative light and electron microscopy (cryo-CLEM), with cryo-visible light fluorescent microscopy (cryo-VLFM) providing the specificity by tagging macromolecules or structures and cryo-electron tomography (cryo-ET) for the structural details at molecular level. The large resolution gap between these techniques, however, is limiting this correlative workflow as cryo-ET targets are often smaller than the resolution limit of cryo-VLFM. Here we introduce cryo-soft X-ray tomography (cryo-SXT) as an intermediate step that can compensate for the partial view caused by the lost cellular material due to FIB-milling and limited resolution of cryo-VLFM by providing invaluable cellular context information in 3D to the cryo-ET dataset within an integrated workflow. This work shows that X-ray and electron imaging are not mutually exclusive, creating opportunities for further correlative imaging strategies.

Despite its rarity, vulvar cancer retains a high potential for reducing the quality of life of patients and poses serious challenges for doctors. One of the central tasks of modern oncology is to improve efficiency of methods of early detection and monitoring of vulvar cancer. The most important instrument in solving this problem is diagnostic cytology based on the analysis of cellular material obtained directly from the area of the suspected lesion. The cytologic method is the most important diagnostic tool due to its high sensitivity and specificity, allowing to confidently recognize early signs of the transformation of normal epithelium into cancerous tissue.

The mesonephric kidney of fish retains its capacity for nephrogenesis and regeneration throughout the organism's lifetime. It is hypothesized that the extracellular matrix (ECM) of trout plays a pivotal role in supporting the proliferation and differentiation of resident cells. The ECM is a complex three-dimensional network of macromolecules secreted by cells during early development and is essential for regulating key cellular functions, including adhesion, migration, proliferation, differentiation, and survival. This study aimed to evaluate the effects of decellularized trout kidney ECM on the growth and differentiation of mouse renal tubular epithelial cells invitro. In this study, trout kidneys were decellularized and bleached using a combination of sodium dodecyl sulfate (SDS), Triton X-100, and a bleaching buffer. The effectiveness of decellularization was evaluated using quantitative assays and histological staining. The resulting ECM was then coated onto culture dishes at concentrations of 100 and 150 μg/cm2. To assess potential cytotoxicity, an indirect extraction assay was performed. Subsequently, human bone marrow-derived mesenchymal stem cells (hMSCs) were seeded onto the decellularized ECM. Cell viability and proliferation were evaluated using the MTS assay [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] and the CFSE (carboxyfluorescein succinimidyl ester) assay, respectively, and results were compared to an uncoated control group. Finally, gene expression analysis of hMSCs was conducted using quantitative reverse transcription polymerase chain reaction (qRT-PCR). The decellularization of fish renal ECM was successfully achieved. Quantitative and qualitative analyses confirmed a significant reduction in DNA content in the decellularized tissue compared to native samples, indicating effective removal of cellular material. At the same time, key ECM components such as collagen and sulfated glycosaminoglycans (sGAGs) were preserved to a substantial extent. Specifically, collagen content decreased from 5.736 μg/mg in native tissue to 4.284 μg/mg in the decellularized ECM, while sGAG content showed a slight decrease from 0.9855 μg/mg to 0.9400 μg/mg, reflecting minimal degradation during processing. Cytotoxicity assessment revealed no toxic effects associated with the decellularized matrices. Furthermore, a significant increase in the proliferation of human mesenchymal stem cells (hMSCs) was observed on day 7 compared to the uncoated control group, suggesting the bioactivity and compatibility of the decellularized trout kidney ECM.

Extracellular vesicles (EVs) have become pivotal in clinical therapeutics, although no consensus exists on optimal production methods. Here, an innovative approach and associated device are introduced that employ hydrodynamic stress to form and stimulate stem cell spheroids. The method involves culturing cellular material in vessels equipped with internal obstacles, subjected to rotational motion at varying speeds and in alternating reversing modes. It achieves EV yields 10 to 20 times greater than those obtained with traditional static culture methods, offering a robust and high-output method compatible with large-scale EV manufacturing. This approach not only streamlines spheroid formation and subsequent EV release into a single, integrated process but also ensures the generation of EVs with enhanced biological activity and a distinctive protein cargo signature. Notably, hydrodynamic stimulation enriched EVs with ectosome- and mitochondria-associated proteins, suggesting distinct biogenesis pathways. Additionally, these EVs demonstrated superior functionality in promoting wound healing, angiogenesis, and anti-inflammatory responses, underscoring their therapeutic potential.

Accurate preoperative assessment of thyroid nodules remains challenging, particularly in indeterminate cytological categories. Integrating molecular testing into cytology could improve diagnostic precision, enable timely intervention, and support better risk stratification and patient management. This proof-of-concept study evaluated the feasibility of performing molecular testing on fine-needle aspiration cytology (FNAC) samples processed on CytoMatrix, a three-dimensional synthetic scaffold designed to capture and preserve cellular material. Thirty-three thyroid FNAC specimens were processed on CytoMatrix, and cytological diagnoses were mirrored to the 2023 Bethesda System for Reporting Thyroid Cytopathology and correlated with final histopathology. DNA was extracted from paraffin-embedded CytoMatrix sections and analyzed for the BRAF V600E mutation. Adequate DNA for molecular testing was obtained in 30 of 33 cases (90%), and BRAF V600E mutations were detected in three papillary thyroid carcinoma samples. DNA adequacy and yield were consistent across Bethesda III–V categories, with insufficiency limited to low-cellularity Bethesda III cases. CytoMatrix enables reliable DNA recovery and targeted molecular testing without compromising cytological evaluation. This integrated cytomolecular workflow provides a feasible approach for combining cytological and molecular data in thyroid FNAC, supporting personalized and timely diagnostic management.