Tumorigenesis mechanism and application strategy of the MDCK cell line: A systematic review.

Influenza is an acute respiratory infectious disease caused by influenza virus that seriously endangers people's health. Influenza vaccination is the most effective means to prevent influenza virus infection and its serious complications. MDCK cells are considered to be superior to chicken embryos for the production of influenza vaccines, but the tumorigenicity of cells is a concern over the theoretical possibility of the risk of adverse events. The theoretical risks need to be adequately addressed if public confidence in programs of immunization are to be maintained. In this paper, studies of the tumorigenic potential of cell lines, with MDCK cells as an example, published since 2010 are reviewed. The mechanism of tumorigenicity of MDCK cells is discussed with reference to cell heterogeneity and epithelial to mesenchymal transition (EMT). Understanding the mechanism of the acquisition of a tumorigenic phenotype by MDCK cells might assist in estimating potential risks associated with using tumorigenic cell substrates for vaccine production.

Automated analysis of rabbit knee calcified cartilage morphology using micro-computed tomography and deep learning.

Structural dynamics of calcified cartilage (CC) are poorly understood. Conventionally, CC structure is analyzed using histological sections. Micro-computed tomography (µCT) allows for three-dimensional (3D) imaging of mineralized tissues; however, the segmentation between bone and mineralized cartilage is challenging. Here, we present state-of-the-art deep learning segmentation for µCT images to assess 3D CC morphology. The sample includes 16 knees from 12 New Zealand White rabbits dissected into osteochondral samples from six anatomical regions: lateral and medial femoral condyles, lateral and medial tibial plateaus, femoral groove, and patella (n = 96). The samples were imaged with µCT and processed for conventional histology. Manually segmented CC from the images was used to train segmentation models with different encoder-decoder architectures. The models with the greatest out-of-fold evaluation Dice score were selected. CC thickness was compared across 24 regions, co-registered between the imaging modalities using Pearson correlation and Bland-Altman analyses. Finally, the anatomical CC thickness variation was assessed via a Linear Mixed Model analysis. The best segmentation models yielded average Dice of 0.891 and 0.807 for histology and µCT segmentation, respectively. The correlation between the co-registered regions was strong (r = 0.897, bias = 21.9 µm, standard deviation = 21.5 µm). Finally, both methods could separate the CC thickness between the patella, femoral, and tibial regions (p < 0.001). As a conclusion, the proposed µCT analysis allows for ex vivo 3D assessment of CC morphology. We demonstrated the biomedical relevance of the method by quantifying CC thickness in different anatomical regions with a varying mean thickness. CC was thickest in the patella and thinnest in the tibial plateau. Our method is relatively straightforward to implement into standard µCT analysis pipelines, allowing the analysis of CC morphology. In future research, µCT imaging might be preferable to histology, especially when analyzing dynamic changes in cartilage mineralization. It could also provide further understanding of 3D morphological changes that may occur in mineralized cartilage, such as thickening of the subchondral plate in osteoarthritis and other joint diseases.

Molecular Mechanisms and Therapeutic Potential of miR-493 in Cancer.

MicroRNAs (miRNAs) are highly conserved short noncoding RNAs with the capacity of regulating gene expression posttranscriptionally. In this process, miRNAs partially bind complementary sites of target mRNAs. Among miRNAs, miR-493 performs important functions under diverse physiological conditions and participates in different pathogeneses, including pancreatic cancer, gastric cancer, and breast cancer. Differential expression of miR-493 plays a vital role in the generation, metastasis, and recurrence of tumors. Above all, increasing evidence indicates that miR-493 inhibits the generation and development of tumors by activating the Wnt/Β-catenin, Wnt/PCP, MEK/ERK, or PI3K/AKT signaling pathway, suggesting possibilities for miR-493 as an effective adjuvant cancer therapy. In this review, we discuss and summarize the biological mechanisms of miR-493 and its potential in cancer therapy. This review may provide a better understanding of the biological functions of miR-493 in tumors and provide important clues to cancer treatment.

Moderate tibia axial loading promotes discordant response of bone composition parameters and mechanical properties in a hindlimb unloading rat model.

OBJECTIVES: The purpose of the present study was to characterize the dynamic alterations of bone composition parameters and mechanical properties to disuse and mechanical intervention. METHODS: A tail suspension hindlimb unloading model and an in vivo axial tibia loading model in rats were used. A moderate mechanical loading that was capable of engendering 800 µÎµ tibia strain was applied to the right tibia of rats in both control and hindlimb unloading group across 28 days of the experimental period. The contralateral tibia served as control. RESULTS: Hindlimb unloading led to bone loss in tibia from day 14. Bone mineral density, mineral content and mechanical properties responded differently with microstructure to disuse in timing course. Mechanical loading of 800 µÎµ tibia strain failed to alter the bone of the control group, but minimized the detrimental effects of unloading by completely prohibiting the decrease of bone mineral content and main mechanical properties after 28 days. Less obvious influence of mechanical loading on bone microstructure was found. CONCLUSIONS: The moderate mechanical loading is not able to stimulate the mechanical response of healthy tibia, but indeed lead to discordant recovery of bone composition parameters and mechanical properties.

Response of Osteoblasts to the Stimulus of Fluid Flow.

Bone is an important porous tissue that supports the body, maintains calcium and phosphate homeostasis, protects vital organs, and houses bone marrow. The interaction between hydrostatic pressure and fluid phase, solid phase, cells, and vascular in bone makes bone inevitably bear baseline levels of fluid flow. Fluid flow plays an important role in regulating the proliferation, differentiation, distribution, and apoptosis of osteoblasts in bone. The effect of fluid flow on osteoblasts is dependent on time, velocity, and type. Some response of osteoblasts to fluid flow is closely related to the soluble factors secreted by the osteoblasts themselves or other types of bone cells. When the response is disordered, related bone diseases such as osteoporosis, osteoarthritis, and abnormal osteogenesis probably happen. In this article we review the current progress in the study of the response of osteoblasts to the direct and indirect stimulus of fluid flow and their roles in osteogenesis and related bone diseases.

3D printed strontium-zinc-phosphate bioceramic scaffolds with multiple biological functions for bone tissue regeneration.

Calcium phosphate (CaP) bioceramics are broadly employed for bone regeneration due to their excellent biocompatibility and osteoconductivity. However, they are not capable of repairing healing-impaired bone defects such as defects with conditions of ischemia or infection due to restricted bioactivities. In this study, we synthesized single-phased strontium-zinc-phosphate (SZP, SrZn2(PO4)2) bioceramics via a solution combustion method and further fabricated SZP scaffolds using a three-dimensional (3D) printing technique. Compared to 3D printed ß-tricalcium phosphate (ß-TCP) scaffolds, the 3D printed SZP scaffolds presented comparable porosity, compressive strength, and Young's modulus, but increased ability of osteogenesis, angiogenesis, immunomodulation and anti-bacterial activity. Specifically, 3D printed SZP scaffolds not only led to significantly higher osteogenic differentiation of MC3T3-E1 cells and pro-angiogenesis of human umbilical vein endothelial cells (HUVECs) directly or through macrophage-mediated immunomodulation, but also inhibited the growth of Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). The in vivo study of the rat cranial bone defect model further confirmed better vascularized bone regeneration in 3D-printed SZP scaffolds. These findings indicate that the proposed 3D-printed SZP scaffolds might be a versatile candidate for bone tissue engineering.

3D-printed Sr2ZnSi2O7 scaffold facilitates vascularized bone regeneration through macrophage immunomodulation.

Biomaterial-based bone grafts are emerged as an effective strategy for the treatment of large bone defects, especially for the scaffolds with enhanced osteogenic and angiogenic bioactivities. However, most studies focused on the direct interactions between scaffolds and bone-related cells such as osteoblasts and endothelial cells, and ignored the effects of material-triggered immunomodulation and the subsequent immune-regulated bone regeneration process. In this study, we developed a silicate bioceramic (Sr2ZnSi2O7, SZS) scaffold with well-defined pore structures using a three-dimensional (3D) printing technique. The prepared scaffolds were biodegradable, and the released bioactive ions were beneficial for immunomodulation, which stimulated macrophages to release more pro-healing cytokines and less pro-inflammatory cytokines. The obtained scaffold/macrophage conditioned medium further promoted the proliferation and osteogenic differentiation of a murine preosteoblast cell line (MC3T3-E1), as well as the angiogenic activity of human umbilical vein endothelial cells (HUVECs). Moreover, the in vivo experiments of critical-sized calvarial defects in rats revealed that the 3D printed SZS scaffolds could facilitate more vascularized bone regeneration than the 3D printed ß-tricalcium phosphate (ß-TCP, a typical clinically used bioceramic) scaffolds, suggesting that the 3D-printed SZS scaffolds hold the potential as implantable biomaterials with favorable osteoimmunomodulation for bone repair.

Changes in subchondral bone structure and mechanical properties do not substantially affect cartilage mechanical responses - A finite element study.

Subchondral bone structure has been observed to change in osteoarthritis (OA). However, it remains unclear how the early-stage OA changes affect the mechanics (stresses and strains) of the osteochondral unit. In this study, we aim to characterize the effect of subchondral bone structure and mechanical properties on the osteochondral unit mechanics. A 3-D finite element model of the osteochondral unit was constructed based on a rabbit femoral condyle µCT data and subjected to creep loading in indentation. Trabecular bone volume fraction, subchondral bone plate thickness, and equilibrium modulus were varied (including experimentally observed changes in early OA) to characterize the effect of these parameters on the osteochondral unit mechanics. At the end of the creep phase, the maximum principal strain at the bone surface of the cartilage-bone interface was decreased by 50% when the trabecular bone volume fraction was reduced from 48% to 28%. The maximum principal stress at the same location was decreased by 36% when plate thickness was reduced by 100 µm (-31%). In cartilage, small changes in the mechanics were seen near the cartilage-bone interface with a considerably thinner (-31%) plate. The changes in trabecular bone volume fraction, subchondral bone thickness and plate equilibrium modulus did not substantially affect the cartilage mechanics. Our results suggest that experimentally observed changes that occur in the subchondral bone structure in early OA have a minimal effect on cartilage mechanics under creep indentation loading; clear changes in the cartilage mechanics were seen only with an unrealistically soft subchondral bone plate.

Early changes in osteochondral tissues in a rabbit model of post-traumatic osteoarthritis.

Concurrent osteoarthritic (OA) manifestations in bone and cartilage are poorly known. To shed light on this issue, this study aims to investigate changes in subchondral bone and articular cartilage at two time points after anterior cruciate ligament transection (ACLT) in a rabbit model. 2 (N = 16) and 8 (N = 10) weeks after ACLT, the subchondral bone structure, cartilage thickness, Osteoarthritis Research Society International (OARSI) score, fixed charged density (FCD), and collagen orientation angle were analyzed. OA related changes were evaluated by comparing the ACLT to the contralateral (C-L) and control knees. Already 2 weeks after ACLT, higher trabecular number in the medial femoral condyle and femoral groove, greater OARSI score in the femoral condyles, and thinner trabeculae in the lateral tibial plateau and femoral groove were observed in ACLT compared to C-L knees. Only minor changes of cartilage collagen orientation in the femoral condyles and femoral groove and smaller FCD in the femoral condyles, medial tibial plateau, femoral groove and patella were observed. 8 weeks post-ACLT, the surgical knees had thinner subchondral plate and trabeculae, and smaller trabecular bone volume fraction in most of the knee locations. OARSI score was greater in the femoral condyle and lateral tibial plateau cartilage. FCD loss was progressive only in the femoral condyle, femoral groove, and patellar cartilage, and minor changes of cartilage collagen orientation angle were present in the femoral condyles, femoral groove, and lateral tibial plateau. We conclude that ACLT induces progressive subchondral bone loss, during which proteoglycan loss occurs followed by their partly recovery, as indicated by FCD results.

Experimental mechanical strain measurement of tissues.

Strain, an important biomechanical factor, occurs at different scales from molecules and cells to tissues and organs in physiological conditions. Under mechanical strain, the strength of tissues and their micro- and nanocomponents, the structure, proliferation, differentiation and apoptosis of cells and even the cytokines expressed by cells probably shift. Thus, the measurement of mechanical strain (i.e., relative displacement or deformation) is critical to understand functional changes in tissues, and to elucidate basic relationships between mechanical loading and tissue response. In the last decades, a great number of methods have been developed and applied to measure the deformations and mechanical strains in tissues comprising bone, tendon, ligament, muscle and brain as well as blood vessels. In this article, we have reviewed the mechanical strain measurement from six aspects: electro-based, light-based, ultrasound-based, magnetic resonance-based and computed tomography-based techniques, and the texture correlation-based image processing method. The review may help solving the problems of experimental and mechanical strain measurement of tissues under different measurement environments.

Potential role of miR-139-5p in cancer diagnosis, prognosis and therapy.

MicroRNAs (miRNAs) are short non-coding RNAs that regulate the expression of protein-coding genes by partially binding to specific target sites of mRNAs. miRNAs perform important functions in complicated cellular biological processes and their abnormal expression is involved in various disorders, including cancer. Among the miRNAs, differential expression of miR-139-5p serves a significant role in tumorigenesis, metastasis and recurrence, thus suggesting that it may potentially be used as a promising biomarker for cancer diagnosis, prognosis and therapy. miR-139-5p is expected to serve as a biomarker to eventually be implemented in a clinical setting. In the present review, we focus on the importance of miR-139-5p in cancer, summarize the association between miR-139-5p expression level and diagnosis and prognosis, and discuss the potential therapeutic implications for the future.