BCL11A Expression in Breast Cancer.

B-cell leukemia/lymphoma 11A (BCL11A) is a transcription factor that regulates the expression of genes involved in cell division or apoptosis. A link between high BCL11A expression and a worse prognosis has been demonstrated in patients with various cancers. The aim of this study was to investigate the expression pattern of BCL11A in breast cancer (BC) cases and mastopathy samples and to correlate the results with the clinicopathological data. The expression of the BCL11A protein was investigated using immunohistochemistry (IHC) on 200 cases of BC and 13 mastopathy samples. The level of BCL11A mRNA was determined using real-time PCR in 22 cases of BC and 6 mastopathy samples. The expression of BCL11A was also examined at the protein and mRNA levels in BC cell lines. A higher expression level of BCL11A in BC cases was shown compared to mastopathy samples. The expression level of BCL11A in BC cases and in the studied cell lines decreased with the increasing grade of histological malignancy (G). It was also negatively correlated with the primary tumor size. A significantly lower expression of BCL11A was found in BC that did not express estrogen or progesterone receptors and in triple-negative cases. The results of our research suggest that BCL11A may be relevant in the development of BC.

The Expression of Testin, Ki-67 and p16 in Cervical Cancer Diagnostics.

Testin is a protein expressed in normal human tissues, being responsible, with other cytoskeleton proteins, for the proper functioning of cell−cell junction areas and focal adhesion plaques. It takes part in the regulation of actin filament changes during cell spreading and motility. Loss of heterozygosity in the testin-encoding gene results in altered protein expression in many malignancies, as partly described for cervical cancer. The aim of our study was the assessment of the immunohistochemical (IHC) expression of testin in cervical cancer and its analysis in regard to clinical data as well the expression of the Ki-67 antigen and p16 protein. Moreover, testin expression was assessed by Western blot (WB) in commercially available cell lines. The IHC analysis disclosed that the expression of testin inversely correlated with p16 (r = −0.2104, p < 0.0465) and Ki-67 expression (r = −0.2359, p < 0.0278). Moreover, weaker testin expression was observed in cancer cases vs. control ones (p < 0.0113). The WB analysis of testin expression in the cervical cancer cell lines corresponded to the IHC results and showed a weaker expression compared to that in the control cell line. When we compared the expression of testin in cervical cancer cell lines, we found a weaker expression in HPV-negative cell lines. In summary, we found that the intensity of testin expression and the number of positive cells inversely correlated with the expression of Ki-67 (a marker of proliferation) and p16 (a marker of cell cycle dysregulation). This study shows that the combined assessment of testin, Ki-67 and p16 expression may improve cervical cancer diagnostics.

Targeting SOX18 Transcription Factor Activity by Small-Molecule Inhibitor Sm4 in Non-Small Lung Cancer Cell Lines.

The transcription factor SOX18 has been shown to play a crucial role in lung cancer progression and metastasis. In this study, we investigated the effect of Sm4, a SOX18 inhibitor, on cell cycle regulation in non-small cell lung cancer (NSCLC) cell lines LXF-289 and SK-MES-1, as well as normal human lung fibroblast cell line IMR-90. Our results demonstrated that Sm4 treatment induced cytotoxic effects on all three cell lines, with a greater effect observed in NSCLC adenocarcinoma cells. Sm4 treatment led to S-phase cell accumulation and upregulation of p21, a key regulator of the S-to-G2/M phase transition. While no significant changes in SOX7 or SOX17 protein expression were observed, Sm4 treatment resulted in a significant upregulation of SOX17 gene expression. Furthermore, our findings suggest a complex interplay between SOX18 and p21 in the context of lung cancer, with a positive correlation observed between SOX18 expression and p21 nuclear presence in clinical tissue samples obtained from lung cancer patients. These results suggest that Sm4 has the potential to disrupt the cell cycle and target cancer cell growth by modulating SOX18 activity and p21 expression. Further investigation is necessary to fully understand the relationship between SOX18 and p21 in lung cancer and to explore the therapeutic potential of SOX18 inhibition in lung cancer.

Label-Free Quantitative Phase Imaging Reveals Spatial Heterogeneity of Extracellular Vesicles in Select Colon Disorders.

Three-dimensional (3D) imaging and quantitative analysis of extracellular vesicles (EVs) remain largely unexplored, mainly because of limitations in detection techniques. In this study, EVs from patients diagnosed with colorectal cancer (CRC) and ulcerative colitis were examined. To investigate the spatial heterogeneity and 3D refractive index (RI) distribution of single EVs, a label-free digital holographic tomography technique was used at a submicrometer spatial resolution. The presented image-processing algorithms were used in quantitative analysis with digital staining and 3D visualization, the determination of the EV size distribution and extraction of fractions with different RIs. Reconstructed 3D RI distributions revealed variations in the spatial heterogeneity of EVs related to tissue specificity, such as CRC, normal colonic mucosa, and ulcerative colitis, as well as the isolation procedures used. The RI values of EVs isolated from solid tissues of frozen CRC samples were also dependent on the tumor grade and cancer cell proliferation. The simultaneous examination of cell culture models confirmed the association of the RI of EVs with the tumor grade. 3D-RI data analysis generates new perspectives with the optical, contact-free, label-free examination of the individual EVs. Depending on the specific tissue and isolation method, EVs exhibit significant spatial heterogeneity. The optical parameters of single EVs enabled their classification into two unique subgroups with different RI values.

Are Biogenic and Pyrogenic Mesoporous SiO2 Nanoparticles Safe for Normal Cells?

Silicon dioxide, in the form of nanoparticles, possesses unique physicochemical properties (size, shape, and a large surface to volume ratio). Therefore, it is one of the most promising materials used in biomedicine. In this paper, we compare the biological effects of both mesoporous silica nanoparticles extracted from Urtica dioica L. and pyrogenic material. Both SEM and TEM investigations confirmed the size range of tested nanoparticles was between 6 and 20 nanometers and their amorphous structure. The cytotoxic activity of the compounds and intracellular ROS were determined in relation to cells HMEC-1 and erythrocytes. The cytotoxic effects of SiO2 NPs were determined after exposure to different concentrations and three periods of incubation. The same effects for endothelial cells were tested under the same range of concentrations but after 2 and 24 h of exposure to erythrocytes. The cell viability was measured using spectrophotometric and fluorimetric assays, and the impact of the nanoparticles on the level of intracellular ROS. The obtained results indicated that bioSiO2 NPs, present higher toxicity than pyrogenic NPs and have a higher influence on ROS production. Mesoporous silica nanoparticles show good hemocompatibility but after a 24 h incubation of erythrocytes with silica, the increase in hemolysis process, the decrease in osmotic resistance of red blood cells, and shape of erythrocytes changed were observed.

The role of YKL-40 in a cancerous process.

YKL-40 is a secretory protein secreted among others by tumor cells and tumor-associated macrophages. Due to the structural homology this protein was classified to chitinases family CLP (chitinase - like protein) and to 18 of glycosyl hydrolase family, but it has no catalytic function. Elevated levels of YKL-40 in serum is observed in the inflammatory diseases of various aetiology and in cancers, such as breast, ovarian, colon or lung. The results of many studies suggest a significant relationship of YKL-40 with progression of cancer: incidence of metastases, shorter relapse-free survival and shorter overall survival. It is believed that YKL-40 may be a prognostic factor of cancer development and the patient's response to the applied therapy. Elevated levels of protein in serum of cancer patients may play a role in angiogenesis, proliferation and migration of tumor cells. Probably the mechanism of this phenomenon is the result of YKL-40 action by FAK and PI3K/AKT signaling pathways. Results obtained so far are largely based on an analysis of the YKL-40 level in the patients' serum and on the assessment of changes in the expression level of this protein in studies in vitro and in vivo.

Low-vacuum SEM imaging and viability test of L929 cells exposed to a Euro 6 diesel exhaust gas mixture in a BAT-CELL chamber in comparison with hydrocarbons emission.

Exhaust emissions, which count among the most common causes of premature death worldwide, can cause irreversible changes in cells, leading to their damage or degeneration. In this research, L929 line cells were observed after exposure in the BAT-CELL chamber to exhaust gases emitted from a Euro 6 compression-ignition engine. Real road traffic conditions were simulated, taking into account air resistance while driving at speeds of 50 km/h, 120 km/h and idling engine. Morphological analysis of the cells was performed using an environmental scanning electron microscope. It has been observed that diesel exhaust fumes can cause inflammation, which can induce apoptosis or leads to necrotic cell death. The impact of the vehicle exhaust gases can inhibit cell proliferation by almost three times. Moreover, a correlation has been observed between the speed of the inflammatory reaction in cells and the presence of specific hydrocarbon compounds that determine the toxicity of exhaust gases. Research has shown that the toxicity of the emitted exhaust gases has been the highest at the driving speed of 120 km/h. In order to reduce the harmful effects of exhaust emissions, ecological alternatives and the supplementation of legal provisions regarding the compounds subject to limitation are necessary.

Three-Dimensional-Printed Modular Titanium Alloy Plates for Osteosynthesis of the Jawbone.

BACKGROUNDS: The titanium-aluminum-vanadium alloy (Ti-6Al-4V) is frequently used in implantology due to its biocompatibility. The use of 3D printing enables the mechanical modification of implant structures and the adaptation of their shape to the specific needs of individual patients. METHODS: The titanium alloy plates were designed using the 3D CAD method and printed using a 3D SLM printer. Qualitative tests were performed on the material surface using a microcomputed tomography scanner. The cytotoxicity of the modular titanium plates was investigated using the MTT assay on the L929 cell line and in direct contact with Balb/3T3 cells. Cell adhesion to the material surface was evaluated with hFOB1.19 human osteoblasts. Microbial biofilm formation was investigated on strains of Lactobacillus rhamnosus, Staphylococcus epidermidis, Streptococcus mutans and Candida albicans using the TTC test and scanning electron microscopy (SEM). RESULTS: The surface analysis showed the hydrophobic nature of the implant. The study showed that the titanium plates had no cytotoxic properties. In addition, the material surface showed favorable properties for osteoblast adhesion. Among the microorganisms tested, the strains of S. mutans and S. epidermidis showed the highest adhesion capacity to the plate surface, while the fungus C. albicans showed the lowest adhesion capacity. CONCLUSIONS: The manufactured modular plates have properties that are advantageous for the implantation and reduction in selected forms of microbial biofilm. Three-dimensional-printed modular titanium plates were investigated in this study and revealed the potential clinical application of this type of materials, regarding lack of cytotoxicity, high adhesion properties for osteoblasts and reduction in biofilm formation. The 3D CAD method allows us to personalise the shape of implants for individual patients.

New insights into the role of tetraspanin 6, 7, and 8 in physiology and pathology.

BACKGROUND: The tetraspanin (TSPAN) family comprises 33 membrane receptors involved in various physiological processes in humans. Tetrasapanins are surface proteins expressed in cells of various organisms. They are localised to the cell membrane by four transmembrane domains (TM4SF). These domains bind several cell surface receptors and signalling proteins to tetraspanin-enriched lipid microdomains (TERM or TEM). Tetraspanins play a critical role in anchoring many proteins. They also act as a scaffold for cell signalling proteins. AIM: To summarise how tetraspanins 6, 7 and 8 contribute to the carcinogenesis process in different types of cancer. METHODS: To provide a comprehensive review of the role of tetraspanins 6, 7 and 8 in cancer biology, we conducted a thorough search in PubMed, Embase and performed manual search of reference list to collect and extract data. DISCUSSION: The assembly of tetraspanins covers an area of approximately 100-400 nm. Tetraspanins are involved in various biological processes such as membrane fusion, aggregation, proliferation, adhesion, cell migration and differentiation. They can also regulate integrins, cell surface receptors and signalling molecules. Tetraspanins form direct bonds with proteins and other members of the tetraspanin family, forming a hierarchical network of interactions and are thought to be involved in cell and membrane compartmentalisation. Tetraspanins have been implicated in cancer progression and have been shown to have multiple binding partners and to promote cancer progression and metastasis. Clinical studies have documented a correlation between the level of tetraspanin expression and the prediction of cancer progression, including breast and lung cancer. CONCLUSIONS: Tetraspanins are understudied in almost all cell types and their functions are not clearly defined. Fortunately, it has been possible to identify the basic mechanisms underlying the biological role of these proteins. Therefore, the purpose of this review is to describe the roles of tetraspanins 6, 7 and 8.

The Effect of Hypoxia on Irisin Expression in HL-1 Cardiomyocytes.

BACKGROUND/AIM: Cardiovascular diseases (CVD) are the leading cause of death worldwide. In 2019, 523 million people were diagnosed with CVD, with 18.6 million deaths. Improved treatment and diagnostics could reduce CVD's impact. Irisin (Ir) is crucial for heart function and may be a biomarker for heart attack. Ir is a glycoprotein with sugar residues attached to its protein structure. This glycosylation affects Ir stability, solubility, and receptor interactions on target cells. Its secondary structure includes a fibronectin type III domain, essential for its biological functions. Ir helps cardiomyocytes to respond to hypoxia and protects mitochondria. The aim of the study was to determine the FNDC5 gene expression level and the Ir level in HL-1 cardiomyocytes subjected to hypoxia. MATERIALS AND METHODS: We examined the effect of hypoxia on the expression levels of the FNDC5 gene and those of Ir in mouse cardiomyocytes of the HL-1 cell line. Real-time PCR (RT-PCR) was used to estimate the expression levels of the FNDC5 gene. Western blot and immunofluorescence methods were used to analyze the Ir protein levels. RESULTS: Analyses showed an increased Ir level in HL-1 cardiomyocytes in response to hypoxia. This is the first study to confirm the presence of Ir in HL-1 cells. CONCLUSION: The observed increase in Ir expression in murine cardiomyocytes is associated with the hypoxic environment and can be potentially used to diagnose hypoxia and CVD.

Inhibition of CHI3L1 decreases N-cadherin and VCAM-1 levels in glioblastoma.

Background: (1)The protein CHI3L1 supports cancer development in several ways, including stimulation of angiogenesis and invasion as well as immunomodulatory effects. These properties make it a potential target for the development of targeted therapies in precision medicine. In this context, the particular potential of CHI3L1 inhibition could be considered in glioblastoma multiforme (GBM), whose tumors exhibit high levels of angiogenesis and increased CHI3L1 expression. This study aims to investigate whether inhibition of CHI3L1 in spheroids used as a GBM model affects the mechanisms of invasiveness. Methods: (2)We analyzed the interactions between CHI3L1 and the inhibitor G721-0282 in molecular docking (in silico) and infrared spectroscopy. Uptake of G721-0282 in GBM spheroids was measured using a label-free physical cytometer. Changes in E-, N- and VE-cadherins, VCAM-1 and EGFR were analyzed by immunohistochemical reactions, Western blot and ddPCR methods in U-87 MG cells and GBM spheroids consisting of U-87 MG glioblastoma cells, HMEC-1 endothelial cells and macrophages. Results: (3)A direct interaction between CHI3L1 and G721-0282 was observed. G721-0282 decreased N-cadherins and VCAM-1 in GBM spheroids, but the changes in the 2D model of U-87 MG glioblastoma cells were different. Conclusion: (4)Inhibition of CHI3L1 has the potential to reduce the invasiveness of GBM tumors. The proposed 3D model of GBM spheroids is of great significance for investigating changes in membrane proteins and the tumor microenvironment.

Impact of RBMS 3 Progression on Expression of EMT Markers.

Epithelial-to-mesenchymal transition (EMT) is a complex cellular process that allows cells to change their phenotype from epithelial to mesenchymal-like. Type 3 EMT occurs during cancer progression. The aim of this study was to investigate the role of RNA-binding motif single-stranded interacting protein 3 (RBMS 3) in the process of EMT. To investigate the impact of RBMS 3 on EMT, we performed immunohistochemical (IHC) reactions on archived paraffin blocks of invasive ductal breast carcinoma (n = 449), allowing us to analyze the correlation in expression between RBMS 3 and common markers of EMT. The IHC results confirmed the association of RBMS 3 with EMT markers. Furthermore, we performed an in vitro study using cellular models of triple negative and HER-2-enriched breast cancer with the overexpression and silencing of RBMS 3. RT-qPCR and Western blot methods were used to detect changes at both the mRNA and protein levels. An invasion assay and confocal microscopy were used to study the migratory potential of cells depending on the RBMS 3 expression. The studies conducted suggest that RBMS 3 may potentially act as an EMT-promoting agent in the most aggressive subtype of breast cancer, triple negative breast cancer (TNBC), but as an EMT suppressor in the HER-2-enriched subtype. The results of this study indicate the complex role of RBMS 3 in regulating the EMT process and present it as a future potential target for personalized therapies and a diagnostic marker in breast cancer.

[Does magnetic stimulation affect wound healing? In vitro studies]. / Czy magnetostymulacja moze miec wplyw na gojenie ran? Badania in vitro.

BACKGROUND: Variable magnetic field of low frequency (200-300 Hz) is one of physical methods used in reducing pain as well as regeneration of bone and soft tissue. In medical literature there are case reports about successful treatment of chronic wound healing with this method. However, there is a lack of research that could explain the mechanism of action of magnetic field in this area. Literature data show that magnetic fields have an influence on cells cultures in vitro. Cells reaction depends on cells line, field parameters and time of exposition. OBJECTIVE: In our study we checked if the magnetic field of 180-195 Hz frequency influences Balb 3T3 cells viability. MATERIAL AND METHODS: This study was conducted on mouse fibroblast Balb 3T3 cells, and the influence of variable magnetic field on cells was checked. Magnetic field was generated by Viofor JPS System Classic (Med&Life). Cells were seeded on 96-well plates. After 24 hours the cells culture was exposed on magnetic fields. Two controls and six groups was included in the study. Two programs generated by Viofor JPS System Classic were chosen: M1P2 and M2P2, as well as two intensities 6 and 12. Groups 1, 2, 5 and 6 were exposed once within two days, groups 3 and 4 were exposed three times a day every hour within two days. Experiment lasted two days and was repeated 3-5 times. RESULTS: Experiment was evaluated with colorimetric MTT test. The test showed influence of magnetic field generated by Viofor JPS System Classic on viability of Balb 3T3 cells. Three from six chosen programs resulted in the increase of viability, compare to control. The control was taken as 100%. In groups 139%, 128%, 108% and 92% of viability was noted. Results were statisticaly significant in four groups (p < 0.05, Student's t test). CONCLUSIONS: The influence of magnetic fields generated by Viofor JPS System Classic (Med&Life) on mouse fibroblast Balb 3T3 cells was noted. Results suggest potential beneficial effect of this physical method on chronic wound treatment.

[New directions of research related to chronic wound healing]. / Nowe kierunki badan zwiazane z gojeniem przewleklych ran.

Optimal nutrition, immunological state and psychological condition play an important role in the process of chronic wound healing. Infections caused by pathogens resistant to commonly used antibiotics additionally complicate and disturb regeneration of wounds. As part of the treatment, modern wound dressings are used, for example designed on the basis of alginates, dextranomers, hydrogels, hydrofiber, polyurethanes foams, hydrocolloids and liquids for wound debridement such us 0.9% NaCl, the PWE liquid, Ringer's liquid, octenidine. Owing to their features, treatment in accordance with TIME concept could be realized, because they provide moisture wound bed, protection against contamination, gas exchange, protection of wound edges and infection control. Repairing process in chronic wounds is dependent on blood flow in tissues, which may be insufficient. The result is a permanent hypoxia. Natural occurring antioxidants are becoming more crucial in chronic wound treatment. They decrease oxygen radical concentration, increase angiogenesis, reduce inflammatory response, stimulate fibroblasts and keratinocytes proliferation, possess antibacterial properties against chemotherapeutic resistant strains. There are a lot of antioxidants in honey, papaya fruit (Carrica papaia L.), transgenic flax (Linum usitatissimum), and in orange oil (Citrus sinensis), stem of acanthus (Acanthus ebracteatus), leafs of tea (Camellia sinensis). Application of biologically active, natural derived compounds is nowadays a direction of intense in vitro and in vivo research focused on the chronic wound treatment. Results suggest beneficial influence of antioxidant on wound repairing process. Clinical research are needed to state effective influence of natural compound in the chronic wound treatment.

[Influence of the gelatin-alginate matrixes with calcium lactate in plasma coagulation system after implantation in soft tissues]. / Wplyw matryc zelatynowo-alginianowych z mleczanem wapnia na osoczowy uklad krzepniecia po implantacji w tkanki miekkie.

BACKGROUND: Bioresorbable porous substrates from copolymers have their application in tissue engineering to culture tissues in vitro. The advantage of polymers is the production of thermoplastic elements and their ability to biodegrade in a living body. Gelatin, collagen, alginates are part of dressings used for topical administration of the drug. Research was undertaken to achieve a porous gelatin-alginate matrix which could be used in therapy as among others, a carrier for a drug. OBJECTIVES: Evaluation of the impact of modified gelatin-alginate matrix on activation of plasma coagulation in vivo. MATERIAL AND METHODS: Gelatin-alginate matrix cross-linked with calcium ions was implanted in the muscle tissue of a rat. The control group constituted animals not implanted with material, but they passed the operating procedure. Blood samples of plasma coagulation test and control group were collected after 1, 2, 3, 5, 7, 10, 14 days of the procedure. RESULTS: Prolongation of APTT and shortening of PT and TT with the unchanged values of fibrinogen and the count of platelet cells was observed till the 5th day on the basis of the obtained results. Prolongation of APTT with the unchanged values of the remaining parameters of the coagulation system was observed after 7, 10 and 14 days with unchanged values of PT and TT coagulation. CONCLUSIONS: The matrix gelatin-alginate with calcium ions in the biological environment undergoes biodegradation in soft tissues. This process in the initial period influences the activation of the coagulation within the intrinsic and extrinsic system. From the 5th to 14th day the activation of coagulation was observed only in the intrinsic system.

[A method of manufacture and characteristic of surface properties and biological activity of thin-film coatings of Cu-Ti system]. / Sposób wytwarzania oraz charakterystyka wlasciwosci powierzchni i aktywnosci biologicznej cienkowarstwowych powlok ukladu typu Cu-Ti.

BACKGROUND: Biomaterials in the form of thin-film coatings as-deposited on different substrates are nowadays increasingly popular. In particular coatings based on a combination of biocompatible materials (eg. titanium) with metals of high biological activity (eg. copper) have a potentially wide range of applications as active films, intended for various types of medical devices. OBJECTIVES: The aim of this study was to present a method for preparation and analysis of the properties of Cu-Ti thin films, in particular their biological activity in connection with the properties of the surface. MATERIAL AND METHODS: The films were prepared by magnetron sputtering method with the aid of an innovative four-target apparatus. During deposition process two metallic targets (copper and titanium) were sputtered under an argon atmosphere. Material composition of produced coatings was characterized by scanning electron microscope equipped with an adapter for energy dispersive spectroscopy. Moreover, the surface microstructure and roughness of coatings was characterized based on three-dimensional surface profiles, which were obtained with the aid of optical profilometer. The research was also carried out by investigations of surface wettability on the apparatus for measurements of contact angle. Characterization of Cu-Ti surface properties was also expanded by microbiological tests involving Staphylococcus aureus (PCM 2602) bacteria and investigations of cytotoxicity with L 929 (NCTC clone 929) cell line. RESULTS: Measurements have shown that the film was composed of 71% at. Cu and 29% at. Ti. The analysis of surface topography has shown that the surface of Cu-Ti thin film was very uniform with roughness in range of nanometers. It was found that as-deposited film is hydrophilic. Obtained results has shown that as-deposited film had a very good bactericidal properties and it was cytotoxic. This effect was associated with the migration of copper ions, which was the most intense at the edge of the sample. CONCLUSIONS: The results presented in this paper testify that manufactured Cu-Ti thin films may find practical application in the industry as a bioactive coating.

Mechanical properties and osteointegration of the mesh structure of a lumbar fusion cage made by 3D printing.

The currently popular 3D printing makes it possible to produce spatial scaffolds, the main purpose of which is to obtain implants that have favourable mechanical properties to promote cell adhesion. This study aims to prove the influence of changes in selected geometrical parameters of scaffolds, used in intervertebral cages, on the mechanical properties obtained and thus on the osteointegration of the studied constructs with osteoblasts and fibroblasts. The stiffness values and maximum failure force of four modifications to geometric dimensions of the meshes were determined from the intendation test. Adhesion assays were conducted (including gentle pendulum motion) for Balb/3T3 fibroblasts and NHOst osteoblasts. The study revealed that an important geometrical parameter affecting the strength of the mesh is the height (h) of the connection point between arms of successive mesh cells. There was no significant effect of the mesh geometry on the abundance and survival of Balb/3T3 and NHOst cells. At the same time, fibroblasts were more likely to form colonies in the area where there is fusion of mesh cells, as opposed to osteoblasts that were more numerous at vertices of the mesh.

Changes in the scar tissue structure after cesarean section as a result of manual therapy.

BACKGROUND: Available statistical data from 2015 show that 28% of pregnancies in developed countries end in cesarean section (CC). Discomfort associated with the scar after surgery is a common complication. OBJECTIVES: This study aimed to evaluate the changes in the structure of the cesarean scar after the application of a scheme of manual therapy. MATERIAL AND METHODS: The study included 15 women in the treatment group (TG) and 15 in the control group (CG). The scars were evaluated twice at 5-week intervals with the use of quantitative scales: the Vancouver Scar Scale (VSS), the Manchester Scar Scale (MSS) and the Patient and Observer Scar Assessment Scale (POSAS). During each examination, the scar was compared, using the specified criteria, to the physiological skin, i.e., the tissues directly bordering the incision. During therapy, 8 manual techniques were used during a 4-week program consisting of 30-minute sessions 3 times per week. RESULTS: Patients in the TG showed a statistically significant improvement in all of the analyzed characteristics of the scar. A statistically significant difference was also observed between the results obtained during the 2nd examination (after the therapy) in the TG and the CG. CONCLUSIONS: As a result of the therapy, the condition of the scar in the TG significantly improved. Onerous scar-related symptoms were alleviated. The vascularity, hyperpigmentation and distortion of the scar were reduced. The elasticity and pliability of the scar increased, and the height of the scar decreased. The texture, finish and contour of the scar improved. Obtained results suggest that manual therapy of the scar after CC should be a part of the treatment in women during the postpartum period.

Chitin scaffolds derived from the marine demosponge Aplysina fistularis stimulate the differentiation of dental pulp stem cells.

The use of stem cells for tissue regeneration is a prominent trend in regenerative medicine and tissue engineering. In particular, dental pulp stem cells (DPSCs) have garnered considerable attention. When exposed to specific conditions, DPSCs have the ability to differentiate into osteoblasts and odontoblasts. Scaffolds are critical for cell differentiation because they replicate the 3D microenvironment of the niche and enhance cell adhesion, migration, and differentiation. The purpose of this study is to present the biological responses of human DPSCs to a purified 3D chitin scaffold derived from the marine demosponge Aplysina fistularis and modified with hydroxyapatite (HAp). Responses examined included proliferation, adhesion, and differentiation. The control culture consisted of the human osteoblast cell line, hFOB 1.19. Electron microscopy was used to examine the ultrastructure of the cells (transmission electron microscopy) and the surface of the scaffold (scanning electron microscopy). Cell adhesion to the scaffolds was determined by neutral red and crystal violet staining methods. An alkaline phosphatase (ALP) assay was used for assessing osteoblast/odontoblast differentiation. We evaluated the expression of osteogenic marker genes by performing ddPCR for ALP, RUNX2, and SPP1 mRNA expression levels. The results show that the chitin biomaterial provides a favorable environment for DPSC and hFOB 1.19 cell adhesion and supports both cell proliferation and differentiation. The chitin scaffold, especially with HAp modification, isolated from A. fistularis can make a significant contribution to tissue engineering and regenerative medicine.

Multimodal study of CHI3L1 inhibition and its effect on angiogenesis, migration, immune response and refractive index of cellular structures in glioblastoma.

Glioblastoma is one of the most aggressive tumours with a poor response to treatment and a poor prognosis for patients. One of the proteins expressed in glioblastoma tissue is CHI3L1 (YKL-40), which is upregulated and known for its angiogenesis-supporting and pro-tumour immunomodulatory effects in a variety of cancers. In this paper we present the anti-angiogenic, anti-migratory and immunomodulatory effects of the compound G721-0282, an inhibitor of CHI3L1. The inhibitor-induced changes were investigated using conventional techniques as well as the novel label-free digital holographic tomography (DHT), a quantitative phase imaging technique that allows the reconstruction of the refractive index (RI), which is used as an image contrast for 3D visualisation of living cells. DHT allowed digital staining of individual cells and intercellular structures based only on their specific RI. Quantitative spatially resolved analysis of the RI data shows that the concentration of G721-0282 leads to significant changes in the density of cells and their intracellular structures (in particular the cytoplasm and nucleus), in the volume of lipid droplets and in protein concentrations. Studies in the U-87 MG glioblastoma cell line, THP-1 monocytes differentiated into macrophages, human microvascular endothelial cells (HMEC-1) and in the spheroid model of glioblastoma composed of U-87 MG, HMEC-1 and macrophages suggest that inhibition of CHI3L1 may have potential in the antitumour treatment of glioblastoma. In this paper, we also propose a spheroid model for in vitro studies that mimics this type of tumour.