Cells Tissues Organs最新文献

Introduction: Fetal microchimerism could be involved in the regulation of breast cancer oncogenesis. CD34+ cells could be of a particular interest as up to 12% of the CD34+ population in maternal blood are of fetal origin. The aim of this research was to analyze the impact of umbilical cord blood (UCB) CD34+ on MCF-7 and MDA-MB-231 breast cancer cell lines, in order to uncover novel biological mechanisms and suggest novel treatment options for breast cancer.

Methods: UCB CD34+ cells were obtained from healthy women at full-term delivery. Direct cultures were grown with MCF-7 and MDA-MB-231 cells. Proliferation, migration, invasion, and transcriptomic analysis of breast cancer cell lines were compared between cultures exposed and nonexposed to UCB CD34+ cells. Interactions between UCB CD34+ and breast cancer cells were analyzed under fluorescent microscopy. Functional analyses were generated with QIAGEN's Ingenuity Pathway Analysis (IPA) and Gene Set Enrichment Analysis (GSEA).

Results: Direct contact between UCB CD34+ and breast cancer cell lines induced a reduction in the proliferative capacities of MCF-7 and MDA-MB-231 and diminished the migration abilities of MDA-MB-231 cells. In 3D coculture, UCB CD34+ cells were attracted by tumor spheroids and incorporated into tumor cells. These cell-to-cell interactions were responsible for transcriptome modifications coherent with observed functional modifications. Among the cytokines secreted by UCB CD34+, IFN-γ was identified as a potential upstream regulator responsible for the molecular modifications observed in transcriptomic analysis of MCF-7 breast cancer cells exposed to UCB CD34+ cells, as was IL-17A in MDA-MB-231 cells.

Conclusion: Direct cell-to-cell contact induced functional modifications in breast cancer cells. Interactions between UCB CD34+ and breast cancer cells could induce cell fusion and signal transmission via cytokines. Further analysis of direct cell-to-cell interactions should be performed at a molecular level to further understand the potential role of fetal CD34+ cells in breast cancer.

Introduction: Mesenchymal stem cell (MSC)-based therapies have emerged as a promising approach for treating articular cartilage injuries. However, enhancing the chondrogenic differentiation potential of MSCs remains a significant challenge. KDM6B, a histone demethylase that specifically removes H3K27me3 marks, is essential in controlling the maturation of chondrocytes. In this study, we examined how KDM6B influences chondrogenic differentiation in SCAPs and investigated the underlying mechanisms involved.

Methods: SCAPs were utilized. Alcian blue staining, pellet culture, and cell transplantation in rabbit knee cartilage defect models assessed MSC chondrogenic differentiation. Western blot, real-time RT-PCR, and microarray analysis examined the underlying molecular mechanisms.

Results: KDM6B promotes the expression of aggrecan, COL2A1, COL5, glycosaminoglycans, and collagen fibers, while also increasing the COL2/COL1 ratio in SCAPs. In vivo, SCAPs overexpressing KDM6B significantly enhanced the repair and regeneration of knee cartilage and subchondral bone, with higher levels of glycosaminoglycan and COL2 expression observed within the tissue. KDM6B promotes the chondrogenic differentiation potential of SCAPs by repressing HES1. In addition, knockdown of HES1 enhanced the chondrogenic differentiation of SCAPs.

Conclusions: KDM6B enhances the differentiation of SCAPs into chondrocytes and demonstrated its effectiveness in the repair and regeneration of cartilage tissue and subchondral bone in vivo experiments. These findings provide an important foundation for future research on the use of dental tissue-derived stem cells to treat cartilage injuries.

Introduction: The hedgehog signaling pathway plays a crucial role in inducing segment polarity through cell-cell interactions in various metazoans, including arthropods and annelids. However, its involvement in organogenesis and segmentation among lophotrochozoans remains inconsistent. This study aimed to explore the role of the hedgehog gene during gut development in the freshwater leech, Helobdella austinensis.

Methods: Developmental RT-PCR and in situ hybridization were performed to examine the expressions of hedgehog genes. In addition, embryos were treated with cyclopamine (a hedgehog signaling antagonist) and purmorphamine (a Smo agonist) to examine the potential interactions between Helobdella orthologs to hedgehog and two NKL genes: Hau-NK2 and Hau-NK4.

Results: We examined the expressions of four core pathway members - Hedgehog (Hh), Patched (Ptc), Smoothened (Smo), and the downstream transcription factor Gli - spatiotemporally during the embryonic stages of H. austinensis. All four genes were expressed in the developing gut and proboscis during organogenesis but not during the segmentation stage. Additionally, the treatment of embryos with cyclopamine and purmorphamine revealed that NK genes are regulated by hedgehog signaling. Furthermore, NK2 and NK4 were expressed in the developing gut rather than in a segmental stripe pattern.

Conclusion: This study confirms that the hedgehog signaling pathway is associated with gut development in the freshwater leech, H. austinensis. The expression patterns of hedgehog pathway genes and their interaction with NK genes suggest a role of hedgehog signaling in regulating gut development rather than segmentation in the freshwater leeches.

Introduction: Damage-associated molecular patterns (DAMPs) are molecules released in response to tissue or cellular damage to facilitate tissue regeneration. This inflammatory response can occur in sterile environments and is promoted by the release of damaged extracellular components such as the extracellular matrix (ECM). DAMPs have been implicated in various stages of wound healing but have yet to be explicitly utilized for regenerative medicine by leveraging selective modulation of the inflammatory response. With this in mind, we leverage inflammation to drive tissue regeneration by utilizing DAMPs collected from the native ECM, extracellular matrix motifs (mECM).

Methods: Here, mECMs were derived from UV-damaged rat tail collagen I. Fibroblast response to various concentrations and presentation of mECMs was investigated by evaluating changes in viability, proliferation, cell phenotype, and cytokine secretion.

Results: mECMs had reduced intensity in collagen I associated bands, indicating successful fragmentation to lower molecular weights. Soluble (mobile) mECMs induced changes in fibroblast phenotype as indicated by a decrease in proliferation, a decrease in nuclei area, and an increase in the percentage of elongated cells. In addition, mobile mECMs contributed to significant increases in cytokine secretion compared to insoluble (bound) mECMs. Across all experiments, bound mECMs exhibited effects on fibroblasts compared to the collagen control.

Conclusion: Fibroblasts in vitro recognize mECMs, with significant differences observed based on the presentation of these proteins. These data indicate that cryptic regions that are recognized by fibroblasts may be exposed in the mobile version of the mECMs, which lead to a myofibroblast-like phenotype in fibroblasts. This work highlights the potential of DAMPs to serve as immunomodulatory therapeutics for tissue regeneration.

Introduction: To date, there have been no studies conducted on the development of interosseous muscles (IO) in the human hand. This study aimed to investigate the development of these muscles in order to clarify their terminal insertions and their relationship with the metacarpophalangeal joints.

Methods: Serial sections of 25 human specimens (9 embryos and 16 fetuses) between the 7th and 14th weeks of development, sourced from the Collection of the Department of Anatomy and Embryology at UCM Faculty of Medicine, were analyzed bilaterally using a conventional optical microscope.

Results: Our findings revealed that, during the 7th week of development, the metacarpophalangeal interzone mesenchyme extended into the extensor apparatus of the fingers. Furthermore, we observed that the joint capsule and the tendon of the IO derive from the articular interzone mesenchyme. By the end of the 7th week, corresponding to Carnegie stage 21, the myotendinous junction appeared, initiating cavitation of the metacarpophalangeal joint. During the fetal period, the terminal insertions of the IO were identified: both the dorsal interosseous (DI) and palmar interosseous (PI) muscles insert into the metacarpophalangeal joint capsule and establish a connection with the volar plate located at the base of the proximal phalanx and the extensor apparatus. Some muscle fibers also attach to the joint capsule at the level of the proximal synovial cul-de-sac. The functional implications of these findings are discussed within this work.

Conclusion: This study provides the first detailed description of the development of the interosseous muscles in the human hand.

Introduction: The University of Wisconsin (UW) and histidine-tryptophan-ketoglutarate (HTK) are the most popular organ-preservative solutions. Ultrastructure of organelles reflects the functionality of cells, but less is understood about ultrastructural changes of hepatocytes after machine perfusion (MP) with HTK, than with UW solution.

Methods: We evaluated the ultrastructure of hepatocytes preserved in HTK solution during hypothermic (4°C) and midthermic (22°C) MP (HMP and MMP, respectively) temperatures using osmium maceration scanning electron microscopy.

Results: The functional ultrastructure of mitochondria in hepatocytes was maintained immediately after HMP and MMP. After 2 h in a porcine isolated liver reperfusion model (IRM) that simulates transplanted liver grafts, mitochondrial cristae became denser and some large vacuoles that we assumed were autophagosomes were detected in hepatocytes after HMP. Autophagy functions in the suppression of reactive oxygen generation and subsequent apoptosis, and these findings indicated that HMP is more effective than MMP when livers are preserved in HTK solution.

Conclusion: The present findings contradict the previous findings of the UW solution that MMP is more effective than HMP. Thus, various combinations of conditions for MP should be carefully optimized before changing preservatives.

Introduction: The cartilago cordis is a structure present within the cardiac skeleton of some, but not all, vertebrate species. This systematic review compared the presence, structure, and function of the cartilago cordis from published works covering all vertebrate species.

Methods: Literature searches were conducted to obtain information relating to the anatomical location, morphology, prevalence, number of structures, development, and function.

Results: The cartilago cordis was most commonly composed of hyaline cartilage but its location within the cardiac skeleton, anatomical, and histological structure varied between species. The cartilago cordis has not been documented in every vertebrate species, or every individual within each species, but it is present in 68 vertebrates including an amphibian, and some mammals, reptiles, and birds. The function of the cartilago cordis is unknown, but theories have ranged from an adaptive mechanism to support cardiac tissue through to roles in conduction and contraction, especially in areas of high mechanical stress. Possible links between the presence of a cartilago cordis and cardiac pathologies were also identified.

Conclusion: The cartilago cordis varied in prevalence, structure, and location; further research is required to understand the function and development. In addition, it is possible there are more vertebrate species containing cartilago cordis than presently known about given its varying prevalence and sometimes small size.

Introduction: Macrophages exist on a spectrum from pro-inflammatory (M1) to pro-healing (M2). Characterization of macrophage phenotype is important to understand tissue healing and response. The gold standard for assessing macrophage phenotypes is immunocytochemistry (ICC), which stains inducible nitric oxide synthase (iNOS) and arginase (Arg1), the proteins secreted before nitrite and urea production. The ICC method is an endpoint assay, time-consuming, and costly. Therefore, a more effective method to assess the phenotype of macrophages in vitro is needed. Based on the phenotype of the macrophage, the amino acid arginine gets metabolized differently. If arginine is metabolized by M1 macrophages, it produces nitrite, and if it is metabolized by M2 macrophages, it produces urea. A method that leverages arginine metabolism through secreted products (urea and nitrite) has the potential to determine macrophage phenotype in real time.

Methods: Rat bone marrow-derived macrophages were cultured to be naïve or polarized to M1-like or M2-like. The gold standard ICC method was used to determine the intensity of the iNOS and Arg1 staining. Nitrite and urea kits were utilized to measure the concentration of nitrite and urea in the media eluted from macrophages of various phenotypes. Nitrite and urea concentrations were compared to ICC results to validate the new method.

Results: ICC revealed the iNOS staining was significant and 2.5-folds higher in M1-like macrophages and the Arg1 staining was significant and 1.5-folds higher in the M2-like macrophages. The nitrite concentration was significant and 4-folds higher in the M1-like macrophage media, and the urea concentration was significant and 2.5-folds higher in the M2-like macrophage media. A correlation analysis showed that iNOS staining intensity and nitrite concentration levels had a linear correlation as well as Arg1 staining intensity and urea concentration levels.

Conclusion: Data confirm that the determination of nitrite and urea concentration can be utilized to assess macrophage phenotypes.