Cells Tissues Organs最新文献

Fibroblasts are central to variety of homeostatic events such as wound healing and tissue regeneration. However, their pathologic activation is thought to play roles in a variety of diseases not only limited to fibrosis, foreign body reaction, scleroderma but also cancer metastasis. Biophysical properties of the extracellular matrix (ECM) deposited by an activated fibroblast determine whether there is a pro-regenerative or scarring response. Compared to aged fibroblasts, embryonic fibroblasts were shown to deposit a pro-regenerative ECM characterized by early hyaluronic acid (HA) deposition and increased levels of pro-regenerative collagens such as type III collagen. Since HA is also a regulator of collagen organization, we propose that early accumulation of HA by fibroblasts can facilitate a pro-regenerative matrix formation. Given that the molecular weights of HA present in pro-regenerative matrix are higher than synthetic HA, we strategize attracting HA synthesized by fibroblasts. In this study, we used a synthetic peptide sequence known to have affinity to HA as a strategy to instruct fibroblasts to retain HA on the surface. We hypothesized that hyaluronic acid binding peptide (HABP) may instruct fibroblast endogenous HA deposition onto functionalized surfaces. We functionalized silica glass surfaces with HABP using aminoorganosilane mediated chemisorption and screened primary human dermal fibroblasts (hDF) for cell morphology, cytoskeletal arrangement, and alpha-smooth muscle actin (α-SMA) expression. Our results show HABP treated surfaces retain higher levels of HA on silica glass compared to control surfaces on fibroblast derived matrices. Analysis of α-SMA shows increased α-SMA expression on hDFs and increased stress fiber formation. HABP treated surfaces were found to have reduced α-SMA expression. The physical features of collagen fibers deposited by fibroblasts were also organized differently in the presence of HABP. We propose that HABPs are a potentially viable strategy to instruct pro-regenerative fibroblasts and can be used therapeutically to treat fibrotic diseases.

Introduction: Hemolymph nodes, characterized by erythrocyte rosettes, are found in humans and animals, including rats. The cytoarchitectural differences they share with lymph nodes and the spleen are unknown. Herein, we describe the cytoarchitecture of rat hemolymph nodes.

Methods: We performed immunohistochemical analyses with antibodies against CD68, Iba-1, CD3, CD20, and S-100. Hematoxylin and eosin staining was used to compare findings with sections from ordinary lymph nodes and spleen.

Results: Hemolymph nodes exhibited erythrocyte rosettes with macrophages immunopositive for CD68, Iba-1, and CD3, which were rare in the physiologically normal spleen and lymph nodes. Additionally, sinusoidal macrophages often showed close apposition to erythrocytes and mast cells. Accumulation of cells immunoreactive to CD20, a B-lymphocyte marker, was seen only in the germinal centers of ordinary lymph nodes, not in the hemolymph nodes or spleen. Ordinary lymph nodes and spleen showed well-developed reticular configurations of cells with immunoreactivity for S-100, a marker for dendritic cells, unlike hemolymph nodes, suggesting less-developed antigen-presenting ability in the latter. Despite similarities to ordinary lymph nodes and spleen, the direct contact with erythrocytes and mast cells in hemolymph nodes suggests they facilitate direct cell-to-cell communication for macrophages, erythrocytes, and mast cells.

Conclusion: Our findings imply that the hemolymph nodes are a unique immune organ, differing from ordinary lymph nodes and spleen.

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.

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

Members of the carbonic anhydrase gene family, responsible for the reversible hydration of carbon dioxide, participate in several important biological processes including processes involved in fertilization. CAIV has been shown to play a role in sperm cell capacitation and regulation of sperm motility and is present in mature murine placentae. The present study specifically analyses the distribution of CAIV in female reproductive organs and during early placenta development. Immunostaining for CAIV was performed on female reproductive organs (ovary, fallopian tube, uterus, vagina) of non-pregnant mice and on implantation sites of early pregnancy between 4.5 and 9.5 days post coitum (dpc). Sex typing of embryos was performed by PCR using three separated gene combinations for X- and Y-chromosomes, respectively. Additionally, reproductive outcome of CAIV-deficient mice was determined. CAIV is largely absent in the female reproductive organs of non-pregnant mice. Immunostaining for CAIV was present in the blastocyst and in consecutive stages of the developing embryo. In the endometrial epithelium distant from the implantation chamber, CAIV is induced from 8.5 dpc onwards. Moreover, the yolk sac epithelium, the trophoblast giant cells and the labyrinthine compartment of the developing hemochorial placenta shows a strong immunostaining for CAIV. In heterozygous mating, the number of CAIV knockout pups is significantly reduced than was to be expected according to the mendelian rules, while homozygous mating of CAIV knockout mice results in a significant reduction of litter size which is mainly due to a reduced number of female mice born. Since at 9.5 dpc the number of female embryos is rather higher than that of males, the observed reduction of female offspring appears to be due to a defect in placentation after 9.5 dpc. Thus, CAIV seems to be involved in the signaling network of embryo development, implantation, and placentation.

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: Acupuncture has been used for pain management for thousands of years. However, it is largely unclear whether this therapeutic approach can effectively reduce heart failure-associated symptoms, including dyspnea. The hypothesis posited in this study was that acupuncture does indeed aid in the management of such symptoms and was motivated by the following statistics that establish a requisite need for efficient management of dyspnea to improve patient outcomes with heart failure. In 2020, an estimated 6.2 million adults in the USA had a heart failure diagnosis; in 2018, 379,800 death certificates reported heart failure; and the national cost of heart failure in 2012 was approximately USD 30.7 billion.

Methods: The methodology employed to conduct this study involved review of trial data extracted from review of papers pertaining to acupuncture, symptoms of heart failure, and dyspnea, from academic and clinical data repositories subject to various inclusion and exclusion criteria. Of the initial set of 293 studies identified, the resulting inclusion set comprised 30 studies. The analysis conducted revealed that the highest frequency of combined acupuncture points prescribed for the foregoing search criteria were as follows: BL13, BL23, LU9, LU5, Dingchuan, LI4, PC6, and HT7.

Results: A meta-analysis of combined pooled p values for the studies revealed that acupuncture does aid in the management of symptoms of dyspnea and heart failure, subject to various limitations including but not limited to heterogeneity inherent between the studies in the inclusion set that were analyzed. Such limitations underscore the need to restrict generalizations from the conclusions of this study.

Conclusion: The impact and novelty of this research study is its attempt to target the apparent paucity of literature that focuses on the management of dyspnea specifically in the context of heart failure with acupuncture and to bridge the gap of the application of acupuncture research on dyspnea to the cardiovascular context of heart failure. Notwithstanding the meta-analysis undertaken under this review study, further statistical analysis and a pilot study are warranted to consolidate or nullify the results of the research.

Introduction: Bioprinting, using "bio-inks" consisting of living cells, supporting structures, and biological motifs to create customized constructs, is an emerging technique that aims to overcome the challenges of cartilaginous reconstruction of head and neck structures. Several living cell lines and culturing methods have been explored as bio-inks with varying efficacy. Coculture of primary chondrocytes and stem cells (SCs) is one technique well established for degenerative joint disease treatment, with potential for use in expanding chondrocyte populations for bio-inks. This study aimed to evaluate the techniques for coculture of primary chondrocytes and SCs for head and neck cartilage regeneration.

Methods: A literature review was performed through OVID/Web of Science/MEDLINE/BIOSIS Previews/Embase. Studies reporting on chondrocytes and SCs in conjunction with coculture or cartilage regeneration were included. Studies not reporting on findings from chondrocytes/SCs of the head and neck were excluded. Extracted data included cell sources, coculture ratios, and histological, biochemical, and clinical outcomes.

Results: Fifteen studies met inclusion criteria. Auricular cartilage was the most common chondrocyte source (n = 10), then nasal septum (n = 5), articular (n = 1), and tracheal cartilage (n = 1). Bone marrow was the most common SC source (n = 9) then adipose tissue (n = 7). Techniques varied, with coculture ratios ranging from 1:1 to 1:10. All studies reported coculture to be superior to SC monoculture by all outcomes. Most studies reported superiority or equivalence of coculture to chondrocyte monoculture by all outcomes. When comparing clinical outcomes, coculture constructs were equivalent to chondrocyte monoculture in diameter and equivalent or inferior in wet weight and height.

Conclusion: Coculture of primary chondrocytes and SCs is a promising technique for expanding chondrocyte populations, with at least equivalence to chondrocyte monoculture and superior to SC monoculture when seeded at the same chondrocyte densities. However, there remains a lack of consensus regarding the optimal cell sources and coculture ratios.

Introduction: Endothelial cells (EC) can be generated from porcine-induced pluripotent stem cells (piPSC), but poor efficiency in driving EC differentiation hampers their application and efficacy. Additionally, the culture of piPSC-derived EC (piPSC-EC) on three-dimensional (3D) scaffolds has not been fully reported yet. Here, we report a method to improve the generation of EC differentiation from piPSC and to facilitate their culture on 3D scaffolds, providing a potential resource for in vitro drug testing and the generation of tissue-engineered vascular grafts.

Methods: We initiated the differentiation of piPSC into EC by seeding them on laminin 411 and employing a three-stage protocol, which involved the use of distinct EC differentiation media supplemented with CHIR99021, BMP4, VEGF, and bFGF.

Results: piPSC-EC not only expressed EC markers such as CD31, VE-cadherin, and von Willebrand factor (vWF) but also exhibited an upregulation of EC marker genes, including CD31, CD34, VEGFR2, VE-cadherin, and vWF. They exhibited functional characteristics similar to those of porcine coronary artery endothelial cells (PCAEC), such as tube formation and Dil-Ac-LDL uptake. Furthermore, when cultured on 3D scaffolds, piPSC-EC developed a 3D morphology and were capable of forming an endothelial layer and engineering capillary-like networks, though these lacked lumen structures.

Conclusion: Our study not only advances the generation of EC from piPSC through an inhibitor and growth factor cocktail but also provides a promising approach for constructing vascular network-like structures. Importantly, these findings open new avenues for drug discovery in vitro and tissue engineering in vivo.