-
Biology Oct 2023Osteoarthritis is a classical age-related disease, which affects millions of patients worldwide. To further understand the pathophysiology and to develop therapeutic...
INTRODUCTION
Osteoarthritis is a classical age-related disease, which affects millions of patients worldwide. To further understand the pathophysiology and to develop therapeutic strategies for this disease, animal models play a significant role. is an established model for accelerated aging that spontaneously develops spinal deformities. Although the bone properties of are well described, characteristics of the intervertebral discs are still unknown. The aim of this study was to investigate the characteristics of the intervertebral discs of healthy and deformed .
MATERIAL AND METHODS
Intervertebral properties of healthy and deformed were investigated in 8-, 12-, 18- and 21.5-week-old male fish of the GRZ strain. For histological evaluations the fish were decalcified, paraffin-embedded and stained with (1) hematoxylin and eosin, (2) toluidine blue and (3) alcian blue/picrosirius red.
RESULTS
8-week-old and deformed showed spongy-like tissue containing vacuolated notochord cells and a beginning formation of fibrous tissue in the central area. Older healthy fish showed fibrous tissue in the central region and a spongy-like tissue in the peripheral region.
CONCLUSION
Our study revealed age- and disease-related alterations of the vertebral discs in . Further studies should investigate the utility of as a model for degenerative spine diseases.
PubMed: 37887015
DOI: 10.3390/biology12101305 -
Journal of Orthopaedic Translation Mar 2024The musculoskeletal system contains an extensive network of lymphatic vessels. Decreased lymph flow of the draining collecting lymphatics usually occurs in clinic after...
BACKGROUND
The musculoskeletal system contains an extensive network of lymphatic vessels. Decreased lymph flow of the draining collecting lymphatics usually occurs in clinic after traumatic fractures. However, whether defects in lymphatic drainage can affect fracture healing is unclear.
METHODS
To investigate the effect of lymphatic dysfunction on fracture healing, we used a selective VEGFR3 tyrosine kinase inhibitor to treat tibial fractured mice for 5 weeks versus a vehicle-treated control. To ensure successfully establishing deceased lymphatic drainage model for fractured mice, we measured lymphatic clearance by near infrared indocyanine green lymphatic imaging (NIR-ICG) and the volume of the draining popliteal lymph nodes (PLNs) by ultrasound at the whole phases of fracture healing. In addition, hindlimb edema from day 0 to day 7 post-fracture, pain sensation by Hargreaves test at day 1 post-fracture, bone histomorphometry by micro-CT and callus composition by Alcian Blue-Hematoxylin/Orange G staining at day 14 post-fracture, and bone quality by biomechanical testing at day 35 post-fracture were applied to evaluate fracture healing. To promote fracture healing via increasing lymphatic drainage, we then treated fractured mice with anti-mouse podoplanin (PDPN) neutralizing antibody or isotype IgG antibody for 1 week to observe lymphatic drainage function and assess bone repair as methods described above.
RESULTS
Compared to vehicle-treated group, SAR-treatment group significantly decreased lymphatic clearance and the volume of draining PLNs. SAR-treatment group significantly increased soft tissue swelling, and reduced bone volume (BV)/tissue volume (TV), trabecular number (Tb.N), woven bone and biomechanical properties of fracture callus. In addition, anti-PDPN treated group significantly reduced the number of CD41 platelets in PLNs and increased the number of pulsatile lymphatic vessels, lymphatic clearance and the volume of PLNs. Moreover, anti-PDPN treated group significantly reduced hindlimb edema and pain sensation and increased BV/TV, trabecular number (Tb.Th), woven bone and biomechanical properties of fracture callus.
CONCLUSIONS
Inhibition of proper lymphatic drainage function delayed fracture healing. Use of a anti-PDPN neutralizing antibody reduced lymphatic platelet thrombosis (LPT), increased lymphatic drainage and improved fracture healing.
THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE
(1) We demonstrated lymphatic drainage function is crucial for fracture healing. (2) To unblock the lymphatic drainage and prevent the risk of bleeding and mortality by blood thinner, we demonstrated PDPN neutralizing antibody is a novel and safe way forward in the treatment of bone fracture healing by eliminating LPT and increasing lymphatic drainage.
PubMed: 38511124
DOI: 10.1016/j.jot.2024.02.001 -
Frontiers in Endocrinology 2023The purpose of this study was to prospectively evaluate the efficacy of a demineralized dentin matrix (DDM) in decreasing the initial inflammatory response of the...
OBJECTIVE
The purpose of this study was to prospectively evaluate the efficacy of a demineralized dentin matrix (DDM) in decreasing the initial inflammatory response of the gingiva and facilitating the repair and regeneration of soft tissue in alveolar ridge preservation.
METHODS
This clinical study employed a split-mouth design. Fourteen patients with a total of forty-four sites underwent extraction and alveolar ridge preservation (ARP) procedures. A Bilaterally symmetrical extraction operation were conducted on the premolars of each patient. The experimental group received DDM as a graft material for ARP, while the control group underwent natural healing. Within the first month postoperatively, the pain condition, color, and swelling status of the extraction sites were initially assessed at different time points Subsequently, measurements were taken for buccal gingival margin height, buccal-lingual width, extraction socket contour, and the extraction socket area and healing rate were digitally measured. Additionally, Alcian Blue staining was used for histological evaluation of the content during alveolar socket healing.
RESULTS
Both groups experienced uneventful healing, with no adverse reactions observed at any of the extraction sites. The differences in VAS pain scores between the two groups postoperatively were not statistically significant. In the early stage of gingival tissue healing (3 days postoperatively), there were statistically significant differences in gingival condition and buccal gingival margin height between the two groups. In the later stage of gingival tissue healing (7, 14, and 30 days postoperatively), there were statistically significant differences in buccal-lingual width, extraction socket healing area, and healing rate between the two groups. Furthermore, the histological results from Alcian Blue staining suggested that the experimental group may play a significant role in promoting gingival tissue healing, possibly by regulating inflammatory responses when compared to the control group.
CONCLUSION
The application of DDM in alveolar ridge preservation has been found to diminish initial gingival inflammation after tooth extraction. Additionally, it has shown the ability to accelerate early gingival soft tissue healing and preserve its anatomical contour.
CLINICAL TRIAL REGISTRATION
chictr.org.cn, identifier ChiCTR2100050650.
Topics: Humans; Alcian Blue; Alveolar Process; Alveolar Ridge Augmentation; Bicuspid; Gingiva; Pain; Tooth Socket; Prospective Studies
PubMed: 37929019
DOI: 10.3389/fendo.2023.1281649 -
Scientific Reports Sep 2023Smoking has been associated with an increased risk of asthma, lung cancer, cardiovascular diseases, chronic bronchitis, and a massive amount of oxidative stress. The...
Smoking has been associated with an increased risk of asthma, lung cancer, cardiovascular diseases, chronic bronchitis, and a massive amount of oxidative stress. The present study was undertaken to determine the modulatory effects of Holi Basil/Tulsi, (Ocimum sanctum) leaf extract on cigarette smoke-induced pulmonary damage in mice. Cigarette smoke (CS) inhalation increased the levels of pulmonary lipid peroxidation, and reactive oxygen species and decreased the levels of glutathione. Histoarchitectural alterations and enhanced tissue lactate dehydrogenase (LDH) activity in pulmonary tissue was distinctly indicative of damage. Enhanced mucin production was also observed through mucicarmine and Alcian Blue-Periodic Acid Schiff (PAS) staining. Increased expression of MUC5AC was also observed. Alterations in the lung were also evident through FTIR studies. Administration of Ocimum sanctum leaf extract (80 mg/kg b.w) to CS exposed mice ameliorated these alterations to a greater extent. These findings are suggestive of the fact that Ocimum sanctum leaf extract effectively modulated CS-induced deleterious effects on pulmonary tissue.
Topics: Animals; Mice; Rodentia; Cigarette Smoking; Ocimum sanctum; Smoke Inhalation Injury; Lung
PubMed: 37737272
DOI: 10.1038/s41598-022-26152-8 -
Orthopaedic Journal of Sports Medicine Jan 2024Bone-tendon injury is characterized by poor self-healing. It is established that exosomes are favorable for tissue repair and regeneration. However, their effect on...
BACKGROUND
Bone-tendon injury is characterized by poor self-healing. It is established that exosomes are favorable for tissue repair and regeneration. However, their effect on bone-tendon healing has not yet been determined.
PURPOSE
To compare the effectiveness of exosomes derived from adipose-derived mesenchymal stromal cells (ADSC-Exos) and bone marrow-derived mesenchymal stromal cells (BMSC-Exos) on bone-tendon interface healing in murine rotator cuff injury model and explore the underlying mechanisms thereof.
STUDY DESIGN
Controlled laboratory study.
METHODS
A total of 63 male C57BL6 mice with rotator cuff injuries underwent surgery and were randomly assigned to a control group treated without exosomes (n = 21), an ADSC-Exos group (n = 21), or a BMSC-Exos group (n = 21). The mice were sacrificed 4 or 8 weeks after surgery, and tissues were collected for histologic examination and radiographic and biomechanical testing. For exosome tracing in vivo, mice were sacrificed 7 days after surgery. A series of functional assays (radiographic evaluation, proliferation assay, Alizarin Red staining, alkaline phosphatase staining and activity, Alcian blue staining, quantitative polymerase chain reaction analyses, and glycosaminoglycans quantification) were conducted to evaluate the effect of exosomes on the cellular behaviors of the BMSCs in vitro. A statistical analysis of multiple-group comparisons was performed by 1-way analysis of variance, followed by the Bonferroni post hoc test to assess the differences between the 2 groups.
RESULTS
The ADSCs and BMSCs were positive for surface markers CD29 and CD90 and negative for surface markers CD34 and CD45 and could differentiate into osteoblasts, chondrocytes, and adipocytes. Exosomes showed a cup- or sphere-shaped morphology and were positive for CD63 and TGS101. Local injection of ADSC-Exos and BMSC-Exos could recruit BMSCs and promote osteogenesis, chondrogenesis, and bone-tendon healing. In vitro, ADSC-Exos and BMSC-Exos could significantly promote the proliferation, migration, osteogenic differentiation, and chondrogenic differentiation ability of BMSCs. In vivo, ADSC-Exos and BMSC-Exos significantly accelerated bone-tendon injury healing, with no significant statistical difference between them.
CONCLUSION
ADSC-Exos and BMSC-Exos exhibited similar therapeutic effects on bone-tendon healing in our murine animal model.
CLINICAL RELEVANCE
ADSC-Exos and BMSC-Exos may be used to develop a new cell-free therapy method for promoting rotator cuff injury repair.
PubMed: 38188618
DOI: 10.1177/23259671231210304 -
Revista de Gastroenterologia de Mexico... 2023Colorectal cancer (CRC) is one of the most prevalent cancers worldwide, and significantly contributes to cancer-related deaths. Most cases arise from adenomatous polyps....
INTRODUCTION AND AIMS
Colorectal cancer (CRC) is one of the most prevalent cancers worldwide, and significantly contributes to cancer-related deaths. Most cases arise from adenomatous polyps. Biomarkers currently play an important role in tumor progression. Our aim was to identify perivascular mast cells and analyze the expression of laminin-332, MMP-9, and VEGF in cases of adenoma and CRC in humans.
MATERIALS AND METHODS
Patients were selected at the Coloproctology Service and samples were obtained through biopsies. Adenoma and CRC slides were examined, utilizing immunohistochemistry to detect molecules, and were processed, using 1% Alcian Blue (pH 0.5) for mast cell staining.
RESULTS
Higher density of perivascular mast cells was observed in adenomas. Laminin-332 expression revealed basement membrane discontinuity associated with tumor invasion in CRC. MMP-9 immunostaining in adenoma was detected in glandular epithelium and lining epithelium, in areas close to the basement membrane, whereas in CRC, the enzyme was found in the cytoplasm of invasive clusters. VEGF expression was associated with cell atypia in adenoma and in areas of disorganization of the epithelium-connective tissue interface in CRC. VEGF has also been detected in endothelial cells from microvessels.
CONCLUSIONS
We demonstrated the different patterns of perivascular mast cells and molecular expression in colorectal neoplasms. Those analyses favor the recognition of the predisposition to the disease, or its early stage, and have the potential to define the molecular profile of the lesions.
Topics: Humans; Adenoma; Colorectal Neoplasms; Endothelial Cells; Laminin; Mast Cells; Matrix Metalloproteinase 9; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors
PubMed: 35810100
DOI: 10.1016/j.rgmxen.2022.06.004 -
PeerJ 2023Mucus derived from many land snails has been extensively utilised in medicine and cosmetics, but some biological activities of the mucus need to be well documented....
BACKGROUND
Mucus derived from many land snails has been extensively utilised in medicine and cosmetics, but some biological activities of the mucus need to be well documented. Nevertheless, most mucus is obtained from land snails, while mucus from freshwater snails has yet to be attended.
METHODS
This study aims to determine and compare mucus's antioxidant and anti-inflammatory activities from the land snail and the freshwater snail . ABTS, DPPH, reducing power and total antioxidant activity assays were used to evaluate the antioxidant capacity. Inhibition of nitric oxide production in lipopolysaccharide-activated RAW 264.7 cells was performed to determine the anti-inflammatory activity. Additionally, the histochemical analysis of mucous cells in each snail foot was conducted to compare the distribution of mucous cells and types of mucins using periodic acid-Schiff and Alcian blue staining.
RESULTS
Mucus from and exhibited antioxidant and anti-inflammatory activities in different parameters. mucus has higher total antioxidant (44.71 ± 2.11 mg AAE/g) and nitric oxide inhibitory activities (IC = 9.67 ± 0.31 µg/ml), whereas mucus has better-reducing power activity (43.63 ± 2.47 mg AAE/g) and protein denaturation inhibition (IC = 0.60 ± 0.03 mg/ml). Histochemically, both species' dorsal and ventral foot regions contained neutral and acid mucins in different quantities. In the dorsal region, the neutral mucins level in (16.64 ± 3.46%) was significantly higher than that in (11.19 ± 1.50%), while the acid mucins level showed no significant difference between species. Levels of both mucins in the ventral foot region of (15.08 ± 3.97% and 10.76 ± 3.00%, respectively) were significantly higher than those of (2.25 ± 0.48% and 2.71 ± 0.56%, respectively). This study revealed scientific evidence of the biological capacity of mucus from and as well as provided helpful information on the region of the foot which produces effective mucus.
Topics: Animals; Antioxidants; Nitric Oxide; Snails; Mucus
PubMed: 37583916
DOI: 10.7717/peerj.15827 -
Biology of Reproduction Nov 2023The prostate of the koala (Phascolarctos cinereus), and of marsupials more generally, is the primary contributor of seminal fluid, yet comparatively little is known...
The prostate of the koala (Phascolarctos cinereus), and of marsupials more generally, is the primary contributor of seminal fluid, yet comparatively little is known about its microanatomy or biochemistry. This study explored evidence of parenchymal segmentation of the koala prostate. The prostate of three sexually mature koalas were processed for histopathology, histochemistry (Masson's trichrome, Alcian Blue, periodic acid Schiff staining), and immunohistochemistry using basal (tumor protein 63, cytokeratin 14) and luminal (cytokeratin 8/18, prostate specific antigen, androgen receptor) markers. Results confirmed clear segmentation of the koala prostate into three zones, anterior, central, and posterior, characterized by differences in the proportion of glandular tissue, as well as the thickness of collagen fibers; there were also distinct differences in the secretions produced in each zone. Based on immunohistochemistry, the koala prostate showed evidence of both basal proliferative and luminal secretory cells. The ratio of cell types varied across the three segments, with the central segment housing the highest density of basal cells. Globular bodies produced in the anterior zone were shown to possess the same markers as those described for human prostasomes. This study is the first to comprehensively document the marsupial prostate in terms of microanatomy and corresponding immunohistochemistry. While further biochemical analysis, such as proteomics of each segment will better define the relative functions of each tissue, the data presented here are consistent with the hypothesis that the koala prostate potentially represents an example of an ontological stage in the evolutionary differentiation of male eutherian accessory glands.
Topics: Animals; Male; Humans; Phascolarctidae; Prostate; Marsupialia; Immunohistochemistry
PubMed: 37593918
DOI: 10.1093/biolre/ioad098 -
Journal of the European Academy of... Jul 2023Primary cutaneous mucinoses (PCM) are rare diseases characterized by dermal or follicular mucin deposits.
BACKGROUND
Primary cutaneous mucinoses (PCM) are rare diseases characterized by dermal or follicular mucin deposits.
OBJECTIVES
A retrospective study characterizing PCM to compare dermal with follicular mucin to identify its potential origin on a single-cell level.
MATERIAL AND METHODS
Patients diagnosed with PCM between 2010 and 2020 at our department were included in this study. Biopsy specimens were stained using conventional mucin stains (Alcian blue, PAS) and MUC1 immunohistochemical staining. Multiplex fluorescence staining (MFS) was used to investigate which cells were associated with MUC1 expression in select cases.
RESULTS
Thirty-one patients with PCM were included, 14 with follicular mucinosis (FM), 8 with reticular erythematous mucinosis, 2 with scleredema, 6 with pretibial myxedema and one patient with lichen myxedematosus. In all 31 specimens, mucin stained positive for Alcian blue and negative for PAS. In FM, mucin deposition was exclusively found in hair follicles and sebaceous glands. None of the other entities showed mucin deposits in follicular epithelial structures. Using MFS, all cases showed CD4+ and CD8+ T cells, tissue histiocytes, fibroblasts and pan-cytokeratin+ cells. These cells expressed MUC1 at different intensities. MUC1 expression in tissue histiocytes, fibroblasts, CD4+ and CD8+ T cells, and follicular epithelial cells of FM was significantly higher than the same cell types in the dermal mucinoses (p < 0.001). CD8+ T cells were significantly more involved in expression of MUC1 than all other analysed cell types in FM. This finding was also significant in comparison with dermal mucinoses.
CONCLUSION
Various cell types seem to contribute to mucin production in PCM. Using MFS, we showed that CD8+ T cells seem to be more involved in the production of mucin in FM than in dermal mucinoses, which could indicate that mucin in dermal and follicular epithelial mucinoses have different origins.
Topics: Humans; Mucinoses; Mucins; Retrospective Studies; Alcian Blue; Scleromyxedema; Staining and Labeling
PubMed: 36807595
DOI: 10.1111/jdv.18992 -
Frontiers in Immunology 2023The intestinal barrier plays a crucial role in distinguishing foods from toxins. Prostaglandin D (PGD) is one of the lipid-derived autacoids synthesized from cell...
INTRODUCTION
The intestinal barrier plays a crucial role in distinguishing foods from toxins. Prostaglandin D (PGD) is one of the lipid-derived autacoids synthesized from cell membrane-derived arachidonic acid. We previously reported that pharmacological stimulation of PGD receptor, D prostanoid 1 (DP1) attenuated the symptoms of azoxymethane/dextran sodium sulfate-induced colitis and ovalbumin-induced food allergy in mouse models. These observations suggested that DP1 stimulation protects the intestinal barrier. The present study aimed to uncover the effects of DP1 stimulation on intestinal barrier function and elucidate the underlying mechanisms.
MATERIALS AND METHODS
Intestinal permeability was assessed in mice by measuring the transfer of orally administered fluorescein isothiocyanate-dextran (40 kDa) into the blood. The DP1 agonist BW245C (1 mg/kg) was administered 10 min prior to dextran administration. The intestinal permeability was confirmed using the everted sac method. Tight junction integrity was evaluated by measuring the transepithelial electrical resistance (TER) in the human intestinal epithelial cell line Caco-2. Mucus secretion was assessed by observing Alcian Blue-stained intestinal sections.
RESULTS
Pharmacological DP1 stimulation reduced intestinal permeability both and . Immunohistochemical staining showed that DP1 was strongly expressed on the apical side of the epithelial cells. DP1 stimulation did not affect TER but induced mucus secretion from goblet cells. Mucus removal by a mucolytic agent N-acetyl-l-cysteine canceled the inhibition of intestinal permeability by DP1 stimulation.
CONCLUSION
These observations suggest that pharmacological DP1 stimulation decreases intestinal permeability by stimulating mucus secretion.
Topics: Humans; Animals; Mice; Prostaglandins; Dextrans; Prostaglandin D2; Caco-2 Cells; Mucus; Permeability
PubMed: 37942331
DOI: 10.3389/fimmu.2023.1276852