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Journal of Orthopaedic Surgery and... May 2024Muscle wasting frequently occurs following joint trauma. Previous research has demonstrated that joint distraction in combination with treadmill exercise (TRE) can...
Exercise following joint distraction inhibits muscle wasting and delays the progression of post-traumatic osteoarthritis in rabbits by activating PGC-1α in skeletal muscle.
OBJECTIVE
Muscle wasting frequently occurs following joint trauma. Previous research has demonstrated that joint distraction in combination with treadmill exercise (TRE) can mitigate intra-articular inflammation and cartilage damage, consequently delaying the advancement of post-traumatic osteoarthritis (PTOA). However, the precise mechanism underlying this phenomenon remains unclear. Hence, the purpose of this study was to examine whether the mechanism by which TRE following joint distraction delays the progression of PTOA involves the activation of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), as well as its impact on muscle wasting.
METHODS
Quadriceps samples were collected from patients with osteoarthritis (OA) and normal patients with distal femoral fractures, and the expression of PGC-1α was measured. The hinged external fixator was implanted in the rabbit PTOA model. One week after surgery, a PGC-1α agonist or inhibitor was administered for 4 weeks prior to TRE. Western blot analysis was performed to detect the expression of PGC-1α and Muscle atrophy gene 1 (Atrogin-1). We employed the enzyme-linked immunosorbent assay (ELISA) technique to examine pro-inflammatory factors. Additionally, we utilized quantitative real-time polymerase chain reaction (qRT-PCR) to analyze genes associated with cartilage regeneration. Synovial inflammation and cartilage damage were evaluated through hematoxylin-eosin staining. Furthermore, we employed Masson's trichrome staining and Alcian blue staining to analyze cartilage damage.
RESULTS
The decreased expression of PGC-1α in skeletal muscle in patients with OA is correlated with the severity of OA. In the rabbit PTOA model, TRE following joint distraction inhibited the expressions of muscle wasting genes, including Atrogin-1 and muscle ring finger 1 (MuRF1), as well as inflammatory factors such as interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in skeletal muscle, potentially through the activation of PGC-1α. Concurrently, the production of IL-1β, IL-6, TNF-α, nitric oxide (NO), and malondialdehyde (MDA) in the synovial fluid was down-regulated, while the expression of type II collagen (Col2a1), Aggrecan (AGN), SRY-box 9 (SOX9) in the cartilage, and superoxide dismutase (SOD) in the synovial fluid was up-regulated. Additionally, histological staining results demonstrated that TRE after joint distraction reduced cartilage degeneration, leading to a significant decrease in OARSI scores.TRE following joint distraction could activate PGC-1α, inhibit Atrogin-1 expression in skeletal muscle, and reduce C-telopeptides of type II collagen (CTX-II) in the blood compared to joint distraction alone.
CONCLUSION
Following joint distraction, TRE might promote the activation of PGC-1α in skeletal muscle during PTOA progression to exert anti-inflammatory effects in skeletal muscle and joint cavity, thereby inhibiting muscle wasting and promoting cartilage regeneration, making it a potential therapeutic intervention for treating PTOA.
Topics: Animals; Rabbits; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; Osteoarthritis; Muscular Atrophy; Disease Progression; Muscle, Skeletal; Male; Humans; Physical Conditioning, Animal; Female; Disease Models, Animal
PubMed: 38822418
DOI: 10.1186/s13018-024-04803-y -
Pathology International May 2024Clear cell hidradenoma is a rare benign tumor of the breast, its origin and pathogenesis are controversial. We have experienced a case of breast clear cell hidradenoma...
Clear cell hidradenoma is a rare benign tumor of the breast, its origin and pathogenesis are controversial. We have experienced a case of breast clear cell hidradenoma with mastermind like transcriptional coactivator 2 (MAML2) gene rearrangement. The patient found a painless mass with a hard texture in the left breast areola without nipple discharge. Microscopically, the tumor was cystic and solid, locally arranged in a glandular structure, covered by single cuboidal cells; it was composed of clear cells, epidermoid cells, and basaloid cells; there were no necrosis or mitotic figures. Immunohistochemical staining showed that the tumor cells positively expressed low-molecular cytokeratin 7, low-molecular cytokeratins (Cam5.2), high-molecular cytokeratin 5/6, cytokeratin 14, CD117, and p63; and did not express calponin, and smooth muscle myosin heavy chain. The cuboidal cells were positive for SOX10 but negative for p63. Additionally, periodic acid-Schiff reaction showed purple-red granules in the tumor cytoplasm, but Alcian blue staining showed no blue mucus in the cytoplasm. The split signals of MAML2 gene were detected by fluorescence in situ hybridization. Subtle histological and immunophenotypical differences may help to distinguish breast clear cell hidradenoma from common breast tumors. Furthermore, the MAML2 gene rearrangement may be a molecular genetic characteristic of breast clear cell hidradenoma.
PubMed: 38818886
DOI: 10.1111/pin.13455 -
Reproductive Toxicology (Elmsford, N.Y.) Aug 2024Zebrafish embryo assays are used by pharmaceutical and chemical companies as new approach methodologies (NAMs) in developmental toxicity screening. Despite an overall...
Zebrafish embryo assays are used by pharmaceutical and chemical companies as new approach methodologies (NAMs) in developmental toxicity screening. Despite an overall high concordance of zebrafish embryo assays with in vivo mammalian studies, false negative and false positive results have been reported. False negative results in risk assessment models are of particular concern for human safety, as developmental anomalies may be missed. Interestingly, for several chemicals and drugs that were reported to be false negative in zebrafish, skeletal findings were noted in the in vivo studies. As the number of skeletal endpoints assessed in zebrafish is very limited compared to the in vivo mammalian studies, the aim of this study was to investigate whether the sensitivity could be increased by including a skeletal staining method. Three staining methods were tested on zebrafish embryos that were exposed to four teratogens that caused skeletal anomalies in rats and/or rabbits and were false negative in zebrafish embryo assays. These methods included a fixed alizarin red-alcian blue staining, a calcein staining, and a live alizarin red staining. The results showed a high variability in staining intensity of larvae exposed to mammalian skeletal teratogens, as well as variability between control larvae originating from the same clutch of zebrafish. Hence, biological variability in (onset of) bone development in zebrafish hampers the detection of (subtle) treatment-related bone effects that are not picked-up by gross morphology. In conclusion, the used skeletal staining methods did not increase the sensitivity of zebrafish embryo developmental toxicity assays.
Topics: Animals; Zebrafish; Teratogens; Embryo, Nonmammalian; Toxicity Tests; Staining and Labeling; Bone and Bones; Embryonic Development; Fluoresceins; Anthraquinones
PubMed: 38815770
DOI: 10.1016/j.reprotox.2024.108615 -
Zhongguo Zhong Yao Za Zhi = Zhongguo... Apr 2024The study aimed to investigate the therapeutic effects of the n-butanol extract of Pulsatilla Decoction(BEPD) on ulcerative colitis(UC) via the bone morphogenetic...
The study aimed to investigate the therapeutic effects of the n-butanol extract of Pulsatilla Decoction(BEPD) on ulcerative colitis(UC) via the bone morphogenetic protein(BMP) signaling pathway. C57BL/6 mice were divided into six groups: control, model, mesalazine, and BEPD low-, medium-, and high-dose groups. Except for the control group, the rest groups were treated with 3% dextran sulfate sodium(DSS) freely for seven consecutive days to establish the UC mouse model, followed by treatment with different concentrations of BEPD and mesalazine by gavage. The murine body weight and disease activity index(DAI) were recorded. After the mice were sacrificed, their colon tissues were collected for histological analysis. Alcian blue/periodic acid-Schiff(AB/PAS) staining was used to detect the number and mucus secretion status of goblet cells; immunohistochemistry was performed to measure the expression of ki67, cleaved caspase-3, mucin 2(Muc2), and matrix metalloproteinase-9(MMP9) in colon tissues; and immunofluorescence was used to analyze the expression of tight junction proteins in colon tissues, and enzyme linked immunosorbent assay(ELISA) was employed to quantify the levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-1β, and IL-6. Western blot was conducted to evaluate the expression of BMP pathway-related proteins in mouse colon tissues. Quantitative real-time PCR(qRT-PCR) was performed to measure the expression of genes related to goblet cell differentiation in mouse colon tissues. In addition, this study also examined the protective effect and underlying mechanism of BEPD-containing serum on lipopolysaccharide(LPS)-induced barrier damages in LS174T goblet cells in vitro. The results showed that BEPD significantly alleviated UC symptoms in mice, restored goblet cell diffe-rentiation function, promoted Muc2 secretion and tight junction protein expression, and suppressed inflammatory factor secretion while activating the BMP signaling pathway. Therefore, BEPD may exert its therapeutic effects on UC by activating the BMP signaling pathway, providing a new strategy for drug intervention in UC.
Topics: Animals; Signal Transduction; Mice; Colitis, Ulcerative; Mice, Inbred C57BL; Drugs, Chinese Herbal; Male; Pulsatilla; Humans; Bone Morphogenetic Proteins
PubMed: 38812188
DOI: 10.19540/j.cnki.cjcmm.20240202.704 -
Journal of Periodontology May 2024This study evaluated the influence of the adjunctive application of a cross-linked hyaluronic acid (HA) in the treatment of multiple gingival recessions, using a...
Tunnel technique and subepithelial connective tissue graft, with or without cross-linked hyaluronic acid, in the treatment of multiple gingival recessions: 12-month outcomes of a randomized clinical trial.
BACKGROUND
This study evaluated the influence of the adjunctive application of a cross-linked hyaluronic acid (HA) in the treatment of multiple gingival recessions, using a modified coronally advanced tunnel (MCAT) technique and subepithelial connective tissue graft (SCTG) (MCAT+SCTG±HA).
METHODS
A randomized, split-mouth, double-masked comparison of the effects of MCAT+HA+SCTG (test) versus MCAT+SCTG (control) in the treatment of multiple, contralateral gingival recessions with clinical, esthetic, and histological evaluations was carried out. All samples were stained with hematoxylin and eosin, Masson's trichrome, Verhoeff-Van Gieson, and Alcian blue stain for semiquantitative evaluation. The primary outcome variable was 12-month mean root coverage (MRC).
RESULTS
Twenty-four patients with 266 gingival recessions received both control and test treatments (133 recessions per group). 12-month MRC of the MCAT+HA+SCTG group was not significantly different from the MCAT+SCTG group with 84.32%± 34.46% and 85.71%± 36.43%, respectively (p = 0.991). Both treatment modes produced favorable esthetic outcomes (root coverage esthetic score [RES] 9.51± 1.01 tests vs. 9.26± 1.10 controls, p = 0.7292). However, the application of HA improved soft tissue texture (p = 0.0091). The remaining end point measures did not differ significantly between groups. Histological evaluation showed a significantly greater number of elastic fibers and a moderate increase in collagen fiber density in biopsy samples taken from the test sides when compared to the control sides (p = 0.0419 and p = 0.300, respectively).
CONCLUSIONS
MCAT+SCTG is an effective procedure in the treatment of multiple recession Type 1 (RT1) and RT2 recessions. There were no statistically significant differences in evaluated clinical treatment outcomes in the MCAT+HA+SCTG group compared to the MCAT+SCTG group within a period of 12 months. The application of HA increased collagen and elastic fiber density.
PubMed: 38808976
DOI: 10.1002/JPER.24-0093 -
International Journal of Biological... Jun 2024The therapeutic potential of tissue engineering in addressing articular cartilage defects has been a focal point of research for numerous years. Despite its promising...
Simultaneous usage of sulforaphane nanoemulsion and tannic acid in ternary chitosan/gelatin/PEG hydrogel for knee cartilage tissue engineering: In vitro and in vivo study.
The therapeutic potential of tissue engineering in addressing articular cartilage defects has been a focal point of research for numerous years. Despite its promising outlook, a persistent challenge within this domain is the lack of sufficient functional integration between engineered and natural tissues. This study introduces a novel approach that employs a combination of sulforaphane (SFN) nanoemulsion and tannic acid to enhance cartilage tissue engineering and promote tissue integration in a rat knee cartilage defect model. To substantiate our hypothesis, we conducted a series of in vitro and in vivo experiments. The SFN nanoemulsion was characterized using DLS, zeta potential, and TEM analyses. Subsequently, it was incorporated into a ternary polymer hydrogel composed of chitosan, gelatin, and polyethylene glycol. We evaluated the hydrogel with (H-SFN) and without (H) the SFN nanoemulsion through a comprehensive set of physicochemical, mechanical, and biological analyses. For the in vivo study, nine male Wistar rats were divided into three groups: no implant (Ctrl), H, and H-SFN. After inducing a cartilage defect, the affected area was treated with tannic acid and subsequently implanted with the hydrogels. Four weeks post-implantation, the harvested cartilage underwent histological examination employing H&E, safranin O/fast green, alcian blue, and immunohistochemistry staining techniques. Our results revealed that the SFN nanodroplets had an average diameter of 75 nm and a surface charge of -11.58 mV. Moreover, degradation, swelling rates, hydrophilicity, and elasticity features of the hydrogel incorporating SFN were improved. Histopathological analysis indicated a higher production of GAGs and collagen in the H-SFN group. Furthermore, the H-SFN group exhibited superior cartilage regeneration and tissue integration compared to the Ctrl and H groups. In conclusion, the findings of this study suggest the importance of considering cell protective properties in the fabrication of scaffolds for knee cartilage defects, emphasizing the potential significance of the proposed SFN nanoemulsion and tannic acid approach in advancing the field of cartilage tissue engineering.
Topics: Tannins; Animals; Chitosan; Hydrogels; Gelatin; Rats; Emulsions; Cartilage, Articular; Isothiocyanates; Polyethylene Glycols; Male; Tissue Engineering; Sulfoxides; Rats, Wistar; Tissue Scaffolds; Nanoparticles; Polyphenols
PubMed: 38806085
DOI: 10.1016/j.ijbiomac.2024.132692 -
Theriogenology Sep 2024Endometrosis in mares is a disease resulting from chronic inflammation characterized by peri glandular fibrosis. There is no effective treatment so far, which opens the...
Short preconditioning with TGFβ of equine adipose tissue-derived mesenchymal stem cells predisposes towards an anti-fibrotic secretory phenotype: A possible tool for treatment of endometrosis in mares.
Endometrosis in mares is a disease resulting from chronic inflammation characterized by peri glandular fibrosis. There is no effective treatment so far, which opens the door for exploring the use of stem cells as a candidate. Transforming growth factor beta (TGFβ) is crucial for the establishment and progression of fibrosis in mare's endometrosis. We aimed to develop regenerative approaches to treat endometrosis by using mesenchymal stem cells (MSC), for which understanding the effect of TGFβ on exogenous MSC is crucial. We isolated and characterized equine adipose MSC from six donors. Cells were pooled and exposed to 10 ng/ml of TGFβ for 0, 4, and 24 h, after which cells were analyzed for proliferation, migration, mesodermal differentiation, expression of fibrosis-related mRNAs, and prostaglandin E2 secretion. At 24 h of exposition to TGFβ, there was a progressive increase in the contraction of the monolayer, leading to nodular structures, while cell viability did not change. Exposure to TGFβ impaired adipogenic and osteogenic differentiation after 4 h of treatment, which was more marked at 24 h, represented by a decrease in Oil red and Alizarin red staining, as well as a significant drop (p < 0.05) in the expression of key gene regulators of differentiation processes (PPARG for adipose and RUNX2 for osteogenic differentiation). TGFβ increased chondrogenic differentiation as shown by the upsurge in size of the resulting 3D cell pellet and intensity of Alcian Blue staining, as well as the significant up-regulation of SOX9 expression (p < 0.05) at 4 h, which reached a maximum peak at 24 h (p < 0.01), indicative of up-regulation of glycosaminoglycan synthesis. Preconditioning MSC with TGFβ led to a significant increase (p < 0.05) in the expression of myofibroblast gene markers aSMA, COL1A1, and TGFβ at 24 h exposition time. In contrast, the expression of COL3A1 did not change with respect to the control but registered a significant downregulation compared to 4 h (p < 0.05). TGFβ also affected the expression of genes involved in PGE synthesis and function; COX2, PTGES, and the PGE receptor EP4 were all significantly upregulated early at 4 h (p < 0.05). Cells exposed to TGFβ showed a significant upregulation of PGE secretion at 4 h compared to untreated cells (p < 0.05); conversely, at 24 h, the PGE values decreased significantly compared to control cells (p < 0.05). Preconditioning MSC for 4 h led to an anti-fibrotic secretory phenotype, while a longer period (24 h) led to a pro-fibrotic one. It is tempting to propose a 4-h preconditioning of exogenous MSC with TGFβ to drive them towards an anti-fibrotic phenotype for cellular and cell-free therapies in fibrotic diseases such as endometrosis of mares.
Topics: Animals; Horses; Mesenchymal Stem Cells; Female; Adipose Tissue; Horse Diseases; Transforming Growth Factor beta; Cell Differentiation; Fibrosis; Cells, Cultured; Gene Expression Regulation
PubMed: 38805994
DOI: 10.1016/j.theriogenology.2024.05.018 -
PloS One 2024This study aimed to assess the localization of chondroitin sulfate (CS), a primary extracellular matrix component, in the stromal region of endometrial carcinoma (EC).
INTRODUCTION
This study aimed to assess the localization of chondroitin sulfate (CS), a primary extracellular matrix component, in the stromal region of endometrial carcinoma (EC).
METHODS
Immunostaining was performed on 26 endometrial endometrioid carcinoma (EEC) samples of different grades and 10 endometrial serous carcinoma (ESC) samples to evaluate CS localization. This was further confirmed by Alcian Blue (AB) staining as well.
RESULTS
In the G1-EEC samples, CS showed reactivity with fibrovascular stroma, supporting closely packed glandular crowding and papillary structures. As the grade increased, the original interstitial structure was re-established, and the localization of CS in the perigulandular region decreased. In the ESC samples, the thick fibrous strands supporting the papillary architecture showed reactivity with CS; however, the delicate stromal region branching into the narrow region showed poor reactivity. The AB staining results showed similar characteristics to the immunostaining ones.
CONCLUSIONS
The characteristic localization of CS in various EC types was elucidated. The present study provides new information on endometrial stromal assessment.
Topics: Humans; Female; Endometrial Neoplasms; Chondroitin Sulfates; Middle Aged; Carcinoma, Endometrioid; Aged; Immunohistochemistry
PubMed: 38805498
DOI: 10.1371/journal.pone.0304420 -
Veterinary Clinical Pathology Jun 2024A 13-year-old male domestic short-hair cat presented for evaluation of labored breathing, hyporexia, and lethargy. Pertinent initial diagnostics yielded leukocytosis,...
A 13-year-old male domestic short-hair cat presented for evaluation of labored breathing, hyporexia, and lethargy. Pertinent initial diagnostics yielded leukocytosis, characterized by neutrophilia and monocytosis. Numerous small, round, magenta granules were observed within all neutrophils in Wright-Giemsa-stained blood films on the day of presentation and the day thereafter. No other neutrophil morphologic abnormalities were present, making cytoplasmic toxicity highly unlikely. Hyperadrenocorticism was diagnosed based on the lack of suppression in a low-dose dexamethasone suppression test, and without other diagnostics, the cat was discharged on trilostane therapy. Neutrophil granules did not stain with Alcian blue pH 1.0, periodic acid-Schiff (PAS), PAS and Alcian blue pH 2.5, and toluidine blue. Electron microscopy identified no differences in the morphology of the secretory granules or other neutrophil features. Metabolic screening tests of the cat's urine did not identify a genetic metabolic disorder. However, serum α- and β -hexosaminidase (HexA and HexB) activities were 4.3% and 0% of normal controls, respectively, which is supportive of GM2-gangliosidosis, that is, Sandhoff disorder. However, the historical, clinical, and electron microscopy findings did not provide evidence to confirm this genetic defect. To the author's knowledge, this is the first case of magenta-staining granules within neutrophils in a breed other than a Birman, Siamese, or Himalayan.
Topics: Animals; Cats; Male; Neutrophils; Cat Diseases; Cytoplasmic Granules
PubMed: 38797715
DOI: 10.1111/vcp.13356 -
Bioengineering (Basel, Switzerland) May 2024Ascorbic acid (AA) plays a crucial role in both the proliferation and chondrogenic differentiation potential of mesenchymal stem/medicinal signalling cells (MSCs); these...
Modulation of Canine Adipose-Derived Mesenchymal Stem/Medicinal Signalling Cells with Ascorbic Acid: Effect on Proliferation and Chondrogenic Differentiation on Standard Plastic and Silk Fibroin Surfaces.
Ascorbic acid (AA) plays a crucial role in both the proliferation and chondrogenic differentiation potential of mesenchymal stem/medicinal signalling cells (MSCs); these are both key aspects of their general therapeutic use and their increasing use in veterinary medicine. Current immunomodulatory therapies require efficient expansion of MSCs in the laboratory, while emerging tissue regeneration strategies, such as cartilage or bone repair, aim to use differentiated MSCs and modulate the expression of chondrogenic and hypertrophic markers. Our aim was to investigate whether the addition of AA to the growth medium enhances the proliferation of canine adipose-derived MSCs (cAMSCs) grown on standard plastic surfaces and whether it affects chondrogenic differentiation potential on silk fibroin (SF) films. We assessed cell viability with trypan blue and proliferation potential by calculating population doubling. Chondrogenic induction on SF films was assessed by Alcian blue staining and gene expression analysis of chondrogenic and hypertrophic genes. The results showed that growth medium with AA significantly enhanced the proliferation of cAMSCs without affecting cell viability and modulated the expression of chondrogenic and hypertrophic genes of cAMSCs grown on SF films. Our results suggest that AA may be used in growth medium for expansion of cAMSCs and, at the same time, provide the basis for future studies to investigate the role of AA and SF in chondrogenic differentiation of MSCs.
PubMed: 38790380
DOI: 10.3390/bioengineering11050513