scholarly journals Development of Injectable Polydactyly-Derived Chondrocyte Sheets

2021 ◽  
Vol 22 (6) ◽  
pp. 3198
Author(s):  
Shiho Wasai ◽  
Eriko Toyoda ◽  
Takumi Takahashi ◽  
Miki Maehara ◽  
Eri Okada ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cartilage Damage ◽  
Enzyme Linked Immunosorbent Assay ◽  
Invasive Treatment ◽  
Experimental Conditions ◽  
Humoral Factors ◽  
Less Invasive ◽  
Cell Counts ◽  
Invasive Method ◽  
Injection Control

We are conducting a clinical study of the use of allogeneic polydactyly-derived chondrocyte sheets (PD sheets) for the repair of articular cartilage damage caused by osteoarthritis. However, the transplantation of PD sheets requires highly invasive surgery. To establish a less invasive treatment, we are currently developing injectable fragments of PD sheets (PD sheets-mini). Polydactyly-derived chondrocytes were seeded in RepCell™ or conventional temperature-responsive inserts and cultured. Cell counts and viability, histology, enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qPCR), and flow cytometry were used to characterize PD sheets-mini and PD sheets collected from each culture. To examine the effects of injection on cell viability, PD sheets-mini were tested in four experimental conditions: non-injection control, 18 gauge (G) needle, 23G needle, and syringe only. PD sheets-mini produced similar amounts of humoral factors as PD sheets. No histological differences were observed between PD sheets and PD sheets-mini. Except for COL2A1, expression of cartilage-related genes did not differ between the two types of PD sheet. No significant differences were observed between injection conditions. PD sheets-mini have characteristics that resemble PD sheets. The cell viability of PD sheets-mini was not significantly affected by needle gauge size. Intra-articular injection may be a feasible, less invasive method to transplant PD sheets-mini.

In vitro effects of Zinc in soluble and homeopathic formulations on macrophages and astrocytes

Homeopathy ◽  
2017 ◽  
Vol 106 (02) ◽  
pp. 103-113 ◽  
Author(s):  
Clara Bonafini ◽  
Marta Marzotto ◽  
Paolo Bellavite
Keyword(s):  
Nitric Oxide ◽  
Cell Viability ◽  
Soluble Form ◽  
Enzyme Linked Immunosorbent Assay ◽  
Therapeutic Effects ◽  
Experimental Conditions ◽  
Nitric Oxide Release ◽  
Tnf Α ◽  
High Zinc ◽  
Zinc Depletion

Zinc is an important metal in body homeostasis. Zinc in soluble form (Zn2+) and homeopathic Zincum metallicum were tested in macrophages and astrocytes in order to investigate its potential toxic or therapeutic effects. We evaluated cell viability (WST assay), cytokine production such as tumour necrosis factor alpha (TNF-α) and interleukin 10 (IL-10) by enzyme-linked immunosorbent assay (ELISA) and nitric oxide release by Griess reaction. The effect of zinc-depletion and high zinc pre-treatments on the cell adaptation capability was also investigated. In THP-1 macrophage cell line and in human primary macrophages, Zn2+ at sub-toxic doses (30 μM) caused stimulation of TNF-α and IL-10 with different dynamics reaching the maximum peak at the zinc concentration 100 μM, before the cell death. Highest doses (300 μM) impaired dramatically cell vitality. Similar effects on cell viability were obtained also in C6 astrocytes, where Zn2+ slightly increased the nitric oxide release only in cells activated by one of the pro-inflammatory stimuli used in our cellular model (interferon gamma plus TNF-α). Zinc depletion markedly reduced IL-10 production and cell viability. Zincum metallicum did not cause toxicity in any cell type and showed some small stimulation in WST assay that was statistically significant in a few experimental conditions.

scholarly journals CTRP9 protects against MIA-induced inflammation and knee cartilage damage by deactivating the MAPK/NF-κB pathway in rats with osteoarthritis

2020 ◽  
Vol 15 (1) ◽  
pp. 971-980
Author(s):  
Shicheng Zheng ◽  
Jing Ren ◽  
Sihai Gong ◽  
Feng Qiao ◽  
Jinlong He
Keyword(s):  
Synovial Fluid ◽  
Nuclear Factor Kappa B ◽  
Cartilage Damage ◽  
Cartilage Tissue ◽  
Enzyme Linked Immunosorbent Assay ◽  
Nuclear Factor ◽  
Knee Cartilage ◽  
Monosodium Iodoacetate ◽  
Dose Dependent ◽  
Different Doses

AbstractC1q/TNF-related protein 9 (CTRP9), the closest paralog of adiponectin, has been reported to protect against inflammation-related diseases. However, its role in regulating osteoarthritis (OA) has not been fully elucidated. First, a rat model of OA was generated. Furthermore, rats with OA were injected with different doses of recombinant CTRP9 protein (rCTRP9), and the knee cartilage damage was evaluated. Finally, the phosphorylation of p38 and the secretion of matrix metalloproteinases (MMPs) were detected by Western blotting and enzyme-linked immunosorbent assay. Results revealed that CTRP9 was highly expressed in adipose tissue, followed by skeletal muscle and cartilage tissue, and less expressed in liver, kidney and lung. Moreover, the expression of CTRP9 significantly decreased in the monosodium iodoacetate (MIA) group in the knee cartilage and knee synovial fluid, and the contents of interleukin-1β (IL-1β) and IL-6 significantly increased in knee synovial fluid. In addition, rCTRP9 alleviated MIA-induced inflammation, oxidative stress and knee cartilage damage in a dose-dependent way. In addition, rCTRP9 could attenuate the expression of p38MAPK and p-p38 and suppress the expression of nuclear factor-kappa B (NF-κB), p65 and MMPs. Collectively, the results of the present study suggested that CTRP9 alleviates the inflammation of MIA-induced OA through deactivating p38MAPK and NF-κB signaling pathways in rats.

scholarly journals Direct Effect of Septic Plasma in Human Cell Lines Viability

2018 ◽  
Vol 47 (1-3) ◽  
pp. 270-276
Author(s):  
Grazia Maria Virzì ◽  
Chiara Borga ◽  
Chiara Pasqualin ◽  
Silvia Pastori ◽  
Alessandra Brocca ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Cell Viability ◽  
Cell Lines ◽  
Caspase 3 ◽  
Test Group ◽  
Organ Injury ◽  
Enzyme Linked Immunosorbent Assay ◽  
Caspase 9 ◽  
Renal Tubular Cells

Background: Sepsis is a life-threatening condition often associated with a high incidence of multiple organs injury. Several papers suggested the immune response by itself, with the production of humoral inflammatory mediators, is crucial in determining organ injury. However, little is known of how sepsis directly induces organ injury at the cellular levels. To assess this point, we set up an in vitro study to investigate the response of renal tubular cells (RTCs), monocytes (U937) and hepatocytes (HepG2) after 24 h-incubation with septic patients’ plasma. Methods: We enrolled 26 septic patients (“test” group). We evaluated cell viability, apoptosis and necrosis by flow cytometer. Caspase-3,-8,-9 and cytochrome-c concentrations have been analyzed using the Human enzyme-linked immunosorbent assay kit. Results: We found that a decrease of cell viability in all cell lines tested was associated to the increase of apoptosis in RTCs and U937 (p < 0.0001) and increase of necrosis in HepG2 (p < 0.5). The increase of apoptosis in RTCs and U937 cells was confirmed by higher levels of caspase-3 (p < 0.0001). We showed that apoptosis in both RTCs and U937 was triggered by the activation of the intrinsic pathway, as caspase-9 and cytochrome-c levels significantly increased (p < 0.0001), while caspase-8 did not change. This assumption was strengthened by the significant correlation of caspase-9 with both cytochrome-c (r = 0.73 for RTCs and r = 0.69 for U937) and caspase-3 (r = 0.69 for RTCs and r = 0.63 for U937). Conclusion: Humoral mediators in septic patients’ plasma induce apoptosis. This fact suggests that apoptosis inhibitors should be investigated as future strategy to reduce sepsis-induced organ damages.

scholarly journals Temozolomide sensitivity of malignant glioma cell lines – a systematic review assessing consistencies between in vitro studies

BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Michael T. C. Poon ◽  
Morgan Bruce ◽  
Joanne E. Simpson ◽  
Cathal J. Hannan ◽  
Paul M. Brennan
Keyword(s):  
Cell Viability ◽  
Malignant Glioma ◽  
Cell Line ◽  
Cell Lines ◽  
Glioma Cell ◽  
Glioma Cell Line ◽  
In Vitro Studies ◽  
Experimental Conditions ◽  
Glioma Cell Lines

Abstract Background Malignant glioma cell line models are integral to pre-clinical testing of novel potential therapies. Accurate prediction of likely efficacy in the clinic requires that these models are reliable and consistent. We assessed this by examining the reporting of experimental conditions and sensitivity to temozolomide in glioma cells lines. Methods We searched Medline and Embase (Jan 1994-Jan 2021) for studies evaluating the effect of temozolomide monotherapy on cell viability of at least one malignant glioma cell line. Key data items included type of cell lines, temozolomide exposure duration in hours (hr), and cell viability measure (IC50). Results We included 212 studies from 2789 non-duplicate records that reported 248 distinct cell lines. The commonest cell line was U87 (60.4%). Only 10.4% studies used a patient-derived cell line. The proportion of studies not reporting each experimental condition ranged from 8.0–27.4%, including base medium (8.0%), serum supplementation (9.9%) and number of replicates (27.4%). In studies reporting IC50, the median value for U87 at 24 h, 48 h and 72 h was 123.9 μM (IQR 75.3–277.7 μM), 223.1 μM (IQR 92.0–590.1 μM) and 230.0 μM (IQR 34.1–650.0 μM), respectively. The median IC50 at 72 h for patient-derived cell lines was 220 μM (IQR 81.1–800.0 μM). Conclusion Temozolomide sensitivity reported in comparable studies was not consistent between or within malignant glioma cell lines. Drug discovery science performed on these models cannot reliably inform clinical translation. A consensus model of reporting can maximise reproducibility and consistency among in vitro studies.

scholarly journals Abdominal Sacral Colpopexy In Treatment Of Vaginal Vault Prolapse: By Less Invasive Method

2013 ◽  
Vol 25 (1) ◽  
pp. 3-8
Author(s):  
Nahar Nurun ◽  
Molina Rani Kundu ◽  
Naher Akterun
Keyword(s):  
Success Rate ◽  
Vaginal Vault Prolapse ◽  
Vaginal Vault ◽  
Secondary Outcome ◽  
Sacral Colpopexy ◽  
Vault Prolapse ◽  
Abdominal Sacral Colpopexy ◽  
Less Invasive ◽  
Invasive Method ◽  
The Impact

Objective: To assess the outcomes of abdominal sacral colpopexy in less invasive method. Study design: It was a prospective study conducted in Comilla Medical College Hospital and Comilla General Hospital during the period from 2005 to 2009.Method: Thirty women with vaginal vault prolapse were selected by inclusion and exclusion criteria in a consecutive, exhaustive method. Primary outcome measurements were included subjective, objective and patient-determined success rate. Secondary outcome included the impact on bowel, bladder, sexual function and quality of life. Result: Result shows that, vault prolapse is mostly associated with older patients, age more than sixty (66.7%), para >5 (60%) and menopausal women (66.7%). Vault prolapse was mostly associated with cystocele (93.3%), stress incontinence (76.7%) and more common following abdominal hysterectomy (70%). During operation the dissection was less (3-4cm). Average operating time was 54.33/min; average estimated blood loss was 49.17/ml. One patient required blood transfusion, one developed haematoma during surgery, no gut injury or haemodynamic instability developed. Post operatively, no internal haemorrhage, 4 patient developed fever (13.3%), wound infection one (3.3%), UTI 4 patients (13.3%), no voiding difficulty or thromboembolism and one patient developed mesh rejection (3.3%). After one year follow-up success rate was 96.7%. Conclusion: Abdominal sacral colpopexy is a safe and effective method for correction of vaginal vault prolapse. DOI: http://dx.doi.org/10.3329/bjog.v25i1.13723 Bangladesh J Obstet Gynaecol, 2010; Vol. 25(1) : 3-8

A quantitative study of the growth and development of the ventral root in normal and experimental conditions

Development ◽  
1974 ◽  
Vol 32 (3) ◽  
pp. 819-833
Author(s):  
M. C. Prestige ◽  
Margaret A. Wilson
Keyword(s):  
Normal Development ◽  
Cell Number ◽  
Ventral Horn ◽  
Ventral Root ◽  
Limb Bud ◽  
Experimental Conditions ◽  
Collateral Sprouting ◽  
Cell Counts ◽  
Ventral Roots ◽  
Fibre Loss

1. The development of the ventral root (VR) in Xenopus has been studied by electron microscopy. Total fibre counts, and counts of classes of fibres were made from large photomontages of the whole of VR 9 at × 15000. 2. The total number of fibres in the root shows the same pattern of initial rise, peak, and subsequent decline that previous ventral horn (VH) cell counts had shown, The two curves overlay each other initially, but after the decline, there were apparently more cells than fibres. 3. Promyelin and myelin formation was seen at the time of the decline. There was no evidence that dying axons had started to myelinate. 4. In some animals the limb-bud was removed at the time of its first penetration by nerve fibres. The ventral roots developed normally for a week, but thereafter fibre loss was accentuated, advanced and more profound, so that after another week, no fibres were left. In these roots, no promyelin or myelin was formed. 5. In other animals, it was shown that there is no evidence for collateral sprouting in the ventral roots during normal development. 6. It is argued that the axons which die in normal development have already reached the limb-bud. 7. The correspondence between axon and cell number is discussed.

Efficiency and safety of sclerotherapy for recurrent nodular goiter

2020 ◽  
pp. 39-51
Author(s):  
G. V. Rodoman ◽  
I. R. Sumedi ◽  
N. V. Sviridenko ◽  
T. I. Shalaeva ◽  
M. M. Meloyan
Keyword(s):  
Minimally Invasive ◽  
Severe Pain ◽  
Nodular Goiter ◽  
Treatment Method ◽  
Age Group ◽  
Invasive Treatment ◽  
The Third ◽  
Invasive Method ◽  
Functional Autonomy

At present, patients with recurrent nodular goiter account for a significant portion of patients operated on for nodular goiter. At the same time, the comorbid background characteristic of this age group and the technical difficulties of the intervention on cicatricial tissues of the neck cause a high risk of complications of the operation, 3–7 times higher than with primary thyroid interventions. The aim of the study was to evaluate the effectiveness and safety of treatment of recurrent nodular goiter using an alternative minimally invasive method — sclerotherapy. The study included 30 patients previously operated on for nodular goiter. All had 4 courses of sclerotherapy, each included 5 sessions with a frequency of 1 session per week, followed by a follow-up period of 3 months. Polydocanol was used as a sclerosant. The analysis showed that sclerotherapy for recurrent nodular goiter allows all patients to reduce recurrent nodular formations, and in almost a third of cases, complete reduction of the nodes. On average, the decrease in the volume of thyroid residues was 9.6 ± 1.5 ml, and the size of nodular formations decreased by 17.2 ± 1.3 mm (3.7 times — from 23.6 ± 1.4 mm to 6.4 ± 0.7 mm, P <0.001). Nodes more than 3 cm, initially 19 %, ceased to be detected after the third course of sclerotherapy. In all cases, managed to eliminate hormonal imbalances in patients who initially had functional autonomy, as well as signs of compression of the neck organs. At the same time, sclerotherapy of nodules of the thyroid gland using polydocanol as a sclerosant is a safe minimally invasive treatment method, is not accompanied by severe pain and the risk of hypoparathyroidism and laryngeal paresis.

ALK7 Inhibition Protects Osteoblast Cells Against High Glucose-induced ROS Production via Nrf2/HO-1 Signaling Pathway

2021 ◽  
Vol 21 ◽  
Author(s):  
Zhen Zhao ◽  
Yu Lu ◽  
Huan Wang ◽  
Xiang Gu ◽  
Luting Zhu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Viability ◽  
Signaling Pathway ◽  
High Glucose ◽  
Collagen I ◽  
Cell Counting ◽  
Enzyme Linked Immunosorbent Assay ◽  
Heme Oxygenase 1 ◽  
Ros Production ◽  
Alizarin Red

Background: Some studies demonstrated that under high-glucose (HG) condition, osteoblasts develop oxidative stress, which will impair their normal functions. The effects of activin receptor-like kinase 7 (ALK7) silencing on HG-induced osteoblasts remained unclear. Objective: The aim of this study was to explore the effect of ALK7 on HG-induced osteoblasts. Methods: MC3T3-E1 cells were treated with different concentrations of HG (0, 50, 100, 200 and 300mg/dL), and the cell viability was detected using cell counting kit-8 (CCK-8). HG-treated MC3T3-E1 cells were transfected with siALK7 or ALK7 overexpression plasmid or siNrf2, and then the viability and apoptosis were detected by CCK-8 and flow cytometry. The levels of reactive oxygen species (ROS), collagen I and calcification nodule were determined by oxidative stress kits, Enzyme-linked immunosorbent assay and Alizarin red staining. The expressions of NF-E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1) and osteoblast-associated genes were determined by quantitative real-time PCR (qRT-PCR) and Western blot. Results: Cell viability was reduced with HG treatment. Silencing ALK7 inhibited the effect of HG on increasing cell apoptosis and ROS production, reduced cell viability, mineralized nodules, and downregulated collagen I and osteoblast-associated genes expression in MC3T3-E1 cells. ALK7 silencing activated the Nrf2/HO-1 signaling pathway by affecting expressions of HO-1 and Nrf2. ALK7 overexpression had the opposite effects. In addition, siNrf2 partially reversed the effects of ALK7 silencing on HG-induced MC3T3-E1 cells. Conclusion: ALK7 silencing protected osteoblasts under HG condition possibly through activating the Nrf2/HO-1 pathway.

Further studies of neuroangiostrongyliasis (rat lungworm disease) in Australian dogs: 92 new cases (2010–2020) and results for a novel, highly sensitive qPCR assay

Parasitology ◽  
2020 ◽  
pp. 1-9
Author(s):  
Rogan Lee ◽  
Tsung-Yu Pai ◽  
Richard Churcher ◽  
Sarah Davies ◽  
Jody Braddock ◽  
...  
Keyword(s):  
Infectious Disease ◽  
Quantitative Polymerase Chain Reaction ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Cell Counts ◽  
Highly Sensitive ◽  
Elisa Testing ◽  
Rat Lungworm ◽  
Polymerase Chain ◽  
Sydney Australia

Abstract The principal aim of this study was to optimize the diagnosis of canine neuroangiostrongyliasis (NA). In total, 92 cases were seen between 2010 and 2020. Dogs were aged from 7 weeks to 14 years (median 5 months), with 73/90 (81%) less than 6 months and 1.7 times as many males as females. The disease became more common over the study period. Most cases (86%) were seen between March and July. Cerebrospinal fluid (CSF) was obtained from the cisterna magna in 77 dogs, the lumbar cistern in f5, and both sites in 3. Nucleated cell counts for 84 specimens ranged from 1 to 146 150 cells μL−1 (median 4500). Percentage eosinophils varied from 0 to 98% (median 83%). When both cisternal and lumbar CSF were collected, inflammation was more severe caudally. Seventy-three CSF specimens were subjected to enzyme-linked immunosorbent assay (ELISA) testing for antibodies against A. cantonensis; 61 (84%) tested positive, titres ranging from <100 to ⩾12 800 (median 1600). Sixty-one CSF specimens were subjected to real-time quantitative polymerase chain reaction (qPCR) testing using a new protocol targeting a bioinformatically-informed repetitive genetic target; 53/61 samples (87%) tested positive, C T values ranging from 23.4 to 39.5 (median 30.0). For 57 dogs, it was possible to compare CSF ELISA serology and qPCR. ELISA and qPCR were both positive in 40 dogs, in 5 dogs the ELISA was positive while the qPCR was negative, in 9 dogs the qPCR was positive but the ELISA was negative, while in 3 dogs both the ELISA and qPCR were negative. NA is an emerging infectious disease of dogs in Sydney, Australia.

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