scholarly journals Fluorescent Nanodiamonds Enable Long-Term Detection of Human Adipose-Derived Stem/Stromal Cells in an In Vivo Chondrogenesis Model Using Decellularized Extracellular Matrices and Fibrin Glue Polymer

Polymers ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 1391 ◽  
Author(s):  
Yi-Chia Wu ◽  
Ya-Chin Wang ◽  
Wei-Ting Wang ◽  
Hui-Min David Wang ◽  
Hsin-Hung Lin ◽  
...  
Keyword(s):  
Fibrin Glue ◽  
Nude Mice ◽  
Stromal Cells ◽  
Histological Analysis ◽  
Control Group ◽  
Efficient Treatment ◽  
Fluorescent Nanodiamonds ◽  
Experimental Group

Clinically available materials, including allogeneic irradiated costal cartilage and fibrin glue polymer, were used as scaffolds for in vivo chondrogenic differentiation of human adipose-derived stem/stromal cells (hASCs) in the attempt to develop a more efficient treatment over current methods. Current studies include the use of growth-factor stimulation, tissue engineering, and biocompatible materials; however, most methods involve complicated processes and pose clinical limitations. In this report, the xenografts in the experimental group composed of a diced decellularized cartilage extracellular matrix (ECM), hASCs, and fibrin glue polymer were implanted into the subcutaneous layer of nude mice, and the results were compared with two groups of controls; one control group received implantation of decellularized cartilage ECM and fibrin glue polymer, and the other control group received implantation of hASCs mixed with fibrin glue polymer. To evaluate whether hASCs had in vivo chondrogenesis in the xenografts, hASCs were labeled with fluorescent nanodiamonds (FNDs), a biocompatible and photostable nanomaterial, to allow for long-term detection and histological analysis. Increased cellularity, glycosaminoglycan, and collagen deposition were found by the histological examination in the experimental group compared with control groups. With the background-free detection technique and time-gated fluorescence imaging, the numbers and locations of the FND-labeled hASCs could be detected by confocal microscopy. The chondrocyte-specific markers, such as aggrecan and type II collagen, were colocalized with cells containing signals of FNDs which indicated in vivo chondrogenesis of hASCs. Taken together, functional in vivo chondrogenesis of the hASCs could be achieved by clinically available decellularized cartilage ECM and fibrin glue polymer in the nude mice model without in vitro chondrogenic induction. The fluorescent signals of FNDs in hASCs can be detected in histological analysis, such as hematoxylin and eosin staining (H&E staining) without the interference of the autofluorescence. Our study may warrant future clinical applications of the combination of decellular cartilage ECM, fibrin glue polymer, and hASCs for cartilage repair.

Cisplatin Nano-Liposomes Promoting Apoptosis of Retinoblastoma Cells Both In Vivo and In Vitro

2020 ◽  
Vol 12 (4) ◽  
pp. 536-542
Author(s):  
Lijuan Zhao ◽  
Fei Wang ◽  
Wei Fan
Keyword(s):  
Protein Expression ◽  
Nude Mice ◽  
Caspase 3 ◽  
Mrna Level ◽  
Control Group ◽  
Mrna Levels ◽  
Bax Protein ◽  
Experimental Group

This study was established to investigate the effects of cisplatin nano-liposomes on the apoptosis of the human retinoblastoma (RB) cell line Y79 in vitro and in vivo. Y79 cells were cultured and then exposed to Annexin V/PI to test their apoptosis, tested with the Caspase-3 activity detection kit to examine the change in activity of Caspase-3, and subjected to western blotting to test Bcl-2 and Bax protein expression. Y79-cell-transplanted tumor model in nude mice was also established and divided into three groups, with five nude mice in each. Cisplatin nano-liposomes were applied to the experimental group, cisplatin was injected into the control group, while saline was administered to the blank group, after which the nude mice were killed and the tumor was removed. Tumor volumes and weights in the three groups were compared. Nucleic acid extraction from magnetic beads was adopted to extract DNA, RT-PCR was employed to test Bcl-2 and Bax mRNA levels in tumor tissues, and in situ cell death assay kit was applied to test apoptotic cells. In comparison to the cisplatin solution and DMSO groups, the cisplatin liposome group showed higher Y79 apoptotic rate, Caspase-3 activity, and Bax protein expression, and lower Bcl-2 protein expression (all P < 0 05). In comparison with the control and blank groups, the experimental group showed lower tumor volume, weight, and Bcl-2 mRNA level of nude mice. In addition, in comparison with the control group, the experimental group showed higher cellular apoptotic rate and Bax mRNA level. In terms of the clinical effects of cisplatin nano-liposomes on a tumor transplant in nude mice with cervical cancer, they were shown to promote tumor apoptosis.

scholarly journals Administration of Beta-Emitting Anti-CD37 Radioimmunoconjugate Lutetium (177Lu) Lilotomab Satetraxetan as Weekly Multiple Injections Increases Maximum Tolerated Activity in Nude Mice with Non- Hodgkin Lymphoma Xenografts

2018 ◽  
Vol 4 (1) ◽  
pp. 181-190
Author(s):  
Helen Heyerdahl ◽  
Ada H.V. Repetto-Llamazares ◽  
Jostein Dahle
Keyword(s):  
Hodgkin Lymphoma ◽  
Nude Mice ◽  
Separate Experiment ◽  
Growth Delay ◽  
Control Group ◽  
Multiple Dosing ◽  
Multiple Injections ◽  
Non Hodgkin Lymphoma

Lutetium (177Lu) lilotomab satetraxetan (177Lu-lilotomab) is a novel anti-CD37 radioimmunoconjugate (RIC) currently in Phase 2b clinical trial for treatment of non-Hodgkin lymphoma (NHL). The aim of the current study was to investigate tolerability and anti-tumor efficacy of multiple dosing of 177Lu-lilotomab in vivo. Nude mice with subcutaneous (s.c.) NHL (Ramos) xenografts were given 2, 3 or 4 weekly injections of 300 MBq/kg 177Lu-lilotomab or NaCl. Treatment tolerability was assessed by body weight, hematology and histopathological evaluation. A separate experiment in mice without xenografts was performed to collect long term toxicity data. Mice in this study were dosed more conservatively with the intention that all mice should survive until end of experiment and were administered 2 or 4 weekly injections of 200 MBq/kg 177Lu-lilotomab or NaCl. Total accumulated activity of 900 MBq/kg 177Lu-lilotomab given as three weekly injections was tolerated by nude mice with s.c. Ramos xenografts, which is 50 % higher than the maximum tolerated activity (MTA) of a single injection of 530-600 MBq/kg. In the long-term toxicity animals, the highest accumulated activity tested, 800 MBq/kg, was also well tolerated. Radioactivity-dependent transient reduction in white blood cell and platelet counts occurred in all treated groups, with nadir 1-3 weeks after last injection. This toxicity was radioactivity dependent and consistent with histopathological changes in hemolymphopoietic tissues. Significant tumor growth delay measured as time to reach 4 times intial tumor volume was observed in all groups given multiple injections of 300 MBq/kg 177Lu-lilotomab compared to NaCl control group (p<0.001). In conclusion, weekly injections of 177Lu-lilotomab increase the accumulated MTA from 530 to 900 MBq/kg in nude mice, allowing the total injected activity and hence the radiation dose to tumor to be increased without increasing the toxicity to normal tissues.

Lentiviral-Mediated Beclin-1 Overexpression Inhibits Cell Proliferation and Induces Autophagy of Human Esophageal Carcinoma Eca109 Cell Xenograft in Nude Mice

2020 ◽  
Vol 15 (1) ◽  
pp. 70-77
Author(s):  
Junhe Zhang ◽  
Weihua Dong
Keyword(s):  
Growth Rate ◽  
Cell Proliferation ◽  
Esophageal Carcinoma ◽  
Nude Mice ◽  
Beclin 1 ◽  
Control Group ◽  
Vector Group ◽  
Empty Vector ◽  
Empty Vector Group

Background: Esophageal carcinoma is one of the common malignant tumors in digestive tract. BECLIN-1 is a key gene that regulates autophagy, and its abnormal expression may be related with many human tumors. However, the mechanism of BECLIN-1 in esophageal carcinoma remains unknown. Objective: In this study, we explored the effect of BECLIN-1 overexpression on tumor growth in mice with esophageal carcinoma and its mechanism. Methods: Recombined lentiviral vector containing BECLIN-1 was used to transfect human esophageal carcinoma Eca109 cells and establish stable cell line. qRT-PCR was used to detect BECLIN-1 mRNA level in the transfected Eca109 cells, CCK-8 assay was used to detect cell proliferation. Beclin-1, P62 and LC3-II protein expression levels in Eca109 cells were detected using Western blot analysis. Subcutaneous xenograft nude mice model of human esophageal carcinoma was established, and the tumor growths in Beclin-1 group, control group and empty vector group were monitored. Beclin-1 protein expression in vivo was detected by immunohistochemistry. Results: Beclin-1 mRNA and protein were overexpressed in Eca109 cells. Compared with empty vector group, the growth rate of cells transfected with BECLIN-1 decreased significantly. Compared with the control group and empty vector group, the expression level of P62 protein in beclin-1 group was significantly decreased, while the expression level of LC3-II protein was significantly increased. The tumor growth rate in nude mice of Beclin-1 group was significantly lower than that of the control group and empty vector group, and Beclin-1 protein was mainly expressed in Beclin-1 group in vivo. Conclusion: BECLIN-1 can induce autophagy in esophageal carcinoma Eca109 cells, and it can significantly inhibit the growth of esophageal carcinoma.

scholarly journals Effects of Long-Term Supplementation of Bovine Colostrum on the Immune System in Young Female Basketball Players. Randomized Trial

Nutrients ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 118
Author(s):  
Anna Skarpańska-Stejnborn ◽  
Mirosława Cieślicka ◽  
Hanna Dziewiecka ◽  
Sławomir Kujawski ◽  
Anita Marcinkiewicz ◽  
...  
Keyword(s):  
Immune System ◽  
Physical Exercise ◽  
Exercise Program ◽  
Bovine Colostrum ◽  
Control Group ◽  
System Function ◽  
Basketball Players ◽  
Immune System Function ◽  
Experimental Group

An intensive physical exercise program could lead to a decrease in immune system function. Effects of long-term supplementation of bovine colostrum on the response of immune function on physical exercise test in athletes were examined. Twenty-seven elite female basketball players (age 16–19) were randomly assigned to either an experimental group or a control group. Eventually, n = 11 athletes completed intervention in the experimental group (3.2 g bovine colostrum orally twice a day for 24 weeks), while n = 9 athletes in the control group were given a placebo. Before the supplementation, after 3 and 6 months, subjects performed the physical exercise stress test. Before, just after, and 3 h after physical exercise testing, blood was drawn and immune system indicators were examined. Plasma interleukin (IL)-1alpha, IL-2, IL-10, IL-13, tumor necrosis factor (TNF) alpha, creatine kinase (CK MM), immunoglobulin G (IgG), insulin-like growth factor 1 (IGF1), and WBC, lymphocyte (LYM), monocyte (MON), and granulocyte (GRA) were measured. A statistically significant change in IL-10 in response to the exercise program during the supplementation period in both groups was observed (p = 0.01). However, the results of the rest of the comparisons were statistically insignificant (p > 0.05). Contrary to our initial hypothesis, there were no significant effects of bovine supplementation on the dynamics of immune system function indicators.

Follicle development in cryopreserved bitch ovarian tissue grafted to immunodeficient mouse

2012 ◽  
Vol 24 (3) ◽  
pp. 461 ◽  
Author(s):  
L. Commin ◽  
S. Buff ◽  
E. Rosset ◽  
C. Galet ◽  
A. Allard ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Histological Analysis ◽  
Smooth Muscle Actin ◽  
Ovarian Tissue ◽  
Follicular Development ◽  
Slow Freezing ◽  
Angiogenic Factor ◽  
Muscle Actin ◽  
Once Daily

The present study evaluated: (1) in vivo follicular development in canine ovarian tissue after slow freezing and xenotransplantation; and (2) the use of erythropoietin (EPO) as an angiogenic factor to optimise the transplantation procedure. Frozen–thawed ovarian tissue from five bitches was grafted into severe combined immunodeficient (SCID) mice (n = 47) treated with or without EPO (500 IU kg–1, once daily for 3 days) (Groups A and B, respectively) and analysed after 0, 1, 8 or 16 weeks. Follicle grade, follicle density, follicle morphology and stromal cells density were assessed by histological analysis, whereas vascularisation of the graft was quantified by immunohistochemistry with anti-α-smooth muscle actin antibody. Despite a massive loss of follicles after grafting, secondary follicle density was higher at 8 and 16 weeks than at 1 week regardless of EPO treatment. EPO significantly improved early follicle morphology and stromal cell density after 8 weeks and blood vessel density at 16 weeks after transplantation (P < 0.05). Intact secondary follicles with more than three granulosa cells layers were observed 16 weeks after transplantation. The results suggest that canine ovarian tissue can be successfully preserved by our slow-freezing protocol because the tissue showed follicular growth after xenotransplantation. EPO treatment did not lessen the massive loss of follicles after transplantation.

scholarly journals Stromal cells from human long-term marrow cultures are mesenchymal cells that differentiate following a vascular smooth muscle differentiation pathway

Blood ◽  
1993 ◽  
Vol 82 (1) ◽  
pp. 66-76 ◽  
Author(s):  
MC Galmiche ◽  
VE Koteliansky ◽  
J Briere ◽  
P Herve ◽  
P Charbord
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Stromal Cells ◽  
Muscle Cells ◽  
Mesenchymal Cells ◽  
Myoid Cells ◽  
Marrow Cultures

In human long-term marrow cultures connective tissue-forming stromal cells are an essential cellular component of the adherent layer where granulomonocytic progenitors are generated from week 2 onward. We have previously found that most stromal cells in confluent cultures were stained by monoclonal antibodies directed against smooth muscle- specific actin isoforms. The present study was carried out to evaluate the time course of alpha-SM-positive stromal cells and to search for other cytoskeletal proteins specific for smooth muscle cells. It was found that the expression of alpha-SM in stromal cells was time dependent. Most of the adherent spindle-shaped, vimentin-positive stromal cells observed during the first 2 weeks of culture were alpha- SM negative. On the contrary, from week 3 to week 7, most interdigitated stromal cells contained stress fibers whose backbone was made of alpha-SM-positive microfilaments. In addition, in confluent cultures, other proteins specific for smooth muscle were detected: metavinculin, h-caldesmon, smooth muscle myosin heavy chains, and calponin. This study confirms the similarity between stromal cells and smooth muscle cells. Moreover, our results reveal that cells in vivo with the phenotype closest to that of stromal cells are immature fetal smooth muscle cells and subendothelial intimal smooth muscle cells; a cell subset with limited development following birth but extensively recruited in atherosclerotic lesions. Stromal cells very probably derive from mesenchymal cells that differentiate along this distinctive vascular smooth muscle cell pathway. In humans, this differentiation seems crucial for the maintenance of granulomonopoiesis. These in vitro studies were completed by examination of trephine bone marrow biopsies from adults without hematologic abnormalities. These studies revealed the presence of alpha-SM-positive cells at diverse locations: vascular smooth muscle cells in the media of arteries and arterioles, pericytes lining capillaries, myoid cells lining sinuses at the abluminal side of endothelial cells or found within the hematopoietic logettes, and endosteal cells lining bone trabeculae. More or less mature cells of the granulocytic series were in intimate contact with the thin cytoplasmic extensions of myoid cells. Myoid cells may be the in vivo counterpart of stromal cells with the above-described vascular smooth muscle phenotype.

scholarly journals 5-Aminolevulinic Acid-Based Photodynamic Therapy Pretreatment Mitigates Ultraviolet A-Induced Oxidative Photodamage

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Hui Hua ◽  
Jiawei Cheng ◽  
Wenbo Bu ◽  
Juan Liu ◽  
Weiwei Ma ◽  
...  
Keyword(s):  
Aminolevulinic Acid ◽  
Epidermal Keratinocytes ◽  
Control Group ◽  
Hacat Cells ◽  
Ultraviolet A ◽  
Hairless Mice ◽  
Human Epidermal Keratinocytes ◽  
Experimental Group

Aim. To determine whether 5-aminolevulinic acid-based photodynamic therapy (ALA-PDT) is effective in combating ultraviolet A- (UVA-) induced oxidative photodamage of hairless mice skin in vivo and human epidermal keratinocytes in vitro. Methods. In in vitro experiments, the human keratinocyte cell line (HaCaT cells) was divided into two groups: the experimental group was treated with ALA-PDT and the control group was left untreated. Then, the experimental group and the control group of cells were exposed to 10 J/m2 of UVA radiation. ROS, O2− species, and MMP were determined by fluorescence microscopy; p53, OGG1, and XPC were determined by Western blot analysis; apoptosis was determined by flow cytometry; and 8-oxo-dG was determined by immunofluorescence. Moreover, HaCaT cells were also treated with ALA-PDT. Then, SOD1 and SOD2 were examined by Western blot analysis. In in vivo experiments, the dorsal skin of hairless mice was treated with ALA-PDT or saline-PDT, and then, they were exposed to 20 J/m2 UVA light. The compound 8-oxo-dG was detected by immunofluorescence. Conclusion. In human epidermal keratinocytes and hairless mice skin, UVA-induced oxidative damage can be prevented effectively with ALA-PDT pretreatment.

scholarly journals The effect of iodine and strumigens long-term foodborne intake on histometrical parameters of thyroid gland in gimmers

2013 ◽  
Vol 61 (5) ◽  
pp. 1365-1369
Author(s):  
Zdeněk Peksa ◽  
Jan Trávníček ◽  
Roman Konečný ◽  
František Jelínek ◽  
Hana Dušová ◽  
...  
Keyword(s):  
Thyroid Gland ◽  
Dry Matter ◽  
Experimental Period ◽  
Rapeseed Meal ◽  
Iodine Intake ◽  
Control Group ◽  
High Iodine ◽  
Average Size ◽  
Experimental Group

In 2010 and 2011 two experiments on gimmers of Šumava mountain sheep were performed. The first experiment was carried out on 12 animals (experimental period was 11 months); control group was fed with 3 mg I*kg−1 in dry matter per day and experimental group was fed with 5 mg I*kg−1 in dry matter per day. The second experiment lasted 10 months and it was carried out on 12 animals. Feed ration for control group contained 10 mg I*kg DM−1. Feed ration for experimental group contained 10 mg I*kg DM−1 too; moreover it included rapeseed meal and 1 g of sodium nitrate. The aim of the first experiment was focused on impact of high iodine intake on structure of thyroid gland. The aim of the second experiment was to discover effect of strumigens during high iodine intake on structure of thyroid gland. The animals were slaughtered after the experiment and there was executed the dissection of thyroid gland. The samples of thyroid gland were processed during classic paraffin method and dyed with haematoxylin and eosin. For finding of histometrical parameters was used program Leica IM 500 Version 4.0. The length, the width and the area of follicles were measured. The follicles were divided into three groups after this procedure (by the length); in each group were measured 20 thyreocytes. In group with intake 5 mg I*kg DM−1 (experiment from year 2010): higher weight of thyroid (p < 0.01), significant higher (p < 0.05) proportional representation of large and medium follicles, significantly (p < 0.05) higher average size of follicles, demonstrably lower (p < 0.05) height of epithelium were found. There were not found any differences in monitored parameters between the groups from experiment which was carried out during the year 2011. After comparing results from the both experiments significantly higher percentage representation of large follicles a significantly lower representation of small follicles, distinctly higher average size and higher height of epithelium in all size categories, in groups with iodine intake 10 mg I*kg DM−1was found (from second experiment – year 2011).

scholarly journals Piglets produced by transfer of embryos obtained by in vitro fertilization of oocytes matured in vitro with thymosin: A case report

2020 ◽  
Vol 76 (03) ◽  
pp. 6356-2020 ◽  
Author(s):  
KATARZYNA PONIEDZIAŁEK-KEMPNY ◽  
BARBARA GAJDA ◽  
IWONA RAJSKA ◽  
LECHOSŁAW GAJDA ◽  
ZDZISŁAW SMORĄG
Keyword(s):  
Case Report ◽  
In Vitro Fertilization ◽  
Control Group ◽  
First Birth ◽  
Vitro Fertilization ◽  
Research Material ◽  
Preliminary Study ◽  
Experimental Group

The aim of the study was to examine the in vivo viability of in vitro-produced (IVP) porcine embryos obtained from oocytes matured with thymosin. The research material for this study consisted of immature pig oocytes obtained from ovaries after slaughter and ejaculated semen obtained from one boar. The immature oocytes were cultured in vitro until the metaphase II stage in a medium supplemented with thymosin (TMS). The presumptive zygotes obtained were cultured in vitro for 4-40 hours. The presumptive zygotes and 2-4-cell embryos were evaluated in vivo after transferring them to synchronized recipients. After the transfer of embryos from the experimental group into 2 recipients (50 embryos into each gilt) and the transfer of 50 embryos from the control group into 1 recipient, both gilts that had received embryos obtained by in vitro fertilization of oocytes matured with TMS became pregnant and delivered a total of 16 live piglets. After the transfer of embryos from the control group, no pregnancy was achieved. In conclusion, the results of our preliminary study suggest that the maturation of pig oocytes with thymosin supports the in vivo survival of in vitro produced embryos. It is important to note, that this was the first birth of piglets obtained after transfer of IVP embryos in Poland.

scholarly journals Longitudinal impact and effects of booster sessions in a cognitive training program for healthy elderly

2020 ◽  
Author(s):  
Lucas Matias Felix ◽  
Marcela Mansur Alves ◽  
Mariana Teles ◽  
Laura Jamison ◽  
Hudson Golino
Keyword(s):  
Older Adults ◽  
Cognitive Training ◽  
Passive Control ◽  
Control Group ◽  
Longitudinal Impact ◽  
Healthy Older Adults ◽  
Booster Sessions ◽  
Experimental Group

This paper reports the results from a three-years follow-up study to access the long-term efficacy of a cognitive training for healthy older adults and investigates the effects of booster sessions on the cognitive performance of the participants using an innovative analytical approach from information theory. Design: semi-randomized quasi-experimental controlled design. Participants: 50 healthy older adults, (M = 73.3, SD = 7.77) were assigned into an experimental (N = 25; Mean age = 73.9; SD = 8.62) and a passive control group (N = 25; mean age = 72.9; SD = 6.97). Instruments: six subtests of WAIS and two episodic memory tasks. Procedures: the participants were assessed in four occasions: after the end of the original intervention, pre-booster sessions (three years after the original intervention), immediately after the booster sessions and three months after the booster sessions. Results: the repeated measures ANOVA showed that two gains reported in the original intervention were identified in the follow-up: Coding (F(1, 44) = 11.79, MSE = 0.77, p = .001, ηˆG2 = .084) and Picture Completion (F(1, 47) = 10.01, MSE = 0.73, p = .003, ηˆG2 = .060). After the booster sessions, all variables presented a significant interaction between group and time favorable to the experimental group (moderate to high effect sizes). To compare the level of cohesion of the cognitive variables between the groups, an entropy-based metric was used. The experimental group presented a lower level of cohesion in three of the four measurement occasions, suggesting a differential impact of the intervention with immediate and short-term effects, but without long-term effects.

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