Autologous Oxygen-Releasing Nano-Biomimetic Scaffold with Chondrocytes Promotes Joint Repair After Trauma

Our study assesses the role of a scaffold constructed by co-culture of autologous oxygen-releasing biomimetic scaffold (AONS) and chondrocytes in joint repair after trauma. A composite scaffold structure was used and a scaffold constructed of AONS and chondrocytes was transplanted into SD rats to create models of patellar cartilage fracture and hip osteochondral fracture, respectively followed by analysis of cell proliferation by immunofluorescence method, osteogenesis-related gene expression by RT-PCR, chondrocytes apoptosis by TUNEL staining. The blank control group and AONS composite chondrocytes have significant differences in apoptosis and cell proliferation of two fracture types (P <0.05). The autologous oxygen-releasing nanometers at 4 and 8 weeks showed a significant difference in the number of PCNA and TUNEL cells between biomimetic scaffold and chondrocytes in two groups (P < 0.05). The AONS and chondrocytes were effective for two types of fractures at 1, 4 and 8 weeks. The expression of various markers of intrachondral osteogenesis was decreased and the markers of hip osteochondral fracture were increased significantly (P < 0.05). Joint recovery was better than patellar cartilage fractures. The AONS composite chondrocyte scaffold promotes repair of patellar cartilage fractures and hip osteochondral fractures with a better effect on hip osteochondral fractures.

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Cellular Compatibility of a New Tantalum-Niobium Scaffold Material

10.21203/rs.3.rs-39637/v1 ◽

2020 ◽

Author(s):

Jianan Ouyang ◽

Zhenhan Deng ◽

Kang Chen ◽

Jianyi Xiong ◽

Ying Li ◽

...

Keyword(s):

Cell Proliferation ◽

Type I Collagen ◽

Material Surface ◽

Control Group ◽

Type I ◽

Scaffold Material ◽

Rt Pcr ◽

Pcr Assay ◽

Porous Tantalum ◽

Significant Difference

Abstract [Objective] To determine the cellular compatibility of porous tantalum-niobium (Ta-Nb) material. [Method] Rabbit osteoblasts were co-cultured with porous Ta-Nb material. The cell proliferation was detected by CCK-8 method, and the cell adhesion was observed under scanning electron microscope (SEM). The expressions of type-I collagen and osteocalcin were detected by RT-PCR assay. [Results] CCK-8 detection indicated that the cell proliferation on the porous Ta-Nb material showed no difference from that of the control group (P>0.05). SEM revealed that a large amount of cells adhered onto the surface and in the pores of the material. The number of cells on the material surface increased obviously over time. RT-PCR assay showed that with the prolonging of the time of co-culture, the expression of type-I collagen was enhanced (P<0.05), while the osteocalcin expression exhibited no significant difference (P>0.05[Conclusion] Porous Ta-Nb scaffold material can be used to promote the adhesion, growth and differentiation of osteoblasts with satisfactory cellular compatibility.

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Abnormal Osteoblastic Response to Leptin in Patients with Adolescent Idiopathic Scoliosis

Scientific Reports ◽

10.1038/s41598-019-53757-3 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 2

Author(s):

Gene Chi-Wai Man ◽

Elisa Man-Shan Tam ◽

Yi Shun Wong ◽

Vivian Wing-Ying Hung ◽

Zongshan Hu ◽

...

Keyword(s):

Cell Proliferation ◽

Adolescent Idiopathic Scoliosis ◽

Idiopathic Scoliosis ◽

Leptin Receptor ◽

Unknown Etiology ◽

Control Group ◽

Functional Responses ◽

Osteoblastic Activity ◽

Significant Difference

AbstractAdolescent idiopathic scoliosis (AIS) is a complex three-dimensional structural deformity of the spine with unknown etiology. Although leptin has been postulated as one of the etiologic factors in AIS, its effects on osteoblastic activity remain unknown. Herein, we conducted this study to investigate whether there are abnormal functional responses to leptin and abnormal expression of leptin receptor in AIS osteoblasts. In vitro assays were performed with osteoblasts isolated from 12 severe AIS girls and 6 non-AIS controls. The osteoblasts were exposed to different concentrations of leptin (0, 10, 100, 1000 ng/mL). The effects of leptin on cell proliferation, differentiation and mineralization were determined. Protein expressions of leptin receptor (LEP-R) under basal and osteogenic conditions were also evaluated by Western blot. Our results showed that leptin significantly stimulated osteoblasts from non-AIS subjects to proliferate, differentiate and mineralized. However, in the AIS group, the stimulatory effects of leptin on cell proliferation, differentiation, and mineralization were not observed. In addition, no statistically significant difference in the expression of leptin receptor under both basal and osteogenic conditions was found between AIS and control group. In conclusion, these findings might help to explain the low bone mass and deranged bone quality that is clinically associated with AIS girls.

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Cryopreservation Effect on Proliferative and Chondrogenic Potential of Human Chondrocytes Isolated from Superficial and Deep Cartilage

The Open Orthopaedics Journal ◽

10.2174/1874325001206010150 ◽

2012 ◽

Vol 6 (1) ◽

pp. 150-159 ◽

Cited By ~ 18

Author(s):

Emma Muiños-López ◽

Mª Esther Rendal-Vázquez ◽

Tamara Hermida-Gómez ◽

Isaac Fuentes-Boquete ◽

Silvia Díaz-Prado ◽

...

Keyword(s):

Cell Proliferation ◽

Articular Cartilage ◽

Protein Expression ◽

Articular Chondrocytes ◽

Control Group ◽

Proliferative Capacity ◽

Superficial Zone ◽

Healthy Human ◽

Chondrogenic Potential ◽

Significant Difference

Objectives:To compare the proliferative and chondrogenic potential of fresh and frozen chondrocytes isolated from superficial and deep articular cartilage biopsies.Materials and Methodology:The study included 12 samples of fresh and frozen healthy human knee articular cartilage. Cell proliferation was tested at 3, 6 and 9 days. Studies of mRNA quantification, protein expression and immunofluorescence for proliferation and chondrogenic markers were performed.Results:Stimulation of fresh and frozen chondrocytes from both superficial and deep cartilage with fetal bovine serum produced an increase in the proliferative capacity compared to the non-stimulated control group. In the stimulated fresh cells group, the proliferative capacity of cells from the deep biopsy was greater than that from cells from the superficial biopsy (0.046vs0.028, respectively, p<0.05). There was also a significant difference between the proliferative capacity of superficial zone fresh (0.028) and frozen (0.051) chondrocytes (p<0.05).CCND1mRNA and protein expression levels, and immunopositivity forKi67revealed a higher proliferative capacity for fresh articular chondrocytes from deep cartilage. Regarding the chondrogenic potential, stimulated fresh cells showed higherSOX9andCol IIexpression in chondrocytes from deep than from superficial zone (p<0.05,Tstudent test).Conclusions:The highest rate of cell proliferation and chondrogenic potential of fresh chondrocytes was found in cells obtained from deep cartilage biopsies, whereas there were no statistically significant differences in proliferative and chondrogenic capacity between biopsy origins with frozen chondrocytes. These results indicate that both origin and cryopreservation affect the proliferative and chondrogenic potential of chondrocytes.

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THE POTENTIAL OF HEDYOTIS CORYMBOSA (L.) LAMK EXTRACT TO INHIBIT THE PROGRESSIVITY OF ORAL CANCER CELL IN RATS INDUCED WITH BENZOPYRENE

Dentika Dental Journal ◽

10.32734/dentika.v19i1.142 ◽

2016 ◽

Vol 19 (1) ◽

pp. 22-27

Author(s):

Theresia Indah Budhy

Keyword(s):

Cell Proliferation ◽

Treatment Group ◽

Oral Cancer ◽

Cancer Cell ◽

Control Treatment ◽

Control Group ◽

Cancer Cell Proliferation ◽

Control Group Design ◽

Significant Difference ◽

Group 2

Cancer is still ranked as the fifth cause of mortality and morbidityin Indonesia.Hedyotis corymbosa (L.) Lamk has ursolat acid as anti-proliferative cancer cell. This research is aimed to determine the potency of Hedyotis corymbosa (L.) Lamk at different doses, namely 375, 750, and 1500 mg/kg, used as an inhibitor for the progressivity of oral cancer, such as proliferation, angiogenesis, and apoptosis of cancer cells. Post test only control group design was used in this research. There were 24 Rattus novergicus used as research samples. Those were divided into four groups, namely control, treatment group 1 with a dose of 375mg/kg, treatment group 2 with a dose of 750 mg/kg, and treatment group 3 with a dose of 1500mg/kg. Their oral cavity was induced intramusculary by benzopyrene with a dose of 8mg/kg for 4 weeks (twice a week) to create cancer. Hedyotis corymbosa (L.) Lamk was given orally for 10 days. All samples were aclimatitation to perform Histo Pathology Anatomi among groups. Haematoxillin Eosin for proliferation cancer cell and capilary. Immunohistochemistry for expression of caspase3. Data were tabulated andanalyzed statistically by ANOVA. There was significant difference of cancer cell proliferation and capilary between control and treatment groups. The most significant decreasing of cancer cell proliferation was in those samples given with a dose of 750 mg/kg. Meanwhile, the highest apoptosis of caspase3 expression was in those samples given with a dose of 750 mg/kg. It can be concluded that Hedyotis corymbosa (L.) Lamk extract could decrease cancer cell proliferation and capilary as well as could increase apoptosis.

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Cancer-Associated Fibroblasts Regulate Bladder Cancer Invasion and Metabolic Phenotypes through Autophagy

Disease Markers ◽

10.1155/2021/6645220 ◽

2021 ◽

Vol 2021 ◽

pp. 1-8

Author(s):

Dahai Dong ◽

Yu Yao ◽

Jinlei Song ◽

Lijiang Sun ◽

Guiming Zhang

Keyword(s):

Cell Proliferation ◽

Bladder Cancer ◽

Aerobic Glycolysis ◽

Lactate Concentration ◽

Control Group ◽

Cancer Associated Fibroblasts ◽

Metabolic Phenotypes ◽

T24 Cells ◽

Significant Difference

Recently, both cancer-associated fibroblasts (CAFs) and autophagy have been proven to play an important role in tumor development, including bladder cancer (BCa). However, the real mechanisms remain largely unclear. Here, we reconstruct a mimic tumor microenvironment to explore the interaction between CAFs and the BCa cell line T24 using a coculture system. Autophagy in CAFs was induced or inhibited by rapamycin or siRNA, respectively. After coculture with CAFs, T24 cell proliferation, invasion, and aerobic glycolysis were tested in vitro. Rapamycin induced and siAtg5 inhibited autophagy in CAFs. Enhanced autophagy in CAFs promoted cell proliferation and invasion in T24 cells in vitro, while there was no significant difference between the autophagy-inhibited group and the controls. Lactate concentration was elevated in both rapamycin-treated and siAtg5-treated groups compared with the control group. In addition, the expression levels of MCT1, MCT4, HK2, SLC2A1, and MMP-9 were all increased in T24 cells in the autophagy-enhanced group. Our results indicated that CAFs could regulate BCa invasion and metabolic phenotypes through autophagy, providing us with new alternative treatments for BCa in the future.

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Hyperplasia of Wistar rat tongue mucosa due to exposure to cigarette side-stream smoke

Dental Journal (Majalah Kedokteran Gigi) ◽

10.20473/j.djmkg.v52.i3.p133-137 ◽

2019 ◽

Vol 52 (3) ◽

pp. 133

Author(s):

Nurina Febriyanti Ayuningtyas ◽

Grahania Octaviono Mahardika ◽

Bagus Soebadi ◽

Adiastuti Endah Permadiati ◽

Saka Winias ◽

...

Keyword(s):

Cell Proliferation ◽

Treatment Group ◽

Oral Cancer ◽

Epithelial Cell ◽

Cigarette Smoke ◽

Wistar Rats ◽

Control Group ◽

Epithelial Cell Proliferation ◽

Sidestream Smoke ◽

Significant Difference

Background: Hyperplasia, a condition whereby an excessive number of cells are produced due to their uncontrolled division, represents a common symptom of carcinogenesis. Cancer is a physical manifestation of cell malignancy resulting from abnormal proliferation. Globally, oral cancer currently constitutes the sixth largest lethal form of the condition. The most common etiology of oral cancer is tobacco of which cigarettes are the most popular related product. The health risks associated with cigarette smoke not only affect active smokers but also individuals who ingest it passively. Sidestream smoke comes from the lighted end of a burning tobacco product such as a cigarette, pipe or cigar and contains nicotine and many harmful cancer-causing chemicals. Inhaling sidestream smoke increases the risk of lung and other types of cancer. Purpose: The purpose of this study was to understand how sidestream cigarette smoke initiates precancerous changes, in this case hyperplasia, in the oral mucosa epithelium of Wistar rats. Methods: The subjects were divided into three groups, a 4-week treatment group (P1), an 8-week treatment group (P2), and a control group (K), each consisting of ten subjects. The subjects were exposed to a daily two-cigarette dose of smoke. The experiment used a post-test only control group design. All samples were sacrificed during the fourth and eighth weeks. Haematoxylin-eosin staining was performed on the tongues of the Wistar rats to establish the presence of hyperplasia. Data was analyzed using a one-way ANOVA test. Results: After the Wistar rats had been exposed to cigarette smoke, an increased degree of epithelial cell proliferation (hyperplasia) showed a significant difference with a p-value <0.05 during the eighth week. Conclusion: Exposure to cigarette sidestream smoke induces increased epithelial cell proliferation (hyperplasia) in Wistar rats.

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Protective Effect of Mitochondrial ND2 C5178A Gene Mutation on Cell and Mitochondrial Functions

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/4728714 ◽

2021 ◽

Vol 2021 ◽

pp. 1-10

Author(s):

Liuyang Tian ◽

Chao Zhu ◽

Huanwan Yang ◽

Yang Li ◽

Yuqi Liu

Keyword(s):

Cell Proliferation ◽

Gene Mutation ◽

Atp Synthesis ◽

Protein Translation ◽

Control Group ◽

Protective Effects ◽

Mitochondrial Oxidative Stress ◽

Nadh Dehydrogenase Subunit 2 ◽

Mitochondrial Functions ◽

Significant Difference

Background. Mitochondrial NADH dehydrogenase subunit 2 (MT-ND2) m. 5178C>A gene mutation has protective effects against various diseases, but the molecular mechanism is still unclear. In previous study, we found a heteroplasmy level of MT-ND2 m. 5178C>A mutation in normotensive controls. Peripheral blood samples were obtained from essential hypertension individuals carrying the mutation and healthy controls without gene mutation to establish immortalized lymphocyte lines. To investigate the effect of the MT-ND2 m. 5178C>A gene mutation, comparative analyses of the two group cell lines were performed, including measurements of cell proliferation, viability, ATP synthesis, mitochondrial oxidative stress, and oxidative phosphorylation. Results. The cell proliferation rate and viability of the MT-ND2 m. 5178C>A mutant lymphocyte line were higher than those of the control group. Mitochondrial functions of the MT-ND2 m. 5178C>A mutant lymphocyte were increased, including increased ATP synthesis, decreased ROS production, increased mitochondrial membrane potential and Bcl-2 gene transcription and protein translation, decreased Caspase 3/7 activity, and decreased early apoptosis and late apoptosis. The oxygen consumption rate (OCR) of the mutant lymphocyte line was higher than that of the control group, including basal OCR, ATP-linked OCR, maximal OCR, proton leak OCR, and reserve OCR, and there was no significant difference in nonmitochondrial OCR. The activity of Mitochondrial Complex I of the mutant group was increased than that of the control group. Conclusions. The MT-ND2 m. 5178C>A mutation is a protective mutation that may be related to improvement of mitochondrial functions and decrease in apoptosis.

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Differential Gene Expression Involved In Angiogenesis, Metabolism, Cell Proliferation and Self-Renewal and Pluripotency In Myelodysplastic Syndromes (MDS) and Acute Myeloid Leukemia (AML)

Blood ◽

10.1182/blood.v116.21.4646.4646 ◽

2010 ◽

Vol 116 (21) ◽

pp. 4646-4646

Author(s):

Jose F Falantes ◽

Cristina Calderón ◽

Pablo Trujillo ◽

Beatriz Martin-Antonio ◽

Jose González ◽

...

Keyword(s):

Gene Expression ◽

Cell Proliferation ◽

Acute Myeloid Leukemia ◽

Myelodysplastic Syndromes ◽

Myeloid Leukemia ◽

Control Group ◽

Mrna Levels ◽

Self Renewal ◽

Significant Difference ◽

Acute Myeloid

Abstract Abstract 4646 Background Myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) are clonal disorders characterized by clinical and prognostic heterogeneity, mainly explained by different genetic abnormalities among other factors. The role of some key genes involved in disease pathogenesis in both entities remain unclear. Methods and patients Study objective was to analyse differences in gene expression related to angiogenesis, metabolism and cell proliferation, self-renewal and pluripotency in patients (pts) with MDS and AML. Thirty-three bone marrow (BM) samples at diagnosis were analysed and distributed in 4 different groups: control group (n=8), low-risk MDS (LR-MDS:<10% BM blasts; n=15), high-risk MDS (HR-MDS:>10% BM blasts; n=4) and AML (n=6). Total RNA was isolated from BM samples. Genes analysed were: vascular endothelial growth factor (VEGF) for angiogenesis, MYC, macrophage migration inhibitory factor (MIF) and glycogen synthase (GYS) for metabolism and cell-proliferation and Oct4 as transcription factor required to maintain an undifferentiated state for self-renewal and pluripotency. Gene expression was quantified by qRT-PCR in triplicate using ß-actin gene as control. SPSS software (v.16) and Mann-Whitney U-test were applied for statistical analysis (P value ≤.05 was considered significant in all cases). Results After analysis, only mRNA levels of VEGF and MYC showed significant difference between LR-MDS and the control group. However, higher expression of all genes were observed in HR-MDS vs LR-MDS (p≤.05) as shown in table 1. When compared HR-MDS to AML, no difference was observed in VEGF, MYC, MIF and GYS, but significant difference was noticed in mRNA expression of Oct4 in AML samples (p=.032) vs HR-MDS. Globally, gene expression in MDS (pts with LR and HR-MDS) was significant lower than in AML pts in all genes studied as expected. Conclusions Increased expression of VEGF, cMYC, MIF, GYS and OCT4 in HR-MDS vs LR-MDS and in AML vs MDS (global) suggests that these factors may play a relevant role in pathogenesis of both entities. These results point towards a different biological behaviour in less proliferative disease respect advanced stages and AML in different cellular pathways involved in disease progression. They might also justify clinical heterogeneity among patients with MDS and in patients with AML vs MDS as a sole group, and also being responsible for different response to treatment options. Analysis of protein expression is ongoing. Disclosures: No relevant conflicts of interest to declare.

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Levels of Prothrombin Fragment F1+2 in Patients with Hyperhomocysteinemia and a History of Venous Thromboembolism

Thrombosis and Haemostasis ◽

10.1055/s-0038-1665405 ◽

1997 ◽

Vol 78 (05) ◽

pp. 1327-1331 ◽

Cited By ~ 21

Author(s):

Paul A Kyrle ◽

Andreas Stümpflen ◽

Mirko Hirschl ◽

Christine Bialonczyk ◽

Kurt Herkner ◽

...

Keyword(s):

Venous Thromboembolism ◽

Thrombin Generation ◽

Oral Anticoagulant Therapy ◽

Oral Anticoagulants ◽

Control Group ◽

Prothrombin Fragment ◽

Hemostatic System ◽

Significant Difference ◽

System Activation ◽

One Year

SummaryIncreased thrombin generation occurs in many individuals with inherited defects in the antithrombin or protein C anticoagulant pathways and is also seen in patients with thrombosis without a defined clotting abnormality. Hyperhomocysteinemia (H-HC) is an important risk factor of venous thromboembolism (VTE). We prospectively followed 48 patients with H-HC (median age 62 years, range 26-83; 18 males) and 183 patients (median age 50 years, range 18-85; 83 males) without H-HC for a period of up to one year. Prothrombin fragment Fl+2 (Fl+2) was determined in the patient’s plasma as a measure of thrombin generation during and at several time points after discontinuation of secondary thromboprophylaxis with oral anticoagulants. While on anticoagulants, patients with H-HC had significantly higher Fl+2 levels than patients without H-HC (mean 0.52 ± 0.49 nmol/1, median 0.4, range 0.2-2.8, versus 0.36 ± 0.2 nmol/1, median 0.3, range 0.1-2.1; p = 0.02). Three weeks and 3,6,9 and 12 months after discontinuation of oral anticoagulants, up to 20% of the patients with H-HC and 5 to 6% without H-HC had higher Fl+2 levels than a corresponding age- and sex-matched control group. 16% of the patients with H-HC and 4% of the patients without H-HC had either Fl+2 levels above the upper limit of normal controls at least at 2 occasions or (an) elevated Fl+2 level(s) followed by recurrent VTE. No statistical significant difference in the Fl+2 levels was seen between patients with and without H-HC. We conclude that a permanent hemostatic system activation is detectable in a proportion of patients with H-HC after discontinuation of oral anticoagulant therapy following VTE. Furthermore, secondary thromboprophylaxis with conventional doses of oral anticoagulants may not be sufficient to suppress hemostatic system activation in patients with H-HC.

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Viscosity and Red Cell Deformity in Arterial Disease

10.1055/s-0038-1665789 ◽

1979 ◽

Author(s):

G Cella ◽

H de Haas ◽

M Rampling ◽

V Kakkar

Keyword(s):

Vascular Disease ◽

Blood Viscosity ◽

Whole Blood ◽

Arterial Disease ◽

Plasma Viscosity ◽

Control Group ◽

Fibrinogen Concentration ◽

Red Cell ◽

Whole Blood Viscosity ◽

Significant Difference

Haemorrheological factors have been shown to be affected in many kings of vascular disease. The present study was undertaken to correlate these factors in normal subjects and patients suffering from peripheral arterial disease. Twenty-two patients were investigated; they had moderate or severe intermittent claudication, extent of disease being confirmed by aorto-arteriography and ankle-systolic pressure studies. Twenty-five controls with no symptoms or signs of arterial disease were selected with comparable age and sex distribution. Whole blood viscosity was measured at shear rates of 230 secs-1 and 23 secs-lat 37°c using a Wells Brookfield cone plate microvisco meter. Plasma viscosity was also measured in an identical manner. Erythrocyte flexibility was measured by centrifuge technique and fibrinogen concentration as well as haematocrit by standard techniques. The fibrinogen concentration appeared to be the only significant parameter; the mean concentration in patients with peripheral vascular disease of 463 ± 73mg/l00ml in the control group ( < 0.05). Although whole blood viscosity was high in patients, when corrected to a common haematocrit, there was no significant difference between patients and controls. The same megative correlation was found for plasma viscosity. The red cell flexibility was found to be increased in patients as compared to the control group, but this effect appeared to be simply proportional to the fibrinogen concentration.

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