scholarly journals Preliminary In Vitro Evaluation of Chitosan–Graphene Oxide Scaffolds on Osteoblastic Adhesion, Proliferation, and Early Differentiation

2020 ◽  
Vol 21 (15) ◽  
pp. 5202 ◽  
Author(s):  
Sonia How Ming Wong ◽  
Siew Shee Lim ◽  
Timm Joyce Tiong ◽  
Pau Loke Show ◽  
Hayyiratul Fatimah Mohd Zaid ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Graphene Oxide ◽  
Cell Adhesion ◽  
Chitosan Solution ◽  
Early Differentiation ◽  
Chitosan Matrix ◽  
Chitosan Scaffolds ◽  
Diphenyl Tetrazolium Bromide ◽  
Differentiation Marker

An ideal scaffold should be biocompatible, having appropriate microstructure, excellent mechanical strength yet degrades. Chitosan exhibits most of these exceptional properties, but it is always associated with sub-optimal cytocompatibility. This study aimed to incorporate graphene oxide at wt % of 0, 2, 4, and 6 into chitosan matrix via direct blending of chitosan solution and graphene oxide, freezing, and freeze drying. Cell fixation, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, alkaline phosphatase colorimetric assays were conducted to assess cell adhesion, proliferation, and early differentiation of MG63 on chitosan–graphene oxide scaffolds respectively. The presence of alkaline phosphatase, an early osteoblast differentiation marker, was further detected in chitosan–graphene oxide scaffolds using western blot. These results strongly supported that chitosan scaffolds loaded with graphene oxide at 2 wt % mediated cell adhesion, proliferation, and early differentiation due to the presence of oxygen-containing functional groups of graphene oxide. Therefore, chitosan scaffolds loaded with graphene oxide at 2 wt % showed the potential to be developed into functional bone scaffolds.

The influence of titanium surfaces treated by alkalis on macrophage and osteoblast-like cell adhesion and gene expression in vitro

RSC Advances ◽  
2015 ◽  
Vol 5 (99) ◽  
pp. 81378-81387 ◽  
Author(s):  
Ting Ma ◽  
Xi-Yuan Ge ◽  
Sheng-Nan Jia ◽  
Xi Jiang ◽  
Yu Zhang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Alkaline Phosphatase ◽  
Cell Adhesion ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Related Gene ◽  
Titanium Surfaces

The effect of alkali-treated titanium surfaces on inflammation-related gene expression of macrophages and alkaline phosphatase activity of osteoblast-like cells.

scholarly journals Gradient Chitosan Hydrogels Modified with Graphene Derivatives and Hydroxyapatite: Physiochemical Properties and Initial Cytocompatibility Evaluation

2020 ◽  
Vol 21 (14) ◽  
pp. 4888
Author(s):  
Karolina Kosowska ◽  
Patrycja Domalik-Pyzik ◽  
Małgorzata Sekuła-Stryjewska ◽  
Sylwia Noga ◽  
Joanna Jagiełło ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Photoelectron Spectroscopy ◽  
Mechanical Analysis ◽  
Biological Properties ◽  
Human Umbilical Cord ◽  
Tissue Engineering Scaffolds ◽  
X Ray ◽  
Chitosan Matrix ◽  
Graphene Derivatives

In this study, we investigated preparation of gradient chitosan-matrix hydrogels through a novel freezing–gelling–thawing method. The influence of three types of graphene family materials (GFM), i.e., graphene oxide (GO), reduced graphene oxide (rGO), and poly(ethylene glycol) grafted graphene oxide (GO-PEG), as well as hydroxyapatite (HAp) on the physicochemical and biological properties of the composite hydrogels was examined in view of their potential applicability as tissue engineering scaffolds. The substrates and the hydrogel samples were thoroughly characterized by X-ray photoelectron spectroscopy, X-ray diffractometry, infrared spectroscopy, digital and scanning electron microscopy, rheological and mechanical analysis, in vitro chemical stability and bioactivity assays, as well as initial cytocompatibility evaluation with human umbilical cord Wharton’s jelly mesenchymal stem cells (hUC-MSCs). We followed the green-chemistry approach and avoided toxic cross-linking agents, using instead specific interactions of our polymer matrix with tannic acid, non-toxic physical cross-linker, and graphene derivatives. It was shown that the most promising are the gradient hydrogels modified with GO-PEG and HAp.

scholarly journals Fabrication and Characteristics of Chitosan Sponge as a Tissue Engineering Scaffold

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Takeshi Ikeda ◽  
Kahori Ikeda ◽  
Kouhei Yamamoto ◽  
Hidetaka Ishizaki ◽  
Yuu Yoshizawa ◽  
...  
Keyword(s):  
Tensile Strength ◽  
Porous Scaffold ◽  
Chitosan Solution ◽  
Spongiform Encephalopathy ◽  
Porous Chitosan ◽  
Chitosan Scaffolds ◽  
Controlled Freezing ◽  
The Relationship ◽  
Releasing Time

Cells, growth factors, and scaffolds are the three main factors required to create a tissue-engineered construct. After the appearance of bovine spongiform encephalopathy (BSE), considerable attention has therefore been focused on nonbovine materials. In this study, we examined the properties of a chitosan porous scaffold. A porous chitosan sponge was prepared by the controlled freezing and lyophilization of different concentrations of chitosan solutions. The materials were examined by scanning electron microscopy, and the porosity, tensile strength, and basic fibroblast growth factor (bFGF) release profiles from chitosan sponge were examined in vitro. The morphology of the chitosan scaffolds presented a typical microporous structure, with the pore size ranging from 50 to 200 μm. The porosity of chitosan scaffolds with different concentrations was approximately 75–85%. A decreasing tendency for porosity was observed as the concentration of the chitosan increased. The relationship between the tensile properties and chitosan concentration indicated that the ultimate tensile strength for the sponge increased with a higher concentration. The in vitro bFGF release study showed that the higher the concentration of chitosan solution became, the longer the releasing time of the bFGF from the chitosan sponge was.

Strontium-substituted hydroxyapatite grown on graphene oxide nanosheet-reinforced chitosan scaffold to promote bone regeneration

2020 ◽  
Vol 8 (16) ◽  
pp. 4603-4615 ◽  
Author(s):  
Tingting Wu ◽  
Binglin Li ◽  
Wanshun Wang ◽  
Lingling Chen ◽  
Zhan Li ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Bone Regeneration ◽  
Chitosan Scaffold ◽  
Alp Activity ◽  
Chitosan Matrix ◽  
Graphene Oxide Nanosheet ◽  
Chitosan Scaffolds

SrHA grown on GO nanosheet-reinforced chitosan scaffolds promoted the bone regeneration as SrHA and GO had good osteoconductivity and high ALP activity while the chitosan matrix played a role in antibacteria.

Study on Degradation Behavior and Biocompatibility of Polymethyl Methacrylate/Mineralized Collagen/Mg–Ca Alloy Composite Material

2020 ◽  
Vol 10 (2) ◽  
pp. 139-150
Author(s):  
Yuan-Xin Shen ◽  
Xi-Rao Sun ◽  
Cheng-Yue Wang ◽  
Jing-Xin Yang ◽  
Jia-Xin Bao
Keyword(s):  
Weight Loss ◽  
Alkaline Phosphatase ◽  
Cell Adhesion ◽  
Orthopedic Implants ◽  
Dip Coating ◽  
Alizarin Red ◽  
Pmma Bone Cement ◽  
Degradation Test ◽  
Mineralized Collagen

In this study, we composited mineralized collagen and magnesium-calcium alloy by freeze-drying, followed by dip-coating PMMA bone cement to enhance the composite of mineralized collagen and magnesium-calcium alloy. In vitro degradation test was performed to observe the pH and weight loss of the material. The contact angle test was used to detect the hydrophilicity of the material. Subsequently, MC3T3-E1 were used to assess cell biocompatibility In vitro by cell adhesion, cytotoxicity, alkaline phosphatase, alizarin red staining, and cytoskeleton. The results showed that the pH changes of the PMMA/NHAC/Mg–Ca was slower than that of the Mg–Ca , and the weight loss rate at 7 d and 14 d were lower than that of the Mg–Ca (P < 0.05) in degradation test. Wettability experiment showed that PMMA/NHAC/Mg–Ca was a hydrophilic material and Mg–Ca was a hydrophobic material (P < 0.05). In vitro cell experiments, the PMMA/NHAC/Mg–Ca had more cell adhesion than Mg–Ca and more synapses were connected to others. In the cytotoxicity experiment, the cell proliferation lever of PMMA/NHAC/Mg–Ca was higher than that of Mg–Ca at each time point (P < 0.05). In the 7 d alkaline phosphatase experiment, the PMMA/NHAC/Mg–Ca showed higher ALP activity than the Mg–Ca (P < 0.05), and in the alizarin red experiment at 14 d and 28 d, there were more obvious calcified nodules and mineralized area. After 1 day of culture in the PMMA/NHAC/Mg–Ca extract, the cells showed a clearer and more complete cytoskeletal structure and better cell morphology. In conclusion, PMMA/NHAC/Mg–Ca orthopedic implants had a better hydrophilicity, cytotoxicity and osteogenic ability, besides with a slower rate of degradation, and could be implanted in animals for further research, which were expected to be used for the repair of clinical bone defects.

Osteogenic capability of strontium and icariin-loaded TiO2 nanotube coatings in vitro and in osteoporotic rats

2021 ◽  
pp. 088532822199799
Author(s):  
Ye Zhu ◽  
Tianxia Zheng ◽  
Li-ming Wen ◽  
Ren Li ◽  
Yan-bo Zhang ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Alkaline Phosphatase ◽  
Cell Adhesion ◽  
Titanium Implants ◽  
Ovariectomized Rats ◽  
Alp Activity ◽  
Ti Alloys ◽  
Bone Integration ◽  
Cell Adhesion And Proliferation

Titanium (Ti) and Ti alloys are widely used biomaterials, but they lack osteogenic capability for rapid bone integration. To improve osseointegration of Ti implants, TiO2 nanotubes were prepared using the anodizing oxidation technique, and strontium (Sr) combined with icariin (ICA) was loaded on TiO2 nanotube coatings. Cell adhesion and proliferation of MC3T3-E1 cells, alkaline phosphatase (ALP) activity, mineralization of extracellular matrix, and bone formation around titanium implants in ovariectomized rats, were examined separately. The results showed that compared with pure Ti, TiO2 and Sr-loaded TiO2 coatings, the coatings loaded with both Sr and ICA showed better effect on cell adhesion and proliferation, higher ALP activity and more red-stained mineralized nodules. Furthermore, more bone was formed around implants loaded with both Sr and ICA in osteoporotic rats. Therefore, coating with Sr and ICA is valuable for clinical application to strengthen the osseointegration of titanium implants, especially in osteoporotic patients.

In Situ Localization of PPAR Gamma and Uncoupling Protein in Mouse Embryo Sections Using Digoxigenin-Labeled Riboprobes

1996 ◽  
Vol 54 ◽  
pp. 32-33
Author(s):  
C. Jennermann ◽  
S. A. Kliewer ◽  
D. C. Morris
Keyword(s):  
Alkaline Phosphatase ◽  
Room Temperature ◽  
Uncoupling Protein ◽  
Ppar Gamma ◽  
Nuclear Hormone Receptor ◽  
Full Length ◽  
Northern Analysis ◽  
Peroxisome Proliferator

Peroxisome proliferator-activated receptor gamma (PPARg) is a member of the nuclear hormone receptor superfamily and has been shown in vitro to regulate genes involved in lipid metabolism and adipocyte differentiation. By Northern analysis, we and other researchers have shown that expression of this receptor predominates in adipose tissue in adult mice, and appears first in whole-embryo mRNA at 13.5 days postconception. In situ hybridization was used to find out in which developing tissues PPARg is specifically expressed.Digoxigenin-labeled riboprobes were generated using the Genius™ 4 RNA Labeling Kit from Boehringer Mannheim. Full length PPAR gamma, obtained by PCR from mouse liver cDNA, was inserted into pBluescript SK and used as template for the transcription reaction. Probes of average size 200 base pairs were made by partial alkaline hydrolysis of the full length transcripts. The in situ hybridization assays were performed as described previously with some modifications. Frozen sections (10 μm thick) of day 18 mouse embryos were cut, fixed with 4% paraformaldehyde and acetylated with 0.25% acetic anhydride in 1.0M triethanolamine buffer. The sections were incubated for 2 hours at room temperature in pre-hybridization buffer, and were then hybridized with a probe concentration of 200μg per ml at 70° C, overnight in a humidified chamber. Following stringent washes in SSC buffers, the immunological detection steps were performed at room temperature. The alkaline phosphatase labeled, anti-digoxigenin antibody and detection buffers were purchased from Boehringer Mannheim. The sections were treated with a blocking buffer for one hour and incubated with antibody solution at a 1:5000 dilution for 2 hours, both at room temperature. Colored precipitate was formed by exposure to the alkaline phosphatase substrate nitrobluetetrazoliumchloride/ bromo-chloroindlylphosphate.

Electron microscopic study of the membrane glycoproteins in the rat kidney after cisplatin treatment

1989 ◽  
Vol 47 ◽  
pp. 912-913
Author(s):  
S.K. Aggarwal
Keyword(s):  
Alkaline Phosphatase ◽  
Rat Kidney ◽  
Light And Electron Microscopy ◽  
Kidney Tissue ◽  
Membrane Glycoproteins ◽  
Electron Microscopic ◽  
Before And After ◽  
Interstrand Cross Links ◽  
Cross Links

The proposed primary mechanism of action of the anticancer drug cisplatin (Cis-DDP) is through its interaction with DNA, mostly through DNA intrastrand cross-links or DNA interstrand cross-links. DNA repair mechanisms can circumvent this arrest thus permitting replication and transcription to proceed. Various membrane transport enzymes have also been demonstrated to be effected by cisplatin. Glycoprotein alkaline phosphatase was looked at in the proximal tubule cells before and after cisplatin both in vivo and in vitro for its inactivation or its removal from the membrane using light and electron microscopy.Outbred male Swiss Webster (Crl: (WI) BR) rats weighing 150-250g were given ip injections of cisplatin (7mg/kg). Animals were killed on day 3 and day 5. Thick slices (20-50.um) of kidney tissue from treated and untreated animals were fixed in 1% buffered glutaraldehyde and 1% formaldehyde (0.05 M cacodylate buffer, pH 7.3) for 30 min at 4°C. Alkaline phosphatase activity and carbohydrates were demonstrated according to methods described earlier.

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