scholarly journals Lysophosphatidic Acid Analogue rather than Lysophosphatidic Acid Promoted the Bone Formation In Vivo

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Zi-Li Yu ◽  
Bin-Fang Jiao ◽  
Zu-Bing Li
Keyword(s):  
Bone Formation ◽  
Lysophosphatidic Acid ◽  
Histological Analysis ◽  
Alizarin Red S ◽  
Porous Scaffolds ◽  
Micro Ct ◽  
Micro Computed Tomography ◽  
Alizarin Red ◽  
Lipid Molecule

Lysophosphatidic acid (LPA), a bioactive lipid molecule, has recently emerged as physiological and pathophysiological regulator in skeletal biology. Here we evaluate the effects of LPA on bone formation in vivo in murine femoral critical defect model. Primary femoral osteoblasts were isolated and treated with osteogenic induction conditional media supplemented with 20 μM LPA or LPA analogue. Mineralized nodules were visualized by Alizarin Red S staining. Forty-five C57BL/6 mice underwent unilateral osteotomy. The femoral osteotomy gap was filled with porous scaffolds of degradable chitosan/beta-tricalcium phosphate containing PBS, LPA, or LPA analogue. 2, 5, and 10 weeks after surgery, mice were sacrificed and femurs were harvested and prepared for Micro-Computed Tomography (Micro-CT) and histological analysis. Alizarin Red S staining showed that LPA and LPA analogue significantly enhanced the mineral deposition in osteoblasts. Micro-CT 3D reconstruction images and HE staining revealed that significantly more newly formed bone in osteotomy was treated with LPA analogue when compared to control and LPA group, which was verified by histological analysis and biomechanical characterization testing. In summary, our study demonstrated that although LPA promotes mineralized matrix formation in vitro, the locally administrated LPA was not effective in promoting bone formation in vivo. And bone formation was enhanced by LPA analogue, administrated locally in vivo. LPA analogue was a potent stimulating factor for bone formation in vivo due to its excellent stability.

Abstract 452: Longitudinal Visualization of Calcification Genesis and Growth in vivo : Novel Implications for Plaque Vulnerability

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Joshua D Hutcheson ◽  
Claudia Goettsch ◽  
Brett Pieper ◽  
Tan Pham ◽  
Jung Choi ◽  
...  
Keyword(s):  
Cardiovascular Risk ◽  
Aortic Root ◽  
Atherogenic Diet ◽  
Alizarin Red S ◽  
Arterial Calcification ◽  
Micro Computed Tomography ◽  
Alizarin Red ◽  
Histological Sections ◽  
Over Time

Background: Clinical evidence links arterial calcification and cardiovascular risk. Fibrous cap microcalcifications can promote atherosclerotic plaque failure, and large calcifications can stabilize the plaque. Therefore, calcification morphology can determine cardiovascular morbidity, but temporal patterns of calcific mineral deposition and growth remain unknown. Results: Apolipoprotein E-deficient ( Apoe-/- ) mice on an atherogenic diet develop plaque calcification. Longitudinal studies were performed using two different fluorescent calcium tracers injected intravenously into Apoe-/- mice: calcein injection following 18 weeks of atherogenic diet (n=7) and alizarin red S injection into the same mice 1 (n=4) or 3 (n=3) weeks later. Imaging green (calcein) and red (alizarin red S) fluorescence provided snapshots of aortic calcification at 18, 19, and 21 weeks. Observations within histological sections revealed green microcalcifications at 18 weeks embedded within alizarin red stained larger calcifications that were formed by 19 weeks (a). These data demonstrate that microcalcifications present at the start of calcification become the core of the larger calcifications that develop over time. Serial histological sections from aortic root to arch (b) were digitally reconstructed into 3D volumes (c) to reveal total calcific burden and localization within the aortic wall (d). Total calcification volume increased at a significant rate of 6.0x10 6 μm 3 per week (R 2 =0.99, p=0.007) and progressed from aortic arch to aortic root over time (p<0.001). Observations closely match calcification morphologies found by micro-computed tomography of human coronary arteries. Conclusion: Temporal and spatial understanding arterial calcification growth is crucial given the link between mineral morphology and cardiovascular risk, and these techniques provide a method for testing therapeutic approaches to control calcification morphology over time in situ .

The In Vivo Inhibition of Bone Formation by Alizarin Red S

1964 ◽  
Vol 46 (3) ◽  
pp. 493-508 ◽  
Author(s):  
WILLIAM H. HARRIS ◽  
DOROTHY F. TRAVIS ◽  
ULF FRIBERG ◽  
ERIC RADIN
Keyword(s):  
Bone Formation ◽  
Alizarin Red S ◽  
Alizarin Red

scholarly journals Biomimetic reduced graphene oxide coated collagen scaffold for in situ bone regeneration

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sajad Bahrami ◽  
Nafiseh Baheiraei ◽  
Mostafa Shahrezaee
Keyword(s):  
Graphene Oxide ◽  
Reduced Graphene Oxide ◽  
Cranial Bone ◽  
Drying Methods ◽  
Porous Scaffolds ◽  
Alizarin Red ◽  
Reduced Graphene ◽  
Coated Collagen

AbstractA variety of bone-related diseases and injures and limitations of traditional regeneration methods require new tissue substitutes. Tissue engineering and regeneration combined with nanomedicine can provide different natural or synthetic and combined scaffolds with bone mimicking properties for implantation in the injured area. In this study, we synthesized collagen (Col) and reduced graphene oxide coated collagen (Col-rGO) scaffolds, and we evaluated their in vitro and in vivo effects on bone tissue repair. Col and Col-rGO scaffolds were synthesized by chemical crosslinking and freeze-drying methods. The surface topography, and the mechanical and chemical properties of scaffolds were characterized, showing three-dimensional (3D) porous scaffolds and successful coating of rGO on Col. The rGO coating enhanced the mechanical strength of Col-rGO scaffolds to a greater extent than Col scaffolds by 2.8 times. Furthermore, Col-rGO scaffolds confirmed that graphene addition induced no cytotoxic effects and enhanced the viability and proliferation of human bone marrow-derived mesenchymal stem cells (hBMSCs) with 3D adherence and expansion. Finally, scaffold implantation into rabbit cranial bone defects for 12 weeks showed increased bone formation, confirmed by Hematoxylin–Eosin (H&E) and alizarin red staining. Overall, the study showed that rGO coating improves Col scaffold properties and could be a promising implant for bone injuries.

scholarly journals Additive-manufactured Ti-6Al-4 V/Polyetheretherketone composite porous cage for Interbody fusion: bone growth and biocompatibility evaluation in a porcine model

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Pei-I Tsai ◽  
Meng-Huang Wu ◽  
Yen-Yao Li ◽  
Tzu-Hung Lin ◽  
Jane S. C. Tsai ◽  
...  
Keyword(s):  
Bone Formation ◽  
Bone Growth ◽  
Lumbar Fusion ◽  
High Porosity ◽  
Ti Alloy ◽  
Micro Ct ◽  
Micro Computed Tomography ◽  
Intervertebral Fusion ◽  
Porous Ti ◽  
Peek Composite

Abstract Background We developed a porous Ti alloy/PEEK composite interbody cage by utilizing the advantages of polyetheretherketone (PEEK) and titanium alloy (Ti alloy) in combination with additive manufacturing technology. Methods Porous Ti alloy/PEEK composite cages were manufactured using various controlled porosities. Anterior intervertebral lumbar fusion and posterior augmentation were performed at three vertebral levels on 20 female pigs. Each level was randomly implanted with one of the five cages that were tested: a commercialized pure PEEK cage, a Ti alloy/PEEK composite cage with nonporous Ti alloy endplates, and three composite cages with porosities of 40, 60, and 80%, respectively. Micro-computed tomography (CT), backscattered-electron SEM (BSE-SEM), and histological analyses were performed. Results Micro-CT and histological analyses revealed improved bone growth in high-porosity groups. Micro-CT and BSE-SEM demonstrated that structures with high porosities, especially 60 and 80%, facilitated more bone formation inside the implant but not outside the implant. Histological analysis also showed that bone formation was higher in Ti alloy groups than in the PEEK group. Conclusion The composite cage presents the biological advantages of Ti alloy porous endplates and the mechanical and radiographic advantages of the PEEK central core, which makes it suitable for use as a single implant for intervertebral fusion.

scholarly journals Longitudinal in vivo evaluation of bone regeneration by combined measurement of multi-pinhole SPECT and micro-CT for tissue engineering

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Philipp S. Lienemann ◽  
Stéphanie Metzger ◽  
Anna-Sofia Kiveliö ◽  
Alain Blanc ◽  
Panagiota Papageorgiou ◽  
...  
Keyword(s):  
Computed Tomography ◽  
High Resolution ◽  
Bone Formation ◽  
Bone Regeneration ◽  
Bone Defects ◽  
Histological Evaluation ◽  
Biological Processes ◽  
Micro Ct ◽  
Pinhole Spect

Abstract Over the last decades, great strides were made in the development of novel implants for the treatment of bone defects. The increasing versatility and complexity of these implant designs request for concurrent advances in means to assess in vivo the course of induced bone formation in preclinical models. Since its discovery, micro-computed tomography (micro-CT) has excelled as powerful high-resolution technique for non-invasive assessment of newly formed bone tissue. However, micro-CT fails to provide spatiotemporal information on biological processes ongoing during bone regeneration. Conversely, due to the versatile applicability and cost-effectiveness, single photon emission computed tomography (SPECT) would be an ideal technique for assessing such biological processes with high sensitivity and for nuclear imaging comparably high resolution (<1 mm). Herein, we employ modular designed poly(ethylene glycol)-based hydrogels that release bone morphogenetic protein to guide the healing of critical sized calvarial bone defects. By combined in vivo longitudinal multi-pinhole SPECT and micro-CT evaluations we determine the spatiotemporal course of bone formation and remodeling within this synthetic hydrogel implant. End point evaluations by high resolution micro-CT and histological evaluation confirm the value of this approach to follow and optimize bone-inducing biomaterials.

scholarly journals The Protective Effect of Cordycepin On Alcohol-Induced Osteonecrosis of the Femoral Head

2017 ◽  
Vol 42 (6) ◽  
pp. 2391-2403 ◽  
Author(s):  
Yi-Xuan Chen ◽  
Dao-Yu Zhu ◽  
Zheng-Liang Xu ◽  
Jun-Hui Yin ◽  
Xiao-Wei Yu ◽  
...  
Keyword(s):  
Femoral Head ◽  
Protective Effect ◽  
Immunohistochemical Staining ◽  
Critical Role ◽  
Micro Ct ◽  
Bone Mesenchymal Stem Cells ◽  
Western Blots ◽  
Alizarin Red ◽  
Alp Activity

Background: Alcohol abuse is known to be a leading risk factor for atraumatic osteonecrosis of the femoral head (ONFH), in which the suppression of osteogenesis plays a critical role. Cordycepin benefits bone metabolism; however, there has been no study to determine its effect on osteonecrosis. Methods: Human bone mesenchymal stem cells (hBMSCs) were identified by multi-lineage differentiation. Alkaline phosphatase (ALP) activity, RT-PCR, western blots, immunofluorescent assay and Alizarin red staining of BMSCs were evaluated. A rat model of alcohol-induced ONFH was established to investigate the protective role of cordycepin against ethanol. Hematoxylin & eosin (H&E) staining and micro-computerized tomography (micro-CT) were performed to observe ONFH. Apoptosis was assessed by TdT-mediated dUTP nick end labeling (TUNEL). Immunohistochemical staining was carried out to detect OCN and COL1. Results: Ethanol significantly suppressed ALP activity, decreased gene expression of OCN and BMP2, lowered levels of RUNX2 protein, and reduced immunofluorescence staining of OCN and COL1 and calcium formation of hBMSCs. However, these inhibitory effects were attenuated by cordycepin co-treatment at concentrations of 1 and 10 µg/mL Moreover, it was revealed that the osteo-protective effect of cordycepin was associated with modulation of the Wnt/β-catenin pathway. In vivo, by micro-CT, TUNEL and immunohistochemical staining of OCN and COL1, we found that cordycepin administration prevented alcohol-induced ONFH. Conclusion: Cordycepin treatment to enhance osteogenesis may be considered a potential therapeutic approach to prevent the development of alcohol-induced ONFH.

scholarly journals Additive-Manufactured Ti-6Al-4V/Polyetheretherketone Composite Porous Cage for Interbody Fusion: Bone Growth and Biocompatibility Evaluation in a Porcine Model

2020 ◽  
Author(s):  
Pei-I Tsai ◽  
Meng-Huang Wu ◽  
Yen-Yao Li ◽  
Tzu-Hung Lin ◽  
Jane SC Tsai ◽  
...  
Keyword(s):  
Bone Formation ◽  
Bone Growth ◽  
Lumbar Fusion ◽  
High Porosity ◽  
Ti Alloy ◽  
Micro Ct ◽  
Micro Computed Tomography ◽  
Intervertebral Fusion ◽  
Porous Ti ◽  
Peek Composite

Abstract Background: We developed a porous Ti alloy/PEEK composite interbody cage by utilizing the advantages of polyetheretherketone (PEEK) and titanium alloy (Ti alloy) in combination with additive manufacturing technology. Methods: Porous Ti alloy/PEEK composite cages were manufactured using various controlled porosities. Anterior intervertebral lumbar fusion and posterior augmentation were performed at three vertebral levels on 20 female pigs. Each level was randomly implanted with one of the five cages that were tested: a commercialized pure PEEK cage, a Ti alloy/PEEK composite cage with nonporous Ti alloy endplates, and three composite cages with porosities of 40%, 60%, and 80%, respectively. Micro-computed tomography (CT), backscattered-electron SEM (BSE-SEM), and histological analyses were performed.Results: Micro-CT and histological analyses revealed improved bone growth in high-porosity groups. Micro-CT and BSE-SEM demonstrated that structures with high porosities, especially 60% and 80%, facilitated more bone formation inside the implant but not outside the implant. Histological analysis also showed that bone formation was higher in Ti alloy groups than in the PEEK group. Conclusion: The composite cage presents the biological advantages of Ti alloy porous endplates and the mechanical and radiographic advantages of the PEEK central core, which makes it suitable for use as a single implant for intervertebral fusion.

scholarly journals Endogenous Production of n-3 Polyunsaturated Fatty Acids Promotes Fracture Healing in Mice

2017 ◽  
Vol 2017 ◽  
pp. 1-6 ◽  
Author(s):  
Yuhui Chen ◽  
He Cao ◽  
Dawei Sun ◽  
Changxin Lin ◽  
Liang Wang ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Fracture Healing ◽  
Histological Analysis ◽  
Callus Formation ◽  
Healing Process ◽  
Proximal Femoral Fracture ◽  
Fracture Repair ◽  
Micro Ct ◽  
Micro Computed Tomography ◽  
X Ray

Bone fracture is a global healthcare issue for high rates of delayed healing and nonunions. Although n-3 polyunsaturated fatty acid (PUFA) is considered as a beneficial factor for bone metabolism, only few studies till date focused on the effects of n-3 PUFAs on fracture healing. In this study, we investigated the effect of endogenous n-3 PUFAs on fracture healing by measuring femur fracture repair in bothfat-1transgenic mice and WT mice. Proximal femoral fracture model was established infat-1transgenic mice and WT mice, respectively, and then the fracture was analyzed by using X-ray, micro-computed tomography (micro-CT), and histological assessment at 7, 14, 21, 28, and 35 days after fixation. The results showed that compared with WT mice,fat-1mice exhibited acceleration in fracture healing through radiographic and histological analysis (18–21 days versus 21–28 days postfracture). Meanwhile, X-ray and micro-CT analysis that showed better remodeling callus formation were in thefat-1group compared to WT group. Furthermore, histological analysis revealed that endogenous n-3 PUFAs promoted local endochondral ossification and accelerated the remodeling of calcified calluses after fracture. In conclusion, the present study indicated that endogenously produced n-3 PUFAs promote fracture healing process and accelerate bone remodeling in mice, and supplementation of n-3 PUFAs was positively associated with fracture healing.

scholarly journals Intraosteal Behavior of Porous Scaffolds: The mCT Raw-Data Analysis as a Tool for Better Understanding

Symmetry ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 532 ◽  
Author(s):  
Parrilla-Almansa ◽  
González-Bermúdez ◽  
Sánchez-Sánchez ◽  
Meseguer-Olmo ◽  
Martínez-Cáceres ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Mathematical Analysis ◽  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Hounsfield Unit ◽  
In Vivo Studies ◽  
Porous Scaffolds ◽  
Micro Computed Tomography ◽  
Radiological Analysis

The aim of the study is to determine the existing correlation between high-resolution 3D imaging technique obtained through Micro Computed Tomography (mCT) and histological-histomorphometric images to determine in vivo bone osteogenic behavior of bioceramic scaffolds. A Ca-Si-P scaffold ceramic doped and non-doped (control) with a natural demineralized bone matrix (DBM) were implanted in rabbit tibias for 1, 3, and 5 months. A progressive disorganization and disintegration of scaffolds and bone neoformation occurs, from the periphery to the center of the implants, without any differences between histomorphometric and radiological analysis. However, significant differences (p < 0.05) between DMB-doped and non-doped materials where only detected through mathematical analysis of mCT. In this way, average attenuation coefficient for DMB-doped decreased from 0.99 ± 0.23 Hounsfield Unit (HU) (3 months) to 0.86 ± 0.32 HU (5 months). Average values for non-doped decreased from 0.86 ± 0.25 HU (3 months) to 0.66 ± 0.33 HU. Combination of radiological analysis and mathematical mCT seems to provide an adequate in vivo analysis of bone-implanted biomaterials after surgery, obtaining similar results to the one provided by histomorphometric analysis. Mathematical analysis of Computed Tomography (CT) would allow the conducting of long-term duration in vivo studies, without the need for animal sacrifice, and the subsequent reduction in variability.

Radiographic and Micro—Computed Tomographic Imaging of Lipopolysaccharide-Mediated Bone Resorption

2009 ◽  
Vol 118 (5) ◽  
pp. 391-396 ◽  
Author(s):  
Robert Nason ◽  
Dong H. Lee ◽  
Jae Y. Jung ◽  
Richard A. Chole
Keyword(s):  
Bone Resorption ◽  
Chronic Otitis Media ◽  
Plain Radiography ◽  
Micro Ct ◽  
Micro Computed Tomography ◽  
Gram Negative ◽  
Successful Model ◽  
Computed Tomographic ◽  
Myeloid Differentiation Factor

Objectives: Chronic otitis media and cholesteatomas cause hearing loss as a result of bony erosion. This bone resorption is known to be more aggressive when cholesteatomas become infected. The most common organism isolated from both diseases is the gram-negative bacterium Pseudomonas aeruginosa. Lipopolysaccharide (LPS), a major virulence factor found in the gram-negative bacterial cell wall, is well known to incite inflammatory bone resorption. The mechanisms underlying this process, however, are poorly understood. In this study, we developed a mouse model of calvarial osteolysis in which resorption was reliably imaged by plain radiography and micro–computed tomography (micro-CT). Methods: A murine calvarial model was developed to study bone resorption induced by P aeruginosa LPS. Calvariae from wild-type and knockout mice used in this model were imaged by plain radiography and micro-CT. Results: A high degree of correlation between plain radiography and micro-CT was identified (R2 = 0.8554). Furthermore, maximal LPS-induced bone resorption required functioning toll-like receptor (TLR) 2, TLR4, and myeloid differentiation factor 88 (MyD88). Conclusions: We have developed a successful model of inflammatory osteolysis in which plain radiography can reliably delineate induced bone resorption. In vivo, we have shown that P aeruginosa LPS signals via TLR2, as well as TLR4 through MyD88.

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