Evaluation of 5% Na2O-Incorporated Calcium Metaphosphate as a Scaffold for Tissue-Engineered Bone Regeneration

2006 ◽  
Vol 309-311 ◽  
pp. 985-988 ◽  
Author(s):  
J.H. Yoon ◽  
J.T. Kim ◽  
Eui Kyun Park ◽  
Shin Yoon Kim ◽  
Chang Kuk You ◽  
...  
Keyword(s):  
Compressive Strength ◽  
Bone Regeneration ◽  
Average Pore Size ◽  
Ectopic Bone Formation ◽  
Cellular Reaction ◽  
Average Pore ◽  
Cellular Attachment ◽  
Ectopic Bone ◽  
Osteogenic Effect

As a part of the effort to develop a suitable scaffold for tissue-engineered bone regeneration, we modified calcium metaphosphate (CMP) ceramic with Na20 and evaluated its efficiency as a scaffold. We incorporate 5% Na20 into pure CMP and prepare for an average pore size of 250 or 450 µm average pore sizes. The incorporation of 5% Na2O caused reduced compressive strength and there was no change in biodegradability. The in vitro cellular attachment and proliferation rate, however, were slightly improved. The 5% Na2O-incorporated macroporous CMP ceramic-cell constructs treated with Emdogain induced ectopic bone formation more effectively than those without Emdogain treatment. These results suggest that the incorporation of 5% Na2O into pure CMP is not effective for improving the physical characteristics of pure CMP but it is positive for improving the cellular reaction and osteogenic effect with the addition of Emdogain.

Influence of Emdogain® Treatment onto K2O Incorporated Calcium Metaphosphate on Osteogenic Activation

2005 ◽  
Vol 284-286 ◽  
pp. 631-634
Author(s):  
Suk Young Kim ◽  
Chang Kuk You ◽  
Jae Ho Jeong ◽  
Eui Kyun Park ◽  
Shin Yoon Kim ◽  
...  
Keyword(s):  
Bone Formation ◽  
Physical Properties ◽  
Bone Regeneration ◽  
Tissue Regeneration ◽  
Ectopic Bone Formation ◽  
Bone Tissue Regeneration ◽  
Osteoblastic Cells ◽  
Average Pore ◽  
Ectopic Bone ◽  
Cell Scaffold

As a part of the efforts to develop a suitable scaffold optimizing bone regeneration that has similar physical properties to bone, we modified calcium metaphosphate (CMP) ceramics with K2O and evaluated their efficiency as a scaffold for tissue engineered bone tissue regeneration. Macroporous CMP ceramics modified by incorporation of 5% K2O to improve biodegradability were prepared to have 250 and 450 µm average pore sizes, respectively. The modified CMP ceramics were cultured with mouse primary calvarial osteoblastic cells in osteogenic media for 2 weeks and these cell-CMP ceramic constructs with or without Emdogain treatment were implanted in the SCID mice subcutaneous pouches. After 1, 2, and 3 weeks, the degree of ectopic bone formation was evaluated. The modified macroporous CMP ceramic-cell constructs treated with Emdogain induced ectopic bone formation, whereas the modified CMP ceramic-cell constructs without Emdogain treatment induced no ectopic bone formation. This result suggests that the Emdogain treatment on cell-scaffold constructs for tissue engineered bone regeneration may be effective for osteogenic activation of attached cells.

Successful Osteoinduction by Cell-Macroporous Biphasic HA-TCP Ceramic Matrix

2005 ◽  
Vol 288-289 ◽  
pp. 245-248 ◽  
Author(s):  
Hong In Shin ◽  
K.H. Kim ◽  
Inn Kyu Kang ◽  
Kyung Sik Oh
Keyword(s):  
Bone Formation ◽  
Bone Regeneration ◽  
Osteogenic Differentiation ◽  
Cellular Proliferation ◽  
Ceramic Matrix ◽  
Ectopic Bone Formation ◽  
Inflammatory Reactions ◽  
Cell Matrix ◽  
Cellular Attachment ◽  
Ectopic Bone

To improve the potential of osteogenic repair, we developed macroporous biphasic hydroxyapatite-tricalcium phosphate (HA-TCP) ceramic and evaluated its efficiency as a scaffold for tissue engineered bone regeneration, which allows for appropriate cellular attachment and proliferation with osteogenic differentiation by evaluating ectopic bone formation ability after the implantation of cell-matrix construct in the skid mice subcutaneous pouches for 3 weeks. The macroporous biphasic HA-TCP ceramic matrix, with an average porosity of 86% and 200 µm mean pore size, provided favorable conditions for the attachment of cultured bone marrow derived osteoblastic cells along its inner surfaces in a filed up pattern and the active proliferation of them. The implanted cell-matrix constructs in the subcutaneous pouches induced favorable ectopic bone formation without any remarkable inflammatory reactions. These findings suggest that the biphasic HA-TCP ceramic matrix with macroporous structure has excellent biocompatibility, and that it allows for favorable cellular attachment with the acceleration of cellular proliferation and osteogenic differentiation support as well. Thus, with the controlled biodegradability, the biphasic HA-TCP ceramic may be a promising scaffold for tissue engineered bone regeneration technology.

scholarly journals Phosphate graphene as an intrinsically osteoinductive scaffold for stem cell-driven bone regeneration

2019 ◽  
Vol 116 (11) ◽  
pp. 4855-4860 ◽  
Author(s):  
Anne M. Arnold ◽  
Brian D. Holt ◽  
Leila Daneshmandi ◽  
Cato T. Laurencin ◽  
Stefanie A. Sydlik
Keyword(s):  
Bone Formation ◽  
Bone Regeneration ◽  
Bone Cells ◽  
Standard Of Care ◽  
Ectopic Bone Formation ◽  
Bone Morphogenetic Protein 2 ◽  
Ectopic Bone ◽  
Human Bone Morphogenetic Protein

Synthetic, resorbable scaffolds for bone regeneration have potential to transform the clinical standard of care. Here, we demonstrate that functional graphenic materials (FGMs) could serve as an osteoinductive scaffold: recruiting native cells to the site of injury and promoting differentiation into bone cells. By invoking a Lewis acid-catalyzed Arbuzov reaction, we are able to functionalize graphene oxide (GO) to produce phosphate graphenes (PGs) with unprecedented control of functional group density, mechanical properties, and counterion identity. In aqueous environments, PGs release inducerons, including Ca2+ and PO43−. Calcium phosphate graphene (CaPG) intrinsically induces osteogenesis in vitro and in the presence of bone marrow stromal cells (BMSCs), can induce ectopic bone formation in vivo. Additionally, an FGM can be made by noncovalently loading GO with the growth factor recombinant human bone morphogenetic protein 2 (rhBMP-2), producing a scaffold that induces ectopic bone formation with or without BMSCs. The FGMs reported here are intrinsically inductive scaffolds with significant potential to revolutionize the regeneration of bone.

Formation of Calcium Phosphate-Ellagic Acid Composites by Layer by Layer Assembly for Cellular Attachment to Osteoblasts

2012 ◽  
Vol 13 ◽  
pp. 1-17
Author(s):  
Ipsita A. Banerjee ◽  
Karl R. Fath ◽  
Nako Nakatsuka ◽  
Nazmul H. Sarker ◽  
Ipsita A. Banerjee
Keyword(s):  
Calcium Phosphate ◽  
Ellagic Acid ◽  
Liquid Crystalline ◽  
Average Pore Size ◽  
Layer By Layer ◽  
Average Pore ◽  
Assembly Method ◽  
Cellular Attachment ◽  
New Strategy

The quest for new biomaterials to serve as cell scaffolds for applications in tissue engineering is of prime importance. In this work, we investigated microfiber assemblies of Ellagic Acid (EA), a plant polyphenol to serve as scaffolds for attachment and proliferation of osteoblasts. The advantage of Ellagic Acid self-assembling system is its intrinsic ability to order into multiple layers due to its capability to form liquid crystalline assemblies. We prepared ellagic acid-microfiber composites by the layer-by-layer (LBL) assembly method, where collagen (COL), poly-Arginine (poly-R), and calcium phosphate nanocrystals were coated on the surface of ellagic acid microfibers. The attachment of the various layers was confirmed by various spectroscopic and microscopic methods. The samples were found to be porous with an average pore size of 600 nm. The formed microconjugates were biodegradable and supported the growth of human fetal osteoblast (hFOB) cells in vitro. Our findings suggest that this system not only promotes initial cell adhesion but also can be utilized to deliver the vital biological molecule ellagic acid to cells at the scaffold interface and displays a new strategy for the design of biomaterials.

Novel phytochemical Cissus quadrangularis extract–loaded chitosan/Na-carboxymethyl cellulose–based scaffolds for bone regeneration

2018 ◽  
Vol 33 (6) ◽  
pp. 629-646 ◽  
Author(s):  
Sedef Tamburaci ◽  
Ceren Kimna ◽  
Funda Tihminlioglu
Keyword(s):  
Cell Culture ◽  
Bone Regeneration ◽  
Bone Tissue ◽  
Carboxymethyl Cellulose ◽  
Average Pore Size ◽  
In Vitro Cytotoxicity ◽  
Bone Tissue Regeneration ◽  
Average Pore ◽  
Cissus Quadrangularis

Medicinal plants are attracting considerable interest as a potential therapeutic agent for bone tissue regeneration. Cissus quadrangularis L. is also a medicinal plant known with its osteogenic activity. In this study, a phytochemical scaffold was produced by incorporating Cissus quadrangularis with chitosan/Na-carboxymethyl cellulose blend by lyophilization technique. The effect of Cissus quadrangularis loading on the mechanical, morphological, chemical, and degradation properties as well as in vitro cytotoxicity, cell proliferation, and differentiation of the composites was investigated. Scanning electron microscopy images showed that porous Cissus quadrangularis–loaded scaffolds were obtained with an average pore size of 148–209 µm which is appropriate for bone regeneration. Cissus quadrangularis incorporation enhanced the compression modulus of scaffolds from 76 to 654 kPa. In vitro cell culture results indicated that Cissus quadrangularis/chitosan/Na-carboxymethyl cellulose scaffolds provided a favorable substrate for the osteoblast adhesion, proliferation, and mineralization. Results supported the osteoinductive property of the Cissus quadrangularis extract–incorporated scaffolds even without osteogenic media supplement. Cissus quadrangularis extract increased the alkaline phosphatase activity of the SaOS-2 cells on scaffolds on 7th and 14th days of incubation. The investigation of characterization and cell culture studies suggest that Cissus quadrangularis–loaded osteoinductive Cissus quadrangularis/chitosan/Na-carboxymethyl cellulose scaffold can serve as a potential biomaterial for bone tissue engineering applications.

scholarly journals Cobalt-doped bioceramic scaffolds fabricated by 3D printing show enhanced osteogenic and angiogenic properties for bone repair

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Jungang Li ◽  
Chaoqian Zhao ◽  
Chun Liu ◽  
Zhenyu Wang ◽  
Zeming Ling ◽  
...  
Keyword(s):  
Compressive Strength ◽  
3D Printing ◽  
Bone Regeneration ◽  
Doping Concentration ◽  
Artificial Bone ◽  
Bone Defect Repair ◽  
Co Doping ◽  
Defect Repair ◽  
Co Doped

Abstract Background The bone regeneration of artificial bone grafts is still in need of a breakthrough to improve the processes of bone defect repair. Artificial bone grafts should be modified to enable angiogenesis and thus improve osteogenesis. We have previously revealed that crystalline Ca10Li(PO4)7 (CLP) possesses higher compressive strength and better biocompatibility than that of pure beta-tricalcium phosphate (β-TCP). In this work, we explored the possibility of cobalt (Co), known for mimicking hypoxia, doped into CLP to promote osteogenesis and angiogenesis. Methods We designed and manufactured porous scaffolds by doping CLP with various concentrations of Co (0, 0.1, 0.25, 0.5, and 1 mol%) and using 3D printing techniques. The crystal phase, surface morphology, compressive strength, in vitro degradation, and mineralization properties of Co-doped and -undoped CLP scaffolds were investigated. Next, we investigated the biocompatibility and effects of Co-doped and -undoped samples on osteogenic and angiogenic properties in vitro and on bone regeneration in rat cranium defects. Results With increasing Co-doping level, the compressive strength of Co-doped CLP scaffolds decreased in comparison with that of undoped CLP scaffolds, especially when the Co-doping concentration increased to 1 mol%. Co-doped CLP scaffolds possessed excellent degradation properties compared with those of undoped CLP scaffolds. The (0.1, 0.25, 0.5 mol%) Co-doped CLP scaffolds had mineralization properties similar to those of undoped CLP scaffolds, whereas the 1 mol% Co-doped CLP scaffolds shown no mineralization changes. Furthermore, compared with undoped scaffolds, Co-doped CLP scaffolds possessed excellent biocompatibility and prominent osteogenic and angiogenic properties in vitro, notably when the doping concentration was 0.25 mol%. After 8 weeks of implantation, 0.25 mol% Co-doped scaffolds had markedly enhanced bone regeneration at the defect site compared with that of the undoped scaffold. Conclusion In summary, CLP doped with 0.25 mol% Co2+ ions is a prospective method to enhance osteogenic and angiogenic properties, thus promoting bone regeneration in bone defect repair.

Fabrication and characterization of bovine hydroxyapatite-gelatin-alendronate scaffold cross-linked by glutaraldehyde for bone regeneration

2021 ◽  
Vol 32 (4) ◽  
pp. 555-560
Author(s):  
Samirah ◽  
Aniek Setiya Budiatin ◽  
Ferdiansyah Mahyudin ◽  
Junaidi Khotib
Keyword(s):  
Compressive Strength ◽  
Bone Regeneration ◽  
Mtt Assay ◽  
Local Treatment ◽  
Swelling Ratio ◽  
High Concentration ◽  
Proliferation And Differentiation ◽  
Bovine Hydroxyapatite

Abstract Objectives Alendronate are widely used in the treatment of bone disorders characterized by inhibit osteoclast-mediated bone resorption such as Paget’s disease, fibrous dysplasia, myeloma, bone metastases and osteoporosis. In recent studies alendronate improves proliferation and differentiation of osteoblasts, thereby facilitating for bone regeneration. The disadvantages of this class are their poor bioavailability and side effects on oral and intravenous application such as stomach irritation and osteonecrosis in jaw. Thus, local treatment of alendronate is needed in order to achieve high concentration of drug. Bovine hydroxyapatite-gelatin scaffold with alendronate was studied. Glutaraldehyde was used as cross-linking agent, increase the characteristics of this scaffold. The objectives of this study were to manufacture and characterize alendronate scaffold using bovine hydroxyapatite-gelatin and crosslinked by glutaraldehyde. Methods Preparation of cross-linked bovine hydroxyapatite-gelatin and alendronate scaffold with different concentration of glutaraldehyde (0.00, 0.50, 0.75, and 1.00%). The scaffolds were characterized for compressive strength, porosity, density, swelling ratio, in vitro degradation, and cytotoxicity (the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay, shorted as MTT assay). Results Bovine hydroxyapatite-gelatin-alendronate scaffold cross-linked with glutaraldehyde showed lower density than without glutaraldehyde. As glutaraldehyde concentration increased, porosity also increased. Eventually, it reduced compressive strength. Swelling ratio and in vitro degradation was negatively dependent on glutaraldehyde concentration. In addition, the scaffold has a good safety by MTT assay. Conclusions Bovine hydroxyapatite-gelatin-alendronate scaffold was fabricated with various concentrations of glutaraldehyde. The presence of glutaraldehyde on bovine hydroxyapatite-gelatin-alendronate is safe and suitable candidate scaffold for bone regeneration.

scholarly journals 3579 The role of CypD/mPTP during osteogenic differentiation - a potential target to prevent bone loss

2019 ◽  
Vol 3 (s1) ◽  
pp. 24-24
Author(s):  
Rubens Sautchuk ◽  
Brianna H. Shares ◽  
Roman A. Eliseev
Keyword(s):  
Bone Loss ◽  
Bone Formation ◽  
Mitochondrial Function ◽  
Ectopic Bone Formation ◽  
Regulatory Mechanisms ◽  
Mrna Levels ◽  
Prevent Bone Loss ◽  
Ectopic Bone

OBJECTIVES/SPECIFIC AIMS: The study aims to further investigate how cyclophilin D (CypD), the key mPTP opening regulator, affects BMSCs fate and to determine potential regulatory mechanisms involved in CypD regulation during osteogenesis. METHODS/STUDY POPULATION: We evaluated CypD mRNA expression in mouse BMSCs and in osteogenic-like (OL) cells during the course of OB differentiation. CypD protein level was also probed. Moreover, BMSCs had their mPTP activity recorded during osteoinduction. We further analyzed the effect of CypD genetic deletion on osteogenesis in vitro and in vivo. For our in vivo model, we performed the ectopic bone formation assay to asses differences in ossicle formation when CypD KO BMSCs were transplanted compared to wild type littermate BMSCs. In our in vitro model, we transfected OL cells with either CypD gain of function or CypD loss of function vector and measured their osteogenic differentiation potential. Additionally, we treated BMSCs with CypD inhibitor and compare to non-treated BMSCs for mineralization level. To determine potential regulatory mechanisms involved in CypD regulation, we analyzed the CypD gene (Ppif) promoter for potential transcription factor (TF) binding sites and found multiple Smad-binding elements within this promoter. Smads (Smad1, 5, 8) are TFs downstream from Bone Morphogenic Protein (BMP) signaling pathway that transmit cell differentiation signaling, and exert either activating or inhibitory effects on a variety of genes. We also transfect OL cells with Smad1 vector and analyzed for CypD mRNA levels. RESULTS/ANTICIPATED RESULTS: - Our data showed that CypD mRNA levels decreased in both primary cells and OL cells at day 7 and day 14 in osteogenic media. - Osteogenic induction also decreased mPTP activity. - In vivo ectopic bone formation assay showed increased ossicle fo DISCUSSION/SIGNIFICANCE OF IMPACT: Our data suggest that downregulation of CypD increases OB differentiation due to improved OxPhos activity led by mPTP closure. Our results corroborate reports of CypD downregulation and mPTP closure during neuronal differentiation in developing rat brains as well as in cardiomyocyte differentiation in developing mouse hearts. Our studies also suggest a yet unknown mechanism linking differentiation signaling with mitochondrial function – BMP/Smad mediated downregulation of CypD transcription. As initially mentioned, in a previous study, our lab showed that CypD KO mice present higher mitochondrial function and osteogenicity in aged BMSCs and less osteoporosis burden. Taken together, these results suggest that CypD can be a potential target to prevent bone loss in aging.

scholarly journals Hedgehog Signaling Inhibition by Smoothened Antagonist BMS-833923 Reduces Osteoblast Differentiation and Ectopic Bone Formation of Human Skeletal (Mesenchymal) Stem Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Nihal AlMuraikhi ◽  
Nuha Almasoud ◽  
Sarah Binhamdan ◽  
Ghaydaa Younis ◽  
Dalia Ali ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bone Formation ◽  
Osteoblast Differentiation ◽  
Global Gene Expression ◽  
Ectopic Bone Formation ◽  
Ectopic Bone ◽  
In Vitro Mineralization ◽  
Global Gene Expression Profiling

Background. Hedgehog (Hh) signaling is essential for osteoblast differentiation of mesenchymal progenitors during endochondral bone formation. However, the critical role of Hh signaling during adult bone remodeling remains to be elucidated. Methods. A Smoothened (SMO) antagonist/Hedgehog inhibitor, BMS-833923, identified during a functional screening of a stem cell signaling small molecule library, was investigated for its effects on the osteoblast differentiation of human skeletal (mesenchymal) stem cells (hMSC). Alkaline phosphatase (ALP) activity and Alizarin red staining were employed as markers for osteoblast differentiation and in vitro mineralization capacity, respectively. Global gene expression profiling was performed using the Agilent® microarray platform. Effects on in vivo ectopic bone formation were assessed by implanting hMSC mixed with hydroxyapatite-tricalcium phosphate granules subcutaneously in 8-week-old female nude mice, and the amount of bone formed was assessed using quantitative histology. Results. BMS-833923, a SMO antagonist/Hedgehog inhibitor, exhibited significant inhibitory effects on osteoblast differentiation of hMSCs reflected by decreased ALP activity, in vitro mineralization, and downregulation of osteoblast-related gene expression. Similarly, we observed decreased in vivo ectopic bone formation. Global gene expression profiling of BMS-833923-treated compared to vehicle-treated control cells, identified 348 upregulated and 540 downregulated genes with significant effects on multiple signaling pathways, including GPCR, endochondral ossification, RANK-RANKL, insulin, TNF alpha, IL6, and inflammatory response. Further bioinformatic analysis employing Ingenuity Pathway Analysis revealed significant enrichment in BMS-833923-treated cells for a number of functional categories and networks involved in connective and skeletal tissue development and disorders, e.g., NFκB and STAT signaling. Conclusions. We identified SMO/Hedgehog antagonist (BMS-833923) as a powerful inhibitor of osteoblastic differentiation of hMSC that may be useful as a therapeutic option for treating conditions associated with high heterotopic bone formation and mineralization.

A Simple but Effective Demineralizing Method for Osteogenic Allograft Preparation

2005 ◽  
Vol 288-289 ◽  
pp. 249-252
Author(s):  
Hong In Shin ◽  
H.M. Ryoo ◽  
Suk Young Kim
Keyword(s):  
Bone Formation ◽  
Preparation Method ◽  
Ectopic Bone Formation ◽  
Short Treatment ◽  
H2o2 Treatment ◽  
Tissue Samples ◽  
Light Microscopic Observation ◽  
Ectopic Bone ◽  
Pathologic Findings ◽  
Osteogenic Effect

Bone regeneration with demineralized bone preparations has demonstrated its potential in grafting procedures in surgical disciplines of both medicine and dentistry. To improve the effectiveness in preparation and osteogenic regeneration, we developed a simple and rapid demineralizing method for osteogenic allograft preparation and evaluated its osteogenic effect using an ectopic bone formation assay. The rat diaphyseal cortical bones were demineralized in heated 0.6N HCl at 60° C for 10 to 30 mins using a controlled-heat ultrasonic cleaner, washed with sterilized distill water, and dehydrated with graded alcohol after 30% H2O2 treatment. The prepared grafts were implanted in rat dorsal subcutaneous pouches for 1-3 weeks and then the harvested tissue samples were prepared for routine light microscopic observation. The allografts demineralized with 60°C HCl for more than 20 mins in an ultrasonic condition were completely demineralized and effectively induced ectopic formation without specific pathologic findings. These findings suggest that demineralization with 60°C HCl for around 20 mins using a controlled-heat ultrasonic cleaner and dehydration with graded alcohol after short treatment with 30% H2O2 is a very simple but effective osteogenic allograft preparation method with minimal antigenicity and sterilizing effects.

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