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Bone Tissue Engineering (bone + tissue_engineering)
Selected AbstractsPreparation and Characterization of Gallium Releasing 3-D Alginate Coated 45S5 Bioglass® Based Scaffolds for Bone Tissue Engineering,ADVANCED ENGINEERING MATERIALS, Issue 7 2010Viviana Mourińo Abstract The aim of this work was to develop functional three-dimensional scaffolds based on Bioglass® derived glass,ceramics for bone tissue engineering (TE) with prophylaxis effect against infections. The highly porous scaffolds were fabricated by the foam replication technique and then coated with sodium alginate crosslinked with Ga3+. It was confirmed that the polymer coating did not affect the interconnectivity of the scaffold pore structure. The resulting composite scaffold exhibited antibacterial effect and improved mechanical properties as well as high bioactivity. The novel Ga3+ loaded 45S5 Bioglass®-based scaffolds coated with alginate are promising candidates for bone TE applications. [source] Hydroxyapatite/SiO2 Composites via Freeze Casting for Bone Tissue Engineering,ADVANCED ENGINEERING MATERIALS, Issue 11 2009Silke Blindow Freeze casting is a fabrication method that allows producing near-net-shaped ceramics with variable porosity. Hydroxyapatite (HA) was modified by the addition of different amounts of SiO2 nanoparticles during freeze cast preparation. The addition of SiO2 introduced a partial phase transformation of HA to , -tricalcium phosphate and improved the form stability due to less shrinkage after sintering. The impact of surface roughness of pure HA ceramics and the influence of SiO2 introduction during freeze casting on adhesion, proliferation, and differentiation of human osteoblast-like cells (MG-63) was investigated. While both cell attachment and proliferation of smooth pressed HA was significantly enhanced compared to rough freeze cast HA, the addition of SiO2 improved the cell numbers of the latter. The expression of cell differentiation markers osteocalcin and collagen I was found to be supported by rough surfaces (Ra,=,5,6,µm) in particular on ceramics containing SiO2 [source] Physical and Biological Properties of a Novel Hydrogel Composite Based on Oxidized Alginate, Gelatin and Tricalcium Phosphate for Bone Tissue Engineering,ADVANCED ENGINEERING MATERIALS, Issue 12 2007K. Cai A novel hydrogel composite is reported in this study, which was derived from oxidized alginate, gelatin and tricalcium phosphate (TCP). The physical and biological properties of these hydrogel composites prepared with oxidized sodium alginate with different oxidation degrees were investigated. The drug delivery potential of this hydrogel composite as a carrier was evaluated by using Vitamin B2 as a model drug as well. An in vitro investigation with encapsulation of osteoblast revealed that these composites were biocompatible. This hydrogel composite presented here may be utilized for the fabrication of potential injectable systems for tissue engineering, drug delivery and other medical applications. [source] Bone tissue engineering in a critical size defect compared to ectopic implantations in the goatJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 3 2004Moyo C. Kruyt Abstract Since the application of the autologous bone graft, the need for an alternative has been recognized. Tissue engineering (TE) of bone by combining bone marrow stromal cells (BMSCs) with a porous scaffold, is considered a promising technique. In this study we investigated the potential of tissue engineered bone to heal a critical sized defect in the goat. Orthotopic bone formation was compared to ectopic bone formation in comparable constructs. TE constructs were prepared from goat BMSCs and porous biphasic calcium phosphate ceramic scaffolds. These constructs and scaffolds without cells were implanted paired in critical sized iliac wing defects. Comparable samples were implanted intramuscularly. After 9 (n = 7) and 12 (n = 8) weeks implantation, the samples were analyzed histomorphometrically. After 9-weeks implantation in the iliac wing defect, significantly more bone apposition was found in the TE condition. After 12 weeks, the defects were almost completely filled with bone, but no significant advantage of TE was determined anymore. This contrasted with the intramuscular samples where TE implants showed significantly more bone at both time points. In conclusion, bone TE is feasible in critical sized defects. However, when appropriate osteoconductive/inductive materials are applied the effect of cell seeding may be temporary. © 2003 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved. [source] Pulsed electromagnetic fields affect osteoblast proliferation and differentiation in bone tissue engineeringBIOELECTROMAGNETICS, Issue 7 2007Ming-Tzu Tsai Abstract Bone tissue engineering is an interdisciplinary field involving both engineers and cell biologists, whose main purpose is to repair bone anatomical defects and maintain its functions. A novel system that integrates pulsed electromagnetic fields (PEMFs) and bioreactors was applied to bone tissue engineering for regulating osteoblast proliferation and differentiation in'vitro. Osteoblasts were acquired from the calvaria of newborn Wistar rats and isolated after sequential digestion. Poly(DL -lactic-co-glycolic acid) (PLGA) scaffolds were made by the solvent merging/particulate leaching method. Osteoblasts were seeded into porous PLGA scaffolds with 85% porosity and cultured in bioreactors for the 18-day culture period. Cells were exposed to PEMF pulsed stimulation with average (rms) amplitudes of either 0.13, 0.24, or 0.32 mT amplitude. The resulting induced electric field waveform consisted of single, narrow 300 µs quasi-rectangular pulses with a repetition rate of 7.5'Hz. The results showed that PEMF stimulation for 2 and 8 h at .13 mT increased the cell number on days 6 and 12, followed by a decrease on day 18 using 8 h stimulation. However, ALP activity was decreased and then increased on days 12 and 18, respectively. On the other hand, PEMF-treated groups (irrespective of the stimulation time) at 0.32 mT inhibited cell proliferation but enhanced ALP activity during the culture period. These findings suggested that PEMF stimulation with specific parameters had an effect on regulating the osteoblast proliferation and differentiation. This novel integrated system may have potential in bone tissue engineering. Bioelectromagnetics 28:519,528, 2007. © 2007 Wiley-Liss, Inc. [source] Positively Charged Material Surfaces Generated by Plasma Polymerized Allylamine Enhance Vinculin Mobility in Vital Human Osteoblastss,ADVANCED ENGINEERING MATERIALS, Issue 8 2010Henrike Rebl Abstract Several studies suggest that the modification of an implant surface by chemical means plays an important role in bone tissue engineering. Previously we have shown that osteoblast cell adhesion and spreading can strongly be increased by a positively charged surface. Cell adhesion and migration are two vital processes that are completely dependent on coordinated formation of focal adhesions. Changes in the organization of the actin cytoskeleton and the focal adhesions are essential for numerous cellular processes including cell motility and tissue morphogenesis. We examined the mobility of the cytoskeletally associated protein vinculin on functionalized surfaces using plasma polymerized allylamine (PPAAm), a homogenous plasma polymer layer with randomly distributed amino groups. In living, GFP,vinculin transfected osteoblastic cells we determined a significant increase in vinculin mobility and vinculin contact length on PPAAm compared to collagen I coated surfaces during the initial adhesion phase. We suggest that positive charges control the cell physiology which seems to be dominant over the integrin receptor binding to collagen I. The results emphasize the role of the surface charge for the design of artificial scaffolds in bone repair. [source] Monodisperse Mesoporous Silica Spheres Inside a Bioactive Macroporous Glass,Ceramic Scaffold,ADVANCED ENGINEERING MATERIALS, Issue 7 2010Renato Mortera In the field of bone tissue engineering, monosized MCM-41 spheres have been incorporated inside a bioactive glass,ceramic macroporous scaffold belonging to the SiO2CaOK2O (SCK) system so obtaining a multiscale hierarchical composite. The MCM-41-SCK system was prepared by dipping the glass,ceramic scaffold into the MCM-41 synthesis solution and was characterized by means of XRD, micro-XRD, N2 sorption and scanning electron microscopy. The MCM-41 spheres inside the scaffold are highly uniform in diameter, as those synthesized in powder form. The adsorption capacity of the composite toward ibuprofen is three times higher than that of the MCM-41-free scaffold, because of the presence of the ordered mesoporous silica. Also the release behavior in SBF at 37,°C is strongly affected by the presence of MCM-41 inside the scaffold macropores. [source] Preparation and Characterization of Gallium Releasing 3-D Alginate Coated 45S5 Bioglass® Based Scaffolds for Bone Tissue Engineering,ADVANCED ENGINEERING MATERIALS, Issue 7 2010Viviana Mourińo Abstract The aim of this work was to develop functional three-dimensional scaffolds based on Bioglass® derived glass,ceramics for bone tissue engineering (TE) with prophylaxis effect against infections. The highly porous scaffolds were fabricated by the foam replication technique and then coated with sodium alginate crosslinked with Ga3+. It was confirmed that the polymer coating did not affect the interconnectivity of the scaffold pore structure. The resulting composite scaffold exhibited antibacterial effect and improved mechanical properties as well as high bioactivity. The novel Ga3+ loaded 45S5 Bioglass®-based scaffolds coated with alginate are promising candidates for bone TE applications. [source] Development of Biomimetic Chitosan-Based Hydrogels Using an Elastin-Like Polymer,ADVANCED ENGINEERING MATERIALS, Issue 1-2 2010Joaquim S. Barbosa Chitosan and an elastin-like polymer, containing a specific osteoconductive sequence in the primary structure, have been combined to obtain bioactive injectable systems with enhanced mechanical properties and hydrogels. Obtained results indicate that the combination of such polymers may be very promising in the development of biomaterials for minimal invasive orthopaedic reconstructive applications or in bone tissue engineering. The figure shows a thermo-sensitive hydrogel, with a gelation point under physiological temperature. [source] Novel Rice-shaped Bioactive Ceramic Nanoparticles (Adv. Eng.ADVANCED ENGINEERING MATERIALS, Issue 5 2009Mater. The cover of Advanced Biomaterials shows Rice-shaped bioactive ceramic nanoparticles with 70 nm in average diameter and around 200 nm in length were produced by an improved sol-gel method. In comparison to most traditional bioactive glass/ceramic materials this novel bioactive ceramic contains a significant lower quantity of silicon and higher content of phosphorous. In vitro bioactivity test showed that this new class of materials can induce the deposition of an apatite layer from SBF solution, having potential to be used in both conventional orthopedic applications or in bone tissue engineering when incorporated in composite scaffolds. More information can be found in the article of J. F. Mano et al. on page B25. [source] Novel Rice-shaped Bioactive Ceramic Nanoparticles,ADVANCED ENGINEERING MATERIALS, Issue 5 2009Zhongkui Hong Rice-shaped bioactive ceramic nanoparticles of 70 nm average diameter and around 200 nm length were produced by an improved sol-gel method. In comparison to most traditional bioactive glass/ceramic materials, this novel bioactive ceramic contains a significant lower quantity of silicon and higher content of phosphorous. In vitro bioactivity tests showed that this new class of materials can induce the deposition of an apatite layer from simulated body fluid, having the potential to be used in both conventional orthopedic applications or in bone tissue engineering when incorporated in composite scaffolds. [source] Chemical characteristics and cytocompatibility of collagen-based scaffold reinforced by chitin fibers for bone tissue engineeringJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 2 2006Xiaoming Li Abstract Chitin is a kind of seemly material to match PLLA for a scaffold, which may create an appropriate environment for the regeneration of tissues. In this study, we prepared and evaluated a new nano-hydroxyapatite/collagen/PLLA (nHACP) scaffold reinforced by chitin fibers for bone-tissue engineering. The chitin fibers were crosslinked with PLLA by dicyclohexylcarbodimide (DCC). The chemical characteristics were evaluated by Fourier transformed infrared (FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS). The mechanical strength was measured by compressive tests. The fibers, crosslinked with PLLA, could enhance the compressive strength of the scaffold about four times. Human marrow mesenchymal stem cells (MSCs) culture showed that the reinforced nHACP scaffolds were more cytocompatible than that without reinforcement. The crosslinks hardly affected the cytocompatibility of the reinforced scaffolds. The results suggested that the reinforced scaffolds (DCC crosslinked) might be a promising candidate for bone-tissue engineering. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006 [source] Formation of three-dimensional cell/polymer constructs for bone tissue engineering in a spinner flask and a rotating wall vessel bioreactorJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 1 2002Vassilios I. Sikavitsas Abstract The aim of this study is to investigate the effect of the cell culture conditions of three-dimensional polymer scaffolds seeded with rat marrow stromal cells (MSCs) cultured in different bioreactors concerning the ability of these cells to proliferate, differentiate towards the osteoblastic lineage, and generate mineralized extracellular matrix. MSCs harvested from male Sprague,Dawley rats were culture expanded, seeded on three-dimensional porous 75:25 poly(D,L -lactic- co -glycolic acid) biodegradable scaffolds, and cultured for 21 days under static conditions or in two model bioreactors (a spinner flask and a rotating wall vessel) that enhance mixing of the media and provide better nutrient transport to the seeded cells. The spinner flask culture demonstrated a 60% enhanced proliferation at the end of the first week when compared to static culture. On day 14, all cell/polymer constructs exhibited their maximum alkaline phosphatase activity (AP). Cell/polymer constructs cultured in the spinner flask had 2.4 times higher AP activity than constructs cultured under static conditions on day 14. The total osteocalcin (OC) secretion in the spinner flask culture was 3.5 times higher than the static culture, with a peak OC secretion occurring on day 18. No considerable AP activity and OC secretion were detected in the rotating wall vessel culture throughout the 21-day culture period. The spinner flask culture had the highest calcium content at day 14. On day 21, the calcium deposition in the spinner flask culture was 6.6 times higher than the static cultured constructs and over 30 times higher than the rotating wall vessel culture. Histological sections showed concentration of cells and mineralization at the exterior of the foams at day 21. This phenomenon may arise from the potential existence of nutrient concentration gradients at the interior of the scaffolds. The better mixing provided in the spinner flask, external to the outer surface of the scaffolds, may explain the accelerated proliferation and differentiation of marrow stromal osteoblasts, and the localization of the enhanced mineralization on the external surface of the scaffolds. © 2002 Wiley Periodicals, Inc. J Biomed Mater Res 62: 136,148, 2002 [source] Strategies for Directing the Differentiation of Stem Cells Into the Osteogenic Lineage In Vitro,JOURNAL OF BONE AND MINERAL RESEARCH, Issue 9 2004Boon Chin Heng Abstract A major area in regenerative medicine is the application of stem cells in bone reconstruction and bone tissue engineering. This will require well-defined and efficient protocols for directing the differentiation of stem cells into the osteogenic lineage, followed by their selective purification and proliferation in vitro. The development of such protocols would reduce the likelihood of spontaneous differentiation of stem cells into divergent lineages on transplantation, as well as reduce the risk of teratoma formation in the case of embryonic stem cells. Additionally, such protocols could provide useful in vitro models for studying osteogenesis and bone development, and facilitate the genetic manipulation of stem cells for therapeutic applications. The development of pharmokinetic and cytotoxicity/genotoxicity screening tests for bone-related biomaterials and drugs could also use protocols developed for the osteogenic differentiation of stem cells. This review critically examines the various strategies that could be used to direct the differentiation of stem cells into the osteogenic lineage in vitro. [source] The polymine spermine regulates osteogenic differentiation in adipose stem cells,JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 5a 2008G.S. Tjabringa Abstract For bone tissue engineering, it is important that mesenchymal stem cells (MSCs) differentiate into osteoblasts. To develop a method for differentiation of adipose tissue-derived mesenchymal stem cells (AT-MSCs) along the osteogenic lineage, we studied the effect of polyamines, which are organic cations implicated in bone growth and development, on differentiation of AT-MSCs. Treatment of goat-derived AT-MSCs with 1,25-dihydroxyvitamin-D3 (1,25(OH)2D3), which stimulates osteogenic differentiation, for 7 days induced gene expression of the polyamine-modulated transcription factor-1 (PMF-1) and spermidine/spermine N (1)-acetyltransferase (SSAT), which are both involved in polyamine metabolism, suggesting that polyamines are involved in osteogenic differentiation of AT-MSCs. Furthermore, treatment of AT-MSCs with the polyamine spermine-regulated gene expression of runx-2, a transcription factor involved in early stages of osteogenic differentiation, and that of osteopontin, a bone matrix protein expressed in later stages of osteogenic differentiation. Runx-2 gene expression was increased 4 and 14 days after a short 30 min. treatment with spermine, while osteopontin gene expression was only increased 4 days after spermine treatment. Finally, alkaline phosphatase activity, which is intimately involved in the formation of extracellular matrix of bone, was increased 4 weeks after the 30 min.-spermine treatment of AT-MSCs. In conclusion, this study shows for the first time that the polyamine spermine regulates differentiation of AT-MSCs along the osteogenic lineage, which can be used as a new method for differentiation of AT-MSCs along the osteogenic lineage. Therefore, polyamines may constitute a promising tool for bone tissue engineering approaches using AT-MSCs, such as a one-step surgical procedure for spinal interbody fusion. [source] Genetic marking with the ,LNGFR-gene for tracing goat cells in bone tissue engineering,JOURNAL OF ORTHOPAEDIC RESEARCH, Issue 4 2004M. C. Kruyt Abstract The use of bone marrow derived stromal cells (BMSC's) for bone tissue engineering has gained much attention as an alternative for autologous bone grafting. Little is known however, about the survival and differentiation of the cells, especially in the clinical application. The aim of this study was to develop a method to trace goat BMSC's in vivo. We investigated retroviral genetic marking, which allows stable expression of the label with cell division. Goat BMSC's were subjected to an amphotropic envelope containing a MoMuLV-based vector expressing the human low affinity nerve growth factor receptor (,LNGFR). Labeling efficiency and effect on the cells were analyzed. Furthermore, transduced cells were seeded onto porous ceramic scaffolds, implanted subcutaneously in nude mice and examined after successive implantation periods. Flow cytometry indicated a transduction efficiency of 40,60%. Immunohistochemistry showed survival and subsequent bone formation of the gene-marked cells in vivo. Besides, marked cells were also found in cartilage and fibrous tissue. These findings indicate the maintenance of the precursor phenotype following gene transfer as well as the ability of the gene to be expressed following differentiation. We conclude that retroviral gene marking with ,LNGFR is applicable to trace goat BMSC's in bone tissue engineering research. © 2003 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved. [source] Self-hardening calcium phosphate composite scaffold for bone tissue engineering,JOURNAL OF ORTHOPAEDIC RESEARCH, Issue 3 2004Hockin H. K. Xu Abstract Calcium phosphate cement (CPC) sets in situ to form solid hydroxyapatite, can conform to complex cavity shapes without machining, has excellent osteoconductivity, and is able to be resorbed and replaced by new bone. Therefore, CPC is promising for craniofacial and orthopaedic repairs. However, its low strength and lack of macroporosity limit its use. This study investigated CPC reinforcement with absorbable fibers, the effects of fiber volume fraction on mechanical properties and macroporosity, and the cytotoxicity of CPC,fiber composite. The rationale was that large-diameter absorbable fibers would initially strengthen the CPC graft, then dissolve to form long cylindrical macropores for colonization by osteoblasts. Flexural strength, work-of-fracture (toughness), and elastic modulus were measured vs. fiber volume fraction from 0% (CPC Control without fibers) to 60%. Cell culture was performed with osteoblast-like cells, and cell viability was quantified using an enzymatic assay. Flexural strength (mean ± SD; n == 6) of CPC with 60% fibers was 13.5 ± 4.4 MPa, three times higher than 3.9 ± 0.5 MPa of CPC Control. Work-of-fracture was increased by 182 times. Long cylindrical macropores 293 ± 46 ,m in diameter were created in CPC after fiber dissolution, and the CPC,fiber scaffold reached a macroporosity of 55% and a total porosity of 81%. The new CPC,fiber formulation supported cell adhesion, proliferation and viability. The method of using large-diameter absorbable fibers in bone graft for mechanical properties and formation of long cylindrical macropores for bone ingrowth may be applicable to other tissue engineering materials. Published by Elsevier Ltd. on behalf of Orthopuedic Research Society. © 2003 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved. [source] A novel synthetic peptide vector system for optimal gene delivery to bone marrow stromal cellsJOURNAL OF PEPTIDE SCIENCE, Issue 3 2007Pan Haitao Abstract A 23-amino acid, bifunctional, integrin-targeted synthetic peptide was evaluated for ex vivo gene delivery to rabbit bone marrow stromal cells (BMSCs). The peptide (K)16GRGDSPC consists of an amino terminal domain of 16 lysines for electrostatic binding of DNA, and a 7-amino acid integrin-binding domain at the carboxyl terminal. PcDNA3-EGFP plasmids were transfected into BMSCs by (K)16GRGDSPC and the positive cells gave out a bright green fluorescence. High levels of gene delivery of pcDNA3-TGF-,1 plasmids were obtained with 2 to 4 µg/ml DNA concentration, with (K)16GRGDSPC at an optimal peptide: DNA w/w ratio of 3:1, with a required exposure time of more than 4 h but shorter than 24 h for BMSC exposure to the peptide/DNA complexes with completely absent serum in the initial stage; with 100 µM chloroquine and at least 8 h exposure for BMSC exposure to chloroquine; with a fusogenic peptide at an optimal (K)16GRGDSPC/DNA/fusogenic peptide w/w ratio of 3:1:5; and with Lipofectamine 2000 at an optimal (K)16GRGDSPC/DNA/Lipofectamine 2000 w/w ratio of 3:1:2 at a constant DNA concentration of 2 µg/ml. Chloroquine, the fusogenic peptide and Lipofectamine 2000 all significantly promoted gene delivery, but chloroquine was more effective than the fusogenic peptide and had obvious synergistic effects with Lipofectamine 2000. Under optimal conditions, TGF-,1 gene was transfected into BMSCs without observable toxicity, and the stable expression was examined by RT-PCR and Western blot analysis. The stable transgenic cells showed obvious bands. This novel synthetic peptide, providing a new way for the use of polylysine and RGD motif in DNA vector system, is potentially well suited to ex vivo gene delivery to BMSCs for experimental and clinical applications in the field of bone tissue engineering. Copyright © 2006 European Peptide Society and John Wiley & Sons, Ltd. [source] The effect of PKC activation and inhibition on osteogenic differentiation of human mesenchymal stem cellsJOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, Issue 5 2010Jun Liu Abstract Human mesenchymal stem cells (hMSCs) are being considered for several areas of clinical therapy, due to their multipotent nature. For instance, osteogenic hMSCs are applied in bone tissue engineering, but current differentiation protocols need further optimization before they can be clinically applied. Protein kinase C (PKC) family members have been implicated in bone metabolism, which prompted us to use a pharmaceutical approach to manipulate PKC signalling in hMSCs. Inhibition of PKC resulted in a dose-dependent inhibition of dexamethasone-induced osteogenic differentiation. Surprisingly, PKC activation using phorbol 12-myristate 13-acetate (PMA) also resulted in inhibition of osteogenesis, although we observed that inhibition was more pronounced at low than at high concentrations of PMA. Furthermore, we observed that inhibition of PKC, blocked alkaline phosphatase (ALP, an early marker of osteogenic differentiation) expression, whereas inhibition of the conventional PKC subfamily and PKCµ using Gö6976 resulted in an induction of ALP activity, collagen (I) expression and mineralization. In conclusion, inhibition of the conventional PKCs/PKCµ and activation of PKC, could further benefit osteogenic differentiation of hMSCs in vitro and in vivo, which is currently under investigation. Copyright © 2009 John Wiley & Sons, Ltd. [source] Skeletal tissue engineering using embryonic stem cellsJOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, Issue 3 2010Jojanneke M. Jukes Abstract Various cell types have been investigated as candidate cell sources for cartilage and bone tissue engineering. In this review, we focused on chondrogenic and osteogenic differentiation of mouse and human embryonic stem cells (ESCs) and their potential in cartilage and bone tissue engineering. A decade ago, mouse ESCs were first used as a model to study cartilage and bone development and essential genes, factors and conditions for chondrogenesis and osteogenesis were unravelled. This knowledge, combined with data from the differentiation of adult stem cells, led to successful chondrogenic and osteogenic differentiation of mouse ESCs and later also human ESCs. Next, researchers focused on the use of ESCs for skeletal tissue engineering. Cartilage and bone tissue was formed in vivo using ESCs. However, the amount, homogeneity and stability of the cartilage and bone formed were still insufficient for clinical application. The current protocols require improvement not only in differentiation efficiency but also in ESC-specific hurdles, such as tumourigenicity and immunorejection. In addition, some of the general tissue engineering challenges, such as cell seeding and nutrient limitation in larger constructs, will also apply for ESCs. In conclusion, there are still many challenges, but there is potential for ESCs in skeletal tissue engineering. Copyright © 2009 John Wiley & Sons, Ltd. [source] State of the art and future directions of scaffold-based bone engineering from a biomaterials perspectiveJOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, Issue 4 2007Dietmar Werner Hutmacher Abstract Scaffold-based bone tissue engineering aims to repair/regenerate bone defects. Such a treatment concept involves seeding autologous osteogenic cells throughout a biodegradable scaffold to create a scaffold,cell hybrid that may be called a tissue-engineered construct (TEC). A variety of materials and scaffolding fabrication techniques for bone tissue engineering have been investigated over the past two decades. This review aims to discuss the advances in bone engineering from a scaffold material point of view. In the first part the reader is introduced to the basic principles of bone engineering. The important properties of the biomaterials and the scaffold design in the making of tissue engineered bone constructs are discussed in detail, with special emphasis placed on the new material developments, namely composites made of synthetic polymers and calcium phosphates. Advantages and limitations of these materials are analysed along with various architectural parameters of scaffolds important for bone tissue engineering, e.g. porosity, pore size, interconnectivity and pore-wall microstructures. Copyright © 2007 John Wiley & Sons, Ltd. [source] Materials in particulate form for tissue engineering.JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, Issue 2 2007Abstract Materials in particulate form have been the subjects of intensive research in view of their use as drug delivery systems. While within this application there are still issues to be addressed, these systems are now being regarded as having a great potential for tissue engineering applications. Bone repair is a very demanding task, due to the specific characteristics of skeletal tissues, and the design of scaffolds for bone tissue engineering presents several difficulties. Materials in particulate form are now seen as a means of achieving higher control over parameters such as porosity, pore size, surface area and the mechanical properties of the scaffold. These materials also have the potential to incorporate biologically active molecules for release and to serve as carriers for cells. It is believed that the combination of these features would create a more efficient approach towards regeneration. This review focuses on the application of materials in particulate form for bone tissue engineering. A brief overview of bone biology and the healing process is also provided in order to place the application in its broader context. An original compilation of molecules with a documented role in bone tissue biology is listed, as they have the potential to be used in bone tissue engineering strategies. To sum up this review, examples of works addressing the above aspects are presented. Copyright © 2007 John Wiley & Sons, Ltd. [source] A Study on Biomineralization Behavior of N -Methylene Phosphochitosan ScaffoldsMACROMOLECULAR BIOSCIENCE, Issue 10 2004Yu Ji Yin Abstract Summary: Biomimetic growth of calcium phosphate over natural polymer may be an effective approach to constituting an organic/inorganic composite scaffold for bone tissue engineering. In this work, N -methylene phosphochitosan (NMPCS) was prepared via formaldehyde addition and condensation with phosphoric acid in a step that allowed homogeneous modification without obvious deterioration in chitosan (CS) properties. The NMPCS obtained was characterized by using FT-IR and elemental analysis. The macroporous scaffolds were fabricated through a freeze-drying technique. A comparative study on NMPCS and CS scaffold biomimetic mineralization was carried out in different media, i.e, a simulated body fluid (SBF) or alternative CaCl2 and Na2HPO4 solutions respectively. Apatite formation within NMPCS and CS scaffolds was identified with FT-IR, scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) and X-ray diffractometery (XRD). The results revealed alternate soaking of the scaffolds in CaCl2 and Na2HPO4 solutions was better than soaking in SBF solution alone in relation to apatite deposition on the scaffold pore walls. Biomineralization provides an approach to improve nature derived materials, e.g., chitosan derivative NMPCS properties e.g., compressive modulus, etc. SEM image of a NMPCS/apatite composite scaffold. [source] Porous silicon as a cell interface for bone tissue engineeringPHYSICA STATUS SOLIDI (A) APPLICATIONS AND MATERIALS SCIENCE, Issue 5 2007Wei Sun Abstract A novel cell interface has been constructed on porous silicon. We have demonstrated that nano- to macro-scale porous architectures have promising osteoconductive potentials. Macroporous silicon (pore opening 1,2 µm) is especially favorable for osteoblast adhesion, growth, protein synthesis and mineralization. An electronic/optoelectronic controllable medical implant with both scaffolding and drug delivery functions may be created for orthopaedic tissue engineering with this material. (© 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Electrospun poly(L -lactic acid)/hydroxyapatite composite fibrous scaffolds for bone tissue engineering,POLYMER INTERNATIONAL, Issue 2 2010Boontharika Chuenjitkuntaworn Abstract Poly(L -lactic acid) (PLLA) is one of the most studied synthetic biodegradable polymeric materials as a bone graft substitute. Taking into account the osteoconductive property of hydroxyapatite (HAp), we prepared fibrous matrices of PLLA without and with HAp particles in amounts of 0.25 or 0.50% (w/v, based on the volume of the base 15% w/v PLLA solution in 70:30 v/v dichloromethane/tetrahydrofuran). These fibrous matrices were assessed for their potential as substrates for bone cell culture. The presence of HAp in the composite fibre mats was confirmed using energy dispersive X-ray spectroscopy mapping. The average diameters of both neat PLLA and PLLA/HAp fibres, as determined using scanning electron microscopy, ranged between 2.3 and 3.5 µm, with the average spacing between adjacent fibres ranging between 5.7 and 8.5 µm. The porosity of these fibrous membranes was high (ca 97,98%). A direct cytotoxicity evaluation with L929 mouse fibroblasts indicated that the neat PLLA fibre mats released no substance at a level that was toxic to the cells. The presence of HAp particles at 0.50% w/v in the PLLA fibrous scaffolds not only promoted the attachment and the proliferation of MC3T3-E1 mouse pre-osteoblastic cells, but also increased the expression of osteocalcin mRNA and the extent of mineralization after the cells had been cultured on the scaffolds for 14 and 21 days, respectively. The results obtained suggested that the PLLA/HAp fibre mats could be materials of choice for bone tissue engineering. Copyright © 2009 Society of Chemical Industry [source] Bone Morphogenetic Protein-6-loaded Chitosan Scaffolds Enhance the Osteoblastic Characteristics of MC3T3-E1 CellsARTIFICIAL ORGANS, Issue 1 2010Abdullah C. Akman Abstract The purpose of this study is to investigate the convenience of bone morphogenetic protein-6 (BMP-6)-loaded chitosan scaffolds with preosteoblastic cells for bone tissue engineering. MC3T3-E1 cells were seeded into three different groups: chitosan scaffolds, BMP-6-loaded chitosan scaffolds, and chitosan scaffolds with free BMP-6 in culture medium. Tissue-engineered constructs were characterized by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide assay, scanning electron microscopy (SEM), mineralization assay (von Kossa), alkaline phosphatase (ALP) activity, and osteocalcin (OCN) assays. BMP-6-loaded chitosan scaffolds supported proliferation of the MC3T3-E1 mouse osteogenic cells in a similar pattern as the unloaded chitosan scaffolds group and as the chitosan scaffolds with free BMP-6 group. SEM images of the cell-seeded scaffolds revealed significant acceleration of extracellular matrix synthesis in BMP-6-loaded chitosan scaffolds. Both levels of ALP and OCN were higher in BMP-6-loaded chitosan scaffold group compared with the other two groups. In addition, BMP-6-loaded scaffolds showed strong staining in mineralization assays. These findings suggest that BMP-6-loaded chitosan scaffold supports cellular functions of the osteoblastic cells; therefore, this scaffold is considered as a new promising vehicle for bone tissue engineering applications. [source] Effect of Osteogenic Induction on the in Vitro Differentiation of Human Embryonic Stem Cells Cocultured With Periodontal Ligament FibroblastsARTIFICIAL ORGANS, Issue 11 2007Bülend Inanē Abstract:, Osteogenesis is one of the principal components of periodontal tissue development as well as regeneration. As pluripotent cells with unlimited proliferative potential and differentiation ability to all germ layer representatives, embryonic stem cells also hold the promise to become a cell source in bone tissue engineering. Our aim was to investigate osteogenic differentiation potential of human embryonic stem cells (hESCs) under the inductive influence of human periodontal ligament fibroblast (hPDLF) monolayers. After being expanded and characterized morphologically and immunohistochemically, hESCs (HUES-9) were cocultured with hPDLFs for 28 days. Two groups were established: (i) osteogenic induction group with ascorbic acid, ,-glycerophosphate, and dexamethasone containing hESC differentiation medium; and (ii) spontaneous differentiation group cultured in hESC differentiation medium. Morphological shift in cells was analyzed under an inverted microscope, and immunohistochemistry was performed on fixed specimens at days 1 and 28 using antibodies against alkaline phosphatase, osteonectin, osteopontin, bone sialoprotein (BSP), and osteocalcin (OSC). Reverse transcription,polymerase chain reaction was utilized for the detection of octameric binding protein-4, BSP, and OSC expression at mRNA level. Mineralization was assessed using alizarin red, and the surface topology shift in colonies was demonstrated with scanning electron microscopy. Results indicate the feasibility of osteogenic differentiation of hESCs in coculture, and suggest a role of periodontal ligament fibroblasts in their differentiation patterns. Advances in the field could allow for potential utilization of hESCs in periodontal tissue engineering applications involving regeneration of bone in periodontal compartment lost as a result of destructive periodontal diseases. [source] Pulsed electromagnetic fields affect osteoblast proliferation and differentiation in bone tissue engineeringBIOELECTROMAGNETICS, Issue 7 2007Ming-Tzu Tsai Abstract Bone tissue engineering is an interdisciplinary field involving both engineers and cell biologists, whose main purpose is to repair bone anatomical defects and maintain its functions. A novel system that integrates pulsed electromagnetic fields (PEMFs) and bioreactors was applied to bone tissue engineering for regulating osteoblast proliferation and differentiation in'vitro. Osteoblasts were acquired from the calvaria of newborn Wistar rats and isolated after sequential digestion. Poly(DL -lactic-co-glycolic acid) (PLGA) scaffolds were made by the solvent merging/particulate leaching method. Osteoblasts were seeded into porous PLGA scaffolds with 85% porosity and cultured in bioreactors for the 18-day culture period. Cells were exposed to PEMF pulsed stimulation with average (rms) amplitudes of either 0.13, 0.24, or 0.32 mT amplitude. The resulting induced electric field waveform consisted of single, narrow 300 µs quasi-rectangular pulses with a repetition rate of 7.5'Hz. The results showed that PEMF stimulation for 2 and 8 h at .13 mT increased the cell number on days 6 and 12, followed by a decrease on day 18 using 8 h stimulation. However, ALP activity was decreased and then increased on days 12 and 18, respectively. On the other hand, PEMF-treated groups (irrespective of the stimulation time) at 0.32 mT inhibited cell proliferation but enhanced ALP activity during the culture period. These findings suggested that PEMF stimulation with specific parameters had an effect on regulating the osteoblast proliferation and differentiation. This novel integrated system may have potential in bone tissue engineering. Bioelectromagnetics 28:519,528, 2007. © 2007 Wiley-Liss, Inc. [source] Design of cellular porous biomaterials for wall shear stress criterionBIOTECHNOLOGY & BIOENGINEERING, Issue 4 2010Yuhang Chen Abstract The microfluidic environment provided by implanted prostheses has a decisive influence on the viability, proliferation and differentiation of cells. In bone tissue engineering, for instance, experiments have confirmed that a certain level of wall shear stress (WSS) is more advantageous to osteoblastic differentiation. This paper proposes a level-set-based topology optimization method to regulate fluidic WSS distribution for design of cellular biomaterials. The topological boundary of fluid phase is represented by a level-set model embedded in a higher-dimensional scalar function. WSS is determined by the computational fluid dynamics analysis in the scale of cellular base cells. To achieve a uniform WSS distribution at the solid,fluid interface, the difference between local and target WSS is taken as the design criterion, which determines the speed of the boundary evolution in the level-set model. The examples demonstrate the effectiveness of the presented method and exhibit a considerable potential in the design optimization and fabrication of new prosthetic cellular materials for bioengineering applications. Biotechnol. Bioeng. 2010;107:737,746. © 2010 Wiley Periodicals, Inc. [source] A rotating bed system bioreactor enables cultivation of primary osteoblasts on well-characterized sponceram® regarding structural and flow propertiesBIOTECHNOLOGY PROGRESS, Issue 3 2010Kirstin Suck Abstract The development of bone tissue engineering depends on the availability of suitable biomaterials, a well-defined and controlled bioreactor system, and on the use of adequate cells. The biomaterial must fulfill chemical, biological, and mechanical requirements. Besides biocompatibility, the structural and flow characteristics of the biomaterial are of utmost importance for a successful dynamic cultivation of osteoblasts, since fluid percolation within the microstructure must be assured to supply to cells nutrients and waste removal. Therefore, the biomaterial must consist of a three-dimensional structure, exhibit high porosity and present an interconnected porous network. Sponceram®, a ZrO2 based porous ceramic, is characterized in the presented work with regard to its microstructural design. Intrinsic permeability is obtained through a standard Darcy's experiment, while Young's modulus is derived from a two plates stress,strain test in the linear range. Furthermore, the material is applied for the dynamic cultivation of primary osteoblasts in a newly developed rotating bed bioreactor. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010 [source] | |||||||||||||||||||