3D Bioplotter Research Papers

Displaying all papers about Bone Regeneration (100 results)

3D-printed scaffolds of biomineralized hydroxyapatite nanocomposite on silk fibroin for improving bone regeneration

Applied Surface Science 2019 Volumes 467–468, Pages 345-353

In an attempt to fabricate biomimetic bone repair scaffolds and improve bone regeneration point of view, we have three dimensionally printed porous scaffolds with biomineralized hydroxyapatite/silk fibroin nanocomposites. SF/HA composite particles were firstly produced via an in-situ mineral precipitation process when SF molecules were served as templates.. Microscopy observations of SF/HA showed homogeneous morphology and narrowly distributed size. By using sodium alginate (SA) as paste binder, scaffolds with different contents of SF/HA were subsequently 3D-printed under proper conditions. All the scaffolds were porous with 3D interconnected large pores (size ~400 μm) and an overall porosity about 70%, combined with a relative…

Collagen/hydroxyapatite bone grafts manufactured by homogeneous/ heterogeneous 3D printing

Materials Letters 2018 Volume 231, Pages 179-182

This paper presents a new way to obtain collagen/hydroxyapatite (COLL/HA) composite materials by 3D printing. Because of high tendency of segregation of COLL/HA composite materials, printing was done using COLL/Ca2+ gel (even COLL/Ca(OH)2) followed by precipitation of HA and crosslinking of COLL. The HA precipitation occurs simultaneously with crosslinking of COLL molecules, these processes being assured by the presence of glutaraldehyde supplemented PBS solution. By printing with COLL/Ca2+ at acidic pH homogeneity was increased. FTIR spectroscopy and microscopy reveal HA formation as the main inorganic phase these nanoparticles being homogeneously dispersed in the volume. In vitro biocompatibility assays were performed…

Heparin/Poly-L-lysine-coated 3D-printed PLGA scaffolds as drug carriers for local immune modulation in bone regeneration

2018

Immune responses after injury play a critical role in bone regeneration. Initiation of inflammation at early stages of repair triggers tissue formation and remodeling; however, uncontrolled inflammation underlies a catabolic effect on tissues as commonly seen in arthritis where inflammation breaks down tissues and hinders regeneration. Our ultimate goal is to design a novel approach on bone scaffolds for which biodegradable scaffolds are loaded with inflammatory cytokines for local immunomodulation as well as bone regeneration. We employed nanoparticles (NPs) composed of heparin (Hep) and poly-L-lysine (PLL) as cytokine drug carriers adhered on 3D-printed poly(lactic-co-glycolic acid) (PLGA) scaffolds. The entire drug…

3D Printing Nanoscale Bioactive Glass Scaffolds Enhance Osteoblast Migration and Extramembranous Osteogenesis through Stimulating Immunomodulation

Advanced Healthcare Materials 2018 Volume 7, Article 1800361

Bioactive glass (BG) can repair bone defects, however, it is not clear whether BG has the ability for bone augmentation without making any bone defect. Unlike the intramembranous osteogenesis in bone defect repair, the extramembranous osteogenesis occurs outside the cortical bone and the osteoprogenitor cells show the reversed migration. Herein, nanoscale bioactive glass scaffolds (BGSs) are fabricated, and their role and immunomodulation‐related mechanism in the extramembranous osteogenesis are investigated. The in vitro migration and differentiation of calvaria preosteoblasts are studied by culturing with peripheral macrophage‐conditioned medium after stimulating with BGSs. The results indicate that the proinflammatory environment significantly promotes preosteoblast…

Zein regulating apatite mineralization, degradability, in vitro cells responses and in vivo osteogenesis of 3D-printed scaffold of n-MS/ZN/PCL ternary composite

RSC Advances 2018 Volume 8, Pages 18745-18756

Bioactive and degradable scaffolds of nano magnesium silicate (n-MS)/zein (ZN)/poly(caprolactone) (PCL) ternary composites were prepared by 3D-printing method. The results showed that the 3D-printed scaffolds possessed controllable pore structure, and pore morphology, pore size, porosity and pore interconnectivity of the scaffolds can be efficiently adjusted. In addition, the apatite-mineralization ability of the scaffolds in simulated body fluids was obviously improved with the increase of ZN content, in which the scaffold with 20 w% ZN (C20) possessed excellent apatite-mineralization ability. Moreover, the degradability of the scaffolds was significantly enhanced with the increase of ZN content in the scaffolds. The degradation of…

Response of hPDLSCs on 3D printed PCL/PLGA composite scaffolds in vitro

Molecular Medicine Reports 2018 Volume 18 Issue 2, Pages 1335-1344

Three‑dimensional printed (3DP) scaffolds have become an excellent resource in alveolar bone regeneration. However, selecting suitable printable materials remains a challenge. In the present study, 3DP scaffolds were fabricated using three different ratios of poly (ε‑caprolactone) (PCL) and poly‑lactic‑co‑glycolic acid (PLGA), which were 0.1PCL/0.9PLGA, 0.5PCL/0.5PLGA and 0.9PCL/0.1PLGA. The surface characteristics and degradative properties of the scaffolds, and the response of human periodontal ligament stem cells (hPDLSCs) on the scaffolds, were assessed to examine the preferable ratio of PCL and PLGA for alveolar bone regeneration. The results demonstrated that the increased proportion of PLGA markedly accelerated the degradation, smoothed the surface…

3D printed mesoporous bioactive glass/metal-organic framework scaffolds with antitubercular drug delivery

Microporous and Mesoporous Materials 2018 Volume 272, Pages 24-30

Three-dimensional (3D) porous scaffolds with sustained drug delivery are pursued for osteoarticular tuberculosis therapy after surgery. In this study, mesoporous bioactive glass/metal-organic framework (MBG/MOF) scaffolds with sustained antitubercular drug release have been fabricated by 3D printing. The results showed that the MBG/MOF scaffolds possess macropores of ca. 400 μm and enhanced compressive strength of 3–7 MPa, also exhibited good biocompatibility and apatite forming ability in vitro. Furthermore, the drug release rate and pH microenvironment of the MBG/MOF scaffolds could be controlled due to the MOF degradation. These results indicated that the 3D printed MBG/MOF scaffolds are promising for treating osteoarticular tuberculosis.

Vascularization of Natural and Synthetic Bone Scaffolds

Cell transplantation 2018 Volume 27, Issue 8, Pages 1269–1280

Vascularization of engineered bone tissue is critical for ensuring its survival after implantation. In vitro pre-vascularization of bone grafts with endothelial cells is a promising strategy to improve implant survival. In this study, we pre-cultured human smooth muscle cells (hSMCs) on bone scaffolds for 3 weeks followed by seeding of human umbilical vein endothelial cells (HUVECs), which produced a desirable environment for microvasculature formation. The sequential cell-seeding protocol was successfully applied to both natural (decellularized native bone, or DB) and synthetic (3D-printed Hyperelastic “Bone” scaffolds, or HB) scaffolds, demonstrating a comprehensive platform for developing natural and synthetic-based in vitro vascularized…

Three-dimensional printing of β-tricalcium phosphate/calcium silicate composite scaffolds for bone tissue engineering

Bio-Design and Manufacturing 2018 Volume 1, Issue 2, Pages 146-156

Bioactive scaffolds with interconnected porous structures are essential for guiding cell growth and new bone formation. In this work, we successfully fabricated three-dimensional (3D) porous β-tricalcium phosphate (β-TCP)/calcium silicate (CS) composite scaffolds with different ratios by 3D printing technique and further investigated the physiochemical properties, in vitro apatite mineralization properties and degradability of porous β-TCP/CS scaffolds. Moreover, a series of in vitro cell experiments including the attachment, proliferation and osteogenic differentiation of mouse bone marrow stromal cells were conducted to testify their biological performances. The results showed that 3D printed β-TCP/CS scaffolds possessed of controllable internal porous structures and external…

3D-printed IFN-γ-loading calcium silicate-β-tricalcium phosphate scaffold sequentially activates M1 and M2 polarization of macrophages to promote vascularization of tissue engineering bone

Acta Biomaterialia 2018 Volume 71, Pages 96-107

To promote vascularization of tissue-engineered bone, IFN-γ polarizing macrophages to M1 was loaded on 5% calcium silicate/β-tricalcium phosphate (CaSiO3-β-TCP) scaffolds. IFN-γ and Si released from the scaffold were designed to polarize M1 and M2 macrophages, respectively. β-TCP, CaSiO3-β-TCP, and IFN-γ@CaSiO3-β-TCP were fabricated and biocompatibilities were evaluated. Polarizations of macrophages were detected by flow cytometry. Human umbilical vein endothelial cells with GFP were cultured and induced on Matrigel with conditioned culture medium extracted from culture of macrophages loaded on scaffolds for evaluating angiogenesis. Four weeks after the scaffolds were subcutaneously implanted into C57B1/6, vascularization was evaluated by visual observation, hematoxylin and…

3D Bio-Plotted Tricalcium Phosphate/Zirconia Composite Scaffolds to Heal Large Size Bone Defects

Molecular & Cellular Biomechanics 2017 Vol. 14, No. 2, pages 125-136

β-TCP-Zirconia scaffolds with different architectures were fabricated by means of 3D-Bioplotting in order to enhance the mechanical and in-vitro ability of the scaffold to heal large size bone defects. In the present study scaffold architecture with different strand orientations (0o-90o, 0o-45o-135o-180o, 0o-108o-216o and 0o-72o-144o-36o-108o) were fabricated, characterized and evaluated for mechanical strength and cell proliferation ability. β-TCP powder (25µm) and PVA (Polyvinyl Alcohol) was acquired from Fisher Scientific, India. Zirconia (18 to 32 µm) was procured from Lobachemie, India. In brief 7.5%, PVA in distilled water was used as a binder and was mixed with 10 grams of (70/30) TCP-Zirconia…

3D printing of hybrid biomaterials for bone tissue engineering: Calcium-polyphosphate microparticles encapsulated by polycaprolactone

Acta Biomaterialia 2017 Volume 64, Pages 377-388

Here we describe the formulation of a morphogenetically active bio-ink consisting of amorphous microparticles (MP) prepared from Ca2+ and the physiological inorganic polymer, polyphosphate (polyP). Those MP had been fortified by mixing with poly-ε-caprolactone (PCL) to allow 3D-bioprinting. The resulting granular PCL/Ca-polyP-MP hybrid material, liquefied by short-time heating to 100 °C, was used for the 3D-printing of tissue-like scaffolds formed by strands with a thickness of 400 µm and a stacked architecture leaving ≈0.5 mm2-sized open holes enabling cell migration. The printed composite scaffold turned out to combine suitable biomechanical properties (Young’s modulus of 1.60 ± 0.1 GPa; Martens hardness of 153 ± 28 MPa), matching those of cortical…

Imaging stem cell distribution, growth, migration, and differentiation in 3-D scaffolds for bone tissue engineering using mesoscopic fluorescence tomography

Biotechnology and Bioengineering 2017 Volume 115, Issue 1, Pages 257-265

Regenerative medicine has emerged as an important discipline that aims to repair injury or replace damaged tissues or organs by introducing living cells or functioning tissues. Successful regenerative medicine strategies will likely depend upon a simultaneous optimization strategy for the design of biomaterials, cell-seeding methods, cell-biomaterial interactions and molecular signaling within the engineered tissues. It remains a challenge to image three-dimensional (3-D) structures and functions of the cell-seeded scaffold in mesoscopic scale (>2∼3 mm). In this study, we utilized angled fluorescence laminar optical tomography (aFLOT), which allows depth-resolved molecular characterization of engineered tissues in 3-D to investigate cell viability, migration and…

Prevascularization of 3D printed bone scaffolds by bioactive hydrogels and cell co-culture

Journal of Biomedical Materials Research Part B: Applied Biomaterials 2017 Volume 106B, Pages 1788–1798

Vascularization is a fundamental prerequisite for large bone construct development and remains one of the main challenges of bone tissue engineering. Our current study presents the combination of 3D printing technique with a hydrogel-based prevascularization strategy to generate prevascularized bone constructs. Human adipose derived mesenchymal stem cells (ADMSC) and human umbilical vein endothelial cells (HUVEC) were encapsulated within our bioactive hydrogels, and the effects of culture conditions on in vitro vascularization were determined. We further generated composite constructs by forming 3D printed polycaprolactone/hydroxyapatite scaffolds coated with cell-laden hydrogels and determined how the co-culture affected vascularization and osteogenesis. It was demonstrated…

Evaluation of 3D-Printed Polycaprolactone Scaffolds Coated with Freeze-Dried Platelet-Rich Plasma for Bone Regeneration

Materials 2017 Volume 10, Issue 7, Article 831

Three-dimensional printing is one of the most promising techniques for the manufacturing of scaffolds for bone tissue engineering. However, a pure scaffold is limited by its biological properties. Platelet-rich plasma (PRP) has been shown to have the potential to improve the osteogenic effect. In this study, we improved the biological properties of scaffolds by coating 3D-printed polycaprolactone (PCL) scaffolds with freeze-dried and traditionally prepared PRP, and we evaluated these scaffolds through in vitro and in vivo experiments. In vitro, we evaluated the interaction between dental pulp stem cells (DPSCs) and the scaffolds by measuring cell proliferation, alkaline phosphatase (ALP) activity,…

Tailorable Surface Morphology of 3D Scaffolds by Combining Additive Manufacturing with Thermally Induced Phase Separation

Macromolecular Rapid Communications 2017 Volume 38, Article 1700186

The functionalization of biomaterials substrates used for cell culture is gearing towards an increasing control over cell activity. Although a number of biomaterials have been successfully modified by different strategies to display tailored physical and chemical surface properties, it is still challenging to step from 2D substrates to 3D scaffolds with instructive surface properties for cell culture and tissue regeneration. In this study, additive manufacturing and thermally induced phase separation are combined to create 3D scaffolds with tunable surface morphology from polymer gels. Surface features vary depending on the gel concentration, the exchanging temperature, and the nonsolvent used. When preosteoblasts…

Short-term hypoxic preconditioning promotes prevascularization in 3D bioprinted bone constructs with stromal vascular fraction derived cells

RSC Advances 2017 Volume 7, Pages 29312-29320

Reconstruction of complex, craniofacial bone defects often requires autogenous vascularized bone grafts, and still remains a challenge today. In order to address this issue, we isolated the stromal vascular fraction (SVF) from adipose tissues and maintained the phenotypes and the growth of endothelial lineage cells within SVF derived cells (SVFC) by incorporating an endothelial cell medium. We 3D bioprinted SVFC within our hydrogel bioinks and conditioned the constructs in either normoxia or hypoxia. We found that short-term hypoxic conditioning promoted vascularization-related gene expression, whereas long-term hypoxia impaired cell viability and vascularization. 3D bioprinted bone constructs composed of polycaprolactone/hydroxyapatite (PCL/HAp) and…

Fabrication and Evaluation of Electrospun, 3D-Bioplotted, and Combination of Electrospun/3D-Bioplotted Scaffolds for Tissue Engineering Applications

BioMed Research International 2017 Volume 2017, Article ID 6956794

Electrospun scaffolds provide a dense framework of nanofibers with pore sizes and fiber diameters that closely resemble the architecture of native extracellular matrix. However, it generates limited three-dimensional structures of relevant physiological thicknesses. 3D printing allows digitally controlled fabrication of three-dimensional single/multimaterial constructs with precisely ordered fiber and pore architecture in a single build. However, this approach generally lacks the ability to achieve submicron resolution features to mimic native tissue. The goal of this study was to fabricate and evaluate 3D printed, electrospun, and combination of 3D printed/electrospun scaffolds to mimic the native architecture of heterogeneous tissue. We assessed their…

Polycaprolactone-and polycaprolactone/ceramic-based 3D-bioplotted porous scaffolds for bone regeneration: A comparative study

Materials Science and Engineering: C 2017 Volume 79, Issue 1, Pages 326–335

One of the critical challenges that scaffolding faces in the organ and tissue regeneration field lies in mimicking the structure, and the chemical and biological properties of natural tissue. A high-level control over the architecture, mechanical properties and composition of the materials in contact with cells is essential to overcome such challenge. Therefore, definition of the method, materials and parameters for the production of scaffolds during the fabrication stage is critical. With the recent emergence of rapid prototyping (RP), it is now possible to create three-dimensional (3D) scaffolds with the essential characteristics for the proliferation and regeneration of tissues, such…

Three dimensional printing of calcium sulfate and mesoporous bioactive glass scaffolds for improving bone regeneration in vitro and in vivo

Scientific Reports 2017 Volume 7, Article number: 42556

In the clinic, bone defects resulting from infections, trauma, surgical resection and genetic malformations remain a significant challenge. In the field of bone tissue engineering, three-dimensional (3D) scaffolds are promising for the treatment of bone defects. In this study, calcium sulfate hydrate (CSH)/mesoporous bioactive glass (MBG) scaffolds were successfully fabricated using a 3D printing technique, which had a regular and uniform square macroporous structure, high porosity and excellent apatite mineralization ability. Human bone marrow-derived mesenchymal stem cells (hBMSCs) were cultured on scaffolds to evaluate hBMSC attachment, proliferation and osteogenesis-related gene expression. Critical-sized rat calvarial defects were applied to investigate the…

3D printing of pearl/CaSO4 composite scaffolds for bone regeneration

Journal of Materials Chemistry B 2017 Volume 6, Issue 3, Pages 499-509

The development of biomaterials with high osteogenic ability for fast osteointegration with a host bone is of great interest. In this study, pearl/CaSO4 composite scaffolds were fabricated using three-dimensional (3D) printing, followed by a hydration process. The pearl/CaSO4 scaffolds showed uniform interconnected macropores (∼400 μm), high porosity (∼60%), and enhanced compressive strength. With CaSO4 scaffolds as a control, the biological properties of the pearl/CaSO4 scaffolds were evaluated in vitro and in vivo. The results showed that the pearl/CaSO4 scaffolds possessed a good apatite-forming ability and stimulated the proliferation and differentiation of rat bone mesenchymal stem cells (rBMSCs), as well as…

Three-dimensional printing of cerium-incorporated mesoporous calcium-silicate scaffolds for bone repair

Journal of Materials Science 2016 Volume 51, Issue 2, Pages 836-844

In the study, we developed hierarchical composite scaffolds by 3D printing technique with mesoporous CaSiO3 containing controlled amounts of Ce substitution in Ca–Si system. The scaffolds were porous with 3D interconnected large pores (size ~400 μm) and an overall porosity above 70 %, combined with a relative high compressive strength (~7 MPa). These properties are essential for enhancing bone ingrowth in tissue engineering. The in vitro biological properties of apatite formation, cell proliferation, and differentiation were characterized on CeO2-MCS scaffolds and MCS scaffolds. Results indicated that CeO2-MCS scaffolds induced similar apatite deposition and cell attachment of human bone marrow stromal…

Hyperelastic “bone”: A highly versatile, growth factor–free, osteoregenerative, scalable, and surgically friendly biomaterial

Science Translational Medicine 2016 Volume 8, Issue 358, Pages 358ra127

Despite substantial attention given to the development of osteoregenerative biomaterials, severe deficiencies remain in current products. These limitations include an inability to adequately, rapidly, and reproducibly regenerate new bone; high costs and limited manufacturing capacity; and lack of surgical ease of handling. To address these shortcomings, we generated a new, synthetic osteoregenerative biomaterial, hyperelastic “bone” (HB). HB, which is composed of 90 weight % (wt %) hydroxyapatite and 10 wt % polycaprolactone or poly(lactic-co-glycolic acid), could be rapidly three-dimensionally (3D) printed (up to 275 cm3/hour) from room temperature extruded liquid inks. The resulting 3D-printed HB exhibited elastic mechanical properties (~32…

Osteogenic Differentiation of Three-Dimensional Bioprinted Constructs Consisting of Human Adipose-Derived Stem Cells In Vitro and In Vivo

PloS One 2016 Volume 11, Issue 6, e0157214

Here, we aimed to investigate osteogenic differentiation of human adipose-derived stem cells (hASCs) in three-dimensional (3D) bioprinted tissue constructs in vitro and in vivo. A 3D Bio-plotter dispensing system was used for building 3D constructs. Cell viability was determined using live/dead cell staining. After 7 and 14 days of culture, real-time quantitative polymerase chain reaction (PCR) was performed to analyze the expression of osteogenesis-related genes (RUNX2, OSX, and OCN). Western blotting for RUNX2 and immunofluorescent staining for OCN and RUNX2 were also performed. At 8 weeks after surgery, osteoids secreted by osteogenically differentiated cells were assessed by hematoxylin-eosin (H&E) staining,…

Accelerated vascularization of tissue engineering constructs in vivo by preincubated co-culture of aortic fragments and osteoblasts

Biochemical Engineering Journal 2016 Volume 105, Part A, Pages 230–241

There is an urgent critical need for the development of clinically relevant tissue-engineered large bone substitutes that can promote early vascularization after transplantation. To promote rapid blood vessel growth in the engineered tissue, we preincubated aortic fragments, as well as, co-cultures of aortic fragments and osteoblast-like cells in matrigel-filled PLGA scaffolds before implantation into the dorsal skinfold chambers of balb/c mice. Despite an acceptable and low inflammatory response, preincubated aortic fragments accelerate early angiogenesis of tissue-engineered constructs; the angiogenesis was found to occur faster than that observed in previous studies. Thus, the time-period for achieving a denser microvascular network could…

[Osteogenesis of human adipose-derived mesenchymal stem cells-biomaterial mixture in vivo after 3D bio-printing]

Journal of Peking University. Health Sciences 2016 Volume 48, Issue 1, Pages 45-50

To construct human adipose-derived mesenchymal stem cells (hASCs)-biomaterial mixture 3D bio-printing body and detect its osteogenesis in vivo, and to establish a guideline of osteogenesis in vivo by use of 3D bio-printing technology preliminarily.P4 hASCs were used as seed cells, whose osteogenic potential in vitro was tested by alkaline phosphatase (ALP) staining and alizarin red staining after 14 d of osteogenic induction. The cells were added into 20 g/L sodium alginate and 80 g/L gelatin mixture (cell density was 1×10(6)/mL), and the cell-sodium alginate-gelatin mixture was printed by Bioplotter 3D bio-printer (Envision company, Germany), in which the cells’survival rate was…

Material design and photo-regulated hydrolytic degradation behavior of tissue engineering scaffolds fabricated via 3D fiber deposition

Journal of Materials Chemistry B 2016 Volume 5, Pages 329-340

An ideal behavior of a tissue engineering scaffold is that it degrades and reshapes at a rate that matches the formation of new tissues. However, this ideal situation may not occur as the scaffold often undergoes too slow or too fast degradation. To test the promise of the active control of scaffold degradation, in this work, a photo/water dual-degradable porous scaffold was designed and fabricated using a 3D fiber deposition (3DF) system from a linear biopolymer (named PLANB) that combined the o-nitrobenzyl linkages and hydrolysable ester bone in the polymer chains. The chemical structure, molecular weight and polydispersity of PLANB…

The effect of calcium sulfate incorporation on physiochemical and biological properties of 3D-printed mesoporous calcium silicate cement scaffolds

Microporous and Mesoporous Materials 2016 Volume 241, Issue 15, Pages 11–20

Development of 3D porous scaffolds with proper mechanical strength is crucial in bone tissue engineering. In this study, calcium sulfate hemihydrate (CSH) cement was functionally incorporated into mesoporous calcium silicate (MCS) through a 3D printing technique in order to improve the scaffold strength. Compared to printed MCS scaffolds, the characterizations revealed that 20% CSH incorporation had enhanced their compressive strength by 2 times via 4 weeks’ hydration. Furthermore, CSH incorporation prevented the fast pH value rise and achieved a balanced degradation rate. SEM observations showed a good apatite formation on the surfaces of both MCS and MCS/CSH scaffolds. Cellular experiments…

Three-dimensional printing of tricalcium silicate/mesoporous bioactive glass cement scaffolds for bone regeneration

Journal of Materials Chemistry B 2016 Volume 4, Pages 7452-7463

Bone defects, particularly large bone defects resulting from infections, trauma, surgical resection or genetic malformations, maintain a significant challenge for clinicians. In this study, the tricalcium silicate/mesoporous bioactive glass (C3S/MBG) cement scaffolds were successfully fabricated for the first time by 3D printing with a curing process, which combined the hydraulicity of C3S with the excellent biological property of MBG together. The C3S/MBG scaffolds exhibited 3D interconnected macropores (~400μm), high porosity (~70%), enhanced mechanical strength (>12MPa) and excellent apatite mineralization ability. Human bone marrow-derived mesenchymal stem cells (hBMSCs) were cultured on the scaffolds to evaluate their cell responses, and the results…

3D silicon doped hydroxyapatite scaffolds decorated with Elastin-like Recombinamers for bone regenerative medicine

Acta Biomaterialia 2016 Volume 45, Pages 349–356

The current study reports on the manufacturing by rapid prototyping technique of three-dimensional (3D) scaffolds based on silicon substituted hydroxyapatite with Elastin-like Recombinamers (ELRs) functionalized surfaces. Silicon doped hydroxyapatite (Si-HA), with Ca10(PO4)5.7(SiO4)0.3(OH)1.7h0.3 nominal formula, was surface functionalized with two different types of polymers designed by genetic engineering: ELR-RGD that contain cell attachment specific sequences and ELR-SNA15/RGD with both hydroxyapatite and cells domains that interact with the inorganic phase and with the cells, respectively. These hybrid materials were subjected to in vitro assays in order to clarify if the ELRs coating improved the well-known biocompatible and bone regeneration properties of calcium…

Fabrication and characterization of bioactive glass/alginate composite scaffolds by a self-crosslinking processing for bone regeneration

RSC Advances 2016 Volume 6, Pages 91201-91208

The aim of this study was to synthesize and characterize self-crosslinked bioactive glass/alginate composite scaffolds, as a kind of potential biomaterial for bone regeneration. The scaffolds were fabricated through a self-crosslinking process of alginate by bioactive glass microspheres provided Ca2+ completely, without any organic solvent, crosslinking agent or binder. The microstructure, mechanical properties, apatite-forming ability, ionic release, adhesion, proliferation and ALP activity of human bone marrow-derived mesenchymal stem cells (hBMSCs) of the scaffolds were evaluated. The results showed that uniform films could be obtained on the surface as well as abundant of crosslinking bridges in the interior of scaffolds. The…

In vitro colonization of stratified bioactive scaffolds by pre-osteoblast cells

Acta Biomaterialia 2016 Volume 44, Issue 15, Pages 73–84

Mesoporous bioactive glass-polycaprolactone (MBG-PCL) scaffolds have been prepared by robocasting, a layer by layer rapid prototyping method, by stacking of individual strati. Each stratus was independently analyzed during the cell culture tests with MC3T3-E1 preosteblast-like cells. The presence of MBG stimulates the colonization of the scaffolds by increasing the cell proliferation and differentiation. MBG-PCL composites not only enhanced pre-osteoblast functions but also allowed cell movement along its surface, reaching the upper stratus faster than in pure PCL scaffolds. The cells behavior on each individual stratus revealed that the scaffolds colonization depends on the chemical stimuli supplied by the MBG dissolution…

Multi‐and mixed 3D‐printing of graphene‐hydroxyapatite hybrid materials for complex tissue engineering

Journal of Biomedical Materials Research Part A 2016 Volume 105, Issue 1, Pages 274–283

With the emergence of 3D-printing (3DP) as a vital tool in tissue engineering and medicine, there is an ever growing need to develop new biomaterials that can be 3D-printed and also emulate the compositional, structural, and functional complexities of human tissues and organs. In this work, we probe the 3D-printable biomaterials spectrum by combining two recently established functional 3D-printable particle-laden biomaterial inks: one that contains hydroxyapatite microspheres (Hyperelastic Bone, HB) and another that contains graphene nanoflakes (3D-Graphene, 3DG). We demonstrate that not only can these distinct, osteogenic and neurogenic inks be co-3D-printed to create complex, multi-material constructs, but that composite…

A new printable and durable N,O-carboxymethyl chitosan–Ca2+–polyphosphate complex with morphogenetic activity

Journal of Materials Chemistry B 2015 Volume 3, Issue 8, Pages 1722-1730

Biomimetic materials have been gaining increasing importance in tissue engineering since they may provide regenerative alternatives to the use of autologous tissues for transplantation. In the present study, we applied for bioprinting of a functionalized three-dimensional template, N,O-carboxymethyl chitosan (N,O-CMC), mimicking the physiological extracellular matrix. This polymer, widely used in tissue engineering, has been provided with functional activity by integration of polyphosphate (polyP), an osteogenically acting natural polymer. The two polymers, N,O-CMC and polyP, are linked together via Ca2+ bridges. This N,O-CMC + polyP material was proven to be printable and durable. The N,O-CMC + polyP printed layers and tissue…

3D fibre deposition and stereolithography techniques for the design of multifunctional nanocomposite magnetic scaffolds

Journal of Materials Science: Materials in Medicine 2015 Volume 26, Issue 250, Pages 250ff

Magnetic nanocomposite scaffolds based on poly(ε-caprolactone) and poly(ethylene glycol) were fabricated by 3D fibre deposition modelling (FDM) and stereolithography techniques. In addition, hybrid coaxial and bilayer magnetic scaffolds were produced by combining such techniques. The aim of the current research was to analyse some structural and functional features of 3D magnetic scaffolds obtained by the 3D fibre deposition technique and by stereolithography as well as features of multimaterial scaffolds in the form of coaxial and bilayer structures obtained by the proper integration of such methods. The compressive mechanical behaviour of these scaffolds was investigated in a wet environment at 37…

3D-printed hierarchical scaffold for localized isoniazid/rifampin drug delivery and osteoarticular tuberculosis therapy

Acta Biomaterialia 2015 Volume 16, Pages 145–155

After surgical treatment of osteoarticular tuberculosis (TB), it is necessary to fill the surgical defect with an implant, which combines the merits of osseous regeneration and local multi-drug therapy so as to avoid drug resistance and side effects. In this study, a 3D-printed macro/meso-porous composite scaffold is fabricated. High dosages of isoniazid (INH)/rifampin (RFP) anti-TB drugs are loaded into chemically modified mesoporous bioactive ceramics in advance, which are then bound with poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) through a 3D printing procedure. The composite scaffolds show greatly prolonged drug release time compared to commercial calcium phosphate scaffolds either in vitro or in vivo….

Three-dimensional printed strontium-containing mesoporous bioactive glass scaffolds for repairing rat critical-sized calvarial defects

Acta Biomaterialia 2015 Volume 12, Pages 270–280

The development of a new generation of biomaterials with high osteogenic ability for fast osseointegration with host bone is being intensively investigated. In this study, we have fabricated three-dimensional (3-D) strontium-containing mesoporous bioactive glass (Sr-MBG) scaffolds by a 3-D printing technique. Sr-MBG scaffolds showed uniform interconnected macropores (∼400 μm), high porosity (∼70%) and enhanced compressive strength (8.67 ± 1.74 MPa). Using MBG scaffolds as a control, the biological properties of Sr-MBG scaffolds were evaluated by apatite-forming ability, adhesion, proliferation, alkaline phosphatase activity and osteogenic gene expression of osteoblast-like cells MC3T3-E1. Furthermore, Sr-MBG scaffolds were used to repair critical-sized rat calvarial…

The preliminary performance study of the 3D printing of a tricalcium phosphate scaffold for the loading of sustained release anti-tuberculosis drugs

Journal of Materials Science 2015 Volume 50, Issue 5, Pages 2138-2147

In the surgical treatment of tuberculosis of the bones, excision of the lesion site leaves defects in the bone structure. Recent research has shown benefits for bone tissue support, such as tricalcium phosphate, as regrowth materials. These biocompatible engineering materials have good bone inductivity and biologic mechanical performance. The goal of this study was to evaluate the use of 3D printing, a new technology, to design and build 3-dimensional support structures for use in grafting at lesion sites and for use in embedding the sustained release anti-tuberculosis drugs Rifampin and Isoniazid and determine the in vivo performance of these structures….

Glasses in bone regeneration: A multiscale issue

Journal of Non-Crystalline Solids 2015 Volume 432, Part A, Pages 9–14

3D scaffolds based in mesoporous bioactive glasses (MBGs) are being widely investigated to use in bone tissue engineering (TE) applications. These scaffolds are often obtained by rapid prototyping (RP) and exhibit an array of interconnected pores in a hierarchy of sizes. The ordered mesopore network (around 4 nm in diameter) is optimal for the adsorption and release of bone inductor biomolecules, and the arrangement of macropores over 100 μm facilitates the bone cell ingrowths and angiogenesis. Nevertheless MBG composition can be varied almost infinitely at the atomic scale by including in the glass network oxides of inorganic elements with a…

Fabrication of novel Si-doped Hydroxyapatite/Gelatine scaffolds by rapid prototyping for drug delivery and bone regeneration

Acta Biomaterialia 2015 Volume 15, Pages 200–209

Porous 3-D scaffolds consisting of gelatine and Si-doped hydroxyapatite were fabricated at room temperature by rapid prototyping. Microscopic characterization revealed a highly homogeneous structure, showing the pre-designed porosity (macroporosity) and a lesser in-rod porosity (microporosity). The mechanical properties of such scaffolds are close to those of trabecular bone of the same density. The biological behavior of these hybrid scaffolds is greater than that of pure ceramic scaffolds without gelatine, increasing pre-osteoblastic MC3T3-E1 cell differentiation (matrix mineralization and gene expression). Since the fabrication process of these structures was carried out at mild conditions, an antibiotic (vancomycin) was incorporated in the slurry…

3D-printed dimethyloxallyl glycine delivery scaffolds to improve angiogenesis and osteogenesis

Biomaterials Science 2015 Voulme 3, Issue 8, Pages 1236-1244

Angiogenesis–osteogenesis coupling processes are vital in bone tissue engineering. Normal biomaterials implanted in bone defects have issues in the sufficient formation of blood vessels, especially in the central part. Single delivery of vascular endothelial growth factors (VEGF) to foci in previous studies did not show satisfactory results due to low loading doses, a short protein half-life and low efficiency. Development of a hypoxia-mimicking microenvironment for cells by local prolyl-4-hydroxylase inhibitor release, which can stabilize hypoxia-inducible factor 1α (HIF-1α) expression, is an alternative method. The aim of this study was to design a dimethyloxallyl glycine (DMOG) delivering scaffold composed of mesoporous…

Three-dimensionally plotted MBG/PHBHHx composite scaffold for antitubercular drug delivery and tissue regeneration

Journal of Materials Science: Materials in Medicine 2015 Volume 26, Issue 102, 102ff

A suitable drug-loaded scaffold that can postoperatively release an antituberculosis drug efficiently in a lesion area and help repair a bone defect is very important in the clinical treatment of bone tuberculosis (TB). In this study, a composite drug-loaded cylindrical scaffold was prepared by using three-dimensional printing technology in combination with the mesoporous confinement range, surface chemical groups, and gradual degradation of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate). This achieves the slow release of a drug for as long as possible. We implanted the drug-loaded compound scaffold into New Zealand rabbits’ femur defect model to study the in vivo drug release performance and osteogenic ability….

Hyperthermia Induced in Magnetic Scaffolds for Bone Tissue Engineering

IEEE Transactions on Magnetics 2014 Volume 50, Issue 11, Pages 1-7

The design and fabrication of advanced biocompatible and bioresorbable materials able to mimic the natural tissues present in the human body constitutes an important challenge in regenerative medicine. The size-dependent properties that materials exhibit at the nanoscale as a consequence of their higher surface-to-volume ratio have opened a wide range of opportunities for applications in almost every imaginable field. In this regard, the incorporation of magnetic nanoparticles (MNPs) into biocompatible scaffold formulations provides final materials with additional multifunctionality and reinforced mechanical properties for bone tissue engineering applications. In addition to the biological implications due to their magnetic character (i.e., magnetic…

3D-Printed Magnetic Fe3O4/MBG/PCL Composite Scaffolds with Multifunctionality of Bone Regeneration, Local Anticancer Drug Delivery and Hyperthermia

Journal of Materials Chemistry B 2014 Volume 2, Issue 43, Pages 7583-7595

In this study, three-dimensional (3D) magnetic Fe3O4 nanoparticles containing mesoporous bioactive glass/polycaprolactone (Fe3O4/MBG/PCL) composite scaffolds have been fabricated by the 3D-printing technique. The physiochemical properties, in vitro bioactivity, anticancer drug delivery, mechanical strength, magnetic heating ability and cell response of Fe3O4/MBG/PCL scaffolds were systematically investigated. The results showed that Fe3O4/MBG/PCL scaffolds had uniform macropores of 400 μm, high porosity of 60% and excellent compressive strength of 13–16 MPa. The incorporation of magnetic Fe3O4 nanoparticles into MBG/PCL scaffolds did not influence their apatite mineralization ability but endowed excellent magnetic heating ability and significantly stimulated proliferation, alkaline phosphatase (ALP) activity, osteogenesis-related gene…

Accelerating the early angiogenesis of tissue engineering constructs in vivo by the use of stem cells cultured in matrigel

Journal of Biomedical Materials Research Part A 2014 Volume 102, Issue 6, Pages 1652–1662

In tissue engineering research, generating constructs with an adequate extent of clinical applications remains a major challenge. In this context, rapid blood vessel ingrowth in the transplanted tissue engineering constructs is the key factor for successful incorporation. To accelerate the microvascular development in engineered tissues, we preincubated osteoblast-like cells as well as mesenchymal stem cells or a combination of both cell types in Matrigel-filled PLGA scaffolds before transplantation into the dorsal skinfold chambers of balb/c mice. By the use of preincubated mesenchymal stem cells, a significantly accelerated angiogenesis was achieved. Compared with previous studies that showed a decisive increase of…

Biocalcite, a multifunctional inorganic polymer: Building block for calcareous sponge spicules and bioseed for the synthesis of calcium phosphate-based bone

Beilstein Journal of Nanotechnology 2014 Volume 5, Pages 610–621

Calcium carbonate is the material that builds up the spicules of the calcareous sponges. Recent results revealed that the calcium carbonate/biocalcite-based spicular skeleton of these animals is formed through an enzymatic mechanism, such as the skeleton of the siliceous sponges, evolutionarily the oldest animals that consist of biosilica. The enzyme that mediates the calcium carbonate deposition has been identified as a carbonic anhydrase (CA) and has been cloned from the calcareous sponge species Sycon raphanus. Calcium carbonate deposits are also found in vertebrate bones besides the main constituent, calcium phosphate/hydroxyapatite (HA). Evidence has been presented that during the initial phase…

Effect of Bioglass on Growth and Biomineralization of SaOS-2 Cells in Hydrogel after 3D Cell Bioprinting

PloS One 2014 Volume 9, Issue 11, Article e112497

We investigated the effect of bioglass (bioactive glass) on growth and mineralization of bone-related SaOS-2 cells, encapsulated into a printable and biodegradable alginate/gelatine hydrogel. The hydrogel was supplemented either with polyphosphate (polyP), administered as polyP•Ca2+-complex, or silica, or as biosilica that had been enzymatically prepared from ortho-silicate by silicatein. These hydrogels, together with SaOS-2 cells, were bioprinted to computer-designed scaffolds. The results revealed that bioglass (nano)particles, with a size of 55 nm and a molar ratio of SiO2∶CaO∶P2O5 of 55∶40∶5, did not affect the growth of the encapsulated cells. If silica, biosilica, or polyP•Ca2+-complex is co-added to the cell-containing alginate/gelatin…

Three-dimensional printing of strontium-containing mesoporous bioactive glass scaffolds for bone regeneration

Acta Biomaterialia 2014 Volume 10, Issue 5, Pages 2269–2281

In this study, we fabricated strontium-containing mesoporous bioactive glass (Sr-MBG) scaffolds with controlled architecture and enhanced mechanical strength using a three-dimensional (3-D) printing technique. The study showed that Sr-MBG scaffolds had uniform interconnected macropores and high porosity, and their compressive strength was ∼170 times that of polyurethane foam templated MBG scaffolds. The physicochemical and biological properties of Sr-MBG scaffolds were evaluated by ion dissolution, apatite-forming ability and proliferation, alkaline phosphatase activity, osteogenic expression and extracelluar matrix mineralization of osteoblast-like cells MC3T3-E1. The results showed that Sr-MBG scaffolds exhibited a slower ion dissolution rate and more significant potential to stabilize the…

Additive effect of mesenchymal stem cells and VEGF to vascularization of PLGA scaffolds

Microvascular Research 2013 Volume 90, Pages 71–79

Bone marrow derived mesenchymal stem cells (bmMSCs) are widely used for the generation of tissue engineering constructs, since they can differentiate into different cell types occurring in bone tissues. Until now their use for the generation of tissue engineering constructs is limited. All cells inside a tissue engineering construct die within a short period of time after implantation of the construct because vascularization and establishment of connections to the recipient circulatory system is a time consuming process. We therefore compared the influences of bmMSC, VEGF and a combination of both on the early processes of vascularization, utilizing the mice skinfold…

Combining technologies to create bioactive hybrid scaffolds for bone tissue engineering

Biomatter 2013 Volume 3, Issue 2, e23705

Combining technologies to engineer scaffolds that can offer physical and chemical cues to cells is an attractive approach in tissue engineering and regenerative medicine. In this study, we have fabricated polymer-ceramic hybrid scaffolds for bone regeneration by combining rapid prototyping (RP), electrospinning (ESP) and a biomimetic coating method in order to provide mechanical support and a physico-chemical environment mimicking both the organic and inorganic phases of bone extracellular matrix (ECM). Poly(ethylene oxide terephthalate)-poly(buthylene terephthalate) (PEOT/PBT) block copolymer was used to produce three dimensional scaffolds by combining 3D fiber (3DF) deposition, and ESP, and these constructs were then coated with a…

Advanced composites for hard-tissue engineering based on PCL/organic–inorganic hybrid fillers: From the design of 2D substrates to 3D rapid prototyped scaffolds

Polymer Composites 2013 Volume 34, Issue 9, Pages 1413–1417

The bioactivity of sol–gel synthesized poly(ε-caprolactone) (PCL)/TiO2 or poly(ε-caprolactone)/ZrO2 particles was already known. In designing innovative 2D composite substrates for hard-tissue engineering, the possibility to embed PCL/TiO2 or PCL/ZrO2 hybrid fillers into a PCL matrix was previously proposed. In the present study, the potential of 3D fiber-deposition technique to design morphologically controlled scaffolds consisting of PCL reinforced with PCL/TiO2 or PCL/ZrO2 hybrid fillers was demonstrated. Finite element analysis was initially carried out on 2D substrates to find a correlation between the previously obtained results from the small punch test and the Young’s modulus of the materials, whilst mechanical and biological…

Physiologic load-bearing characteristics of autografts, allografts, and polymer-based scaffolds in a critical sized segmental defect of long bone: an experimental study

International Journal of Nanomedicine 2013 Volume 8, Pages 1637-1643

Background To address the challenge of treating critical sized intercalary defects, we hypothesized that under physiologic cyclic loading, autografts, allografts, and scaffolds loaded with and without human mesenchymal stem cells (hMSCs) would have different biomechanical characteristics. Methods Using a rat femoral defect model, 46 rats were assigned to four groups, ie, autograft (n = 12), allograft (n = 10), scaffold (n = 13), and scaffold with hMSCs (n = 11). The scaffold groups used a 5 mm segment of scaffold composed of 80% poly-ε-caprolactone and 20% hydroxyapatite. Rats were sacrificed 4 months postoperatively, and the repairs were assessed radiographically and…

Monolithic and assembled polymer–ceramic composites for bone regeneration

Acta Biomaterialia 2013 Volume 9, Issue 3, Pages 5708–5717

The rationale for the use of polymer–ceramic composites for bone regeneration stems from the natural composition of bone, with collagen type I and biological apatite as the main organic and inorganic constituents, respectively. In the present study composite materials of PolyActive™ (PA), a poly(ethylene oxide terephthalate)/poly(butylene terephtalate) co-polymer, and hydroxyapatite (HA) at a weight ratio of 85:15 were prepared by rapid prototyping (RP) using two routes. In the first approach pre-extruded composite filaments of PA–HA were processed using three-dimensional fibre deposition (3DF) (conventional composite scaffolds). In the second approach PA scaffolds were fabricated using 3DF and combined with HA pillars…

Poly(ester-urethane) scaffolds: effect of structure on properties and osteogenic activity of stem cells

Journal of Tissue Engineering and Regenerative Medicine 2013 Volume 9, Issue 8, Pages 930–942

The present study aimed to investigate the effect of structure (design and porosity) on the matrix stiffness and osteogenic activity of stem cells cultured on poly(ester-urethane) (PEU) scaffolds. Different three-dimensional (3D) forms of scaffold were prepared from lysine-based PEU using traditional salt-leaching and advanced bioplotting techniques. The resulting scaffolds were characterized by differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), mercury porosimetry and mechanical testing. The scaffolds had various pore sizes with different designs, and all were thermally stable up to 300 °C. In vitro tests, carried out using rat bone marrow stem cells (BMSCs) for bone tissue…

Systematic analysis of injectable materials and 3D rapid prototyped magnetic scaffolds: from CNS applications to soft and hard tissue repair/regeneration

Procedia Engineering 2013 Volume 59, Pages 233–239

Over the past years, polymer-based materials have attracted research interest in the field of tissue repair and regeneration. As reported in literature, different injectable systems have been proposed, trying to reduce surgical invasiveness. In a first step of the current research, the rheological and functional features of injecatble hydrogel-based materials for central nervous system applications or soft tissue regeneration (collagen/PEG semi-IPNs) as well as for hard tissue engineering (alginate/iron-doped hydroxyapatite) were evaluated. Then, the study was also devoted to the development of 3D nanocomposite poly(ɛ- caprolactone)/iron-doped hydroxyapatite scaffolds for bone tissue engineering, providing a preliminary approach to assess magnetic attraction…

Benefits of Spine Stabilization with Biodegradable Scaffolds in Spinal Cord Injured Rats

Tissue Engineering Part C: Methods 2013 Volume 19, Issue 2, 101-108

Spine stabilization upon spinal cord injury (SCI) is a standard procedure in clinical practice, but rarely employed in experimental models. Moreover, the application of biodegradable biomaterials for this would come as an advantage as it would eliminate the presence of a nondegradable prosthesis within the vertebral bone. Therefore, in the present work, we propose the use of a new biodegradable device specifically developed for spine stabilization in a rat model of SCI. A 3D scaffold based on a blend of starch with polycaprolactone was implanted, replacing delaminated vertebra, in male Wistar rats with a T8-T9 spinal hemisection. The impact of…

Hierarchical Fibrillar Scaffolds Obtained by Non-conventional Layer-By-Layer Electrostatic Self-Assembly

Advanced Healthcare Materials 2013 Volume 2, Issue 3, pages 422–427

A new application of layer-by-layer assembly is presented, able to create nano/micro fibrils or nanocoatings inside 3D scaffolds using non-fibrillar polyelectrolytes for tissue-engineering applications. This approach shows promise for developing advanced scaffolds with controlled nano/micro environments, and nature and architectures similar to the natural extracellular matrix, leading to improved biological performance.

Design and preparation of biocompatible zwitterionic hydroxyapatite

Journal of Materials Chemistry B 2013 Volume 11, Issue 1, Pages 1595-1606

This study reports the design and preparation of zwitterionic nanocrystalline hydroxyapatite (HA) capable of inhibiting bacterial adhesion while allowing osteoblast cell colonization. The surface functionalization of HA powders was carried out by post-synthesis grafting of 3-aminopropyltriethoxysilane (APTES) and carboxyethylsilanetriol sodium salt (CES) as amine and carboxylate precursors, respectively. The successful functionalization of HA surfaces was assessed by elemental chemical analysis, FTIR, 29Si, 31P and 13C solid state CP/MAS NMR and ζ-potential measurements, and the zwitterionic nature of the synthesized HA was proved through the presence of –NH3+/–COO− pairs on the material surfaces. With the aim of evaluating the feasibility of…

Mesoporous bioactive scaffolds prepared with cerium-, gallium- and zinc-containing glasses

Acta Biomaterialia 2013 Volume 9, Issue 1, Pages 4836–4844

Mesoporous bioactive glass scaffolds (MBG_Scs), based on 80% SiO2–15% CaO–5% P2O5 (in mol.%) mesoporous sol–gel glasses substituted with Ce2O3, Ga2O3 (both 0.2% or 1.0%) and ZnO (0.4% or 2.0%), were synthesized by combination of evaporation-induced self-assembly and rapid prototyping techniques. Cerium, gallium and zinc trace elements were selected because of their inherent beneficial biological properties. Fabricated scaffolds were characterized and compared with unsubstituted scaffold (B_Sc). All of them contained well interconnected ultralarge pores (pores >400 μm) ideal for vascular ingrowth and proliferation of cells. Macropores of size 100–400 μm were present inside the scaffolds. In addition, low-angle X-ray diffraction showed…

An in vivo study on the effect of scaffold geometry and growth factor release on the healing of bone defects

Journal of Tissue Engineering and Regenerative Medicine 2013 Volume 7, Issue 9, Pages 687–696

The hypothesis of this study was that the extent of bone regeneration could be enhanced by using scaffolds with appropriate geometry, and that such an effect could be further increased by mimicking the natural timing of appearance of bone morphogenetic proteins BMP-2 and BMP-7 after fracture. Bioplotted poly(ε-caprolactone) (PCL) disks with four different fibre organizations were used to study the effect of 3D scaffold architecture on the healing of bone defects in a rat pelvis model. Moreover, one PCL construct was further modified by introducing a nanoparticulate sequential BMP-2/BMP-7 delivery system into this scaffold. Scaffolds and functionalized construct along with…

In‐vivo behavior of Si‐hydroxyapatite/polycaprolactone/DMB scaffolds fabricated by 3D printing

Journal of Biomedical Materials Research Part A 2013 Volume 101A, Issue 7, Pages 2038–2048

Scaffolds made of polycaprolactone and nanocrystalline silicon-substituted hydroxyapatite have been fabricated by 3D printing rapid prototyping technique. To asses that the scaffolds fulfill the requirements to be considered for bone grafting applications, they were implanted in New Zealand rabbits. Histological and radiological studies have demonstrated that the scaffolds implanted in bone exhibited an excellent osteointegration without the interposition of fibrous tissue between bone and implants and without immune response after 4 months of implantation. In addition, we have evaluated the possibility of improving the scaffolds efficiency by incorporating demineralized bone matrix during the preparation by 3D printing. When demineralized bone…

A biomimetic growth factor delivery strategy for enhanced regeneration of iliac crest defects

Biomedical Materials 2013 Volume 8, Number 4, 045009

The importance of provision of growth factors in the engineering of tissues has long been shown to control the behavior of the cells within the construct and several approaches were applied toward this end. In nature, more than one type of growth factor is known to be effective during the healing of tissue defects and their peak concentrations are not always simultaneous. One of the most recent strategies includes the delivery of a combination of growth factors with the dose and timing to mimic the natural regeneration cascade. The sequential delivery of bone morphogenetic proteins BMP-2 and BMP-7 which are…

Peripheral mineralization of a 3D biodegradable tubular construct as a way to enhance guidance stabilization in spinal cord injury regeneration

Journal of Materials Science: Materials in Medicine 2012 Volume 23, Issue 11, Pages 2821-2830

Spinal cord injuries (SCI) present a major challenge to therapeutic development due to its complexity. Combinatorial approaches using biodegradable polymers that can simultaneously provide a tissue scaffold, a cell vehicle, and a reservoir for sustained drug delivery have shown very promising results. In our previous studies we have developed a novel hybrid system consisting of starch/poly-e-caprolactone (SPCL) semi-rigid tubular porous structure, based on a rapid prototyping technology, filled by a gellan gum hydrogel concentric core for the regeneration within spinal-cord injury sites. In the present work we intend to promote enhanced osteointegration on these systems by pre-mineralizing specifically the external…

Degradable amorphous scaffolds with enhanced mechanical properties and homogeneous cell distribution produced by a three‐dimensional fiber deposition method

Journal of Biomedical Materials Research Part A 2012 Volume 100A, Issue 10, Pages 2739-2749

The mechanical properties of amorphous, degradable, and highly porous poly(lactide-co-caprolactone) structures have been improved by using a 3D fiber deposition (3DF) method. Two designs of 3DF scaffolds, with 45° and 90° layer rotation, were printed and compared with scaffolds produced by a salt-leaching method. The scaffolds had a porosity range from 64% to 82% and a high interconnectivity, measured by micro-computer tomography. The 3DF scaffolds had 8–9 times higher compressive stiffness and 3–5 times higher tensile stiffness than the salt-leached scaffolds. There was a distinct decrease in the molecular weight during printing as a consequence of the high temperature. The…

Three-Dimensional Poly(ε-caprolactone) Bioactive Scaffolds with Controlled Structural and Surface Properties

Biomacromolecules 2012 Volume 13, Issue 11, Pages 3510-3521

The requirement of a multifunctional scaffold for tissue engineering capable to offer at the same time tunable structural properties and bioactive interface is still unpaired. Here we present three-dimensional (3D) biodegradable polymeric (PCL) scaffolds with controlled morphology, macro-, micro-, and nano-mechanical performances endowed with bioactive moieties (RGD peptides) at the surface. Such result was obtained by a combination of rapid prototyping (e.g., 3D fiber deposition) and surface treatment approach (aminolysis followed by peptide coupling). By properly designing process conditions, a control over the mechanical and biological performances of the structure was achieved with a capability to tune the value of…

A route toward the development of 3D magnetic scaffolds with tailored mechanical and morphological properties for hard tissue regeneration: Preliminary study

Virtual and Physical Prototyping 2011 Volume 6, Issue 4, Pages 189-195

A basic approach toward the design of three-dimensional (3D) rapid prototyped magnetic scaffolds for hard-tissue regeneration has been proposed. In particular, 3D scaffolds consisting of a poly(ε-caprolactone) (PCL) matrix and iron oxide (Fe3O4) or iron-doped hydroxyapatite (FeHA) nanoparticles were fabricated through a 3D fibre deposition technique. As a first approach, a polymer to nanoparticle weight ratio of 90/10 (wt/wt) was used. The effect of the inclusion of both kinds of nanoparticles on the mechanical, magnetic, and biological performances of the scaffolds was studied. The inclusion of Fe3O4 and FeHA nanoparticles generally improves the modulus and the yield stress of the…

A basic approach toward the development of nanocomposite magnetic scaffolds for advanced bone tissue engineering

Jounal of Applied Polymer Science 2011 Volume 122, Issue 6, Pages 3599-3605

Magnetic scaffolds for bone tissue engineering based on a poly(ε-caprolactone) (PCL) matrix and iron oxide (Fe3O4) magnetic nanoparticles were designed and developed through a three-dimensional (3D) fiber-deposition technique. PCL/Fe3O4 scaffolds were characterized by a 90/10 w/w composition. Tensile and magnetic measurements were carried out, and nondestructive 3D imaging was performed through microcomputed tomography (Micro-CT). Furthermore, confocal analysis was undertaken to investigate human mesenchymal stem cell adhesion and spreading on the PCL/Fe3O4 nanocomposite fibers. The results suggest that nanoparticles mechanically reinforced the PCL matrix; the elastic modulus and the maximum stress increased about 10 and 30%, respectively. However, the maximum strain…

Comparably accelerated vascularization by preincorporation of aortic fragments and mesenchymal stem cells in implanted tissue engineering constructs

Journal of Biomedical Materials Research Part A 2011 Volume 97A, Issue 4, Pages 383-394

The demanding need for tissue replacement resulted in manifold approaches for the construction of different tissues. One common problem which hampers the clinical usage of tissue engineering constructs is a limited vascularization. In an attempt to accelerate the vascularization of tissue engineering constructs we compared the usage of bone marrow mesenchymal stem cells (bmMSCs) and fragments derived from the aorta in vivo. Tissue engineering constructs composed of PLGA scaffolds containing Matrigel (n = 8), aortic fragments embedded in Matrigel (n = 8), bmMSCs embedded in Matrigel (n = 8), and aortic fragments embedded in Matrigel combined with bmMSCs (n =…

Calvaria bone chamber-A new model for intravital assessment of osseous angiogenesis

Journal of Biomedical Materials Research Part A 2011 Volume 99A, Issue 2, pages 151-157

The faith of tissue engineered bone replacing constructs depends on their early supply with oxygen and nutrients, and thus on a rapid vascularization. Although some models for direct observation of angiogenesis are described, none of them allows the observation of new vessel formation in desmal bone. Therefore, we developed a new chamber model suitable for quantitative in vivo assessment of the vascularization of bone substitutes by intravital fluorescence microscopy. In the parietal calvaria of 32 balb/c mice a critical size defect was set. Porous 3D-poly(L-lactide-co-glycolide) (PLGA)-blocks were inserted into 16 osseous defects (groups 3 and 4) while other 16 osseous…

A Novel Approach for Studying Microcirculation in Bone Defects by Intravital Fluorescence Microscopy

Tissue Engineering Part C: Methods 2011 Volume 17, Issue 12, Pages 1151-1159

Angiogenic and inflammatory responses to biodegradable scaffolds were previously studied using the dorsal skinfold chamber for testing different scaffold materials. In this model, the angiogenic response originates from the soft tissue of the skin. Herein, we introduce a new model that allows the study of developing microcirculation of bone defects for testing tissue-engineered constructs. A bone defect was prepared in the femur of Balb/c mice by inserting a pin for intramedullary fixation, and a custom-made observation window fixed over the defect allowed constant observation. This study included three different groups: empty defect (control), defect filled with porous poly(l-lactide-co-glycolide), and beta-tricalcium-phosphate…

Three-dimensional plotted scaffolds with controlled pore size gradients: Effect of scaffold geometry on mechanical performance and cell seeding efficiency

Acta Biomaterialia 2011 Volume 7, Issue 3, Pages 1009-1018

Scaffolds produced by rapid prototyping (RP) techniques have proved their value for tissue engineering applications, due to their ability to produce predetermined forms and structures featuring fully interconnected pore architectures. Nevertheless, low cell seeding efficiency and non-uniform distribution of cells remain major limitations when using such types of scaffold. This can be mainly attributed to the inadequate pore architecture of scaffolds produced by RP and the limited efficiency of cell seeding techniques normally adopted. In this study we aimed at producing scaffolds with pore size gradients to enhance cell seeding efficiency and control the spatial organization of cells within the…

Comparison of the osteoblastic activity conferred on Si-doped hydroxyapatite scaffolds by different osteostatin coatings

Acta Biomaterialia 2011 Volume 7, Issue 10, Pages 3555-3562

Parathyroid hormone-related protein (107-111) (osteostatin) induces osteogenic effects in osteoblasts in vitro and in regenerating bone in mice and rabbits. In this study we used osteoblastic MC3T3-E1 cell cultures to evaluate and compare the bioactivity of this peptide either adsorbed or covalently bound (by its C-terminus) to Si-doped hydroxyapatite (Si-HA) scaffolds after organic (-NH2) functionalization. By these means osteostatin can be locally released or kept anchored to the scaffold surface. This was confirmed by chemical analysis and by testing the efficiency of osteostatin-loaded Si-HA scaffolds (placed in Transwell chambers) in healing a scratch wound in mouse pluripotent mesenchymal C3H10T1/2 cells….

Preparation of 3-D scaffolds in the SiO2–P2O5 system with tailored hierarchical meso-macroporosity

Acta Biomaterialia 2011 Volume 7, Issue 3, Pages 1265-1273

Herein we report for the first time the synthesis of three-dimensional scaffolds in the binary system SiO2–P2O5 exhibiting different scales of porosity: (i) highly ordered mesopores with diameters of ca. 4 nm; (ii) macropores with diameters in the 30–80 μm range with interconnections of ca. 2–4 and 8–9 μm; and (iii) ultra-large macropores of ca. 400 μm. The hierarchical porosity of the resulting scaffolds makes them suitable for bone tissue engineering applications. The chemical nature and mesoporosity of these matrices would allow these scaffolds to act as local controlled delivery systems of biologically active molecules, such as certain drugs to…

Comparison of bacterial adhesion and cellular proliferation on newly developed three-dimensional scaffolds manufactured by rapid prototyping technology

Journal of Biomedical Materials Research Part A 2011 Volume 98A, Issue 2, pages 303-311

Scaffolds used in the field of tissue engineering should facilitate the adherence, spreading, and ingrowth of cells as well as prevent microbial adherence. For the first time, this study simultaneously deals with microbial and tissue cell adhesion to rapid prototyping-produced 3D-scaffolds. The cell growth of human osteosarcoma cells (CAL-72) over a time period of 3-11 days were examined on three scaffolds (PLGA, PLLA, PLLA-TCP) and compared to the adhesion of salivary microorganisms and representative germs of the oral flora (Porphyromonas gingivalis, Prevotella nigrescens, Candida albicans, Enterococcus faecalis, Streptococcus mutans, and Streptococcus sanguinis). Scanning electron microscopy (SEM), cell proliferation measurements, and…

Prolongated Survival of Osteoblast-Like Cells on Biodegradable Scaffolds by Heat Shock Preconditioning

Tissue Engineering Part A 2011 Volume 17, Issue 15-16, Pages 1935-1943

The implantation of tissue-engineered constructs leads to hypoxic and physical stress to the seeded cells until they were reached by a functional microvascular system. Preconditioning of cells with heat shock induced heat shock proteins, which can support the cells to survive a subsequent episode of stress that would otherwise be lethal. Preconditioning of tissue-engineered constructs resulted in significantly higher number of surviving osteoblast-like cells (OLC). At the 6th and 10th day, angiogenic response was found comparative to poly(L-lactide-co-glycolide) (PLGA) scaffolds vitalized with either unconditioned or preconditioned OLC. However, they were significantly enhanced compared with the nonvitalized collagen-labeled PLGA scaffolds. This…

Immobilization and bioactivity evaluation of FGF-1 and FGF-2 on powdered silicon-doped hydroxyapatite and their scaffolds for bone tissue engineering

Journal of Materials Science: Materials in Medicine 2011 Volume 22, Issue 2, pp 405-416

Fibroblast growth factors (FGFs) are polypeptides that control the proliferation and differentiation of various cell types including osteoblasts. FGFs are also strong inducers of angiogenesis, necessary to obtain oxygen and nutrients during tissue repair. With the aim to incorporate these desirable FGF biological properties into bioceramics for bone repair, silicon substituted hydroxyapatites (Si-HA) were used as materials to immobilize bioactive FGF-1 and FGF-2. Thus, the binding of these growth factors to powdered Si-HA and Si-HA scaffolds was carried out efficiently in the present study and both FGFs maintained its biological activity on osteoblasts after its immobilization. The improvement of cell…

An approach in developing 3D fiber‐deposited magnetic scaffolds for tissue engineering

AIP Conference Proceedings 2010 1255, 420

Scaffolds should possess suitable properties to play their specific role. In this work, the potential of 3D fiber deposition technique to develop multifunctional and well‐defined magnetic poly(ε‐caprolactone)/iron oxide scaffolds has been highlighted, and the effect of iron oxide nanoparticles on the biological and mechanical performances has been assessed.

Accelerated Angiogenic Host Tissue Response to Poly(L-Lactide-co-Glycolide) Scaffolds by Vitalization with Osteoblast-like Cells

Tissue Engineering Part A 2010 Volume 16, Issue 7, Pages 2265-2279

Background: Bone substitutes should ideally promote rapid vascularization, which could be accelerated if these substitutes were vitalized by autologous cells. Although adequate engraftment of porous poly(L-lactide-co-glycolide) (PLGA) scaffolds has been demonstrated in the past, it has not yet been investigated how vascularization is influenced by vitalization or, more precisely, by seeding PLGA scaffolds with osteoblast-like cells (OLCs). For this reason, we conducted an in vivo study to assess host angiogenic and inflammatory responses after the implantation of PLGA scaffolds vitalized with isogeneic OLCs. Materials and Methods: OLCs were seeded on collagen-coated PLGA scaffolds that were implanted into dorsal skinfold chambers…

Effects of VEGF loading on scaffold-confined vascularization

Journal of Biomedical Materials Research Part A 2010 Volume 95A, Issue 3, Pages 783-792

Adequate vascularization of tissue-engineered constructs remains a major challenge in bone grafting. In view of this, we loaded ß-tricalcium-phosphate (ß-TCP) and porous poly(L-lactide-co-glycolide) (PLGA) scaffolds via collagen coating with vascular endothelial growth factor (VEGF) and studied whether the VEGF loading improves scaffold angiogenesis and vascularization. Dorsal skinfold chambers were implanted into 48 balb/c mice, which were assigned to 6 groups (n = 8 each). Uncoated (controls), collagen-coated, and additionally VEGF-loaded PLGA and ß-TCP scaffolds were inserted into the chambers. Angiogenesis, neovascularization, and leukocyte-endothelial cell interaction were analyzed repeatedly during a 14-day observation period using intravital fluorescence microscopy. Furthermore, VEGF release…

Development and Characterization of a Novel Hybrid Tissue Engineering-Based Scaffold for Spinal Cord Injury Repair

Tissue Engineering Part A 2010 Volume: 16 Issue 1, Pages 45-54

Spinal cord injury (SCI) represents a significant health and social problem, and therefore it is vital to develop novel strategies that can specifically target it. In this context, the objective of the present work was to develop a new range of three-dimensional (3D) tubular structures aimed at inducing the regeneration within SCI sites. Up to six different 3D tubular structures were initially developed by rapid prototyping: 3D bioplotting–based on a biodegradable blend of starch. These structures were then further complemented by injecting Gellan Gum, a polysaccharide-based hydrogel, in the central area of structures. The mechanical properties of these structures were…

Effect of scaffold architecture and BMP-2/BMP-7 delivery on in vitro bone regeneration

Journal of Materials Science: Materials in Medicine 2010 Volume 21, Issue 11, Pages 2999-3008

The aim of this study was to develop 3-D tissue engineered constructs that mimic the in vivo conditions through a self-contained growth factor delivery system. A set of nanoparticles providing the release of BMP-2 initially followed by the release of BMP-7 were incorporated in poly(ε-caprolactone) scaffolds with different 3-D architectures produced by 3-D plotting and wet spinning. The release patterns were: each growth factor alone, simultaneous, and sequential. The orientation of the fibers did not have a significant effect on the kinetics of release of the model protein BSA; but affected proliferation of bone marrow mesenchymal stem cells. Cell proliferation…

The effect of scaffold architecture on properties of direct 3D fiber deposition of porous Ti6Al4V for orthopedic implants

Journal of Biomedical Materials Research Part A 2010 Volume 92A, Issue 1, pages 33-42

3D porous Ti6Al4V scaffolds were directly fabricated by a rapid prototyping technology, 3D fiber deposition (3DF). In this study, scaffolds with different structures were fabricated by changing fiber spacing and fiber orientation. The influence of different architectures on mechanical properties and permeability of the scaffold were investigated. Mechanical analysis revealed that compressive strength and E-modulus increase with decreasing the porosity. Permeability measurements showed that not only the total porosity but also the porous structure can influence the permeability. 3DF was found to provide good control and reproducibility of the desired degree of porosity and the 3D structure. Results of this…

Bone repair by cell-seeded 3D-bioplotted composite scaffolds made of collagen treated tricalciumphosphate or tricalciumphosphate-chitosan-collagen hydrogel or PLGA in ovine critical-sized calvarial defects

Journal of Biomedical Materials Research Part B: Applied Biomaterials 2010 Volume 93B, Issue 2, Pages 520-530

The aim of this study was to investigate the osteogenic effect of three different cell-seeded 3D-bioplotted scaffolds in a ovine calvarial critical-size defect model. The choice of scaffold-materials was based on their applicability for 3D-bioplotting and respective possibility to produce tailor-made scaffolds for the use in cranio-facial surgery for the replacement of complex shaped boneparts. Scaffold raw-materials are known to be osteoinductive when being cell-seeded [poly(L-lactide-co-glycolide) (PLGA)] or having components with osteoinductive properties as tricalciumphosphate (TCP) or collagen (Col) or chitosan. The scaffold-materials PLGA, TCP/Col, and HYDR (TCP/Col/chitosan) were cell-seeded with osteoblast-like cells whether gained from bone (OLB) or from…

Tissue Formation and Vascularization in Anatomically Shaped Human Joint Condyle Ectopically in Vivo

Tissue Engineering Part A 2009 Volume 15, Issue 12, Pages 3923-3930

Scale-up of bioengineered grafts toward clinical applications is a challenge in regenerative medicine. Here, we report tissue formation and vascularization of anatomically shaped human tibial condyles ectopically with a dimension of 20 × 15 × 15 mm3. A composite of poly-ɛ-caprolactone and hydroxyapatite was fabricated using layer deposition of three-dimensional interlaid strands with interconnecting microchannels (400 μm) and seeded with human bone marrow stem cells (hMSCs) with or without osteogenic differentiation. An overlaying layer (1 mm deep) of poly(ethylene glycol)-based hydrogel encapsulating hMSCs or hMSC-derived chondrocytes was molded into anatomic shape and anchored into microchannels by gel infusion. After 6 weeks of subcutaneous implantation in athymic rats,…

Consequences of seeded cell type on vascularization of tissue engineering constructs in vivo

Microvascular Research 2009 Volume 78, Issue 2, Pages 180-190

Implantation of tissue engineering constructs is a promising technique to reconstruct injured tissue. However, after implantation the nutrition of the constructs is predominantly restricted to vascularization. Since cells possess distinct angiogenic potency, we herein assessed whether scaffold vitalization with different cell types improves scaffold vascularization. 32 male balb/c mice received a dorsal skinfold chamber. Angiogenesis, microhemodynamics, leukocyte–endothelial cell interaction and microvascular permeability induced in the host tissue after implantation of either collagen coated poly (l-lactide-co-glycolide) (PLGA) scaffolds (group 4), additionally seeded with osteoblast-like cells (OLCs, group 1), bone marrow mesenchymal stem cells (bmMSCs, group 2) or a combination of OLCs…

3D polycaprolactone scaffolds with controlled pore structure using a rapid prototyping system

Journal of Materials Science: Materials in Medicine 2009 Volume 20, Issue 1, Pages 229-234

Designing a three-dimensional (3-D) ideal scaffold has been one of the main goals in biomaterials and tissue engineering, and various mechanical techniques have been applied to fabricate biomedical scaffolds used for soft and hard tissue regeneration. Scaffolds should be biodegradable and biocompatible, provide temporary support for cell growth to allow cell adhesion, and consist of a defined structure that can be formed into customized shapes by a computer-aided design system. This versatility in preparing scaffolds gives us the opportunity to use rapid prototyping devices to fabricate polymeric scaffolds. In this study, we fabricated polycaprolactone scaffolds with interconnecting pores using a…

Nucleation and growth of biomimetic apatite layers on 3D plotted biodegradable polymeric scaffolds: Effect of static and dynamic coating conditions

Acta Biomaterialia 2009 Volume 5, Issue 5, Pages 1626-1638

Apatite layers were grown on the surface of newly developed starch/polycaprolactone (SPCL)-based scaffolds by a 3D plotting technology. To produce the biomimetic coatings, a sodium silicate gel was used as nucleating agent, followed by immersion in a simulated body fluid (SBF) solution. After growing a stable apatite layer for 7 days, the scaffolds were placed in SBF under static, agitated (80 strokes min−1) and circulating flow perfusion (Q = 4 ml min−1; tR = 15 s) for up to 14 days. The materials were characterized by scanning electron microscopy/energy dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy and thin-film X-ray diffraction….

Hierarchical starch‐based fibrous scaffold for bone tissue engineering applications

Journal of Tissue Engineering and Regenerative Medicine 2009 Volume 3, Issue 1, Pages 37-42

Fibrous structures mimicking the morphology of the natural extracellular matrix are considered promising scaffolds for tissue engineering. This work aims to develop a novel hierarchical starch-based scaffold. Such scaffolds were obtained by a combination of starch-polycaprolactone micro- and polycaprolactone nano-motifs, respectively produced by rapid prototyping (RP) and electrospinning techniques. Scanning electron microscopy (SEM) and micro-computed tomography analysis showed the successful fabrication of a multilayer scaffold composed of parallel aligned microfibres in a grid-like arrangement, intercalated by a mesh-like structure with randomly distributed nanofibres (NFM). Human osteoblast-like cells were dynamically seeded on the scaffolds, using spinner flasks, and cultured for 7…

3D Plotted PCL Scaffolds for Stem Cell Based Bone Tissue Engineering

Macromolecular Symposia 2008 Volume 269, Issue 1, Pages 92-99

The ability to control the architecture and strength of a bone tissue engineering scaffold is critical to achieve a harmony between the scaffold and the host tissue. Rapid prototyping (RP) technique is applied to tissue engineering to satisfy this need and to create a scaffold directly from the scanned and digitized image of the defect site. Design and construction of complex structures with different shapes and sizes, at micro and macro scale, with fully interconnected pore structure and appropriate mechanical properties are possible by using RP techniques. In this study, RP was used for the production of poly(ε-caprolactone) (PCL) scaffolds….

Dynamic Co-Seeding of Osteoblast and Endothelial Cells on 3D Polycaprolactone Scaffolds for Enhanced Bone Tissue Engineering

Journal of Bioactive and Compatible Polymers 2008 Volume 23, No. 3, Pages 227-243

Tissue engineered scaffolds must have an organized and repeatable microstructure which enables cells to assemble in an ordered matrix that allows adequate nutriental perfusion. In this work, to evaluate the reciprocal cell interactions of endothelial and osteoblast-like cells, human osteoblast-like cells (MG63) and Human Umbilical Vein Endothelial Cells (HUVEC) were co-seeded onto 3D geometrically controlled porous poly(ε-caprolactone) (PCL) and cultured by means of a rotary cell culture system (RCCS-4DQ). In our dynamic co-culture system, the lack of significant enhancement of osteoblast ALP activity and ECM production indicated that the microgravity conditions of the rotary system affected the cells by favoring…

Vascularization and biocompatibility of scaffolds consisting of different calcium phosphate compounds

Journal of Biomedical Materials Research Part A 2008 Volume 86A, Issue 4, Pages 1002-1011

Scaffolds for tissue engineering of bone should mimic bone matrix and promote vascular ingrowth. Whether synthetic hydroxyapatite and acellular dentin, both materials composed from calcium phosphate, fulfill these material properties has not been studied yet. Therefore, we herein studied in vivo the host angiogenic and inflammatory response to these biomaterials. Porous scaffolds of hydroxyapatite and isogeneic acellular dentin were implanted into the dorsal skinfold chamber of balb/c mice. Additional animals received perforated implants of isogeneic calvarial bone displaying pores similar in size and structure to those of both scaffolds. Chambers of animals without implants served as controls. Angiogenesis and neovascularization…

Incorporation of growth factor containing Matrigel promotes vascularization of porous PLGA scaffolds

Journal of Biomedical Materials Research Part A 2008 Volume 85A, Issue 2, Pages 397-407

In tissue engineering, rapid ingrowth of blood vessels into scaffolds is a major prerequisite for the survival of three-dimensional tissue constructs. In the present study, we investigated whether the vascularization of implanted poly-D,L-lactic-co-glycolic acid (PLGA) scaffolds may be accelerated by incorporation of Matrigel. For this purpose, we investigated in the aortic ring assay the proangiogenic properties of growth factor reduced Matrigel (GFRM) and growth factor containing Matrigel (GFCM), which were then incorporated into the pores of PLGA scaffolds. Subsequently, we analyzed vascularization, biocompatibility, and incorporation of these scaffolds during 14 days after implantation into dorsal skinfold chambers of balb/c mice…

Improvement of Vascularization of PLGA Scaffolds by Inosculation of In Situ-Preformed Functional Blood Vessels With the Host Microvasculature

Annals of Surgery 2008 Volume 248 - Issue 6 - Pages 939-948

Objective: We analyzed, in vivo, whether the establishment of blood supply to implanted scaffolds can be accelerated by inosculation of an in situ-preformed microvascular network with the host microvasculature. Background: A rapid vascularization is crucial for the survival of scaffold-based transplanted tissue constructs. Methods: Poly-lactic-glycolic acid scaffolds were implanted into the flank of balb/c or green fluorescent protein (GFP)-transgenic mice for 20 days to create in situ a new microvascular network within the scaffolds. The prevascularized scaffolds were then transferred into the dorsal skinfold chamber of isogeneic recipient mice. Nonvascularized poly-lactic-glycolic acid scaffolds served as controls. Vascularization, blood perfusion, and…

Biological performance in goats of a porous titanium alloy-biphasic calcium phosphate composite

Biomaterials 2007 Volume 28, Issue 29, Pages 4209-4218

In this study, porous 3D fiber deposition titanium (3DFT) and 3DFT combined with porous biphasic calcium phosphate ceramic (3DFT+BCP) implants, both bare and 1 week cultured with autologous bone marrow stromal cells (BMSCs), were implanted intramuscularly and orthotopically in 10 goats. To assess the dynamics of bone formation over time, fluorochrome markers were administered at 3, 6 and 9 weeks and the animals were sacrificed at 12 weeks after implantation. New bone in the implants was investigated by histology and histomorphometry of non-decalcified sections. Intramuscularly, no bone formation was found in any of the 3DFT implants, while a very limited…

Comparative in vitro study of the cell proliferation of ovine and human osteoblast‐like cells on conventionally and rapid protot yping produced scaffolds tailored for application as potential bone replacement material

Journal of Biomedical Materials Research Part A 2007 Volume 83A, Issue 4, pages 1154-1164

Reconstruction of bone defects in the field of craniomaxillofacial surgery is a relevant problem. In regenerative medicine, autologous bone is not available sufficiently. The full replacement of autologous bone grafts is required. A promising research field is the bone engineering. Especially the application of rapid prototyping (RP) enables new perspectives concerning the scaffold design. The aim of the study was to compare scaffolds produced by RP-technology (native and plasma-coated PLGA-scaffolds) with conventionally produced scaffolds (agar plates with hydroxyapatite and hyaluronic acid coated agar plates with hydroxyapatite) relating to proliferation, adhesion, and morphology of osteoblasts to get knowledge about the application…

Bone ingrowth in porous titanium implants produced by 3D fiber deposition

Biomaterials 2007 Volume 28, Issue 18, Pages 2810-2820

3D fiber deposition is a technique that allows the development of metallic scaffolds with accurately controlled pore size, porosity and interconnecting pore size, which in turn permits a more precise investigation of the effect of structural properties on the in vivo behavior of biomaterials. This study analyzed the in vivo performance of titanium alloy scaffolds fabricated using 3D fiber deposition. The titanium alloy scaffolds with different structural properties, such as pore size, porosity and interconnecting pore size were implanted on the decorticated transverse processes of the posterior lumbar spine of 10 goats. Prior to implantation, implant structure and permeability were…

Porous Ti6Al4V scaffold directly fabricating by rapid prototyping: Preparation and in vitro experiment

Biomaterials 2006 Volume 27, Issue 8, Pages 1223-1235

Three-dimensional (3D) fiber deposition (3DF), a rapid prototyping technology, was successfully directly applied to produce novel 3D porous Ti6Al4V scaffolds with fully interconnected porous networks and highly controllable porosity and pore size. A key feature of this technology is the 3D computer-controlled fiber depositing of Ti6Al4V slurry at room temperature to produce a scaffold, consisting of layers of directionally aligned Ti6Al4V fibers. In this study, the Ti6Al4V slurry was developed for the 3D fiber depositing process, and the parameters of 3D fiber depositing were optimized. The experimental results show how the parameters influence the structure of porous scaffold. The potential…

Angiogenic and inflammatory response to biodegradable scaffolds in dorsal skinfold chambers of mice

Biomaterials 2006 Volume 27, Issue 29, Pages 5027-5038

For tissue engineering, scaffolds should be biocompatible and promote neovascularization. Because little is known on those specific properties, we herein studied in vivo the host angiogenic and inflammatory response after implantation of commonly used scaffold materials. Porous poly(l-lactide-co-glycolide) (PLGA) and collagen–chitosan–hydroxyapatite hydrogel scaffolds were implanted into dorsal skinfold chambers of balb/c mice. Additional animals received cortical bone as an isogeneic, biological implant, while chambers of animals without implants served as controls. Angiogenesis and neovascularization as well as leukocyte–endothelial cell interaction and microvascular permeability were analyzed over 14 day using intravital fluorescence microscopy. PLGA scaffolds showed a slight increase in leukocyte…

Fabrication of soft and hard biocompatible scaffolds using 3D-Bioplotting

Virtual Modelling and Rapid Manufacturing-Advanced Research in Virtual and Rapid Prototyping 2005 London, England: Taylor & Francis Group, 97-102

In Tissue Engineering and bone reconstruction, alongside the choice of materials, the scaffold design is of great importance. Three dimensional structures not only permit the tuning of chemical and mechanical properties, but they can also copy the outer form of the required bone or cartilaginous structures. While new processes that create such 3D scaffolds by means of Rapid Prototyping have been developed, they are still restricted to a limited type of materials. At the Freiburger Materialforschungszentrum, our group has developed a new process called 3D BioplottingTM. Most kinds of polymers and biopolymers can be used for the fabrication of 3D…

Biofunctional rapid prototyping for tissue‐engineering applications: 3D bioplotting versus 3D printing

Journal of Polymer Science Part A: Polymer Chemistry 2004 Volume 42, Issue 3, Pages 624-638

Two important rapid-prototyping technologies (3D Printing and 3D Bioplotting) were compared with respect to the computer-aided design and free-form fabrication of biodegradable polyurethane scaffolds meeting the demands of tissue-engineering applications. Aliphatic polyurethanes were based on lysine ethyl ester diisocyanate and isophorone diisocyanate. Layer-by-layer construction of the scaffolds was performed by 3D Printing, that is, bonding together starch particles followed by infiltration and partial crosslinking of starch with lysine ethyl ester diisocyanate. Alternatively, the 3D Bioplotting process permitted three-dimensional dispensing and reactive processing of oligoetherurethanes derived from isophorone diisocyanate, oligoethylene oxide, and glycerol. The scaffolds were characterized with X-ray microtomography, scanning…