Three-dimensional printing of polycaprolactone/hydroxyapatite bone tissue engineering scaffolds mechanical properties and biological behavior (original) (raw)

Fabrication of three-dimensional polycaprolactone/hydroxyapatite tissue scaffolds and osteoblast-scaffold interactions in vitro

Biomaterials, 2007

Computer-aided tissue-engineering approach was used to develop a novel precision extrusion deposition (PED) process to directly fabricate Polycaprolactone (PCL) and composite PCL/hydroxyapatite (PCL-HA) tissue scaffolds. The process optimization was carried out to fabricate both PCL and PCL-HA (25% concentration by weight of HA) with a controlled pore size and internal pore structure of the 01/901 pattern. Two groups of scaffolds having 60% and 70% porosity and with pore sizes of 450 and 750 mm, respectively, were evaluated for their morphology and compressive properties using scanning electron microscopy (SEM) and mechanical testing. Our results suggested that inclusion of HA significantly increased the compressive modulus from 59 to 84 MPa for 60% porous scaffolds and from 30 to 76 MPa for 70% porous scaffolds. In vitro cell-scaffolds interaction study was carried out using primary fetal bovine osteoblasts to assess the feasibility of scaffolds for bone tissue-engineering application. The cell proliferation and differentiation were calculated by Alamar Blue assay and by determining alkaline phosphatase activity. The osteoblasts were able to migrate and proliferate over the cultured time for both PCL as well as PCL-HA scaffolds. Our study demonstrated the viability of the PED process to the fabricate PCL and PCL-HA composite scaffolds having necessary mechanical property, structural integrity, controlled pore size and pore interconnectivity desired for bone tissue engineering. r

Solid Free-Form Fabrication-Based PCL/HA Scaffolds Fabricated with a Multi-head Deposition System for Bone Tissue Engineering

Journal of Biomaterials Science, Polymer Edition, 2010

In this study, we fabricated polycaprolactone/hydroxyapatite (PCL/HA) scaffolds with a multi-head deposition system, a solid free-form fabrication technology that was developed in our previous study. The bone regeneration potential of the scaffolds was compared with that of PCL scaffolds fabricated with the same system. The fabricated scaffolds had a pore size of 400 µm and a porosity of 66.7%. The PCL/HA scaffolds had higher mechanical strength and modulus than the PCL scaffolds. To compare the osteogenic potential, the two types of scaffolds were seeded with rat osteoblasts and cultured in vitro or implanted subcutaneously into athymic mice. The cells cultured on PCL/HA scaffolds expressed higher levels of osteopontin and osteonectin, both of which are osteogenic proteins. The PCL/HA scaffolds resulted in larger bone area and calcium deposition in the implants compared to the PCL scaffolds.

Biological behavior of bone cells on two types of hydroxyapatite incorporated polymer scaffold

2009

The gold standard for bone loss treatment is autograft but the problem is insufficient bony source. Therefore we used bone tissue engineering concept to develop biodegradable composite materials for repairing bony defect. First we constructed hydroxyapatite (HAp) incorporated poly (L-lactic acid) (PLLA) by electrospinning technique. We determined morphology and mechanical properties of these scaffold by scanning electron microscope and universal testing machine respectively. For biological testing, we evaluated cytotoxicity and bone biological marker gene expression. From the results we found the increasing of osteocalcin mRNA expression and mineralization in MC3T3-E1, a mouse pre-osteoblastic cell line, culturing on PLLA/HAp scaffolds. The results from mechanical properties and biological responses indicated that the fibrous PLLA/HAp could be the material of choice for use in bone regeneration. The PLLA/HAp scaffolds had satisfiable results and electrospinning technique could const...

3D Printed Poly(?-Caprolactone)/Hydroxyapatite Scaffolds for Bone Tissue Engineering: A Comparative Study on Composite Preparation by Melt Blending or Solvent Casting Techniques and Influence of Bioceramic Content on Scaffold Properties

Bone tissue engineering has been developed in the past decades, with the engineering of bone substitutes on the vanguard of this regenerative approach. PCL based scaffolds are fairly applied for bone regeneration, and several composites have been incorporated, as to improve the devices’ mechanical properties and tissue ingrowth. In this study, HA was incorporated on PCL based scaffolds with two different proportions, 80:20 and 60:40. Devices were produced with two different techniques, SC and MB, and further investigated with regards to their mechanical characteristics and in vitro cytocompatibility. Results show the MB devices to present more promising mechanical properties, along with the incorporation of HA. The latter is also related to an increase in osteogenic activity and promotion. Overall, this study suggest PCL:HA scaffolds to be promising candidates for bone tissue engineering, particularly when produced by the MB method.

Pellet-Based Fused Filament Fabrication (FFF)-Derived Process for the Development of Polylactic Acid/Hydroxyapatite Scaffolds Dedicated to Bone Regeneration

Materials

Scaffolds can be defined as 3D architectures with specific features (surface properties, porosity, rigidity, biodegradability, etc.) that help cells to attach, proliferate, and to differentiate into specific lineage. For bone regeneration, rather high mechanical properties are required. That is why polylactic acid (PLA) and PLA/hydroxyapatite (HA) scaffolds (10 wt.%) were produced by a peculiar fused filament fabrication (FFF)-derived process. The effect of the addition of HA particles in the scaffolds was investigated in terms of morphology, biological properties, and biodegradation behavior. It was found that the scaffolds were biocompatible and that cells managed to attach and proliferate. Biodegradability was assessed over a 5-month period (according to the ISO 13781-Biodegradability norm) through gel permeation chromatography (GPC), differential scanning calorimetry (DSC), and compression tests. The results revealed that the presence of HA in the scaffolds induced a faster and ...

Poly(lactide-co-glycolide)/hydroxyapatite composite scaffolds for bone tissue engineering

Biomaterials, 2006

Biodegradable polymer/bioceramic composite scaffolds can overcome the limitations of conventional ceramic bone substitutes such as brittleness and difficulty in shaping. However, conventional methods for fabricating polymer/bioceramic composite scaffolds often use organic solvents (e.g., the solvent casting and particulate leaching (SC/PL) method), which might be harmful to cells or tissues. Furthermore, the polymer solutions may coat the ceramics and hinder their exposure to the scaffold surface, which may decrease the likelihood that the seeded osteogenic cells will make contact with the bioactive ceramics. In this study, a novel method for fabricating a polymer/nano-bioceramic composite scaffold with high exposure of the bioceramics to the scaffold surface was developed for efficient bone tissue engineering. Poly(D,L-lactic-co-glycolic acid)/nano-hydroxyapatite (PLGA/HA) composite scaffolds were fabricated by the gas forming and particulate leaching (GF/PL) method without the use of organic solvents. The GF/PL method exposed HA nanoparticles at the scaffold surface significantly more than the conventional SC/PL method does. The GF/PL scaffolds showed interconnected porous structures without a skin layer and exhibited superior enhanced mechanical properties to those of scaffolds fabricated by the SC/PL method. Both types of scaffolds were seeded with rat calvarial osteoblasts and cultured in vitro or were subcutaneously implanted into athymic mice for eight weeks. The GF/PL scaffolds exhibited significantly higher cell growth, alkaline phosphatase activity, and mineralization compared to the SC/PL scaffolds in vitro. Histological analyses and calcium content quantification of the regenerated tissues five and eight weeks after implantation showed that bone formation was more extensive on the GF/PL scaffolds than on the SC/PL scaffolds. Compared to the SC/PL scaffolds, the enhanced bone formation on the GF/PL scaffolds may have resulted from the higher exposure of HA nanoparticles at the scaffold surface, which allowed for direct contact with the transplanted cells and stimulated the cell proliferation and osteogenic differentiation. These results show that the biodegradable polymer/ bioceramic composite scaffolds fabricated by the novel GF/PL method enhance bone regeneration compared with those fabricated by the conventional SC/PL method. r

Development of an osteoconductive PCL-PDIPF-hydroxyapatite composite scaffold for bone tissue engineering

Journal of tissue engineering and regenerative medicine, 2011

Runx-2 were detected when MC3T3E1 osteoblasts were induced to differentiate and mineralize on HAP-polymer scaffolds. In conclusion, a novel biocompatible HAP-Blend composite with uniform dispersion of semi-nano HAP particles and good interphase compatibility has been prepared successfully. The development of HAP-Blend composite, with improved physical, mechanical and osteoinductive properties, may potentially be used in bone tissue-engineering applications.

Hexagonal pore geometry and the presence of hydroxyapatite enhance deposition of mineralized bone matrix on additively manufactured polylactic acid scaffolds

Materials Science and Engineering: C, 2021

Additive manufacturing (AM) has revolutionized the design of regenerative scaffolds for orthopaedic applications, enabling customizable geometric designs and material compositions that mimic bone. However, the available evidence is contradictory with respect to which geometric designs and material compositions are optimal. There is a lack of studies that systematically compare different pore sizes and geometries in conjunction with the presence or absence of calcium phosphates. We therefore evaluated the physicochemical and biological properties of additively manufactured scaffolds based on polylactic acid (PLA) in combination with hydroxyapatite (HA). HA was either incorporated in the polymeric matrix or introduced as a coating, yielding 15 and 2% wt., respectively. Pore sizes of the scaffolds varied between 200 and 450 μm and were shaped either triangularly or hexagonally. All scaffolds supported the adhesion, proliferation and differentiation of both primary mouse osteoblasts and osteosarcoma cells up to four weeks, with only small differences in the production of alkaline phosphatase (ALP) between cells grown on different pore geometries and material compositions. However, mineralization of the PLA scaffolds was substantially enhanced in the presence of HA, either embedded in the PLA matrix or as a coating at the surface level, and by larger hexagonal pores. In conclusion, customized HA/PLA composite porous scaffolds intended for the repair of critical size bone defects were obtained by a cost-effective AM method. Our findings indicate that the analysis of osteoblast adhesion and differentiation on experimental scaffolds alone is inconclusive without the assessment of mineralization, and the effects of geometry and composition on bone matrix deposition must be carefully considered in order to understand the regenerative potential of experimental scaffolds.

A Proposed Fabrication Method of Novel PCL-GEL-HAp Nanocomposite Scaffolds for Bone Tissue Engineering Applications

Advanced Composites Letters, 2010

In this study, poly(∊-caprolactone) (PCL), gelatin (GEL) and nanocrystalline hydroxyapatite (HAp) was applied to fabricate novel PCL-GEL-HAp nanaocomposite scaffolds through a new fabrication method. With the aim of finding the best fabrication method, after testing different methods and solvents, the best method and solvents were found, and the nanocomposites were prepared through layer solvent casting combined with freeze-drying. Acetone and distillated water were used as the PCL and GEL solvents, respectively. The mechanical test showed that the increasing of the PCL weight through the scaffolds caused the improvement of the final nanocomposite mechanical behavior due to the increasing of the ultimate stress, stiffness and elastic modulus (8 MPa for 0% wt PCL to 23.5 MPa for 50% wt PCL). The biomineralization investigation of the scaffolds revealed the formation of bone-like apatite layers after immersion in simulated body fluid (SBF). In addition, the in vitro cytotoxity of the ...

Engineering new bone tissuein vitro on highly porous poly(?-hydroxyl acids)/hydroxyapatite composite scaffolds

Journal of Biomedical Materials Research, 2001

Engineering new bone tissue with cells and a synthetic extracellular matrix (scaffolding) represents a new approach for the regeneration of mineralized tissues compared with the transplantation of bone (autografts or allografts). In the present work, highly porous poly(L-lactic acid) (PLLA) and PLLA/hydroxyapatite (HAP) composite scaffolds were prepared with a thermally induced phase separation technique. The scaffolds were seeded with osteoblastic cells and cultured in vitro. In the pure PLLA scaffolds, the osteoblasts attached primarily on the outer surface of the polymer. In contrast, the osteoblasts penetrated deep into the PLLA/HAP scaffolds and were uniformly distributed. The osteoblast survival percentage in the PLLA/HAP scaffolds was superior to that in the PLLA scaffolds. The osteoblasts proliferated in both types of the scaffolds, but the cell number was always higher in the PLLA/HAP composite scaffolds during 6 weeks of in vitro cultivation. Bone-specific markers (mRNAs encoding bone sialoprotein and osteocalcin) were expressed more abundantly in the PLLA/HAP composite scaffolds than in the PLLA scaffolds. The new tissue increased continuously in the PLLA/HAP composite scaffolds, whereas new tissue formed only near the surface of pure PLLA scaffolds. These results demonstrate that HAP imparts osteoconductivity and the highly porous PLLA/ HAP composite scaffolds are superior to pure PLLA scaffolds for bone tissue engineering.