Comparative Efficacies of Collagen-Based 3D Printed PCL/PLGA/β-TCP Composite Block Bone Grafts and Biphasic Calcium Phosphate Bone Substitute for Bone Regeneration.
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ABSTRACT:
SUBMITTER: Hwang KS
PROVIDER: S-EPMC5506921 | biostudies-other | 2017 Apr
REPOSITORIES: biostudies-other
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Project description: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 (??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 and increased the wettability of the hybrid scaffold. Furthermore, the flow cytometry and Cell Counting Kit?8 assay revealed that the adhesion and proliferation of hPDLSCs were markedlyincreased on the 0.5PCL/0.5PLGA and 0.1PCL/0.9PLGA scaffolds. Additionally, the alkaline phosphatase activity detection and reverse?transcription quantitative polymerase chain reaction demonstrated that the hPDLSCs on the 0.5PCL/0.5PLGA scaffold exhibited the best osteogenic capacity. Consequently, PCL/PLGA composite scaffolds may represent a candidate focus for future bone regeneration studies, and the 0.5PCL/0.5PLGA scaffold demonstrated the best bio?response from the hPDLSCs.
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Project description:This work aims at providing guidance through systematic experimental characterization, for the design of 3D printed scaffolds for potential orthopaedic applications, focusing on fused deposition modeling (FDM) with a composite of clinically available polycaprolactone (PCL) and ?-tricalcium phosphate (?-TCP). First, we studied the effect of the chemical composition (0% to 60% ?-TCP/PCL) on the scaffold's properties. We showed that surface roughness and contact angle were respectively proportional and inversely proportional to the amount of ?-TCP, and that degradation rate increased with the amount of ceramic. Biologically, the addition of ?-TCP enhanced proliferation and osteogenic differentiation of C3H10. Secondly, we systematically investigated the effect of the composition and the porosity on the 3D printed scaffold mechanical properties. Both an increasing amount of ?-TCP and a decreasing porosity augmented the apparent Young's modulus of the 3D printed scaffolds. Third, as a proof-of-concept, a novel multi-material biomimetic implant was designed and fabricated for potential disk replacement.
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Project description:The aim of this study was to predefine the pore structure of ?-tricalcium phosphate (?-TCP) scaffolds with different macro pore sizes (500, 750, and 1000 µm), to characterize ?-TCP scaffolds, and to investigate the growth behavior of cells within these scaffolds. The lead structures for directional bone growth (sacrificial structures) were produced from polylactide (PLA) using the fused deposition modeling techniques. The molds were then filled with ?-TCP slurry and sintered at 1250 °C, whereby the lead structures (voids) were burnt out. The scaffolds were mechanically characterized (native and after incubation in simulated body fluid (SBF) for 28 d). In addition, biocompatibility was investigated by live/dead, cell proliferation and lactate dehydrogenase assays. The scaffolds with a strand spacing of 500 µm showed the highest compressive strength, both untreated (3.4 ± 0.2 MPa) and treated with simulated body fluid (2.8 ± 0.2 MPa). The simulated body fluid reduced the stability of the samples to 82% (500), 62% (750) and 56% (1000). The strand spacing and the powder properties of the samples were decisive factors for stability. The fact that ?-TCP is a biocompatible material is confirmed by the experiments. No lactate dehydrogenase activity of the cells was measured, which means that no cytotoxicity of the material could be detected. In addition, the proliferation rate of all three sizes increased steadily over the test days until saturation. The cells were largely adhered to or within the scaffolds and did not migrate through the scaffolds to the bottom of the cell culture plate. The cells showed increased growth, not only on the outer surface (e.g., 500: 36 ± 33 vital cells/mm² after three days, 180 ± 33 cells/mm² after seven days, and 308 ± 69 cells/mm² after 10 days), but also on the inner surface of the samples (e.g., 750: 49 ± 17 vital cells/mm² after three days, 200 ± 84 cells/mm² after seven days, and 218 ± 99 living cells/mm² after 10 days). This means that the inverse 3D printing method is very suitable for the presetting of the pore structure and for the ingrowth of the cells. The experiments on which this work is based have shown that the fused deposition modeling process with subsequent slip casting and sintering is well suited for the production of scaffolds for bone replacement.
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Project description:OBJECTIVE:Vascularization is a critical process during bone regeneration/repair and the lack of tissue vascularization is recognized as a major challenge in applying bone tissue engineering methods for cranial and maxillofacial surgeries. The aim of our study is to fabricate a vascular endothelial growth factor (VEGF)-loaded gelatin/alginate/?-TCP composite scaffold by 3D printing method using a computer-assisted design (CAD) model. METHODS:The paste, composed of (VEGF-loaded PLGA)-containing gelatin/alginate/?-TCP in water, was loaded into standard Nordson cartridges and promptly employed for printing the scaffolds. Rheological characterization of various gelatin/alginate/?-TCP formulations led to an optimized paste as a printable bioink at room temperature. RESULTS:The in vitro release kinetics of the loaded VEGF revealed that the designed scaffolds fulfill the bioavailability of VEGF required for vascularization in the early stages of tissue regeneration. The results were confirmed by two times increment of proliferation of human umbilical vein endothelial cells (HUVECs) seeded on the scaffolds after 10 days. The compressive modulus of the scaffolds, 98±11MPa, was found to be in the range of cancellous bone suggesting their potential application for craniofacial tissue engineering. Osteoblast culture on the scaffolds showed that the construct supports cell viability, adhesion and proliferation. It was found that the ALP activity increased over 50% using VEGF-loaded scaffolds after 2 weeks of culture. SIGNIFICANCE:The 3D printed gelatin/alginate/?-TCP scaffold with slow releasing of VEGF can be considered as a potential candidate for regeneration of craniofacial defects.
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Project description:Healing of contaminated/infected bone defects is a significant clinical challenge. Prevalence of multi-antibiotic resistant organisms has renewed interest in the use of antiseptic silver as an effective, but less toxic antimicrobial with decreased potential for bacterial resistance. In this study, we demonstrated that metallic nanosilver particles (with a size of 20-40nm)-poly(lactic-co-glycolic acid) (PLGA) composite grafts have strong antibacterial properties. In addition, nanosilver particles-PLGA composite grafts did not inhibit adherence, proliferation, alkaline phosphatase activity, or mineralization of ongrowth MC3T3-E1 pre-osteoblasts compared to PLGA controls. Furthermore, nanosilver particles did not affect the osteoinductivity of bone morphogenetic protein 2 (BMP-2). Infected femoral defects implanted with BMP-2 coupled 2.0% nanosilver particles-PLGA composite grafts healed in 12 weeks without evidence of residual bacteria. In contrast, BMP-2 coupled PLGA control grafts failed to heal in the presence of continued bacterial colonies. Our results indicate that nanosilver of defined particle size is bactericidal without discernable in vitro and in vivo cytotoxicity or negative effects on BMP-2 osteoinductivity, making it an ideal antimicrobial for bone regeneration in infected wounds.
| S-EPMC3202602 | biostudies-literature
Project description:The use of lignocellulosic remnants of the açaí agro-business will benefit the environment with a precursor material for biomedical applications. Nanocellulose (NC) allows the biomimetic growth of biphasic ceramics on its surface, with characteristics compatible with bone tissue, including bioactive properties and biocompatibility. In this study, the composites were obtained from açaí tegument (Euterpe Oleracea Mart.) NC using acid hydrolysis. The characterization performed by scanning electron microscopy showed the characteristic crystals of hydroxyapatite (HA) and calcium triphosphate (?-TCP) based on the results of X-ray diffraction, with the peak at 22°, showing the NC nucleation of HA and peak at 17° showing tricalcium phosphate (?-TCP). Fourier transform infrared spectroscopy confirmed the presence of O-H at 3400 cm-1 and C-H at 2900 cm-1, which is characteristic of cellulose; peaks were also observed at 1609 cm-1, verifying the reduction in lignin content. Groups PO?-3 at approximately 1070 cm-1, P-OH at 910?1040 cm-1, and HCO?- at 2450 cm-1 confirmed the formation of HA and ?-TCP. The zeta potential had a range of -11 ± 23.8 mV related to particle size, which had a range of 164.2 × 10-9?4748 × 10-9 m.
| S-EPMC6266682 | biostudies-literature
Project description:Treatment of large bone defects using synthetic scaffolds remain a challenge mainly due to insufficient vascularization. This study is to engineer a vascularized bone graft by integrating a vascularized biomimetic cell-sheet-engineered periosteum (CSEP) and a biodegradable macroporous beta-tricalcium phosphate (β-TCP) scaffold. We first cultured human mesenchymal stem cells (hMSCs) to form cell sheet and human umbilical vascular endothelial cells (HUVECs) were then seeded on the undifferentiated hMSCs sheet to form vascularized cell sheet for mimicking the fibrous layer of native periosteum. A mineralized hMSCs sheet was cultured to mimic the cambium layer of native periosteum. This mineralized hMSCs sheet was first wrapped onto a cylindrical β-TCP scaffold followed by wrapping the vascularized HUVEC/hMSC sheet, thus generating a biomimetic CSEP on the β-TCP scaffold. A nonperiosteum structural cell sheets-covered β-TCP and plain β-TCP were used as controls. In vitro studies indicate that the undifferentiated hMSCs sheet facilitated HUVECs to form rich capillary-like networks. In vivo studies indicate that the biomimetic CSEP enhanced angiogenesis and functional anastomosis between the in vitro preformed human capillary networks and the mouse host vasculature. MicroCT analysis and osteocalcin staining show that the biomimetic CSEP/β-TCP graft formed more bone matrix compared to the other groups. These results suggest that the CSEP that mimics the cellular components and spatial configuration of periosteum plays a critical role in vascularization and osteogenesis. Our studies suggest that a biomimetic periosteum-covered β-TCP graft is a promising approach for bone regeneration.
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Project description:ObjectivesIn recent years, the increase in utilisation of bone substitutes in the reconstruction of bone defects has been fuelled by donor site complications associated with autologous bone harvesting. However the ability of bone substitute to stimulate bone union while maintaining fracture reduction has been a topic of debate. Cerament Bone Void Filler (CBVF) is a novel biphasic and injectable ceramic bone substitute that has high compressive strength and the ability to promote cancellous bone healing.Materials and methodThis is a retrospective study to evaluate the surgical outcome of utilising CBVF in the treatment of depressed metaphyseal bone fractures over a two year period. The patients were followed up for at least six months after surgery and clinical parameters such as wound site complications were collated. Radiographic imaging was evaluated to determine loss of fracture reduction and rate of cement resorption.ResultsThirteen patients with depressed metaphyseal fractures were enrolled, which included: (i) one proximal humerus fracture; (ii) three tibial plateau fractures; and (iii) nine calcaneal fractures. None of the patients showed significant collapse in fracture reduction after six months of follow up. Cement resorption was noted in one patient as early as three weeks after surgery. There were no cases of cement leak or wound site complications.ConclusionCerament Bone Void Filler (CBVF) is a promising bone graft substitute in the management of depressed metaphyseal bone fractures, with the ability to maintain fracture reduction despite cement resorption.
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Project description:Research and development of the ideal artificial bone-substitute materials to replace autologous and allogeneic bones for repairing bone defects is still a challenge in clinical orthopedics. Recently, poly(lactic-co-glycolic acid) (PLGA)-based artificial bone-substitute materials are attracting increasing attention as the benefit of their suitable biocompatibility, degradability, mechanical properties, and capabilities to promote bone regeneration. In this article, we comprehensively review the artificial bone-substitute materials made from PLGA or the composites of PLGA and other organic and inorganic substances, elaborate on their applications for bone regeneration with or without bioactive factors, and prospect the challenges and opportunities in clinical bone regeneration.
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Project description:Biofabrication has been adapted in engineering patient-specific biosynthetic grafts for bone regeneration. Herein, we developed a three-dimensional (3D) high-resolution, room-temperature printing approach to fabricate osteoconductive scaffolds using calcium phosphate cement (CPC). The non-aqueous CPC bioinks were composed of tetracalcium phosphate, dicalcium phosphate anhydrous, and Polyvinyl butyral (PVB) dissolved in either ethanol (EtOH) or tetrahydrofuran (THF). They were printed in an aqueous sodium phosphate bath, which performs as a hardening accelerator for hydroxyapatite formation and as a retainer for 3D microstructure. The PVB solvents, EtOH or THF, affected differently the slurry rheological properties, scaffold microstructure, mechanical properties, and osteoconductivity. Our proposed approach overcomes limitations of conventional fabrication methods, which require high-temperature (>50 °C), low-resolution (>400 μm) printing with an inadequate amount of large ceramic particles (>35 μm). This proof-of-concept study opens venues in engineering high-resolution, implantable, and osteoconductive scaffolds with predetermined properties for bone regeneration.
| S-EPMC8164647 | biostudies-literature