Huifeng Shao

3D Printing Surgical Implants at the clinic: A Experimental Study on Anterior Cruciate Ligament Reconstruction

Desktop three-dimensional (3D) printers (D3DPs) have become a popular tool for fabricating personalized consumer products, favored for low cost, easy operation, and other advantageous qualities. This study focused on the potential for using D3DPs to successfully, rapidly, and economically print customized implants at medical clinics. An experiment was conducted on a D3DP-printed anterior cruciate ligament surgical implant using a rabbit model. A well-defined, orthogonal, porous PLA screw-like scaffold was printed, then coated with hydroxyapatite (HA) to improve its osteoconductivity. As an internal fixation as well as an ideal cell delivery system, the osteogenic scaffold loaded with mesenchymal stem cells (MSCs) were evaluated through both in vitro and in vivo tests to observe bone-ligament healing via cell therapy. The MSCs suspended in Pluronic F-127 hydrogel on PLA/HA screw-like scaffold showed the highest cell proliferation and osteogenesis in vitro. In vivo assessment of rabbit anterior cruciate ligament models for 4 and 12 weeks showed that the PLA/HA screw-like scaffold loaded with MSCs suspended in Pluronic F-127 hydrogel exhibited significant bone ingrowth and bone-graft interface formation within the bone tunnel. Overall, the results of this study demonstrate that fabricating surgical implants at the clinic (fab@clinic) with D3DPs can be feasible, effective, and economical.

Bioactive glass-reinforced bioceramic ink writing scaffolds: sintering, microstructure and mechanical behavior

The densification of pore struts in bioceramic scaffolds is important for structure stability and strength reliability. An advantage of ceramic ink writing is the precise control over the microstructure and macroarchitecture. However, the use of organic binder in such ink writing process would heavily affect the densification of ceramic struts and sacrifice the mechanical strength of porous scaffolds after sintering. This study presents a low-melt-point bioactive glass (BG)-assisted sintering strategy to overcome the main limitations of direct ink writing (extrusion-based three-dimensional printing) and to produce high-strength calcium silicate (CSi) bioceramic scaffolds. The 1% BG-added CSi (CSi-BG1) scaffolds with rectangular pore morphology sintered at 1080 °C have a very small BG content, readily induce apatite formation, and show appreciable linear shrinkage (∼21%), which is consistent with the composite scaffolds with less or more BG contents sintered at either the same or a higher temperature. These CSi-BG1 scaffolds also possess a high elastic modulus (∼350 MPa) and appreciable compressive strength (∼48 MPa), and show significant strength enhancement after exposure to simulated body fluid-a performance markedly superior to those of pure CSi scaffolds. Particularly, the honeycomb-pore CSi-BG1 scaffolds show markedly higher compressive strength (∼88 MPa) than the scaffolds with rectangular, parallelogram, and Archimedean chord pore structures. It is suggested that this approach can potentially facilitate the translation of ceramic ink writing and BG-assisted sintering of bioceramic scaffold technologies to the in situ bone repair.

Simultaneous mechanical property and biodegradation improvement of wollastonite bioceramic through magnesium dilute doping.

The large-area bone defects in head (including calvarial, orbital, and maxillofacial bone) and segmental bone are attracting increased attention in a wide range of clinical departments. A key requirement for the clinical success of the bioactive ceramics is the match of the mechanical behavior of the implants with the specific bone tissue to be filled. This raises the question as to what design strategy might be the best indicators for the balance between mechanical properties and biological performances. Here we go beyond the traditional approaches that use phase conversion or biphasic hybrid; instead, we achieved a simultaneous enhancement of several mechanical parameters and optimalization of biodegradability by using a dilute doping of Mg in a single-phase wollastonite bioceramic. We show that the wollastonite ceramic can be rationally tuned in phase (α or β), mechanical strength (in compression and bending mode), elastic modulus (18-23GPa), and fracture toughness (>3.2MPam(1/2)) through the usage of Mg dopant introduced at precisely defined dilute concentrations (Mg/Ca molar ratio: 1.2-2.1%). Meanwhile, the dilute Mg-doped wollastonite ceramics are shown to exhibit good bioactivity in vitro in SBF but biodegradation in Tris is inversely proportional to Mg content. Consequently, such new highly bioactive ceramics with appreciable strength and toughness are promising for making specific porous scaffolds for enhancing large segmental bone defect and thin-wall bone defect repair.

Systematical Evaluation of Mechanically Strong 3D Printed Diluted magnesium Doping Wollastonite Scaffolds on Osteogenic Capacity in Rabbit Calvarial Defects

Wollastonite (CaSiO3; CSi) ceramic is a promising bioactive material for bone defect repair due to slightly fast degradation of its porous constructs in vivo. In our previous strategy some key features of CSi ceramic have been significantly improved by dilute magnesium doping for regulating mechanical properties and biodegradation. Here we demonstrate that 6 ~ 14% of Ca substituted by Mg in CSi (CSi-Mgx, x = 6, 10, 14) can enhance the mechanical strength (>40 MPa) but not compromise biological performances of the 3D printed porous scaffolds with open porosity of 60‒63%. The in vitro cell culture tests in vitro indicated that the dilute Mg doping into CSi was beneficial for ALP activity and high expression of osteogenic marker genes of MC3T3-E1 cells in the scaffolds. A good bone tissue regeneration response and elastoplastic response in mechanical strength in vivo were determined after implantation in rabbit calvarial defects for 6‒12 weeks. Particularly, the CSi-Mg10 and CSi-Mg14 scaffolds could enhance new bone regeneration with a significant increase of newly formed bone tissue (18 ~ 22%) compared to the pure CSi (~14%) at 12 weeks post-implantation. It is reasonable to consider that, therefore, such CSi-Mgx scaffolds possessing excellent strength and reasonable degradability are promising for bone reconstruction in thin-wall bone defects.

Bone regeneration in 3D printing bioactive ceramic scaffolds with improved tissue/material interface pore architecture in thin-wall bone defect

Three-dimensional (3D) printing bioactive ceramics have demonstrated alternative approaches to bone tissue repair, but an optimized materials system for improving the recruitment of host osteogenic cells into the bone defect and enhancing targeted repair of the thin-wall craniomaxillofacial defects remains elusive. Herein we systematically evaluated the role of side-wall pore architecture in the direct-ink-writing bioceramic scaffolds on mechanical properties and osteogenic capacity in rabbit calvarial defects. The pure calcium silicate (CSi) and dilute Mg-doped CSi (CSi-Mg6) scaffolds with different layer thickness and macropore sizes were prepared by varying the layer deposition mode from single-layer printing (SLP) to double-layer printing (DLP) and then by undergoing one-, or two-step sintering. It was found that the dilute Mg doping and/or two-step sintering schedule was especially beneficial for improving the compressive strength (∼25-104 MPa) and flexural strength (∼6-18 MPa) of the Ca-silicate scaffolds. The histological analysis for the calvarial bone specimens in vivo revealed that the SLP scaffolds had a high osteoconduction at the early stage (4 weeks) but the DLP scaffolds displayed a higher osteogenic capacity for a long time stage (8-12 weeks). Although the DLP CSi scaffolds displayed somewhat higher osteogenic capacity at 8 and 12 weeks, the DLP CSi-Mg6 scaffolds with excellent fracture resistance also showed appreciable new bone tissue ingrowth. These findings demonstrate that the side-wall pore architecture in 3D printed bioceramic scaffolds is required to optimize for bone repair in calvarial bone defects, and especially the Mg doping wollastontie is promising for 3D printing thin-wall porous scaffolds for craniomaxillofacial bone defect treatment.

Systematic investigation of β-dicalcium silicate-based bone cements in vitro and in vivo in comparison with clinically applied calcium phosphate cement and Bio-Oss®

Ca-silicate cements have drawn considerable attention for their potential applications in the field of bone repair due to their excellent bioactivity in vitro. Significant progress with regard to physicochemical properties and optimization of fabrication techniques of this new cement system has been achieved. However, it is unknown whether these so-called bioactive cements could efficiently repair critical-size bone defects in vivo. Herein we systematically investigated the biological performance of a β-dicalcium silicate (β-C2S)-based bone cement in comparison with the clinically used calcium phosphate cement (CPC) and Bio-Oss®. The results indicated that β-C2S-based cement with 25% gypsum exhibited appreciable Ca and Si release and weight loss (∼28%) in Tris buffer within the initial 7 d but then maintained a mild degradation rate during 1–4 weeks. Also, the β-C2S-based cement extracts readily enhanced MC3T3 proliferation at a 25 mg ml−1 level at 4 d and ALP expression at 50–100 mg ml−1 levels at 7 d. For the β-C2S-based group, increased mRNA levels of osteogenic genes, including Collagen I, osteocalcin, special protein 7, and runt-related transcription factor 2, were observed. In particular, histological staining and microCT reconstruction analyses demonstrated that this new cement could significantly enhance new bone regeneration in a critical-sized skull defect model in rabbits compared with CPC and Bio-Oss®. These findings suggest that β-C2S-based biocement is a promising bone implant for bone regeneration and repair due to its excellent biological performance in vitro and in vivo.

The outstanding mechanical response and bone regeneration capacity of robocast dilute magnesium-doped wollastonite scaffolds in critical size bone defects

The regeneration and repair of damaged load-bearing segmental bones require considerable mechanical strength for the artificial implants. The ideal biomaterials should also facilitate the production of porous implants with high bioactivity desirable for stimulating new bone growth. Here we developed a new mechanically strong, highly bioactive dilute magnesium-doped wollastonite (CaSiO3-Mg; CSi-Mg) porous scaffold by the robocasting technique. The sintered scaffolds had interconnected pores 350 µm in size and over 50% porosity with appreciable compressive strength (>110 MPa), 5-10 times higher than those of pure CSi and β-TCP porous ceramics. Extensive in vitro and in vivo investigations revealed that such Ca-silicate bioceramic scaffolds were particularly beneficial for osteogenic cell activity and osteogenic capacity in critical size femoral bone defects. The CSi-Mg porous constructs were accompanied by an accelerated new bone growth (6-18 weeks) and a mechanically outstanding elastoplastic response to finally match the strength (10-15 MPa) of the rabbit femur host bone after 18 weeks, and the material itself experienced mild resorption and apatite-like phase transformation. In contrast, the new bone regeneration in the β-TCP scaffolds was substantially retarded after 6-12 weeks of implantation, and exhibited a low level of mechanical strength (<10 MPa) similar to the pure CSi scaffolds. These results suggest a promising application of robocast CSi-Mg scaffolds in the clinic, especially for the load-bearing bone defects.

45S5 Bioglass analogue reinforced akermanite ceramic favorable for additive manufacturing mechanically strong scaffolds

Calcium–magnesium silicate bioceramics have attracted increased interest in the development of porous scaffolds for bone tissue engineering applications, mainly due to their excellent bioactivity and ability to bond to hard tissue. However, the shaping of these bioceramics into complex porous constructs is challenging, and, especially, conventional high temperature pressureless sintering is not always an effective method to improve their mechanical properties without further compromising their biologically relevant performances. Here we developed a low melting-point bioactive glass (BG)-assisted sintering approach to improve the mechanical properties of akermanite ceramics with and without intentionally manufacturing macroporous structures. The experimental results indicated that the 4 wt% B2O3-containing 45S5 BG analogue could readily reinforce akermanite ceramics at a 20–40 wt% content, and material extrusion 3D-printing followed by a pressureless sintering process could be employed to fabricate high-strength porous scaffolds with compressive strength (∼36 MPa) ten times higher than those of pure akermanite porous ceramics. Moreover, the composite porous ceramics showed slower biodegradation in Tris buffer in vitro and this did not heavily affect the strength of their porous formulation over a long time period (6 weeks). It is proposed that 3D printing followed by an NCS-B-assisted sintering process represents an effective alternative for developing high strength bioceramic scaffolds potentially for the repair of load-bearing segmental bone defects.

Preparation and Characterization of Low Temperature Heat-Treated 45S5 Bioactive Glass-Ceramic Analogues

Abstract The 45S5 Bioglassr and its sintered bioactive glass-ceramic (BGC) have been widely investigated as bone implants, mainly for its ability to bond to hard tissues. However, high temperature treatment is not enough to improve its poor mechanical properties, but compromise its biologically relevant performances. The innovative BGC compositions based on the thermally treated 45S5 Bioglassr were developed by decreasing the P2O5 quantity and adding B2O3 (0-6%) into the Na2O–2CaO–3SiO2- based bioactive glasses (BG). The thermally treated BGCs were fully characterized from the microstructural and mechanical points of view and compared to each other. Their bioactivity and bio-dissolutionwere established by means of in vitro soaking tests. The new B2O3-added 45S5 BG analogues, named NCS-xB, can be transformed to crystalline phase (Na2Ca2Si3O9)-based BGCs of high compactness and bioactivity at a relatively low temperature heat treatment (≤ 900ºC), since their bioactivity is preserved. Our experimental results suggest that the new 45S5 BGC analogues with optimized composition exhibit improved micro- structural and mechanical properties, and are beneficial for making specific products such as porous scaffolds or composites for bone defect repair.

Custom Repair of Mandibular Bone Defects with 3D Printed Bioceramic Scaffolds.

Implanting artificial biomaterial implants into alveolar bone defects with individual shape and appropriate mechanical strength is still a challenge. In this study, bioceramic scaffolds, which can precisely match the mandibular defects in macro and micro, were manufactured by the 3-dimensional (3D) printing technique according to the computed tomography (CT) image. To evaluate the stimulatory effect of the material substrate on bone tissue regeneration in situ in a rabbit mandibular alveolar bone defect model, implants made with the newly developed, mechanically strong ~10% Mg-substituted wollastonite (Ca90%Mg10%SiO3; CSi-Mg10) were fabricated, implanted into the bone defects, and compared with implants made with the typical Ca-phosphate and Ca-silicate porous bioceramics, such as β-tricalcium phosphate (TCP), wollastonite (CaSiO3; CSi), and bredigite (Bred). The initial physicochemical tests indicated that although the CSi-Mg10 scaffolds had the largest pore dimension, they had the lowest porosity mainly due to the significant linear shrinkage of the scaffolds during sintering. Compared with the sparingly dissolvable TCP scaffolds (~2% weight loss) and superfast dissolvable (in Tris buffer within 6 wk) pure CSi and Bred scaffolds (~12% and ~14% weight loss, respectively), the CSi-Mg10 exhibited a mild in vitro biodissolution and moderate weight loss of ~7%. In addition, the CSi-Mg10 scaffolds showed a considerable initial flexural strength (31 MPa) and maintained very high flexural resistance during soaking in Tris buffer. The in vivo results revealed that the CSi-Mg10 scaffolds have markedly higher osteogenic capability than those on the TCP, CSi, and Bred scaffolds after 16 wk. These results suggest a promising potential application of customized CSi-Mg10 3D robocast scaffolds in the clinic, especially for repair of alveolar bone defects.