Development of biodegradable bioactive glass ceramics by DLP printed containing EPCs/BMSCs for bone tissue engineering of rabbit mandible defects.

Bioactive glass ceramics have excellent biocompatibility and osteoconductivity; and can form direct chemical bonds with human bones; thus, these ceramic are considered as "Smart" materials. In this study, we develop a new type of bioactive glass ceramic (AP40mod) as a scaffold containing Endothelial progenitor cells (EPCs) and Mesenchymal stem cells (BMSCs) to repair critical-sized bone defects in rabbit mandibles. For in vitro experiments: AP40mod was prepared by Dgital light processing (DLP) system and the optimal ratio of EPCs/BMSCs was screened by analyzing cell proliferation and ALP activity, as well as the influence of genes related to osteogenesis and angiogenesis by direct inoculation into scaffolds. The scaffold showed suitable mechanical properties, with a Bending strength 52.7 MPa and a good biological activity. Additionally, when EPCs/BMSCs ratio were combined at a ratio of 2:1 with AP40mod, the ALP activity, osteogenesis and angiogenesis were significantly increased. For in vivo experiments: application of AP40mod/EPCs/BMSCs (after 7 days of in vitro spin culture) to repair and reconstruct critical-sized mandible defect in rabbit showed that all scaffolds were successfully accurately implanted into the defect area. As revealed by macroscopically and CT at the end of 9 months, defects in the AP40mod/EPCs/BMSCs group were nearly completely covered by normal bone and the degradation rate was 29.9% compared to 20.1% in the AP40mod group by the 3D reconstruction. As revealed by HE and Masson staining analyses, newly formed blood vessels, bone marrow and collagen maturity were significantly increased in the AP40mod/EPCs/BMSCs group compared to those in the AP40mod group. We directly inoculated cells on the novel material to screen for the best inoculation ratio. It is concluded that the AP40mod combination of EPCs/BMSCs is a promising approach for repairing and reconstructing large load bearing bone defect.

The effect of latency on bone lengthening force and bone mineralization: an investigation using strain gauge mounted on internal distractor device.

BACKGROUND: The purpose of this study was to investigate the effect of latency on the development of bone lengthening force and bone mineralization during mandible distraction osteogenesis. METHODS: Distraction tensions were investigated at different latency period in 36 rabbits using internal unilateral distractor. Strain gauges were prepared and attached to the distractor to directly assess the level of distraction tension during mandible lengthening. The tensile force environment of the mandible of rabbit during distraction was evaluated through in vivo experiments using two gauges. The animals were divided into 3 groups each containing 12 rabbits. Latency periods of 0, 4 and 7 days respectively were observed prior to beginning distraction. The distraction protocol consisted of a lengthening rate of 1 mm once daily for 8 days, followed by a consolidation phase of 2 weeks after which the animals were killed. Biopsies specimens were taken from the distracted area at the end of the distraction period. A non-distracted area of the mandible bone served as control. The specimens were analyzed by scanning electron microscopy to assess the ultrastructural pattern, and the bone mineralization. RESULTS: The resting tension acting on the distraction gap increases through distraction. The 7-day latency groups exhibit higher tension then those of 0-day and 4-days latency groups. Quantitative energy dispersive spectral analysis confirmed that immediate distractions were associated with lower calcium and phosphate atomic weight ratio. CONCLUSION: the latency periods could affect the bone lengthening tension and the bone mineralization process.

Design and fabrication of biomimetic multiphased scaffolds for ligament-to-bone fixation.

Conventional ligament grafts with single material composition cannot effectively integrate with the host bones due to mismatched properties and eventually affect their long-term function in vivo. Here we presented a multi-material strategy to design and fabricate composite scaffolds including ligament, interface and bone multiphased regions. The interface region consists of triphasic layers with varying material composition and porous structure to mimic native ligament-to-bone interface while the bone region contains polycaprolactone (PCL) anchor and microchanneled ceramic scaffolds to potentially provide combined mechanical and biological implant-bone fixation. Finite element analysis (FEA) demonstrated that the multiphased scaffolds with interference value smaller than 0.5 mm could avoid the fracture of ceramic scaffold during the implantation process, which was validated by in-vitro implanting the multiphased scaffolds into porcine joint bones. Pull-out experiment showed that the initial fixation between the multiphased scaffolds with 0.47 mm interference and the host bones could withstand the maximum force of 360.31±97.51 N, which can be improved by reinforcing the ceramic scaffolds with biopolymers. It is envisioned that the multiphased scaffold could potentially induce the regeneration of a new bone as well as interfacial tissue with the gradual degradation of the scaffold and subsequently realize long-term biological fixation of the implant with the host bone.

Design and optimization of the fixing plate for customized mandible implants.

Customized mandible implants are used as the most effective surgical option for the reconstruction of the mandible after resection, and have become more prevalent, especially with the development of reverse engineering and rapid prototyping (RP). The fixing plate is the most important and complicated part; however, improper structures of the fixing plate often cost unnecessary workloads during surgery and might lead to fracture failure eventually. The fillet radius, cross-section, and countersinks distribution of the fixing plate are the three most significant factors to affect the strength of the implant. The fillet radius on the plate-body transition determines the amount of grinding bone and can also affect the strength of the fixing plate. In addition, both the different cross-sections of the fixing plate and the different distributions of the countersinks can influence the strength and anti-bending capacity of the fixing plate. Various structures of the fixing plate have been designed, and theoretical calculations and finite element analysis on its strength have been conducted in this study, and results presented an optimized design of the structure of the fixing plate. Moreover, for validation purposes, several clinical applications were successfully implemented with the optimized structure.

[Fabrication and in vivo implantation of ligament-bone composite scaffolds based on three-dimensional printing technique].

OBJECTIVE: To solve the fixation problem between ligament grafts and host bones in ligament reconstruction surgery by using ligament-bone composite scaffolds to repair the ligaments, to explore the fabrication method for ligament-bone composite scaffolds based on three-dimensional (3-D) printing technique, and to investigate their mechanical and biological properties in animal experiments. METHODS: The model of bone scaffolds was designed using CAD software, and the corresponding negative mould was created by boolean operation. 3-D printing techinique was employed to fabricate resin mold. Ceramic bone scaffolds were obtained by casting the ceramic slurry in the resin mould and sintering the dried ceramics-resin composites. Ligament scaffolds were obtained by weaving degummed silk fibers, and then assembled with bone scaffolds and bone anchors. The resultant ligament-bone composite scaffolds were implanted into 10 porcine left anterior cruciate ligament rupture models at the age of 4 months. Mechanical testing and histological examination were performed at 3 months postoperatively, and natural anterior cruciate ligaments of the right sides served as control. RESULTS: Biomechanical testing showed that the natural anterior cruciate ligament of control group can withstand maximum tensile force of (1 384 +/- 181) N and dynamic creep of (0.74 +/- 0.21) mm, while the regenerated ligament-bone scaffolds of experimental group can withstand maximum tensile force of (370 +/- 103) N and dynamic creep of (1.48 +/- 0.49) mm, showing significant differences (t = 11.617, P = 0.000; t = 2.991, P = 0.020). In experimental group, histological examination showed that new bone formed in bone scaffolds. A hierarchical transition structure regenerated between ligament-bone scaffolds and the host bones, which was similar to the structural organizations of natural ligament-bone interface. CONCLUSION: Ligament-bone composite scaffolds based on 3-D printing technique facilitates the regeneration of biomimetic ligament-bone interface. It is expected to achieve physical fixation between ligament grafts and host bone.

[Application of digital design and three-dimensional printing technique on individualized medical treatment].

OBJECTIVE: To summarize the latest research development of the application of digital design and three-dimensional (3-D) printing technique on individualized medical treatment. METHODS: Recent research data and clinical literature about the application of digital design and 3-D printing technique on individualized medical treatment in Xi'an Jiaotong University and its cooperation unit were summarized, reviewed, and analyzed. RESULTS: Digital design and 3-D printing technique can design and manufacture individualized implant based on the patient's specific disease conditions. And the implant can satisfy the needs of specific shape and function of the patient, reducing dependence on the level of experience required for the doctor. So 3-D printing technique get more and more recognition of the surgeon on the individualized repair of human tissue. Xi'an Jiaotong University is the first unit to develop the commercial 3-D printer and conduct depth research on the design and manufacture of individualized medical implant. And complete technological processes and quality standards of product have been developed. CONCLUSION: The individualized medical implant manufactured by 3-D printing technique can not only achieve personalized match but also meet the functional requirements and aesthetic requirements of patients. In addition, the individualized medical implant has the advantages of accurate positioning, stable connection, and high strength. So 3-D printing technique has broad prospects in the manufacture and application of individualized implant.

[Cartilage repair and subchondral bone reconstruction based on three-dimensional printing technique].

OBJECTIVE: To investigate whether subchondral bone microstructural parameters are related to cartilage repair during large osteochondral defect repairing based on three-dimensional (3-D) printing technique. METHODS: Biomimetic biphasic osteochondral composite scaffolds were fabricated by using 3-D printing technique. The right trochlea critical sized defects (4.8 mm in diameter, 7.5 mm in depth) were created in 40 New Zealand white rabbits (aged 6 months, weighing 2.5-3.5 kg). Biomimetic biphasic osteochondral composite scaffolds were implanted into the defects in the experimental group (n = 35), and no composite scaffolds implantation served as control group (n = 5); the left side had no defect as sham-operation group. Animals of experimental and sham-operation groups were euthanized at 1, 2, 4, 8, 16, 24, and 52 weeks after operation, while animals of control group were sampled at 24 weeks. Subchondral bone microstructural parameters and cartilage repair were quantitatively analyzed using Micro-CT and Wayne scoring system. Correlation analysis and regression analysis were applied to reveal the relationship between subchondral bone parameters and cartilage repair. The subchondral bone parameters included bone volume fraction (BV/TV), bone surface area fraction (BSA/BV), trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular spacing (Tb.Sp). RESULTS: In the experimental group, articular cartilage repair was significantly improved at 52 weeks postoperatively, which was dominated by hyaline cartilage tissue, and tidal line formed. Wayne scores at 24 and 52 weeks were significantly higher than that at 16 weeks in the experimental group (P < 0.05), but no significant difference was found between at 24 and 52 weeks (P > 0.05); the scores of experimental group were significantly lower than those of sham-operation group at all time points (P < 0.05). In the experimental group, new subchondral bone migrated from the surrounding defect to the centre, and subchondral bony plate formed at 24 and 52 weeks. The microstructural parameters of repaired subchondral bone followed a "twin peaks" like discipline to which BV/TV, BSA/BV, and Tb.N increased at 2 and 16 weeks, and then they returned to normal level. The Tb.Sp showed reversed discipline compared to the former 3 parameters, no significant change was found for Tb.Th during the repair process. Correlation analysis showed that BV/TV, BSA/BV, Tb.Th, Tb.N, and Tb.Sp were all related with gross appearance score and histology score of repaired cartilage. CONCLUSION: Subchondral bone parameters are related with cartilage repair in critical size osteochondral repair in vivo. Microstructural parameters of repaired subchondral bone follow a "twin peaks" like discipline (osteoplasia-remodeling-osteoplasia-remodeling) to achieve reconstruction, 2nd week and 16th week are critical time points for subchondral bone functional restoration.

Layer-by-layer micromolding of natural biopolymer scaffolds with intrinsic microfluidic networks.

A three-dimensional (3D) microfluidic network plays an important role in engineering thick organs. However, most of the existing methods are limited to mechanically robust synthetic biomaterials and only planar or simple microfluidic networks have been incorporated into soft natural biopolymers. Here we presented an automatic layer-by-layer micromolding strategy to reproducibly fabricate 3D microfluidic porous scaffolds directly from the aqueous solution of soft natural biopolymers. Process parameters such as the liquid volume for each layer and contact displacement were investigated to produce a structurally stable 3D microfluidic scaffold. Microscopic characterization demonstrated that the microfluidic channels were interconnected in 3D and successfully functioned as a convective pathway to transport a polymer solution. Endothelial cells grew relatively well in the porous microfluidic channels. It is envisioned that this method could provide an alternative way to reproducibly build complex 3D microfluidic networks into extracellular matrix-like scaffolds for the fabrication of soft vascularized organs.

Fabrication of nature-inspired microfluidic network for perfusable tissue constructs.

A microreplication method is presented to transfer nature optimized vascular network of leaf venation into various synthetic matrixes. The biomaterial hydrogel with these microfluidic networks is proven to facilitate the growth of endothelial cells and simultaneously function as convection pathways to transport nutrients and oxygen in a pump-free bioreactor setup, which is crucial for the long-term viability of encapsulated cells.

Ice-template-induced silk fibroin-chitosan scaffolds with predefined microfluidic channels and fully porous structures.

Scaffold-based tissue engineering has made great progress in fabricating relatively simple tissues. One of the major challenges in creating thick complex organs is to achieve sufficient nutrient supply as well as uniform cell distribution in a three-dimensional (3D) scaffold. Here we employed microstructured ice templates to fabricate silk fibroin-chitosan (SF-CS) scaffolds with predefined microfluidic channels, open-pore surface and oriented porous structures. The effects of these structural organizations in ice-template-induced (ITI) scaffolds on nutrient delivery, cell seeding as well as cell growth were well investigated in comparison with that of polydimethylsiloxane-template-induced scaffolds. The ITI scaffolds exhibited better structural properties in promoting mass transport, facilitating uniform cell distribution and growth. The ITI scaffolds uniformly seeded with living cells could be further rolled up to form a thick tissue-engineered construct with predefined microfluidic channels. We envision that our ITI scaffolds can be potentially used to engineer thick prevascularized organs when the oriented porous structures are uniformly seeded with primary cells and the predefined microfluidic channels are incorporated with endothelial cells.

Detection of cytochrome c at biocompatible nanostructured Au-lipid bilayer-modified electrode.

The present work describes the dectection of cytochrome c (cyt c) at biocompatible aurum (Au) nanoparticle-structured supported bilayer lipid membrane (sBLM) modified with anionic sites. Au nanoparticles were directly deposited through sBLM modified with lauric acid (LA) to build a hybrid device of nanoscale electrode array via potential cycling in 10 mM HAuCl4 solution containing 0.1 M KCl. The properties of Au nanoparticle-doped sBLM composite were then characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). Results indicated that Au nanoparticles grew in voids of the sBLM with size around 20 - 30 nm. With SWV, after optimization, the results of the experiments indicate that the currents of cyt c were linear functions of its concentrations over the range from 1.0 x 10(-7) to 3.2 x 10(-6) M and the limit of detection (LOD, S/N = 3) was 5 x 10(-8) M. The influences of several common base pairs, amino acids and metal ions on determination of cyt c via this Au nanoparticle-doped sBLM composite were relatively low in experiments, suggesting the excellent biocompatibility of this detection method.

Individually Prefabricated Prosthesis for Maxilla Reconstuction.

The reconstruction of maxillofacial bone defects by the intraoperative modeling of implants may reduce the predictability of the esthetic result, leading to more invasive surgery and increased surgical time. To improve the maxillofacial surgery outcome, modern manufacturing methods such as rapid prototyping (RP) technology and methods based on reverse engineeing (RE) and medical imaging data are applicable to the manufacture of custom-made maxillary prostheses. After acquisition of data, an individual computer-based 3D model of the bony defect is gernerated. These data are tranferrred into RE software to create the prosthesis using a computer-aided design (CAD) model, which is directed into the RP machine for the production of the physical model. The precise fit of the prosthesis is evaulated using the prosthesis and skull model. The prosthesis is then directly used in investment casting such as "Quick Cast" pattern to produce the titanium model. In the clincical reports presented here, reconstructions of two patients with large maxillary bone defects during the operations, and surgery time was reduced. These cases show that the prefabrication of a prosthesis using modern manufacturing technology is an effective method for maxillofacial defect reconstruction.