An Innovative Design to Enhance Osteoinductive Efficacy and Biomechanical Behavior of a Titanium Dental Implant.

The present study investigated the in vivo bone-forming efficacy of an innovative titanium (Ti) dental implant combined with a collagen sponge containing recombinant human bone morphogenetic protein-2 (BMP-2) in a pig model. Two different concentrations of BMP-2 (20 and 40 µg/mL) were incorporated into collagen sponges and placed at the bottom of Ti dental implants. The investigated implants were inserted into the edentulous ridge at the canine-premolar regions of Lanyu small-ear pigs, which were then euthanized at weeks 1, 2, 4, 8, and 12 post-implantation. Specimens containing the implants and surrounding bone tissue were collected for histological evaluation of their bone-to-implant contact (BIC) ratios and calculation of maximum torques using removal torque measurement. Analytical results showed that the control and BMP-2-loaded implants presented good implant stability and bone healing for all testing durations. After 1 week of healing, the BMP-2-loaded implants with a concentration of 20 µg/mL exhibited the highest BIC ratios, ranging from 58% to 76%, among all groups (p = 0.034). Additionally, they also possessed the highest removal torque values (50.1 ± 1.3 N-cm) throughout the 8-week healing period. The BMP-2-loaded implants not only displayed excellent in vivo biocompatibility but also presented superior osteoinductive performance. Therefore, these findings demonstrate that BMP-2 delivered through a collagen sponge can potentially enhance the early-stage osseointegration of Ti dental implants.

Biomechanical stress distribution of medical inelastic fabrics with different porosity structures.

Clothing fit and pressure comfort play important role in clothing comfort, especially in medical body sculpting clothing (MBSC). In the present study, different body movements (forward bending, side bending, and twisting) were adopted to simulate and investigate the biomechanical stress distribution of the human body with three kinds of porosity inelastic MBSCs through the finite element analysis method. The elastic modulus of the investigated MBSCs was also measured by means of tensile testing. Analytical results showed that in the compression region during body movements, the investigated inelastic MBSCs endured less compression stress, and most of the stress was transmitted to the human body. Moreover, the stresses on the body surface were decreased with the porosity increasing. However, most of the von Mises stresses on the human body were in the desired pressure comfort range. Therefore, these results could provide potential information in the modification of MBSC for medical applications.

An Innovative Biofunctional Composite Hydrogel with Enhanced Printability, Rheological Properties, and Structural Integrity for Cell Scaffold Applications.

The present study was conducted to manipulate various biomaterials to find potential hydrogel formulations through three-dimensional (3D) bioprinting fabrication for tissue repair, reconstruction, or regeneration. The hydrogels were prepared using sodium alginate and gelatin combined with different concentrations of Pluronic F127 (6% (3 g), 8% (4 g), and 10% (5 g)) and were marked as AGF-6%, AGF-8%, and AGF-10%, respectively. The properties of the hydrogels were investigated using a contact angle goniometer, rheometer, and 3D bioprinter. In addition, the osteoblast-like cell line (MG-63) was used to evaluate the cell viability including hydrogels before and after 3D bioprinting. It was found that the ratio of contact angle was lowest at AGF-6%, and the rheological results were higher for all samples of AGF-6%, AGF-8%, and AGF-10% compared with the control sample. The printability indicated that the AGF-6% hydrogel possessed great potential in creating a cell scaffold with shape integrity. Moreover, the live/dead assay also presented the highest numbers of live cells before printing compared with after printing. However, the number of live cells on day 7 was higher than on day 1 before and after printing (** p < 0.01). Therefore, the combination of AGF-6% could be developed as a biofunctional hydrogel formulation for potential tissue regeneration applications.

A Three-Dimensional Bioprinted Copolymer Scaffold with Biocompatibility and Structural Integrity for Potential Tissue Regeneration Applications.

The present study was to investigate the rheological property, printability, and cell viability of alginate−gelatin composed hydrogels as a potential cell-laden bioink for three-dimensional (3D) bioprinting applications. The 2 g of sodium alginate dissolved in 50 mL of phosphate buffered saline solution was mixed with different concentrations (1% (0.5 g), 2% (1 g), 3% (1.5 g), and 4% (2 g)) of gelatin, denoted as GBH-1, GBH-2, GBH-3, and GBH-4, respectively. The properties of the investigated hydrogels were characterized by contact angle goniometer, rheometer, and bioprinter. In addition, the hydrogel with a proper concentration was adopted as a cell-laden bioink to conduct cell viability testing (before and after bioprinting) using Live/Dead assay and immunofluorescence staining with a human corneal fibroblast cell line. The analytical results indicated that the GBH-2 hydrogel exhibited the lowest loss rate of contact angle (28%) and similar rheological performance as compared with other investigated hydrogels and the control group. Printability results also showed that the average wire diameter of the GBH-2 bioink (0.84 ± 0.02 mm (*** p < 0.001)) post-printing was similar to that of the control group (0.79 ± 0.05 mm). Moreover, a cell scaffold could be fabricated from the GBH-2 bioink and retained its shape integrity for 24 h post-printing. For bioprinting evaluation, it demonstrated that the GBH-2 bioink possessed well viability (>70%) of the human corneal fibroblast cell after seven days of printing under an ideal printing parameter combination (0.4 mm of inner diameter needle, 0.8 bar of printing pressure, and 25 °C of printing temperature). Therefore, the present study suggests that the GBH-2 hydrogel could be developed as a potential cell-laden bioink to print a cell scaffold with biocompatibility and structural integrity for soft tissues such as skin, cornea, nerve, and blood vessel regeneration applications.

Three-Dimensional Printing of a Hybrid Bioceramic and Biopolymer Porous Scaffold for Promoting Bone Regeneration Potential.

In this study, we proposed a three-dimensional (3D) printed porous (termed as 3DPP) scaffold composed of bioceramic (beta-tricalcium phosphate (ß-TCP)) and thermoreversible biopolymer (pluronic F-127 (PF127)) that may provide bone tissue ingrowth and loading support for bone defect treatment. The investigated scaffolds were printed in three different ranges of pore sizes for comparison (3DPP-1: 150−200 µm, 3DPP-2: 250−300 µm, and 3DPP-3: 300−350 µm). The material properties and biocompatibility of the 3DPP scaffolds were characterized using scanning electron microscopy, X-ray diffractometry, contact angle goniometry, compression testing, and cell viability assay. In addition, micro-computed tomography was applied to investigate bone regeneration behavior of the 3DPP scaffolds in the mini-pig model. Analytical results showed that the 3DPP scaffolds exhibited well-defined porosity, excellent microstructural interconnectivity, and acceptable wettability (θ < 90°). Among all groups, the 3DPP-1 possessed a significantly highest compressive force 273 ± 20.8 Kgf (* p < 0.05). In vitro experiment results also revealed good cell viability and cell attachment behavior in all 3DPP scaffolds. Furthermore, the 3DPP-3 scaffold showed a significantly higher percentage of bone formation volume than the 3DPP-1 scaffold at week 8 (* p < 0.05) and week 12 (* p < 0.05). Hence, the 3DPP scaffold composed of ß-TCP and F-127 is a promising candidate to promote bone tissue ingrowth into the porous scaffold with decent biocompatibility. This scaffold particularly fabricated with a pore size of around 350 µm (i.e., 3DPP-3 scaffold) can provide proper loading support and promote bone regeneration in bone defects when applied in dental and orthopedic fields.

An Innovative Customized Biomimetic Hydrogel for Drug Screening Application Potential: Biocompatibility and Cell Invasion Ability.

The ability of Pluronic F127 (PF127) conjugated with tetrapeptide Gly-Arg-Gly-Asp (GRGD) as a sequence of Arg-Gly-Asp (RGD) peptide to form the investigated potential hydrogel (hereafter referred to as 3DG bioformer (3BE)) to produce spheroid, biocompatibility, and cell invasion ability, was assessed in this study. The fibroblast cell line (NIH 3T3), osteoblast cell line (MG-63), and human breast cancer cell line (MCF-7) were cultured in the 3BE hydrogel and commercial product (Matrigel) for comparison. The morphology of spheroid formation was evaluated via optical microscopy. The cell viability was observed through cell counting Kit-8 assay, and cell invasion was investigated via Boyden chamber assay. Analytical results indicated that 3BE exhibited lower spheroid formation than Matrigel. However, the 3BE appeared biocompatible to NIH 3T3, MG-63, and MCF-7 cells. Moreover, cell invasion ability and cell survival rate after invasion through the 3BE was displayed to be comparable to Matrigel. Thus, these findings demonstrate that the 3BE hydrogel has a great potential as an alternative to a three-dimensional cell culture for drug screening applications.

Biomimetic Ceramic Composite: Characterization, Cell Response, and In Vivo Biocompatibility.

The present study aimed to synthesize biphasic calcium phosphate ceramics (CaPs) composed of ß-tricalcium phosphate (ß-TCP) and hydroxyapatite (HAp) from the propagated Scleractinian coral and dicalcium phosphate anhydrous using a solid-state reaction followed by heat treatment at a temperature of 1100 °C for 1 h to 7 days. The as-prepared coral and coral-derived biphasic CaPs samples were characterized through scanning electron microscopy, X-ray diffractometry, Fourier transform infrared spectroscopy, and Raman spectroscopy. The cell response of the biphasic CaPs was evaluated by in vitro cytotoxicity assessment using mouse fibroblast (L929) cells. The bilateral femoral defect rabbit model was used to assess the early local reaction of the coral-derived biphasic CaPs bone graft on tissue. The results confirmed that the co-existence of ß-TCP and HAp was formed at 1100 °C for 1 h. The ratio of HA/ß-TCP increased as the heat-treatment time increased. The coral-derived biphasic CaPs comprising 61% HAp and 39% ß-TCP (defined as HT-3) were not cytotoxic. Furthermore, no significant differences in local tissue reaction were observed between the HT-3 sample and autogenous bone. Therefore, the synthesized coral-derived biphasic CaPs is a candidate for bone grafting due to its good biocompatibility.

Toxic mechanisms of Roth801, Canals, microparticles and nanoparticles of ZnO on MG-63 osteoblasts.

ZnO eugenol-based materials are widely used for restoration of caries cavity, apical retrograde filling and root canal sealer. Their effects on apical bone healing await investigation. The toxic mechanisms of ZnO particles and nanoparticles to MG-63 osteoblastic cells were studied. We found the different morphology and size of various particles as observed by scanning electron microscope. Particles of Canals and Roth801 were larger than ZnO-205532 microparticles and ZnO-677450 nanoparticles. Four ZnO particles showed cytotoxicity (>25 µg/ml) as analyzed by MTT. Transmission electron microscope found intracellular vacuoles with particle content. Exposure to ZnO particles induced ROS production and cell cycle arrest as studied by DCF and propidium iodide flow cytometry. ZnO particles activated ATM, ATR, Chk1, Chk2, γ-H2AX, ERK and p38 phosphorylation as detected by immunofluorescent staining and western blotting. The protein expression of cdc2, cyclin B1 and cdc25C were decreased, whereas GADD45α and hemeoxygenase-1 (HO-1) were stimulated. ZnO particles' cytotoxicity to MG63 cells was prevented by N-acetylcysteine (NAC), but not CGK733, AZD7762, U0126 and SB203580. ZnO showed little effect on IL-8 and sICAM-1 secretion. These results indicated that ZnO particles are toxic to osteoblasts. ZnO particles' toxicity were related to ROS, and DNA damage responses, checkpoint kinases, cell cycle arrest, ERK and p38 signaling, but not IL-8 and ICAM-1. These results were useful for materials' development and promote apical healing. Dentists should avoid of extruding ZnO-based sealers excessively over root apex and prevent residual ZnO-based retrograde filling materials in apical area during endodontic practice.

Regulation of the regenerative activity of dental pulp stem cells from exfoliated deciduous teeth (SHED) of children by TGF-ß1 is associated with ALK5/Smad2, TAK1, p38 and MEK/ERK signaling.

Transforming growth factor-ß1 (TGF-ß1) regulates wound healing/regeneration and aging processes. Dental pulp stem cells from human exfoliated deciduous teeth (SHED) are cell sources for treatment of age-related disorders. We studied the effect of TGF-ß1 on SHED and related signaling. SHED were treated with TGF-ß1 with/without pretreatment/co-incubation by SB431542, U0126, 5Z-7-oxozeaenol or SB203580. Sircol collagen assay, 3-(4,5-Dimethylthiazol-2-yl)-2,5- diphenyl tetrazolium bromide (MTT) assay, alkaline phosphatase (ALP) assay, RT-PCR, western blotting and PathScan phospho-ELISA were used to measure the effects. We found that SHED expressed ALK1, ALK3, ALK5, TGF-RII, betaglycan and endoglin mRNA. TGF-ß1 stimulated p-Smad2, p-TAK1, p-ERK, p-p38 and cyclooxygenase-2 (COX-2) protein expression. It enhanced proliferation and collagen content of SHED that were attenuated by SB431542, 5Z-7-oxozeaenol and SB203580, but not U0126. TGF-ß1 (0.5-1 ng/ml) stimulated ALP of SHED, whereas 5-10 ng/ml TGF-ß1 suppressed ALP. SB431542 reversed the effects of TGF-ß1. However, 5Z-7-oxozeaenol, SB203580 and U0126 only reversed the stimulatory effect of TGF-ß1 on ALP. Four inhibitors attenuated TGF-ß1-induced COX-2 expression. TGF-ß1-stimulated TIMP-1 and N-cadherin was inhibited by SB431542 and 5Z-7-oxozeaenol. These results indicate that TGF-ß1 affects SHED by differential regulation of ALK5/Smad2/3, TAK1, p38 and MEK/ERK. TGF-ß1 and SHED could potentially be used for tissue engineering/regeneration and treatment of age-related diseases.

Application of a Promising Bone Graft Substitute in Bone Tissue Regeneration: Characterization, Biocompatibility, and In Vivo Animal Study.

The purpose of the present study was to investigate the effect of local hydroxyapatite (HA) combined with extracted sea cucumber (Stichopus hermanni) collagen as a promising bone graft substitute on bone remodeling. Fourier-transform infrared spectroscopy, X-ray diffractometry, transmission electron microscopy, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and Sprague-Dawley rat model were used to characterize the microstructure, in vitro cytotoxicity, and in vivo bone-healing properties of the investigated biocomposite material. Analytical results found that the hydrothermal reaction-synthesized local HA had a hexagonal close-packed structure. The addition of extracted S. hermanni collagen did not influence the microstructure and functional groups of the local HA. Moreover, the MTT assay indicated that the investigated biocomposite material possessed a good in vitro biocompatibility. The in vivo animal study also revealed that the investigated biocomposite material exhibited the highest number of osteoblasts after 14 days of healing. Therefore, the results demonstrate that the local HA combined with extracted S. hermanni collagen could potentially enhance osteoblast formation in promoting bone healing and regeneration.

Development of silver-containing austenite antibacterial stainless steels for biomedical applications part I: microstructure characteristics, mechanical properties and antibacterial mechanisms.

The as-quenched (AQ) microstructure of the Ag-containing alloys was found to be essentially a mixture of austenite (γ) and Ag phases. The Ag phase precipitates had a face-centered-cubic structure and lattice parameter a = 4.09 Å. When the alloy contained Ag ≥0.2 wt%, the mechanical properties were slightly enhanced because of the precipitate strengthening by the Ag phase precipitates. Moreover, the Ag-containing alloys exhibited ductile fracture after tensile testing. The results of an antibacterial test revealed that the Ag phase precipitates play a key role in the antibacterial mechanism of Ag-containing alloys: Ag(+) ions released from the Ag phase precipitates can kill bacteria. It is suggested that as AISI 316L alloy has an Ag content ≥0.2 wt%, it will have excellent antibacterial properties against both Staphylococcus aureus and Escherichia coli, with an antibacterial rate of nearly 100%.

Cytotoxicity of conventional and modified glass ionomer cements.

Various glass ionomer cements (GICs) and resin-modified GICs are widely used as tooth-colored restorative materials. However, their potential effects on pulp tissues are not fully understood. In this study, the authors compared the toxicity of nine types of GICs on cultured human dental pulp cells. Exposure of pulp cells to GICs for five days led to differential growth inhibition as analyzed by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Exposure of pulp cells to ProTec CEM, Fuji II LC, Compoglass and GC Lining cement for five days decreased the cell numbers to 11%, 12%, 19% and 25%, respectively, of the control. Exposure of pulp cells to Fuji IX, GIC FX and Fuji II SC also decreased cell numbers by 62%, 33% and 24%, respectively. By contrast, Hy-Bond and Fuji I showed only mild suppression on the growth of pulp cells, with 12% and 16% decreased cell numbers. Morphologically, marked retraction and rounding of pulp cells were noted following exposure to GC Lining cement; in addition, cell surface blebbing was noted following exposure to Compoglass, Fuji II LC and ProTec CEM. Exposure of the pulp cells to Fuji II SC and Fuji IX, however, led to decreases in the cell density, with no obvious morphological changes. These results indicate that resin-modified GICs, such as Compoglass, Fuji II LC, ProTec CEM and GC Lining cements, are more toxic to pulp cells than conventional GICs. It is not recommended that resin-modified GICs be directly applied onto dental pulp cells. However, additional in vivo studies are needed to evaluate the potential toxicities of these resin-modified GICs during clinical operative procedures.