In Vitro Evaluation of Calcium Phosphate Bone Cement Composite Hydrogel Beads of Cross-Linked Gelatin-Alginate with Gentamicin-Impregnated Porous Scaffold.

Calcium phosphate bone cement (CPC) is in the form of a paste, and its special advantage is that it can repair small and complex bone defects. In the case of open wounds, tissue debridement is necessary before tissue repair and the subsequent control of wound infection; therefore, CPC composite hydrogel beads containing antibiotics provide an excellent option to fill bone defects and deliver antibiotics locally for a long period. In this study, CPC was composited with the millimeter-sized spherical beads of cross-linked gelatin-alginate hydrogels at the different ratios of 0 (control), 12.5, 25, and 50 vol.%. The hydrogel was impregnated with gentamicin and characterized before compositing with CPC. The physicochemical properties, gentamicin release, antibacterial activity, biocompatibility, and mineralization of the CPC/hydrogel composites were characterized. The compressive strength of the CPC/hydrogel composites gradually decreased as the hydrogel content increased, and the compressive strength of composites containing gentamicin had the largest decrease. The working time and setting time of each group can be adjusted to 8 and 16 min, respectively, using a hardening solution to make the composite suitable for clinical use. The release of gentamicin before the hydrogel beads was composited with CPC varied greatly with immersion time. However, a stable controlled release effect was obtained in the CPC/gentamicin-impregnated hydrogel composite. The 50 vol.% hydrogel/CPC composite had the best antibacterial effect and no cytotoxicity but had reduced cell mineralization. Therefore, the optimal hydrogel beads content can be 25 vol.% to obtain a CPC/gentamicin-impregnated hydrogel composite with adequate strength, antibacterial activity, and bio-reactivity. This CPC/hydrogel containing gentamicin is expected to be used in clinical surgery in the future to accelerate bone regeneration and prevent prosthesis infection after surgery.

In Vitro Evaluation of a Composite Gelatin-Hyaluronic Acid-Alginate Porous Scaffold with Different Pore Distributions for Cartilage Regeneration.

Although considerable achievements have been made in the field of regenerative medicine, since self-repair is not an advanced ability of articular cartilage, the regeneration of osteochondral defects is still a challenging problem in musculoskeletal diseases. Cartilage regeneration aims to design a scaffold with appropriate pore structure and biological and mechanical properties for the growth of chondrocytes. In this study, porous scaffolds made of gelatin, hyaluronic acid, alginate, and sucrose in different proportions of 2 g (SL2) and 4 g (SL4) were used as porogens in a leaching process. Sucrose with particle size ranges of 88-177 µm (Hµ) and 44-74 µm (SHµ) was added to the colloid, and the individually cross-linked hydrogel scaffolds with controllable pore size for chondrocyte culture were named Hµ-SL2, Hµ-SL4, SHµ-SL2 and SHµ-SL4. The perforation, porosity, mechanical strength, biocompatibility, and proliferation characteristics of the hydrogel scaffold and its influence on chondrocyte differentiation are discussed. Results show that the addition of porogen increases the porosity of the hydrogel scaffold. Conversely, when porogens with the same particle size are added, the pore size decreases as the amount of porogen increases. The perforation effect of the hydrogel scaffolds formed by the porogen is better at 88-177 µm compared with that at 44-74 µm. Cytotoxicity analysis showed that all the prepared hydrogel scaffolds were non-cytotoxic, indicating that no cross-linking agent residues that could cause cytotoxicity were found. In the proliferation and differentiation of the chondrocytes, the SHµ-SL4 hydrogel scaffold with the highest porosity and strength did not achieve the best performance. However, due to the compromise between perforation pores, pore sizes, and strength, as well as considering cell proliferation and differentiation, Hµ-SL4 scaffold provided a more suitable environment for the chondrocytes than other groups; therefore, it can provide the best chondrocyte growth environment for this study. The development of hydrogels with customized pore properties for defective cartilage is expected to meet the requirements of the ultimate clinical application.

Effect of pH on the In Vitro Biocompatibility of Surfactant-Assisted Synthesis and Hydrothermal Precipitation of Rod-Shaped Nano-Hydroxyapatite.

Given their wide range of biomedical applications, hydroxyapatite (HA) nanoparticles are an attractive material widely used in many fields. Therefore, a simple, inexpensive, and stable process for the synthesis of HA nanoparticles is necessary to meet current needs. Herein, we studied HA synthesis assisted by four surfactants, namely cation, anion, non-ionic, and zwitterion templates, to verify the synthesis phase, aspect ratio, morphology, and biocompatibility under different environments (i.e., pH 4 and 9) before and after calcination. Results showed that before calcination, the surfactant-free groups could not produce HA but showed an abundant dicalcium phosphate anhydrous (DCPA) phase at pH 4. Except for the anionic group containing a small amount of DCPA, all surfactant-assistant groups presented single-phase HA in acidic and alkaline environments. The diameter of HA synthesized at pH 4 was significantly larger than that of HA synthesized at pH 9, and the effect of aspect ratio changes after calcination was more significant than that before calcination. The uncalcined rod-shaped HA synthesized with a non-ionic template at pH 4 demonstrated excellent cell viability, whereas anionic, cationic, and non-ionic surfactants exhibited biocompatibility only after calcination. At pH 9, non-ionic and uncalcined zwitterion-assisted rod-shaped HA showed excellent biocompatibility. In conclusion, the uncalcined HA rod-shaped nanoparticles synthesized from the non-ionic template at pH 4 and 9 and the zwitterion template at pH 9, as well as all surfactant-assisted HA after calcination, had no cytotoxicity. These tailor-made non-toxic HA types can meet the different requirements of apatite composite materials in biomedical applications.

Effects of Hinokitiol and Dicalcium Phosphate on the Osteoconduction and Antibacterial Activity of Gelatin-Hyaluronic Acid Crosslinked Hydrogel Membrane In Vitro.

Many hydrogel-based crosslinking membranes have been designed and tailored to meet the needs of different applications. The aim of this research is to design a bifunctional hydrogel membrane with antibacterial and osteoconducting properties to guide different tissues. The membrane uses gelatin and hyaluronic acid as the main structure, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride as the crosslinker, hinokitiol as the antibacterial agent, and dicalcium phosphate anhydrous (DCPA) micron particles for osteoconduction. Results show that the hydrogel membrane with added DCPA and impregnated hinokitiol has a fixation index higher than 88%. When only a small amount of DCPA is added, the tensile strength does not decrease significantly. The tensile strength decreases considerably when a large amount of modified DCPA is added. The stress-strain curve shows that the presence of a large amount of hinokitiol in hydrogel membranes results in considerably improved deformation and toughness properties. Each group impregnated with hinokitiol exhibits obvious antibacterial capabilities. Furthermore, the addition of DCPA and impregnation with hinokitiol does not exert cytotoxicity on cells in vitro, indicating that the designed amount of DCPA and hinokitiol in this study is appropriate. After a 14-day cell culture, the hydrogel membrane still maintains a good shape because the cells adhere and proliferate well, thus delaying degradation. In addition, the hydrogel containing a small amount of DCPA has the best cell mineralization effect. The developed hydrogel has a certain degree of flexibility, degradability, and bifunctionality and is superficial. It can be used in guided tissue regeneration in clinical surgery.

Strength and Biocompatibility of Heparin-Based Calcium Phosphate Cement Grafted with Ferulic Acid.

The biomimetic synthesis of carbonated apatites by biomolecule-based templates is a promising way for broadening apatite applications in bone tissue regeneration. In this work, heparin was used as an organic template to prepare uniform carbonate-based apatite nanorods (CHA) and graft ferulic acid (F-CHA) for enhanced bone mineralization. Next, by combining calcium phosphate cement (CPC) with different F-CHA/CPC ratios, a new type of injectable bone cement combined with F-CHA bioactive apatite was developed (CPC + F-CHA). The physicochemical properties, biocompatibility, and mineralization potential of the CPC + F-CHA composites were determined in vitro. The experimental results confirmed the preparation of highly biocompatible CHA and the compatibility of F-CHA with CPC. Although CPC + F-CHA composites with F-CHA (2.5 wt%, 5 wt%, and 10 wt%) showed a significant reduction in compressive strength (CS), compositing CPC with 10 wt% F-CHA yielded a CS suitable for orthopedic repair (CS still larger than 30 MPa). Spectroscopic and phase analyses revealed that the phase of the hydrothermally synthesized CHA product was not modified by the heparin template. Injection and disintegration tests indicated that the CPC + F-CHA composites have good biocompatibility even at 10 wt% F-CHA. D1 osteoprogenitor cells were cultured with the composites for 7 days in vitro, and the CPC + 10%F-CHA group demonstrated significantly promoted cell mineralization compared with other groups. Given these results, the use of over 10% F-CHA in CPC composites should be avoided if the latter is to be applied to load-bearing areas. A stress-shielding device may also be recommended to stabilize these areas. These newly developed biocompatible CPC + F-CHA have great potential as osteoconductive bone fillers for bone tissue engineering.