K-Carrageenan Stimulates Pre-Osteoblast Proliferation and Osteogenic Differentiation: A Potential Factor for the Promotion of Bone Regeneration?

Current cell-based bone tissue regeneration strategies cannot cover large bone defects. K-carrageenan is a highly hydrophilic and biocompatible seaweed-derived sulfated polysaccharide, that has been proposed as a promising candidate for tissue engineering applications. Whether κ-carrageenan can be used to enhance bone regeneration is still unclear. In this study, we aimed to investigate whether κ-carrageenan has osteogenic potential by testing its effect on pre-osteoblast proliferation and osteogenic differentiation in vitro. Treatment with κ-carrageenan (0.5 and 2 mg/mL) increased both MC3T3-E1 pre-osteoblast adhesion and spreading at 1 h. K-carrageenan (0.125–2 mg/mL) dose-dependently increased pre-osteoblast proliferation and metabolic activity, with a maximum effect at 2 mg/mL at day three. K-carrageenan (0.5 and 2 mg/mL) increased osteogenic differentiation, as shown by enhanced alkaline phosphatase activity (1.8-fold increase at 2 mg/mL) at day four, and matrix mineralization (6.2-fold increase at 2 mg/mL) at day 21. K-carrageenan enhanced osteogenic gene expression (Opn, Dmp1, and Mepe) at day 14 and 21. In conclusion, κ-carrageenan promoted MC3T3-E1 pre-osteoblast adhesion and spreading, metabolic activity, proliferation, and osteogenic differentiation, suggesting that κ-carrageenan is a potential osteogenic inductive factor for clinical application to enhance bone regeneration.

Effect of Precursor Deficiency Induced Ca/P Ratio on Antibacterial and Osteoblast Adhesion Properties of Ag-Incorporated Hydroxyapatite: Reducing Ag Toxicity

Ag-containing hydroxyapatite (HA) can reduce risks associated with bacterial infections which may eventually require additional surgical operations to retrieve a failed implant. The biological properties of HA in such applications are strongly affected by its composition in terms of dopants as well as Ca/P stoichiometry, which can be easily controlled by altering processing parameters, such as precursor concentrations. The objective of this in vitro study was to understand the effect of variations in HA precursor solutions on antibacterial properties against Escherichia coli (E. coli) and for promoting osteoblast (bone-forming cell) adhesion on Ag incorporated HA (AgHA) which has not yet been investigated. For this, two groups of AgHAs were synthesized via a precipitation method by adjusting precursor reactants with a stoichiometric value of 1.67, being either (Ca + Ag)/P (Ca-deficient) or Ca/(P + Ag) (P-deficient), and were characterized by XRD, FTIR, and SEM-EDS. Results showed that Ag+ incorporated into the Ca2+ sites was associated with a corresponding OH− vacancy. Additional incorporation of CO32− into PO43− sites occurred specifically for the P-deficient AgHAs. While antibacterial properties increased, osteoblast adhesion decreased with increasing Ag content for the Ca-deficient AgHAs, as anticipated. In contrast, significant antibacterial properties with good osteoblast behavior were observed on the P-deficient AgHAs even with a lower Ag content, owing to carbonated HA. Thus, this showed that by synthesizing AgHA using P-deficient precursors with carbonate substitution, one can keep the antibacterial properties of Ag in HA while reducing its toxic effect on osteoblasts.

Incorporation of heparin/BMP2 complex on GOCS-modified magnesium alloy to synergistically improve corrosion resistance, anticoagulation, and osteogenesis

The in vivo fast degradation and poor biocompatibility are two major challenges of the magnesium alloys in the field of artificial bone materials. In this study, graphene oxide (GO) was first functionalized by chitosan (GOCS) and then immobilized on the magnesium alloy surface, finally the complex of heparin and bone morphogenetic protein 2 was incorporated on the modified surface to synergistically improve the corrosion resistance, anticoagulation, and osteogenesis. Apart from an excellent hydrophilicity after the surface modification, a sustained heparin and BMP2 release over 14 days was achieved. The corrosion resistance of the modified magnesium alloy was significantly better than that of the control according to the results of electrochemical tests. Moreover, the corrosion rate was also significantly reduced in contrast to the control. The modified magnesium alloy not only had excellent anticoagulation, but also can significantly promote osteoblast adhesion and proliferation, upregulate the expression of alkaline phosphatase and osteocalcin, and enhance mineralization. Therefore, the method of the present study can be used to simultaneously improve the corrosion resistance and biocompatibility of the magnesium alloys targeted for the orthopedic applications.

Nano-hydroxyapatite coating synthesized on quasi-fibrillar superstructures of collagen hydrolysate leads to superior osteoblast proliferation when compared to nano-hydroxyapatite synthesized on collagen fibrils

This study had a two-fold objective: To utilize collagen hydrolysate for synthesizing a nanoscale Hydroxyapatite (HA) coating that would act as a superior osteoblast adhesion/proliferation agent compared to collagen-derived HA (C/HA) and to comprehend the significant role played by structural constraints on HA nucleation. Collagen was extracted from pacu skin with a high yield of 65.3% (w/w of tissue). It was digested by collagenase and the hydrolysate (CH) was purified with a high yield of 0.68g/g of collagen. The CH peptides had a mass of 6kDa, a predominant PP-II conformation and formed self-assembling hierarchical structures at physiological pH with dimensions of 842.2±229nm. The HA synthesized on CH (CH/HA) displayed higher yield when compared to C/HA. Structural analysis of CH/HA revealed that the PP-II peptides coiled to form mimic-helical moieties with reduced intermolecular packing distance of 0.9nm. The mimic helices cross-linked to form a vast quasi-fibrillar network that was comparatively smaller than collagen fibrils but exhibited enhanced stability and greater dynamicity. CH/HA displayed intense calcium-carboxyl interactions, sharper diffraction planes, smaller size of 48±6.2nm and a Ca/P ratio closer to 1.69 when compared to C/HA along with displaying serrated edge blooming crystals. Because of the small size, the CH/HA nanocrystals displayed significantly better osteoblast adhesion than C/HA and reduced the doubling time of cells. Overall, the results indicated that CH based nanocomposites displayed suitable morphological characteristics and cellular response for potential application as implant and bone graft coating material.Graphical abstract

Antibacterial Behavior of Chitosan-Sodium Hyaluronate-PEGDE Crosslinked Films

Chitosan is a natural polymer that can sustain not only osteoblast adhesion and proliferation for bone regeneration purposes, but it is also claimed to exhibit antibacterial properties towards several Gram-positive and Gram-negative bacteria. In this study, chitosan was modified with sodium hyaluronate, crosslinked with polyethylene glycol diglycidyl ether (PEGDE) and both osteoblast cytotoxicity and antibacterial behavior studied. The presence of sodium hyaluronate and PEGDE on chitosan was detected by FTIR, XRD, and XPS. Chitosan (CHT) films with sodium hyaluronate crosslinked with PEGDE showed a better thermal stability than pristine hyaluronate. In addition, osteoblast cytocompatibility improved in films containing sodium hyaluronate. However, none of the films exhibit antimicrobial activity against Escherichia coli, Enterococcus faecalis, and Staphylococcus aureus while exhibiting low to mild activity against Salmonella typhimurion.

Biocompatibility and Cellular Behavior of TiNbTa Alloy with Adapted Rigidity for the Replacement of Bone Tissue

In this work, the mechanical and bio-functional behavior of a TiNbTa alloy is evaluated as a potential prosthetic biomaterial used for cortical bone replacement. The results are compared with the reference Ti c.p. used as biomaterials for bone-replacement implants. The estimated mechanical behavior for TiNbTa foams was also compared with the experimental Ti c.p. foams fabricated by the authors in previous studies. A TiNbTa alloy with a 20–30% porosity could be a candidate for the replacement of cortical bone, while levels of 80% would allow the manufacture of implants for the replacement of trabecular bone tissue. Regarding biocompatibility, in vitro TiNbTa, cellular responses (osteoblast adhesion and proliferation) were compared with cell growth in Ti c.p. samples. Cell adhesion (presence of filopodia) and propagation were promoted. The TiNbTa samples had a bioactive response similar to that of Ti c.p. However, TiNbTa samples show a better balance of bio-functional behavior (promoting osseointegration) and biomechanical behavior (solving the stress-shielding phenomenon and guaranteeing mechanical resistance).

Novel Coatings to Minimize Bacterial Adhesion and Promote Osteoblast Activity for Titanium Implants

Titanium nitride (TiN) and silicon carbide (SiC) adhesion properties to biofilm and the proliferation of human osteoblasts were studied. Quaternized titanium nitride (QTiN) was produced by converting the surface nitrogen on TiN to a positive charge through a quaternization process to further improve the antibacterial efficiency. The SiC required a nitridation within the plasma chamber of the surface layer before quaternization could be carried out to produce quaternized SiC (QSiC). The antimicrobial activity was evaluated on the reference strains of Porphyromonas gingivalis for 4 h by fluorescence microscopy using a live/dead viability kit. All the coatings exhibited a lower biofilm coverage compared to the uncoated samples (Ti—85.2%; TiN—24.22%; QTiN—11.4%; SiC—9.1%; QSiC—9.74%). Scanning Electron Microscope (SEM) images confirmed the reduction in P. gingivalis bacteria on the SiC and TiN-coated groups. After 24 h of osteoblast cultivation on the samples, the cell adhesion was observed on all the coated and uncoated groups. Fluorescence images demonstrated that the osteoblast cells adhered and proliferated on the surfaces. TiN and SiC coatings can inhibit the attachment of Porphyromonas gingivalis and promote osteoblast adhesion on the titanium used for implants. These coatings may possess the ability to prevent the development of peri-implantitis and stimulate osteointegration.

High Nanodiamond Content-PCL Composite for Tissue Engineering Scaffolds

Multifunctional scaffolds are becoming increasingly important in the field of tissue engineering. In this research, a composite material is developed using polycaprolactone (PCL) and detonation nanodiamond (ND) to take advantage of the unique properties of ND and the biodegradability of PCL polymer. Different ND loading concentrations are investigated, and the physicochemical properties of the composites are characterized. ND-PCL composite films show a higher surface roughness and hydrophilicity than PCL alone, with a slight decrease in tensile strength and a significant increase in degradation. Higher loading of ND also shows a higher osteoblast adhesion than the PCL alone sample. Finally, we show that the ND-PCL composites are successfully extruded to create a 3D scaffold demonstrating their potential as a composite material for tissue regeneration.