Alginate/gelatin (Alg-Gel) hydrogels have been used experimentally, associated with mesenchymal stromal/stem cells (MSCs), to guide bone tissue formation. One of the main challenges for clinical application is optimizing Alg-Gel stiffness to guide osteogenesis. In this study, we investigated how Alg-Gel stiffness could modulate the dental pulp stem cell (DPSC) attachment, morphology, proliferation, and osteogenic differentiation, identifying the optimal conditions to uncouple osteogenesis from the other cell behaviors. An array of Alg-Gel hydrogels was prepared by casting different percentages of alginate and gelatin cross-linked with 2% CaCl₂. We have selected two hydrogels: one with a stiffness of 11 ± 1 kPa, referred to as “low-stiffness hydrogel”, formed by 2% alginate and 8% gelatin, and the other with a stiffness of 55 ± 3 kPa, referred to as “high-stiffness hydrogel”, formed by 8% alginate and 12% gelatin. Hydrogel analyses showed that the average swelling rates were 20 ± 3% for the low-stiffness hydrogels and 35 ± 2% for the high-stiffness hydrogels. The degradation percentage was 47 ± 5% and 18 ± 2% for the low- and high-stiffness hydrogels, respectively. Both hydrogel types showed homogeneous surface shape and protein (Alg-Gel) interaction with CaCl₂ as assessed by physicochemical characterization. Cell culture showed good adhesion of the DPSCs to the hydrogels and proliferation. Furthermore, better osteogenic activity, determined by ALP activity and ARS staining, was obtained with high-stiffness hydrogels (8% alginate and 12% gelatin). In summary, this study confirms the possibility of characterizing and optimizing the stiffness of Alg-Gel gel to guide osteogenesis in vitro without altering the other cellular properties of DPSCs.

A novel, green route for pre-exfoliation of graphite based on a biodegradable polymer and high-power ultrasound is presented. Candelilla wax (CW), derived from the leaves of the candelilla plant, has been used for the first time as a natural non aqueous medium to induce the pre-exfoliation of expanded graphite (EG) under ultrasonic irradiation in an economical way. The proposed method uses also D-limonene as a natural organic solvent for reducing viscosity and increasing the affinity between the polar groups of EG and candelilla wax, thus improving the intercalation/exfoliation of EG. The quality of dispersion of the nanofiller in the natural wax matrix has been evaluated using multiple techniques. The addition of EG to wax and use of ultrasonic treatment leads to a reduced crystallinity, probably due to restrictions of the molecular movements, improved thermal stability of wax, and to an increased shear thinning exponent, which are all indicative of a high degree of EG dispersion. The ultrasonic dynamic mechanical results suggest a reduction in the cluster size and a better filler dispersion in the wax matrix promoted by polar or chemical reactions between the CW fractions and the graphite stacks, which was observed by XPS analysis. The results were compared to those obtained with paraffin, a synthetic wax, and confirmed the dispersion improvement obtained by using natural wax as a pre-exfoliating medium.

Bone tissue engineering relies on crucial scaffolds for tissue formation and stem cell differentiation. A composite scaffold of alginate‐gelatin effectively supports these processes. This study aims to design a porous alginate‐gelatin hydrogel and assess pore size effects on cell behavior, focusing on morphology, adhesion, and proliferation in distinct osteogenic environments. Hydrogels are prepared using various alginate‐gelatin concentrations: 4% alginate and 6% gelatin (4A6G) or 3% alginate and 5% gelatin (3A5G), cross‐linked with 2% CaCl2. Pore size optimization employs simple freezing and thawing cycles. Scanning electron microscopy reveals varying pore sizes: 340 µm ± 30 µm for 4A6G and 635 µm ± 25 µm for 3A5G. Stiffness measurements indicate significant differences: ≈26.3 kPa ± 0.6 KPa for 4A6G and 21.6 kPa ± 0.2 KPa for 3A5G. Cell interaction studies demonstrate higher adhesion and proliferation rates in larger‐pored hydrogels. Evaluation of bone tissue formation, including RT‐PCR, ALP activity, and ARS staining, reveal superior osteogenic potential in the 3A5G hydrogel compared to 4A6G. In conclusion, the 3A5G hydrogel (3% alginate and 5% gelatin) holds promise for bone tissue regeneration due to its biodegradability and favorable bone‐forming properties.

Alginate-gelatin (Alg-Gel) hydrogels have been used experimentally but not clinically associated with mesenchymal stromal / stem cells (MSCs) to guide bone tissue formation. One of the main challenges for its clinical application is optimizing Alg-Gel stiffness to guide osteogenesis. In this study, we investigated how Alg-Gel stiffness could modulate the dental pulp stem cell (DPSCs) attachment, morphology, proliferation, and osteogenic differentiation, identifying the optimal condition to uncouple osteogenesis from the other cell behaviors. An array of Alg-Gel hydrogels was prepared by casting different percentages of Alg and Gel being crosslinked with 2 % CaCl2. We selected two hydrogels, one with 11± 1 kPa called “low” stiffness and one with 55 ± 3 kPa called “high” stiffness. Hydrogel analyses showed that the average swelling rates were 20 ± 3% for low and 35 ± 2% for high hydrogels. The degradation percentage was 47 ± 5% and 18 ± 2% for low and high hydrogels, respectively. Both hydrogel types showed homogeneous surface shape and pro-tein (Alg-Gel) interaction with CaCl2 as assessed by FTIR-ATR and XPS. Cell culture showed good adhesion of the DPSCs to the hydrogels and proliferation. Furthermore, better osteogenic activity was obtained with high-stiffness hydrogels. In summary, this study confirms the possibility of characterizing and optimizing the stiffness of alginate-gelatin gel to guide osteogenesis in vitro without altering other cellular properties of DPSCs.

Nanoparticles (NP) of 12.7 nm in diameter of the poly(methyl methacrylate (MMA)-co-methacrylic acid (MAA)) copolymer were prepared. ¹³C-NMR results showed a MMA:MAA molar ratio of 0.64:0.36 in the copolymer, which is similar to the poly(MMA-co-MAA) commercially known as the FDA approved Eudragit S100 (0.67:0.33). The NP prepared in this study were loaded at pH 5 with varying amounts (from 0.54 to 6.91%) of doxorubicin (DOX), an antineoplastic drug. ¹H-NMR results indicated the electrostatic interactions between the ionized carboxylic groups of the MAA units in the copolymer and the proton of the glycosidic amine in DOX. Measurements by QLS and TEM indicated that the loading destabilizes the NP, and that for increase stability, they aggregate in a reversible way, forming aggregates with a diameter up to 99.5 nm at a DOX load of 6.91%. The analysis of drug release data at pH 7.4 showed that loaded NP with at least 4.38% DOX release the drug very slowly and follows the Higuchi model; the former suggests that they could remain for long periods in the bloodstream to reach and destroy cancer cells.

Titanium and its alloys are the most widely used metallic materials for bone contact implants. However, despite advances in implant technology, these alloys are still susceptible to post-operative clinical complications such as inflammation, which is often joined by infections and biofilm formation. A number of coatings were studied to overcome the drawbacks of these complications, but the controlled release of bioactive molecules over the first few days and the adhesion of the coating to the substrate remain recognized challenges. Carbon dots and the antibacterial potential of chiral carbon dots (CCDs) were recently reported, and their chirality was identified as a major contribution to the bactericidal effect. This study aimed to achieve a stimuli-responsive medium-term controlled release for up to one month. Two types of chiral carbon dots (CCDs) with distinct functional groups were incorporated into a stable and adherent biodegradable polymer coating, i.e., poly(lactic-co-glycolic acid) (PLGA). To enhance the coating adhesion, the titanium alloy surfaces were pre-treated and activated. The wettability, morphology, and surface composition of the coatings were characterized by contact angle, profilometry, SEM, and XPS, respectively. Coating degradation, adhesion, and CCDs release were studied at physiological pH (7.4) and at an acidic pH characteristic of an inflammatory site (pH 3.0) for up to one month. Their biological performances and blood compatibility were assessed as well. Degradation studies conducted over 28 days revealed a slow mass loss of approximately 10%, with maximum release rates for CCDs-OH and CCDs-NH₂ of 67% and 45% at pH 7.4, respectively. At pH 3.0 an inverse trend was observed with 49% and 59% maximum release after 28 days. Furthermore, the coatings did not exhibit any cytotoxic and hemolytic effects. These findings demonstrate the potential of this approach to providing titanium implants with pH-sensitive controlled release of bioactive CCDs lasting up to one month, which could address key challenges in implant-associated complications.