Dual-crosslinked network of polyacrylamide-carboxymethylcellulose hydrogel promotes osteogenic differentiation in vitro

In our previous study, we successfully designed a dual-crosslinked network hydrogel by introducing the monomers acrylamide (AM), carboxymethylcellulose (CMC), zeolitic imidazolate framework-8 (ZIF-8), and alendronate (Aln). With the simultaneous presentation of physical and chemical crosslinks, the fabricated hydrogel with 10 % concentration of Aln@ZIF-8 (PAM-CMC-10%Aln@ZIF-8) exhibited excellent mechanical characteristics, high Aln loading efficiency (63.83 %), and a slow release period (6 d). These results demonstrate that PAM-CMC-10%Aln@ZIF-8 is a potential carrier for delaying Aln. In this study, we mainly focused on the biocompatibility and osteogenic ability of PAM-CMC-10%Aln@ZIF-8 in vitro, which is a continuation of our previous work. First, this study investigated the biocompatibility of dual-crosslinked hydrogels using calcein-AM/Propidium Iodide and cell counting kit-8. The morphology of rat bone mesenchymal stem cells was assessed using FITC-phalloidin/DAPI and vinculin immunostaining. Finally, osteogenic induction ability in vitro was assessed via alkaline phosphatase expression and alizarin red S staining, which was also confirmed using real-time PCR at the gene level and immunofluorescence at the protein level. The results indicated that the introduction of Aln enabled a dual-crosslinked hydrogel with superior biocompatibility and outstanding osteogenic differentiation ability in vitro, providing a solid foundation for subsequent animal experiments in vivo.