Chitosan Hydrogel Supplemented with Metformin Promotes Neuron–like Cell Differentiation of Gingival Mesenchymal Stem Cells

Abstract

Human gingival mesenchymal stem cells (GMSCs) are derived from migratory neural crest stem cells and have the potential to differentiate into neurons. Metformin can inhibit stem–cell aging and promotes the regeneration and development of neurons. In this study, we investigated the potential of metformin as an enhancer on neuronal differentiation of GMSCs in the growth environment of chitosan hydrogel. The crosslinked chitosan/β–glycerophosphate hydrogel can form a perforated microporous structure that is suitable for cell growth and channels to transport water and macromolecules. GMSCs have powerful osteogenic, adipogenic and chondrogenic abilities in the induction medium supplemented with metformin. After induction in an induction medium supplemented with metformin, Western blot and immunofluorescence results showed that GMSCs differentiated into neuron–like cells with a significantly enhanced expression of neuro–related markers, including Nestin (NES) and β–Tubulin (TUJ1). Proteomics was used to construct protein profiles in neural differentiation, and the results showed that chitosan hydrogels containing metformin promoted the upregulation of neural regeneration–related proteins, including ATP5F1, ATP5J, NADH dehydrogenase (ubiquinone) Fe–S protein 3 (NDUFS3), and Glutamate Dehydrogenase 1 (GLUD1). Our results help to promote the clinical application of stem–cell neural regeneration.

Thermosensitive Injectable Hydrogels for Intra-Articular Delivery of Etanercept for the Treatment of Osteoarthritis

“…Images taken from the freeze-dried hydrogels ( Figure 1 C) revealed that the surface morphology in the CS/PF samples ( Figure 1 C (1)) was compact with the presence of small channels. In contrast, in the CS/PF/BGP hydrogels ( Figure 1 C (2)) an interconnected porous morphology was observed, suggesting that the addition of the gelling agent created micro- and nano-scale pores in the structure [ 35 ]. As previously shown, the gelation time reduced with the addition of BGP because the hydrogen bridge bonds between the CS chains increase and electrostatic interactions arise between BGP phosphate groups and CS amino groups [ 30 , 36 ].…”

“…The peak at 1320 cm −1 was attributed to the C-N stretching in the secondary amide (amide III) [ 43 ]. In the BGP spectrum, two intense signals were detected, with one at 1050 cm −1 related to the asymmetric stretching vibrations of the PO 4 3− group (ν 3 ) and the other at 960 cm −1 related to the symmetrical stretching vibrations (ν 1 ) of the same group [ 35 , 42 , 44 ]. There was another band of lower intensity due to the aliphatic stretching of P-O-C [ 32 ].…”

“…Unlike our observation in the CS/PF hydrogel, we detected a shift towards higher frequencies of the amide II band from 1560 to 1572 cm −1 . This indicates there were interactions between the amino group of the CS and the phosphate group of the BGP due to the electrostatic forces or hydrogen bonds [ 35 , 41 ]. Further evidence of the formation of intermolecular interaction between CS and BGP during the process of CS/PF/GP hydrogel formation was obtained effectively by conducting an analysis of the infrared spectrum of the stretching vibrations of -OH groups and -NH 2 groups.…”

Chitosan nanoparticles for sustained release of metformin and its derived synthetic biopolymer for bone regeneration

“…MHC NPs were not cytotoxic in a CCK-8 assay. Next, we examined whether metformin could enhance the osteogenic differentiation of BMSCs in chitosan hydrogel growth environments ( Cai et al, 2022 ). Alkaline phosphatase activity in BMSCs was significantly increased after 3 days of treatment with the studied MHC NPs.…”

Proteomics and Metabolomics in Biomedicine

“…The correlation between proteome or metabolome changes and drug perturbation was also explored in some of the selected papers. A proteomic snapshot of progressive molecular signatures was carried out in hippocampal mice tissue after pharmacological inhibition of nitric oxide synthase activity [46], whilst a proteomic analysis highlighted the effects of chitosan hydrogel supplemented with metformin on the neural differentiation of stem cells, with the final aim of employing them in the construction of biomimetic scaffolds for disease treatment [47].…”