Thermo-responsive polymeric hydrogels have received great attention in recent years. The current study aimed to fabricate and characterize injectable chitosan/ laponite (CS-L) hydrogels using a hybrid cross-linking method by genipin and β-glycerophosphate (BGP). Fourier transform infrared analysis confirmed the ionic and covalent interactions between the employed materials in the hydrogels. Scanning electron microscope images showed a decrease in the mean pore size from 129 to 83 μm after the incorporation of 1% laponite into the hydrogels. Energy dispersive X-ray analysis proved the uniform distribution of laponite in the hydrogels. Besides, the gelation time of the chitosan/laponite hydrogels declined in comparison to the chitosan hydrogel owing to the presence of abundant hydroxyl groups in the laponite structure. Rheological investigations revealed 600% improvement in the storage modulus after the incorporation of 1% laponite. The compression test results similarly showed that the elastic modulus and compressive strength of CS-L1% were significantly enhanced in comparison with pristine polymeric hydrogel. Non-toxicity and antibacterial properties of the hydrogels demonstrated 95% cell viability and 99% antibacterial activity, respectively. In conclusion, the obtained results confirmed that the introduced hybrid hydrogels are appropriate candidates for cartilage tissue engineering applications.cartilage tissue engineering, chitosan hydrogel, laponite, smart biomaterials
| INTRODUCTIONPolymer hydrogels, with a cross-linked macromolecular structure, have been effectively utilized in tissue regeneration and drug delivery due to their hydrophilicity, biodegradation properties, releasing biochemical factors and simulating cellular milieus. [1][2][3] Injectable hydrogels are considered appropriate scaffolds for bone and cartilage tissue regeneration because of their ability to provide homogenous cell distribution before injection and in situ forming after injection, which results in the complete filling of irregular defects. These scaffolds provide a minimally invasive implantation procedure. 4 Smart nanohybrid hydrogels could endure sol-gel transition when exposed to external stimuli, such as temperature, pH, electrical or ionic strength alteration, leading to in-situ gelation of the injected sol inside the body. 5 Several chemical and physical cross-linking procedures such as the Michael reaction, "Click" reaction, Schiff base reaction, enzymatic and UV reaction have been employed in hydrogels. [6][7][8] Chemical reactions between a cross-linker and a polymer lead to a stable network, while physical interactions such as Van der Waals and hydrogen bonds result in a weaker network. In thermoresponsive hydrogels, the sol-gel transition occurs through temperature alteration. 9