Photo-crosslinkable hydrogel for guided periodontal tissue regeneration

Periodontitis is an inflammatory disease resulting from the presence of oral bacteria biofilm in periodontal tissue, which destroys the tooth-supporting attachment apparatus. Untreated inflammation can spread to the gum tissue and lead, ultimately, to the loosening of the supporting tooth bone, with the risk that the tooth eventually falls.
Guided Tissue Regeneration is a technique based on the application of a barrier membrane designed to prevent colonization of the wound space by epithelial cells from soft tissues. Indeed, these cells, characterized by a faster migration and proliferation rate compared to bone and periodontal ligament cells, could interfere with the regeneration process.
In previously work Struillou et al. demonstrated the benefit effect of silated hydroxypropylmethylcellulose (Si HMPC)-based hydrogel can act as an efficient physical barrier in periodontal defect. Typically, this material is able to form a 3D network through the condensation of silanoate groups at physiological pH. However, a decrease of gelation time is necessary to assure the stability in peripheral part of the wound.
In this project, we developed an injectable photo-crosslinkable membrane based on methacrylated carboxymethyl chitosan (CMCs) and Si HPMC that can be applied as a viscous solution and cured in situ in presence of a photoinitiator system made of riboflavin and triethanolamine. A visible light lamp (λ 420-480 nm), already used in dentistry, was preferred over a UV lamp. The addition of methacrylated polymer increase the stability of the material and increase the mass loss, in order to improve the bioresorption of the membrane.
The chemical grafting of methacrylated carboxymethyl chitosan was characterized by 1H NMR and Infrared Spectroscopy. The gel point of the solution was determined by rheology and remained compatible with a clinical application.
Moreover, the biocompatibility of this biomaterials was tested using murine cells using two assay: Neutral Red assay and MTT Cell Proliferation Assay. The in vitro tests validate the chemical synthesis in a biological point of view. The irradiation on cells and the direct contact with hydrogel doesn’t have an impact on cells viability.
The capability of this material to act as a physical barrier was also evaluated using human gingival fibroblast. The cells were isolated from human gum explant before being put in contact with the hydrogel. After four days of contact no cells invasion was observed in the hydrogel using confocal microscopy.
These preliminary results are quite promising for the development of novel injectable systems for Guided Periodontal Regeneration. In the future work, in vivo assays will be performed in Periodontal defect in a canine model.