Effect And Mechanism Of Berberine Thermosensitive Hydrogel In The Treatment Of Periodontitis

Background:Periodontitis is a chronic inflammatory disease that is the main cause of tooth loss in adults.Due to the anatomical conditions of the teeth such as root bifurcation lesions and the depth of periodontal pockets,the complete elimination of plaque microorganisms is often not possible with basic periodontal therapy alone.Therefore,topical pharmacotherapy has become an important adjunct to the treatment of chronic periodontitis.Drug treatment mainly applies antibiotics and non-steroidal anti-inflammatory drugs(NSAIDs).However,there are some side effects associated with the use of these drugs.In the last two decades,berberine(BBR)has been found to have various pharmacological effects such as hypoglycemic,antitumor,antiarrhythmic,anti-inflammatory,and immunomodulatory effects.However,the efficacy of berberine is greatly limited by its low utilization and non-targeting characteristics.Due to the anatomical irregularity of the periodontal pocket,topical administration is a common form of administration for the treatment of periodontitis.Therefore,the search for safe,cost-effective and efficient drugs and carriers for the treatment of periodontitis is imminent.Objective:The objective of this study was to develop a safe and non-toxic carrier that can effectively release berberine to reduce inflammation in periodontal tissues,and to investigate whether berberine-loaded temperature-sensitive hydrogels can exert anti-inflammatory and osteogenic effects by modulating the phosphatifylinositol-3-kinase/Protein Kinase B(PI3K/AKT)signaling pathway.Methods:1.The temperature-sensitive hydrogel containing berberine was prepared,and the morphology of the hydrogel was observed by scanning electron microscopy(SEM),characterized by Fourier transform infrared spectroscopy(FTIR),and tested for in vitro release using an enzyme marker.2.The cytotoxicity of berberine warming hydrogel on RAW264.7 and MC3T3-E1 cell lines was examined by CCK-8 and live-dead cell staining.Quantitative real-time polymerase chain reaction(q RT-PCR)and western blotting(WB)were performed to detect the in vitro anti-inflammatory and osteogenic effects of the hydrogel in the PI3K/AKT pathway.Results:1.In this experiment,berberine-loaded temperature-sensitive hydrogel was successfully prepared,and the mixed solution could be gelled at 37°C for 3 min.The hydrogel was loose and porous with pore size ranging from 10-130 μm,and the cumulative release of berberine reached89.99% at 21 days.2.CCK-8 assay and live-dead cell staining results indicate that this hydrogel is not biotoxic.3.The qRT-PCR and WB results showed that the m RNA and protein levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were significantly reduced in the berberine-loaded hydrogel group,and the protein phosphorylation levels in the PI3K/AKT pathway were reduced,and LY294002(PI3K inhibitor)could further reduce the m RNA and protein levels of the above-mentioned inflammatory factors,and the protein phosphorylation levels in the PI3K/AKT pathway were further reduced(P<0.05);In the berberine-loaded hydrogel group,alkaline phosphatase(ALP),type I collagen(COL 1),osteocalcin(OCN),and runt-related transcription factor 2(Runx2)m RNA levels were significantly increased,alkaline phosphatase expression was increased,and protein phosphorylation levels in the PI3K/AKT pathway were increased,while LY294002 decreased the m RNA levels of the above osteogenesis-related factors,decreased alkaline phosphatase expression,and protein phosphorylation in the PI3K/AKT pathway levels were reduced(P<0.05).Conclusion:The berberine-loaded temperature-sensitive hydrogel has good morphology,no obvious toxicity,and can achieve slow drug release.The berberine-loaded temperature-sensitive hydrogel may exert anti-inflammatory and osteogenic effects through PI3K/AKT signaling pathway,and may be an effective method for treating periodontitis.