Berberine@NMOF Encapsulated In Hydrogel With ROS Scavenging Function Promotes Diabetic Foot Wound Healing

Objective:Diabetic foot wound is a common chronic refractory wound with limited treatment options.Reducing excess reactive oxygen speciesand inflammatory response on the wound bed is a key strategy for diabetic wound healing.Overexpression of MMP-9 is one of the important reasons for hindering wound healing.In this study,we designed and synthesized a zinc-based metal-organic framework loaded with natural small molecule berberine to form BR@Zn-BTB nanoparticles with the ability of wound pH response.Then hydrogel with ROS scavenging function was used to wrap the nanoparticles.A new hydrogel composite system(BR@Zn-BTB/Gel)was prepared.method:1.An in situ "one-pot" method was used to synthesize BR@Zn-BTB,which utilizes self-assembly of ZnCl2 pairs with H3BTB ligands to load BR into the framework voids.BR@Zn-BTB with high berberine loading was prepared,and its morphology,crystal structure,particle size,drug loading rate,and BR release rate were characterized.The BR@Zn-BTB-encapsulated hydrogels were prepared,and the hydrogel morphology,pores,Zn2+ and BR release rates were identified.2.The degradation of BR@Zn-BTB/Gel in PBS containing H2O2 was observed;the inhibitory effect of the hydrogel on Escherichia coli,Staphylococcus epidermidis and Staphylococcus aureus was detected by the zone of inhibition method.3.Appropriate dosing concentrations were screened using CCK-8.The cytocompatibility of BR@Zn-BTB/Gel was verified by cell death staining.A highglucose fibroblasts and keratinocytes model was constructed,the effect of BR@ZnBTB/Gel on cell migration was verified by scratch assay,and the expressions of MMP9 and TIMP-1 related genes in keratinocytes cells under high-glucose environment were detected by qRT-PCR.Fibroblasts were co-cultured with H2O2,and ROS fluorescence detection kit was used to verify the scavenging effect of BR@Zn-BTB/Ge on H2O2.4.Macrophages were stimulated with LPS to construct an inflammatory macrophage model.After co-culture with the hydrogel composites,the expression of inflammatory factors IL-1β,IL-6 and TNF-α were detected by qRT-PCR.5.The in vitro results showed that the hydrogel composite system could be antibacterial,inhibit inflammation,scavenge ROS,release Zn2+ and BR slowly,and inhibit the overexpression of MMP-9 in keratinocytes under high glucose environment.The in vivo results show that the hydrogel composite system can promote collagen deposition in diabetic wounds,eliminate inflammation,inhibit the expression of MMP9 in wounds,and ultimately promote wound healing in diabetic mice.Results:The in vitro results showed that the hydrogel composite system could be antibacterial,inhibit inflammation,scavenge ROS,release Zn2+ and BR slowly,and inhibit the overexpression of MMP-9 in keratinocytes under high glucose environment.The in vivo results show that the hydrogel composite system can promote collagen deposition in diabetic wounds,inhibit inflammation and the expression of MMP-9 in wounds,and ultimately promote wound healing in diabetic mice.Conclusion:The results will elucidate the biological effects of the BR@Zn-BTB composite hydrogel and obtain a novel metal-organic orthopaedic wound dressing.Finally,it can provide a new method for the treatment of diabetic wounds.