| Objective:In order to solve the problems of complicated preparation methods and poor antibacterial properties of new wound dressings,an injectable hydrogel with good antibacterial and anti-inflammatory properties based on the good antibacterial and anti-inflammatory properties of Lactoferrin（LF）and the shea-thinning properties of Lithium magnesium silicate hydrogel（LMSH）was prepared by a simple method to improve wound healing and to provide new ideas for the design and development of wound dressings.Methods:（1）Preparation of lactoferrin/lithium magnesium silicate hydrogel（LF/LMSH）:LF（20 wt.%）and LMSH（0.3 wt.%）were mixed in different volumes（1:1,2:1,1:2）and named LF/LMSH（1:1）,LF/LMSH（2:1）and LF/LMSH（1:2）,respectively.The hydrogels are formed by the mutual attraction of positive and negative charges between the two.The morphology and structure were characterized using scanning electron microscopy（SEM）and Fourier infrared spectroscopy（FT-IR）,and the shear-thinning behavior of LF/LMSH hydrogels was verified using rheological tests.（2）In vitro antibacterial performance and biocompatibility test of LF/LMSH:The antibacterial performance of different groups of LF/LMSH against Staphylococcus aureus and Escherichia coli were investigated by measuring the inhibition rate;the toxicity effect on L929 cells was studied by MTT reaction colorimetric method and Calcein-AM/PI double staining method.（3）LF/LMSH wound healing effect study: untreated rat wounds were used as the control group and LF,LMSH,LF/LMSH（1:1）,LF/LMSH（2:1）,and LF/LMSH（1:2）as experimental groups.Hydrogels were injected on the wound surface and observed and photographed on days 0,4,7,and 14,respectively,while H＆E staining,Masson staining,CD31,and immunofluorescence staining for inflammatory factors（IL-1β,IL-6,and TNF-α）were used to assess wound healing.Results:（1）Injectable hydrogels were successfully prepared and SEM results showed that the LF/LMSH（1:2）group showed a porous network structure on the surface,and FT-IR results further confirmed the successful composite of the two materials.The rheological results show that the LF/LMSH（1:2）group hydrogel has a significant shear-thinning behavior and meets the injectability requirements of the hydrogel.（2）The results of in vivo antibacterial experiments showed that LF/LMSH had good antibacterial properties against S.aureus and E.coli.The results of the MTT assay showed that the survival rate of L929 cells in each group exceeded 90% at 1,2,and 4 days,indicating that the LF/LMSH has good biocompatibility to L929 cells.The results of the live/dead cell staining showed that the cells in the control and experimental groups gradually increased during the culture cycle and no dead cells appeared,further confirming the results of the MTT experiments.（3）Gross observations of wound healing in rats showed that the LF/LMSH（1:1）,LF/LMSH（2:1）,and LF/LMSH（1:2）groups were effective in promoting healing compared to the positive and negative control groups,with the LF/LMSH（1:2）group having the best healing effect with almost complete closure of the wound at 14 days and a healing rate of 99.1%.The healing rate was 99.1%.Microscopic observation of the tissues also showed that the LF/LMSH（1:2）group hydrogel has good anti-inflammatory ability and can effectively promote collagen deposition and angiogenesis,thus accelerating wound healing.Conclusion:The LF/LMSH injectable hydrogels were prepared with good shear-thinning properties,antibacterial properties,anti-inflammatory properties,and biocompatibility.In a rat whole skin wound model,the LF/LMSH（1:2）group hydrogel can effectively promote wound healing with a 14-day healing rate of 99.1% due to its pro-collagen forming ability,pro-angiogenic ability,and good anti-inflammatory effect,and has potential for application as a wound dressing. |
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