ABSTRACT
Background@#Antibiotic resistance is a significant public health concern around the globe.Antimicrobial peptides exhibit broad-spectrum and efficient antibacterial activity with an added advantage of low drug resistance. The higher water content and 3D network structure of the hydrogels are beneficial for maintaining antimicrobial peptide activity and help to prevent degradation. The antimicrobial peptide released from hydrogels also hasten the local wound healing by promoting epithelial tissue regeneration and granulation tissue formation. @*Objective@#This study aimed at developing sodium alginate based hydrogel loaded with a novel antimicrobial peptide Chol-37(F34-R) and to investigate the characteristics in vitro and in vivo as an alternative antibacterial wound dressing to treat infectious wounds. @*Methods@#Hydrogels were developed and optimized by varying the concentrations of crosslinkers and subjected to various characterization tests like cross-sectional morphology, swelling index, percent water contents, water retention ratio, drug release and antibacterial activity in vitro, and Pseudomonas aeruginosa infected wound mice model in vivo. @*Results@#The results indicated that the hydrogel C proved superior in terms of cross-sectional morphology having uniformly sized interconnected pores, a good swelling index, with the capacity to retain a higher quantity of water. Furthermore, the optimized hydrogel has been found to exert a significant antimicrobial activity against bacteria and was also found to prevent bacterial infiltration into the wound site due to forming an impermeable barrier between the wound bed and external environment. The optimized hydrogel was found to significantly hasten skin regeneration in animal models when compared to other treatments in addition to strong inhibitory effect on the release of pro-inflammatory cytokines (interleukin-1β and tumor necrosis factor-α). @*Conclusions@#Our results suggest that sodium alginate -based hydrogels loaded with Chol-37(F34-R) hold the potential to be used as an alternative to conventional antibiotics in treating infectious skin wounds.
ABSTRACT
Objective:To research on protective immunity of omph DNA vaccine against avian Pasteurella multocida in mice.Methods: The omph gene fragment amplified by PCR from avian Pasteurella multocida was cloned into pMD18-T.Subsequently it was subcloned into the eukaryotic expression vector pcDNA3.1(+),and the recombinant plasmid pOMPH was obtained.Then the recombinant plasmid was trans fected into SP2/O cells in vitro.The transcription and expression of target gene were analyzed by RT-PCR,Westem blot analysis and indirect immunofluorescence.Three groups of BALB/c mice(n=16) named pOMPH,pCDNA3.1(+) and PBS were intramuscularly vaccinated with the recombinant plasmid,control vector and PBS respectively.The serum antibodies were detected by indirect ELISA.The spleen lymphocyte proliferation (SLP) and secreted IFN-γof spleen were tested by MTT.The mice were challenged with virulent of avian Pasteurella multocida on week 2 post the third immunization,the protection rate were counted.Results: RT-PCR,Western blot analysis and indirect immunofluorescence showed that the omph gene could be,transfected into SP2/0 cells in vitro and expressed the target protein.Indirect ELISA showed that the levels of antibodies in pOMPH group were most significantly higher than in the other groups(P＜0.01).Spleen lymphocyte proliferation by MTT assay indicated that the SI value induced with avian Pasteurella multocida Omps in pOMPH group was higher than those in pCDNA3.1 (+) and PBS groups (P＜0.05).The IFN-γexperiments(Double-antibodies-sandwich-ELISA)showed that the levels of IFN-γ induced with Omps in the group of pOMPH was mostly higher than in the other control groups apperent(P＜0.01 ).The protection rate of pOMPH(70%) was better than in the other groups.Conclusion: The omph DNA vaccine against avian Pasteurella multocida had been constructed successfully.The DNA vaccine could enhance the immunity level and the protective effect of the vaccinated mice.Present study may be useful for the development of avian Pasteurella multocida vaccine.
ABSTRACT
Objective:To research on protective immunity of omph DNA vaccine against avian Pasteurella multocida in mice.Methods:The omph gene fragment amplified by PCR from avian Pasteurella multocida was cloned into pMD18-T.Subsequently it was subcloned into the eukaryotic expression vector pcDNA3.1(+),and the recombinant plasmid pOMPH was obtained.Then the recombinant plasmid was transfected into SP2/0 cells in vitro.The transcription and expression of target gene were analyzed by RT-PCR,Western blot analysis and indirect immunofluorescence.Three groups of BALB/c mice(n=16) named pOMPH,pCDNA3.1(+) and PBS were intramuscularly vaccinated with the recombinant plasmid,control vector and PBS respectively.The serum antibodies were detected by indirect ELISA.The spleen lymphocyte proliferation (SLP) and secreted IFN-? of spleen were tested by MTT.The mice were challenged with virulent of avian Pasteurella multocida on week 2 post the third immunization,the protection rate were counted.Results:RT-PCR,Western blot analysis and indirect immunofluorescence showed that the omph gene could be transfected into SP2/0 cells in vitro and expressed the target protein.Indirect ELISA showed that the levels of antibodies in pOMPH group were most significantly higher than in the other groups(P