Preparation And In Vitro Study Of Specific Anti-P.gingivalis Yolk Antibody Liposome

Objective Porphyromonas gingivalis is the main pathogenic bacteria of chronic periodontitis(Porphyromonas gingivalis,Pg).The egg yolk antibody(immunoglobulin of yolk(IgY))is produced by B-lymphocytes and stimulated by specific antigens in birds.Anti-Pg gingivalis egg yolk antibody liposomes(anti-Pg-IgY liposomes)were prepared using two methods,the encapsulation rates of the liposomes were measured,and their in vitro response was evaluated.The effect of liposome was observed,and the liposome morphology was observed under an electron microscope,and the liposome was stored at different temperatures to evaluate its biophysical properties,providing a theoretical basis and new information for clinically assisting the treatment of chronic periodontitis and improving its efficacy treatment method.Methods Anti-Pg-IgY liposomes were prepared using a thin film dispersion method,and then half of the liposomes previously prepared were treated with ultrasound.The BCA method was used to determine the recovery and precision of anti-Pg-IgY to determine the feasibility and repeatability of the experiment.The dynamic light scattering particle size distribution analyzer was used to determine the Zeta potentialand particle size of the anti-Pg-IgY liposomes prepared by the above two methods.After determining the dialysis time,the encapsulation efficiency was measured by dialysis in combination with a microplate reader.The in vitro release rates of pure anti-Pg-IgY and anti-Pg-IgY liposomes in 0.9% physiological saline were investigated,and the quality of the two liposomes was evaluated using a transmission electron microscope.The same liposome solution was divided into two equal parts,one wasplaced at room temperature(25 ?),and the other was stored in a refrigerator at 4?for seven days.The morphological changes of the two were observed under an electron microscope.Results The average recoveries of the three groups for BCA protein determination were 1.045,1.00,and 1.02,respectively,and the precision was 1.29%,which met the methodological requirements.The zeta potential of the liposomes prepared by the thin-film method was(-38.55±0.16)m V(n=3),the average particle size was(82.48±2.3)nm(n=3),and the polydispersity index was(0.313±0.01)(n=3),the zeta potential of the liposomes prepared by ultrasound method was-11.36 m V±5.52 m V,the average particle diameter was 20.6 nm±1.1 nm,and the polydispersity index was0.266±0.034.The encapsulation rate of liposomes prepared by the thin film dispersion method was 40%(n=3),and the encapsulation rate of liposomes prepared by the ultrasonic method was 22%(n=3).In 0.9% saline,anti-Pg-IgY released 95.8% within24 hours,while the cumulative release rate of anti-Pg-IgY liposomes within 4 hours was less than 20%,reached approximately 50% within 24 hours,and the release rate at 168 hours was 85.8%(n=3).Observation of liposome samples stored at different temperatures for 7 days under a transmission electron microscope showed that the shape of the liposomes was relatively complete at 4 ?,while the phospholipid twin layer structure of the liposomes stored at 25? at room temperature had been broken.Conclusions Compared with the ultrasonic method,the anti-Pg-IgY liposomes prepared by the thin-film dispersion method have a higher encapsulation rate,and their particle size is in the nanometer range.The in vitro release experiments performed under the simulated human environment prove that it is expected to achieve a long-cycle sustained release effect in periodontal pockets.