| Background: Endometriosis(EMs),referred to as endometriosis,is a benign disease that is prone to recurrence.Multiple factors,including genetic,environmental,immune,endocrine,and microbial,have been reported to be associated with the development of endometriosis.However,there is currently no consensus on whether these factors alone or in combination affect the pathophysiology of endometriosis.Currently,surgery and hormonal therapy remain the most commonly used clinical treatments for women with endometriosis.Unfortunately,the estimated recurrence rate of endometriosis after surgery or hormone therapy is as high as 21.5% at 2 years and 40-50% at 5 years.In addition,these treatments are associated with adverse side effects,including mood swings,pseudomenopause,decreased bone density,and a high risk of osteoporosis.Therefore,in order to benefit women with endometriosis,there is an urgent need to develop new medical regimens that can effectively suppress endometriosis and minimize adverse effects.Previous studies have observed increased neutrophil infiltration in the systemic circulation and peritoneal fluid of patients with endometriosis,suggesting that neutrophils may play a key role in the treatment of endometriosis.However,the role of neutrophil depletion in the treatment of endometriosis remains controversial,and the design of neutrophil-based therapeutic strategies needs to be further refined.Excitingly,bovine serum albumin(BSA)nanoparticles(NPs)have been reported to carry neutrophils and be delivered to areas of inflammation for intrauterine therapy by neutrophils.Membrane disease offers the possibility.Objective: Based on endometriosis patients,a minimally invasive mouse model of endometriosis was constructed by injection method to observe the proportion of neutrophils in ectopic lesions.By further synthesizing albumin nanoparticles capable of carrying neutrophils,the enrichment of albumin nanoparticles in ectopic lesions was detected by intraperitoneal or intravenous injection.To understand whether glucose starvation therapy is suitable for the treatment of endometriosis,in order to provide a new theoretical basis for the nano-treatment of endometriosis.Methods: By collecting ectopic lesion tissue and eutopic endometrium and endometrial clinical biological samples of matched normal women from the Department of Gynecology of the First Affiliated Hospital of Anhui Medical University who underwent ovarian endometrioma cystectomy,we used the differential distribution of centriocytes in these tissues was analyzed by immunohistochemistry.A minimally invasive mouse model of endometriosis was then constructed using the injection method.Using this model can help us to further explore new strategies for endometriosis treatment.By flow cytometry,we analyzed the proportion of neutrophils in ectopic foci,eutopic endometrium,and eutopic endometrial tissue of control mice in a mouse model of endometriosis.Afterwards,BSA nanoparticles capable of carrying neutrophils in situ were constructed by desolubilizing the BSA solution with ethanol and then cross-linking with glutaraldehyde.The average size of BSA nanoparticles was measured using dynamic light scattering and transmission electron microscopy.At the same time,in this study,BSA nanoparticles combined with fluorescein isothiocyanate(FITC)and indocyanine green(ICG)were prepared respectively,and flow cytometry proved that the nanoparticles carried neutral capacity of granulocytes.In order to understand the in vivo distribution of BSA nanoparticles,this study continued to use fluorescently labeled FITC-BSA nanoparticles and ICG-BSA nanoparticles,which were injected into endometriosis mice by intraperitoneal or intravenous injection.Observe its tissue distribution.In order to clarify the role of neutrophils in the differential distribution of BSA nanoparticles under different injection routes,we first compared the phagocytosis of BSA nanoparticles by neutrophils in various tissues under different injection routes,followed by intraperitoneal injection.The mouse neutrophils were specifically depleted by injection of anti-Ly6 G antibody,and BSA nanoparticles were injected intraperitoneally to observe the tissue distribution of BSA nanoparticles after depletion of neutrophils.Finally,because we found that in the mouse model of endometriosis,we found that the demand for glucose of ectopic foci stromal cells was significantly higher than that of eutopic endometrial stromal cells,so we tried to use glucose starvation therapy for endometriosis.In the treatment of,BSA-GOx nanoparticles loaded with glucose oxidase(GOx)were constructed,and the catalytic activity and cytotoxicity of BSA-GOx nanoparticles were verified.We administered intraperitoneal injection of BSA-GOx nanoparticles to endometriosis mice to observe the anti-endometrial effect of BSA-GOx nanoparticles.Results: This study found a significant increase in neutrophils in ectopic lesions in patients with ovarian endometriosis compared with matched eutopic endometrial or healthy control endometrial tissue.This phenomenon also holds true in a mouse model of endometriosis,where neutrophils in the peritoneal fluid and ectopic lesions of endometriosis mice are significantly different from those in the early post-lesion(day 3)after lesion formation.There was a significant increase compared to the control group,but no significant increase in neutrophils was observed in the eutopic endometrium.On day 14 after lesion formation,the proportion of neutrophils in the peritoneal fluid decreased to a level similar to that in the control group,while the proportion of neutrophils in ectopic lesions remained high.This study constructed BSA nanoparticles with an average size of about 320 nm and showed that FITC-BSA nanoparticles could be internalized by neutrophils.When BSA nanoparticles were injected intraperitoneally,the accumulation of FITC-BSA nanoparticles in ectopic lesions was the highest,which was much higher than that of other major organs(heart,liver,spleen,lung,and kidney),while the ectopic lesions after intravenous injection were very high.Few FITC-BSA nanoparticles were detected.The distribution of ICG-BSA nanoparticles was similar to that of FITC-BSA nanoparticles.Since neutrophils in the peritoneal fluid and ectopic lesions of endometriotic mice can phagocytose large amounts of BSA nanoparticles,we specifically depleted the mice by intraperitoneal injection of anti-Ly6 G antibody 24 h before BSA nanoparticle injection In vivo neutrophils,it was found that depletion of neutrophils significantly reduced the accumulation of BSA nanoparticles in endometriotic lesions,and the ratio of ectopic/eutopic BSA nanoparticles was significantly reduced.Finally,in the mouse model of endometriosis,we found that the demand for glucose of ectopic stromal cells was significantly higher than that of eutopic endometrial stromal cells,so we synthesized BSA-GOx nanoparticles loaded with glucose oxidase and synthesized them.It was proved that the particle has good catalytic activity and cell killing ability(IC50=0.637 ?g/ml),and the BSA nanoparticle platform maintained 41.6% cytotoxicity of GOx.In addition,by intraperitoneal injection of BSA-GOx nanoparticles,we found that BSA-GOx nanoparticles significantly reduced the size of ectopic lesions,and the apoptosis of ectopic lesions was also significantly increased in the BSA-GOx nanoparticles-treated group.After 14 days of dosing,no body weight loss or pathological changes in major organs were observed in the treated mice.In conclusion,intraperitoneal injection of BSA-GOx nanoparticles can produce excellent anti-endometriosis effects via GOx-mediated in situ depletion of glucose.Conclusions:(1)The ectopic lesions of patients with endometriosis are enriched in neutrophils.(2)BSA nanoparticles can accumulate in ectopic lesions,and this phenomenon is mediated by neutrophils.(3)Glucose oxidase can exert good anti-endometriosis performance with the help of bovine serum albumin nanoparticles. |
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