环孢素A纳米晶体联合滚轮微针治疗雄激素性脱发的实验研究OA

Objective To address the problems of poor transdermal permeability of cyclosporine A and limited development of clinical topical preparations,cyclosporine A nanocrystals are prepared,and the efficacy and action mechanism of their combination with roller microneedles in the treatment of androgenetic alopecia are investigated.Methods The anti-solvent precipitation method was employed,with sodium dodecyl sulfate(SDS)used as the surfactant.Single-factor screening was performed to optimize the mass ratio of drug to sur-factant and the volume ratio of organic phase to aqueous phase,and then cyclosporine A nanocrystals were prepared,and their particle size,polydispersity index(PDI)and stability were evaluated.Twenty testosterone-induced C57BL/6 mouse models of androgenetic alopecia were established,and the models were divided into the blank group,the model group,the minoxidil group and the cyclosporine A combined with roller micronee-dle group(cyclosporine A group),with 5 mice in each group.The therapeutic efficacy was evaluated based on the area of regenerated hair.HE staining and Masson staining were applied to observe hair follicle morphology and collagen distribution.Immunofluorescence staining was used to detect the expression of transforming growth factor-β(TGF-β)and Ki67,and Western blot was adopted to detect the protein expression level of Wnt3a.Results The optimized cyclosporine A nanocrystal formulation was as follows:the mass ratio of SDS to cyclosporine A was 1∶2,and the volume ratio of organic phase to aqueous phase was 1∶19.The particle size of the obtained nanocrystals was(165±3)nm,and the PDI was 0.238±0.034,and the particle size distri-bution was uniform.A mouse model of androgenetic alopecia was successfully constructed.Compared with the model group,the minoxidil group and the cyclosporine A group had higher regenerated hair coverage and hair growth score,shorter pigmentation time after depilation,larger numbers of anagen hair follicles and telogen hair follicles,greater skin thickness,dermal layer thickness,Ki67 fluorescence intensity and Wnt3a protein ex-pression level,lower proportion of collagen deposition area,proportion of inflammatory infiltration area and TGF-β fluorescence intensity.In addition,the cyclosporine A group had longer pigmentation time after depila-tion and higher Wnt3a protein expression level than the minoxidil group,and the differences were statistically significant(P<0.05).Conclusion Cyclosporin A nanocrystals with uniform particle size and good stability are successfully prepared.When combined with roller microneedles,they significantly promote hair growth in mice with androgenetic alopecia.The mechanism is related to the inhibition of TGF-β-mediated hair follicle de-generation,upregulation of Wnt3a expression and subsequent activation of the Wnt signaling pathway.