| ObjectiveRheumatoid arthritis(RA)is a refractory and chronic inflammatory disease;meanwhile,osteoarthritis(OA)is the most common disorder of the locomotor system,both of which belong to one of the major geriatric diseases.Sinomenine hydrochloride(SH)is an ideal drug for the treatment of RA and OA.But SH tablets and injections belong to systemic administration.The high plasma concentration of systemically administered SH will potently release histamine in association with the degranulation of mast cells in mammalian tissues,and this could cause rash and gastrointestinal side effects.However,passive diffusion of SH was inefficient because of the stratum corneum(SC).Therefore,electroporation is required to compensate for the low efficiency of SH passive diffusion.In this study,transdermal experiments in vitro and clinical tests were utilized to explore the optimized parameters electroporation of topical delivery for SH.Through the experiments of fluorescence method and pathological analysis,this study revealed the mechanism of electroporation.Highly efficient transdermal delivery by optimized parameters electroporation would be recommended to enhance SH transdermal administration in future clinical application.Methods1.The transdermal drug delivery experiments in vitro were performed utilizing the mice skin that is one of the most common models and miniature pig skin that is similar to human skin.The vertical Franz-type diffusion cells were used to collect the samples obtained from different time-points.The flux ehancement ratio(ER)and cumulative drug permeation amounts(Qn)of SH under different electroporation parameters were investigated.Finally,the optimal electroporation parameters were filtrated out.2.For study on the ER and the Qn of SH in different drug concentrations,transdermal experiments(mice and miniature pig skins)in vitro were performed under the fixed parameters of electroporation.Subsequently,we investigated the relationship between SH administration concentration and SH percutaneous enhancement induced by electroporation.3.HE staining and fluorescent agent substitution were performed to study the mechanism that electroporation promoting SH transdermal delivery.The transformation of mice and miniature pig skin structure was observed after electroporation stimulating or not.Also,the distribution of fluorescent agent in the mice and miniature pig skin was observed.Thus,we studied what skin structure change had appeared after electroporation,and how electroporation promoted the drug permeated into skin.Finally,the promoting permeation mechanism of electroporation was illuminated.4.The optimal parameters electroporation promoting the SH percutaneous administration was used in clinical tests.We studied the concentration of SH in synovial fluid(SF)after electroporation stimulating or not.Also,we compared the improvement rate of clinical symptoms in the test group that combined of celecoxib with electroporation and the positive control group that was treated of celecoxib without electroporation.Finally,we would certifie the clinical efficacy of that the optimal parameters electroporation promoting transdermal administration of SH.Results1.After electroporation,transdermal permeation of SH was increased to 1.9-10.1 folds in mice skins and 1.6-47.1 folds in miniature pig skins compared with the passive diffusion.The results of miniature pig skins were in accordance with that of mice skins:The Qn and ER induced by the electroporation parameters(3 KHz,exponential waveform,and intensity 10)were higher compared with that induced by the other electroporation parameters.And the Qn values increased gradually as the drug concentration(0.383.04 mmol/L)increases.2.In normal skin tissues,the skin structures were tight and integrated,and the cells of the epidermis were tightly arranged,and the edge of cells was hard to distinguish.After the electroporation stimulation,the skin structures were loose,and the intercellular intervals and epidermal cracks increased.NaFluo only distributed in the outmost SC after passive diffusion without electroporation.However,when electroporation was applied on the skins,the stratum spinosum,stratum granulosum and SC of skins were observed to have very strong fluorescence of NaFluo.3.In the test group that combined of celecoxib with electroporation,the improvement percentage of joint movement degree was 94.75 ± 6.84%,and the effective percentage of joint pain relief(VAS)was 79.39 ± 4.63%.Compared with the test group,the above two parameters were lower in positive control group that was treated of celecoxib without electroporation,with the improvement percentage of joint movement degree being 52.41±12.51%and the effective percentage of joint pain relief(VAS)being 62.40±7.54%.Moreover,the improvement percentage of joint movement degree had statistic difference(P<0.05).In the patient SF of the test groups,the maximum and minimum concentrations of SH were approximately 20.84 and 0.39.ng/mL,respectively.The concentration of SH in patient SF samples without electroporation was below limit of quantification.Conclusions1.Electroporation did significantly promote SH transdermal permeation.We found that the electroporation parameter and drug concentration both were one of the key factors for electroporation to promote SH transdermal penetration.In this study,the optimal parameters of electroporation for human were 3 KHz,exponential waveform,and intensity 10.2.Electroporation promotes transdermal permeation by increasing of epidermal cracks,which increase the skin permeability and enhance transdermal delivery SH.3.The transdermal treatment of SH with the optimized electroporation did significantly improve the clinical improvement rate.And electroporation did significantly enhance the SH concentration in the patient SF.Therefore,because of its advantages of improving the efficacy and reducing the side effects,transdermal treatment of SH with the optimized electroporation in this study may be an ideal treatment of arthritic diseases(RA or OA). |
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