Therapeutic Research Of Silk Protein Loading Drugs For Changes In Skin Sensitivity Induced By Phototherapy

Objectives In recent years,ablative laser resurfacing and fractional resurfacing using professional devices have been widely used for facial rejuvenation.Multiple treatments give a more satisfactory effect.However,the extensive and long-term use of laser technologies has also brought new questions to dermatologists: whether long-term repeated laser treatment will change the skin's sensitive condition? Will it cause an attenuating of the epidermis? Will it affect the surface,the microcirculation,and thus damage the skin barrier function? However,current observations of adverse effects of laser technology mainly focus on the pigment abnormalities or scarring.Objective long-term observations of skin condition changes along the treatment are scare,especially in terms of ablative laser's impact on skin sensitivity.Therefore,this study focuses on the widely used course of multiple treatments with the laser devices and its influence on skin's sensitivity.We designed a randomized split-face study with a long-term follow up.Materials and methods The qualified volunteers recruited through strict screening standard respectively treated by three different types of photoelectric pattern.At least 20 people of each group finished 1 year follow-up and received the combination of subjective and objective skin sensitivity evaluation.The objective evaluation mainly adopts noninvasive type physical physical testing equipment.Firstly,through strict split-face experiment,the short term and long term effects of skin microcirculation,skin gloss and oxygen partial pressure,cause by different types of photoelectric treatment in healthy people.Secondly,to explore the relatationship between sensitive skin and the photoelectron therapy,we examined the effects of different type photoelectric treatment on healthy people's current perception threshold.Thirdly,to evaluate comprehensively the basic skin conditions before and after different photoelectron treatment,the parameters such as skin thickness,density,and so on were examined through the different skin of trauma associated testing parameters to explore the sensitive skin possible pathogenesis between sensitive skin and photoelectric treatment.Isolated the human primary skin keratinocytes and endothelial cells,and treated them respectively by three kinds of laser irradiation.Then we detected TRPV1 activation,as well as its downstream related molecular PAR2 and nf-kappa B signaling pathways activated by using Western Blot(WB)method.ELISA method was brought here to detect the secretion of the downstream IL-6,PEG2,ET – 1,exploring the molecular mechanism of photoelectric mechanism underling the sensitivity skin.By inspection of the above two levels found in the process of laser processing related molecular activation,we the molecule channel blockers SB705498.We mixed the SB705498 with the silk protein solution and added HRP/H2O2 catalysis,then the enzyme-crosslinked silk protein hydrogel loading SB705498 were prepared.The instrument of Franz medications in vitro transdermal absorption were used to exmine the drug transdernal diffusion,and then the transdermal drug quantity were detect by high performance liquid chromatography(HPLC)method.Meanwhile,we planted the keratinocytes and endothelial cells on the silk based hydrogel,and used Western Blot(WB)method to detect TRPV1 activation,as well as the axtivation of its downstream related molecular PAR2 and nf-kappa B signaling pathways.ELISA method was used to detect the downstream the secretion of IL-6,PEG2 and ET-1 to verify that the silk protein hydrogel drug delivery can maintain its medicinal properties.Results 1.By noninvasive physical and physiological instrumental testing,after three bait laser treatment,volunteers' skin sensitization increased within one week compared with the control side,the specific performance was transcutaneous water loss(TWEL)value increased,the glossiness decreased and blood flow perfusion in facial skin treatment area increased significantly,after a week the above parameters returned to normal.However,volunteers who received 10 Q-switched Nd: YAG 1064 nm laser treatment sessions and volunteers with 5 intense pulsed light(IPL)treatment sessions showed a decrease in TWEL value,an increase in glossiness,an increase in cutaneous perfusion of blood,wich returned to normal after 3 days.That after the treatment of the latter two makes sensitive skin index tends to decline.2.Western Blot(WB)results showed that compare to the Q-switched Nd: YAG 1064 nm laser group and the intense pulsed light group,the keratinocytes and capillary endothelial cells of Er:YAG laser group were exposed to three kinds of photoelectric devices in vitro respectively.The TRPV1,PAR2,NF-?B expression were significantly increased,while the ELISA results showed that downstream inflammatory mediators secretion was significantly higher than the latter two groups.Explain Er:YAG laser treatment is bliative treatment,tin the early stage,the skin sensitivity was increased because of skin barrier function damage.3.SB705498 is a blocker of the TRPV1 channel receptor.The enzyme-crosslinked silk fibroin hydrogel carrier delivering SB705498 more efficiently than the control carbachine by comparing.Western Blot(WB)confirmed that SB705498 could block the TRPV1 channel and inhibited the expression of PAR2 and NF-?B,and the secretion of IL-6,PEG2,ET-1 and other mediators was also inhibited,which further showed that the silk fibroin hydrogel drug delivery system can effectively maintain the stability of the chemical structure of the drug and thereby maintain its drug-resistance.Concludions This study systematically compared three kinds of widely-used optoelectronic devices in clinic,and evaluated the change of index parameters of subjective and objective skin sensitivities of volunteers after repeated treatments.At the same time,we explored the mechanism of changes in skin sensitivity from the molecular level.By comparing and optimizing the enzyme-crosslinked silk fibroin hydrogel as a sustained-release delivery carrier of the TRPV1 channel blocker SB705498,the sustained release of SB705498 was achieved and the function of blocking the TRPV1 channel was verified.In summary,this study tried a comprehensive set of subjective and objective evaluation of sensitive skin.Targeted blocking of the relevant molecular channels and thus blocking the secretion of neurotrophic mediators of the skin is expected to improve the sensitivity of the skin treated with phototherapy provides a new way of thinking.