A Novel Automatic Fire-acupuncture Instrument And Its Application In Promoting Hair Growth

Hair follicle(HF)is an important accessory organ in skin,which has biological functions such as protecting skin,secreting sebum,regulating body temperature,and transmitting information.Hair follicle injury and aging can cause hair cycle disorder,including hair loss and other hair diseases.Continuous development of injury and/or improper treatment can even lead to skin tumors,which could seriously affect the health,beauty and quality of life.As an external treatment method of traditional Chinese medicine,fire-acupuncture has the characteristics of local stimulation,meridian conduction and whole-body regulation,has been widely applied in the clinical treatment of various diseases.The use of fire-acupuncture for treating hair diseases,mainly through acupuncture skin to achieve the effect of regulating Qi and blood,dredging the meridian,stimulating hair growth from telogen to anagen.Since the traditional fire-acupuncture relay on personal experience,it is difficult to precisely control the needle temperature,the depth and the density of acupuncture,as well as the time of needle retention.To overcome these shortcomings,some fire-acupuncture devices were developed,usually heated by flame or resistance wire.These devices still have defects such as non-standard treatment parameters,low operation efficiency and poor safety,which can’t meet the needs of clinical application.Therefore,a new automatic fire-acupuncture instrument with better performance need to be developed.This study aims at developing a novel automatic fire-acupuncture with high efficiency and stability,controllable parameters and easy operation,and to evaluate its application in promoting hair growth.The main research contents and conclusions are as follows:(1)Design and manufacture of automatic fire-acupunctureWe designed and manufactured a new type of automatic fire-acupuncture.The handheld part adopts a gun-like shape design,which is easy to operate.The heating system adopts electromagnetic heating to control the temperature of needles.The ejection system adopts electromagnetic ejection combined with spring pullback design to control the retention time of the needle in skin and the retention time is controlled at 0.2s.The depth adjustment system uses a scale knob to control the depth of needle in skin,which is accurately adjustable within 0-4mm.The needle can be adjusted to meet treatment needs.The above design,combined with a process control system,can achieve the precise control of needle temperature,acupuncture density,acupuncture depth,and needle retention time.Thus this design has the advantages of simple operation,rapid operation,asepsis,high efficiency,conformability and visualization,which has outstanding clinical application value.(2)Application of automatic fire-acupuncture in promoting hair growth1)Automatic fire-acupuncture promotes hair growthWe firstly evaluated the efficiency of automatic fire-acupuncture on hair growth in mice.3 groups of C57BL/6J mice with hair in telogen were applied automatic fireacupuncture(AFA group),room temperature acupuncture(RTA group)and sham control respectively,after shaving dorsal hair.Macroscopic observation showed “black”appearance of the dorsal skin after 3 times in AFA group,5 times in RTA group.The hair growth score of AFA group was higher than the other two groups.HE staining showed that the hair cycle in AFA group entered early anagen upon 3 times of acupuncture and middle anagen upon 5 times of acupuncture,while the hair follicles remain in early anagen in RTA group upon 5 times of acupuncture.These results demonstrate that automatic fire-acupuncture can promote hair growth by shifting the hair cycle from telogen to anagen.2)Automatic fire-acupuncture promotes hair growth through Wnt/β-catenin pathwayIn order to explore the mechanism of promoting hair growth by automatic fireacupuncture,Wnt/β-catenin signaling pathway was studied by q RT-PCR,Western blot and immunofluorescence staining.q RT-PCR showed that Wnt3 a,Wnt10b and Wnt7 b m RNA significantly increased in the early stage of fire-acupuncture(1-2 times),and continuously increased on a dose dependent manner.β-catenin and Lef-1 m RNA also increased significantly upon 2 times of fire-acupuncture.Cyclin D1 m RNA significantly increased upon 4 times of fire-acupuncture.Western blot showed that the Wnt3 a,β-catenin,Lef-1 and Cyclin D1 protein increased significantly with the times of fireacupuncture.Immunofluorescence staining showed that β-catenin and Lef-1 translocated into the nucleus at 3 times of fire-acupuncture.These results demonstrate that fireacupuncture induces the expression of Wnt signal upon injury,increases the expression level of transcription factors i.e.β-catenin and Lef-1,and induces nuclear translocation of β-catenin,thus activates the expression of the downstream proliferating gene Cyclin D1,leading to the hair cycle shift from the telogen to anagen.To further verify the role of Wnt signaling pathway in promoting hair growth by fire-acupuncture,XAV939 was used to block the Wnt signaling pathway and detected the effect of automatic fire-acupuncture on hair growth.Macroscopic observation showed that the skin color of mice in the XAV939 injection group did not change significantly after 5 times of acupuncture,while the skin "blackened" after 3 times of acupuncture in the control(DMSO)group,indicating the beginning of hair growth.HE staining showed that the hair follicles of the control group had entered anagen after 5 times of acupuncture,while the hair follicles of the XAV939 injection group remain at telogen.Western blot showed that the protein expressions of Wnt3 a,Lef-1 and β-catenin in XAV939 injection group were significantly down-regulated.Immunofluorescence staining showed that there was no nucleus location of β-catenin XAV939 group.These results demonstrate that hair growth is delayed after blocking the Wnt signaling pathway.In summary,this study designed and manufactured a novel automatic fireacupuncture instrument,which has the advantages of high efficiency and stability,adjustable parameters,simple operation and high safety.It is efficient in promoting hair growth,by a potential mechanism of regulating Wnt/β-catenin pathway.The present study laid a foundation for the treatment of hair loss related diseases.