Rhafgf Ethanol Liposome Preparation And Its Combination Of Micro Needle Technology Promotes Research Rhafgf Transdermal Absorption

Research1Preparation Method of Recombinant Human Acidic Fibroblast Growth Factor EthosomesAFGF (acidic Fibroblast Growth Factor) has the bioactivity of promoting the cell proliferation and angiogenesis.However, aFGF has poor stability and short half-life period, it is also easily degradable and sensitive to the temperature and pH,its molecular weight is big and it can’t penetrate skin,which limite the application of aFGF.Objective:In this study, we made ethosomes as a drug carrier for rhaFGF in order to effectively rhaFGF from unstabilizing factors,and promote it through skin.In order to find a simple and feasible way for external use which can break through the obstacles of the skin, penetrate the skin, promote the growth of skin.Methods:rhaFGF ethosomes was prepared by the method of ethanol injection, and were optimized by single-factor test with encapsulating efficiency of as the ethosomes parameter.The physical properties of rhaFGF ethosomes was identified by Cryo transmission electron microscopy.Results:The preparation of rhaFGF ethosomes maximum encapsulating efficiency was (58.82±2.56)%,when the concentration of rhaFGF was0.06mg/mLConclusions:Research indicated, rhaFGF ethosomes had high encapsulating efficiency, when the concentration of rhaFGF was0.06mg/mL. Research2Study of Combination Stratege of Ethosomes and Microneedles Array Technology for Improving the Permeation Rate of rhaFGF Through SkinObjective:To explore a minimally invasive administration method of rhaFGF to promote the permeation rate through skin,and the best time for using rhFGF.Methods:Following treatment of microneedle arrays in skin of rabbit,the distribution and the depth of microporous were visualized using methylene blue staining of the skin and HE staining of cryostat sections;Different groups:Normal skin group, rhaFGF direct coating group, rhaFGF ethosomes direct coating group, microneedle with blank liposome group,microneedle with rhaFGF direct coating group, microneedle with rhaFGF ethosomes direct coating group, intradermal injection group were applied in this research.Distribution of rhaFGF was confirmed by immunofluorescence and immunohistochemical study, rhaFGF concentration of the skin homogenate of rabbit graft mode were analyzed using ELISA.Changing the length and density of the microneedle, the group were divided into long microneedle sparse group, short microneedle sparse group, long microneedle dense group, short microneedle dense group.RhaFGF concentration of the skin homogenate of rabbit graft mode were analyzed using ELISA. Changing the time, when the concentration is certain. Harvesting the skin tissue after15min,30min,45min,60min,120min,180min,240min from the microneedle with rhaFGF ethosomes direct coating group, rhaFGF concentration of the skin tissue homogenate were analyzed using ELISA.Results:After treatment of microneedle arays, the microporous were uniform distribution in skin; the microchannels extended through the epidermis to dermal layer of section. In30min later of administration, Normal skin group, rhaFGF direct coating group, rhaFGF ethosomes direct coating group, Microneedle with blank liposome group the homogenate of the skin tissue showed no significant change, the homogenate of skin tissue showed a significant(P<0.05) increase in concentration of rhaFGF the microneedle with rhaFGF direct coating group, Microneedle with rhaFGF ethosomes direct coating group, intradermal injection group compared with the rhaFGF direct coating group. Changes between microneedle with rhaFGF direct coating group, microneedle with rhaFGF ethosomes direct coating group showed a significant(P<0.05). Changes between Microneedle with rhaFGF ethosomes direct coating group, intradermal injection group showed no significant(P>0.05). Immunofluorescence and immunohistochemical staining of skin for rhaFGF showed that a large number of rhaFGF were arranged equality in dermal tissue of the microneedle with rhaFGF ethosomes direct coating group.The homogenate of skin tissue showed a significant(P<0.05) increase in concentration of rhaFGF the long microneedle dense group and the short microneedle dense group respectively compared with the long microneedle sparse group, the short microneedle sparse group. The homogenate of skin tissue showed no significant(P>0.05) in concentration of rhaFGF the long microneedle dense group and the long microneedle sparse group respectively compared with the short microneedle dense group and the short microneedle sparse group.RhaFGF concentration of the skin tissue homogenate were highest after100min using the drug,when the concentration is certain.Conclusion:Combinating ethosomes with microneedles array technology can get a higher effection through the skin for rhaFGF. It make rhaFGF promote skin proliferation, accelerate skin flap expansion by transdermal delivery systerm becomes possible.They were easy and painless.It maybe a new way to improve the absorption through skin for the macromolecular medicine such as polypeptide and protein.