Study On The Differences Of Facial Skin Aging Among Women Of Different Ages

Objective: The experiment intends to explore the influence of age factors on the aging of female facial skin through the study of the differences in the histopathological characteristics of facial skin and related biological characteristics of dermal fibroblasts,so as to provide a certain experimental basis for delaying aging,and also to The evaluation of the efficacy of new treatment methods provides an evaluation standard.rovides an evaluation standard for the evaluation of new treatment.Methods: 1.A total of 45 samples of fresh and discarded maxillofacial skin tissue specimens removed during mid-face surgery in the Department of Medical Aesthetics and Oral and Maxillofacial Surgery of Suining Central Hospital from January 2019 to December 2020 will be collected.The tissues are divided into histology experiment group and cytology experiment group(the skin tissues of the two groups are from the same part of the same individual).According to the age of the patient,they are divided into low-age group(18-29 years old)and middle-age group(30-49 years old))And senior age group(?50 years old)three age groups.At the same time,the preoperative facial image data and the GIogau light aging classification standard were used to analyze the facial aging.2.Take the histology experimental group skin tissue(n=45 cases,15 cases in each group)as the research object,observe and analyze by hematoxylin-eosin(HE)staining and Masson collagen staining(Image J software)The thickness of the skin epidermis,the length of the nail process,and the area of collagen fibers in the dermis of each age group.3.Taking the skin tissues of the cytology experimental group(n=9 cases,3 cases in each group)as the research object,the facial fibroblast cell lines of women in different age groups were obtained by enzymatic digestion.(1)Collect each constituent fiber cell,make cell slides and perform HE staining,observe the morphology of each group of cells under an optical microscope and collect pictures;(2)Detect the expression of specific protein Vimentin by immunofluorescence;(3)Use CCK-8 method to detect the proliferation of fibroblasts and draw growth and proliferation curves;(4)Detect the cell cycle status of each component fiber by flow cytometry technology;(5)Detect the senescence of fibroblasts by staining with ?-galactosidase(SA-?-Gal);(6)Make cell membranes: take The third-generation fibroblasts were cultured in membrane-forming medium for 14 days and then HE stained.The conditions of each group of membranes were observed under an optical microscope and pictures were collected;(6)RT-q PCR method was used to detect the genes expression of COL-1,COL-3,MMP-1,and MMP-3 of each group.4.Measurement data are expressed as mean±standard deviation,single-factor analysis of variance with multi-sample means is used for comparison between multiple groups,two-sample t-test is used for comparison between two groups,and P<0.05 indicates that the difference is statistically significant.Results: 1.Facial imaging data:with the increase of age,women's facial skin becomes rough,wrinkles and pigment spots appear,pores are enlarged,and the complexion becomes dull.Facial wrinkles mainly appear in the outer canthus and lower eyelid.The low-age group,the middle-age group,and the senior-age group were in accordance with grade I photoaging,grade II photoaging and grade III photoaging in turn.2.Histological experiment results:(1)compared with the low-age group,the thickness of the facial skin epidermis decreased in the middle-aged group and the old-age group,and the difference was statistically significant(P<0.01);at the same time,the length of the facial skin epithelial spikes in the low-age group,middle-age group,and senior-age group were successively Decrease,the difference is statistically significant(P<0.01).Among them,the epithelial spike process is the most obvious in the younger age group,the epidermis-dermis boundary is clear and the connection is tight,there are many fibroblasts in the dermis,and the collagen fibers are abundant and densely arranged.In the middle and senior age groups,the epithelial spikes basically disappeared,the epidermis-dermis boundary is unclear and the connection is poor,the number of dermal fibroblasts is reduced,and the collagen fibers are sparse and broken.(2)Compared with the middle-aged group and the old-age group,the young group had the largest collagen fiber area in the dermis,and the difference was statistically significant(P<0.01);compared with the old group,the middle-aged group had a significant difference in collagen fiber area,and the difference was statistically significant.Academic significance(P<0.01).3.Cytology experiment results:(1)The cells extracted from facial skin tissues are in a long spindle shape,and the immunofluorescence results show that the isolated fibroblasts express vimentin positive.(2)From the young age group to the middle age group and the senior age group,the morphology of fibroblasts gradually changed from a clear and regular long spindle to an irregular flat and cord shape.(3)The experimental results of CCK-8 suggested that the cells grew rapidly and the number of cells increased significantly on the 1st to 3rd day after inoculation;the growth of cells became slower on the 5th to 6th day,and the number of cells gradually entered the plateau phase,which was in line with the general cell.Growth law.Compared with the middle-aged group and the old-age group,the cell proliferation of the low-age group was faster.(4)The cell cycle test results suggest that the cell cycle of facial fibroblasts in each group is mainly in the G0/G1 phase(about 80%+),which indicates that the cells used in the experiment have the ability to grow and proliferate.(5)The results of SA-?-Gal staining and counting showed that: compared with the middle-aged group and the old-age group,the number of senescent cells in the low-age group was less,and the difference was statistically significant(P<0.01);compared with the old-age group,the middle-age group was senescent The number of cells was small,and the difference was statistically significant(P<0.01).(6)The cell membrane sheet results show that: as time increases,the number of fiber cells in each component increases,and the cell membrane sheet gradually forms;the cell arrangement changes from loose and orderly to messy and compact,and there is no space between the cells.Around the 10 th day,cell membrane formation was seen in the younger age group.The results of HE staining of the cell membrane showed that the cell membrane was composed of multiple layers of fibroblasts surrounded by abundant extracellular matrix.(7)The RT-q PCR results showed that the relative expression levels of COL-1,COL-3,MMP-1,and MMP-3 m RNA in each group were different.Compared with the middle-aged group and the high-age group,the low-age group expressed high levels of COL-1,COL-3,and low expression of MMP-1,MMP-3,and the difference was statistically significant(P<0.05).Compared with the low-age group and the middle-age group,the middle-age group expressed high levels of MMP-1 and MMP-3,and the difference was statistically significant(P<0.01).Conclusion: The degree of facial aging of women in different age groups is different,and there are different degrees of differences in the three aspects of epidermal aging,dermal aging and dermal fibroblast aging-related secretory phenotypes.Among them,the changes of the true-epidermal junction and the collagen fibrin density of the dermis are the most correlated with age,which can be used as relevant evaluation indicators for future facial aging research.