The Influence Of PDGF-BB And CTGF On The Proliferation Of Human Skin Fibroblast In Vitro

Objective:Skin aging and minor depression in the face may seriously affect the appearance. Research confirmed that all these problems are related to collagen reduction and structural changes in the dermis.Theoretically speaking.fibroblast is an ideal filling agent,it can effectively improve the wrinkles and tiny depression.This experiment major to evaluate the influence of recombinant human plateled derived growth factor-BB (rhPDGF-BB). recombinant human connective tissue growth factor (rhCTGF) and their synergy on the proliferation of human skin fibroblast(HSF).For supply a variety of methods to rapid proliferation of HSF in vitro,shorten cell culture period and clinical treatment cycle,thereby promoting this technology early into clinical,benefiting the masses.Methods:1,The cell culture and purification of HSF Take the normal foreskin tissue removed from the healthy donors accepted cosmetic surgery,and obtain HSF by tissue adherent method.Differential digestion method was applied to depurate HSF.2,The influence of PDGF-BB and CTGF on the proliferation of HSF in vitro by the MTT assay Take up 4-7 generation fibroblasts which had a good condition,and were vaccinated into several pieces of 96-well culture plates with 2×104/ml,100u/well.These cells were randomly divided into control group and experimental groups.The control group without any kind of growth factor,while the experimental groups were added to different concentrations of rhPDGF-BB, rhCTGF or rhPDGF-BB+rhCTGF.The concentration-time effect was studied at 1d,3d,6d and 9d by the MTT colorimetric method.Results:1-Human skin fibroblasts all grew as adherence in vitro.At first,the cells were round or oval,colony-like growth.and then,gradually scattered and extended as fusiform.When the number of HSF were more enough,they would spread like paliform,swirl or radiation.By the way,passaged to 3 to 4 generations,the purity of HSF increased,showed a typical fibroblast morphology and dominant growth.2,In four test time,compared with control group,the four experimental groups which added different concentrations of rhPDGF-BB,were all have statistically significant differences (P<0.05).Comparison between two experimental groups,and found rhPDGF-BB 100ng/ml group had a significantly higher proliferating effect than other concentrations (P<0.05),while promoting proliferation of rhPDGF-BB O.1ng/ml was weakest (P<0.05). The effect on the proliferation in the 1-9d,0.1-100ng/ml was concentration-time dependent.3,In four test time,compared control group with four experimental groups, differences were statistically significant (P<0.05).Comparison between two experimental groups,the proliferating effect was in a concentration-time dependent manner with the concentration from 0.1ng/ml to 1 OOng/ml in 9 days,except rhCTGF50ng/ml group had a better influence at Id (P<0.05).Besides,contrast 150ng/ml group with 100ng/ml group,the proliferating effect had roughly downward trend.4,There were marked differences when two combined groups compared with control group (P<0.05).To some extentthe proliferation effect of combined groups were also concentration-time depended, and better than used them alone (P<0.05), except the group of rhPDGF-BB O.1ng/ml (P>0.05).In addition,compared rhPDGF-BB 100ng/ml group with rhCTGF 100ng/ml group,the difference was significant,P< 0.05;but there was no obvious difference between rhPDGF-BB 0.1ng/ml group and rhCTGF 10ng/ml group,P>0.05.Conclusion:Both of rhPDGF-BB and rhCTGF can promote the proliferation of HSF in vitro, and the function can be concentration-time depended, and the former is better. The combination of two growth factors has a synergistic action on the promotion of HSF.