Effects of embryonic fibroblasts on the aging-related phenotypes and dysfunction of diabetic ulcer fibro-blasts / 中华内分泌外科杂志

ABSTRACT

Objective To observe the effects of human embryonic fibroblasts ( FBs ) on the biological characteristics of diabetic FBs in vitro , and to explore the possible therapeutic mechanism on diabetic ulcers . Methods Diabetic FBs were isolated and cultured with embryonic FBs (experimental group), adult FBs(control group 1)and alone(control group 2)in transwell chambers.Diabetic FBs growth curves in the coculture systems were described.The cell shape was observed by microscopy with HE staining .On the 4th and 7th day of cocul-ture, diabetic FBs were isolated to measure the expression levels of hydroxyproline and transforming growth factor -beta 1(TGF-β1)in the supernatant of monoculture .The cell senescense was displayed by β-galactosidase histo-chemical staining on the 3th, 5th and 7th day of coculture.Eight patients with diabetic foot ulcers who were can-didates for embryonic FBs constructs after failed traditional treatment were included in the study .The pre and post self-control study was conducted by observing the wound healing process before and after embryonic FBs treat -ment.Results Diabetic FBs in the experimental coculture system displayed more typical pattern and faster grow -ing rate.The level of hydroxyproline and TGF-β1 in the supernatant of diabetic FBs monoculture was significantly higher in the experimental group than in the control group after treated 4 or 7 days ( hydroxyproline4 d, 7 dcompared with control group 1P=0.023,P=0.007;compared with control group 2 P=0.007, P=0.003;TGF-β1 4 d, 7 dcompared with control group 1P=0.000, P=0.000;compared with control group 2 P=0.000, P=0.000).The expression of SA-β-gal tended to be more apparent in control group 1 and 2 than in the experimental group on the 3rd,5th,and 7th day.All of the 8 cases with diabetic ulcers were cured by using em-bryonic FBs constructs .Conclusion Embryonic FBs can promote the proliferation of diabetic FBs , correct cell senescence and increase the production of hydroxyproline and TGF-β1,which maybe enhance the process of dia-betic wound healing .