[The proapoptotic effect of the homogenate of the tissue of different parts of pig's full thickness dermal wounds on cultured fibroblasts].
OBJECTIVE
To observe the proapoptotic effect of the homogenate of different parts of pig's full thickness dermal wounds on cultured fibroblasts.


METHODS
The tissues were dissected from the wound center and sub-neoepithelium separately 15 days after homogenization and sterilization, the specimens stored at -70 degrees C. The forth passage of the fibroblasts were cultured for 16 hours in different culture solutions and were grouped into 7 groups: DMEM containing 5% fetal bovine serum as Group I, DMEM containing 5% homogenate of tissue from wound center as Group II, DMEM containing 5% homogenate of tissue from sub-neoepithelium as Group III, the culture solution of Group I mixed with 10 microg/ml GM6001 in Group IV, with the culturing medium of Group III plus 10 microg/ml GM6001 as Group V, the culture solution of Group II mixed with 10 ng/ml aFGF as Group VII, and the culture solution of Group III mixed with 10 ng/ml aFGF as Group VII. In all groups except Group I, the fibroblasts of the 6 pigs were treated with the homogenate derived from the same animal respectively. After being incubated in Annexin V-FITC and PI, cells were analyzed by Flow Cytometry and the rate of apoptotic cells was acquired. The data were analyzed by SPSS 11.0 using Least-significant Difference test (LSD).


RESULTS
The apoptotic rate of the 7 groups were as follows: 4.39% +/- 0.41% in Group I, 10.98% +/- 1.42% in Group II, 13.47% +/- 1.44% in Group III, 7.2% +/- 0.46% in Group IV, 12.1% +/- 0.85% in Group V, 3.9% +/- 0.63% in Group VI, 9.8% +/- 0.50% in Group VII; there were significant differences between every two groups except Group I and Group VI.


CONCLUSIONS
Homogenate of the tissue derived from the sub-neoepithelium has greater proapoptotic effect than that from the wound center; the proapoptotic effect of homogenate of the tissue both under neoepithelium and in wound center can be significantly alleviated by acid fibroblast growth factor, partly because of MMPs.
Li
Jinqing
J
Key Research Laboratory of Wound Repair of PLA, 304th Clinical Department, General Hospital of PLA, Beijing 100037, P R China.
Fu
Xiaobing
X
Sun
Tongzhu
T
Gu
Shaofeng
S
Du
Yiri
Y
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D020405:Dermis / Q000737:chemistry / Q000293:injuries; D004151:Dipeptides / Q000494:pharmacology*; D005260:Female; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects*; D005434:Flow Cytometry; D008297:Male; D061965:Matrix Metalloproteinase Inhibitors; D011480:Protease Inhibitors / Q000494:pharmacology; D013552:Swine; D014020:Tissue Extracts / Q000494:pharmacology*
D004151:Dipeptides; D061965:Matrix Metalloproteinase Inhibitors; C078131:N-(2(R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl)-L-tryptophan methylamide; D011480:Protease Inhibitors; D014020:Tissue Extracts; D016220:Fibroblast Growth Factor 1