[A comparative study on preparing acellular larynx scaffold between perfusing method and immersing method].
OBJECTIVE
To compare the difference of preparing the acellular larynx scaffold between perfusion method and immersion method, and find better way to make acellular larynx scaffold for tissue engineering.


METHODS
Twenty 6-month-old male New Zealand rabbits, weighing 2.0-2.5 kg, were divided into perfusion group (n=10) and immersion group (n=10) at random. All the larynxes were excised in a sterile fashion. The acellular larynx scaffold was obtained by perfusion method and immersion method respectively, and then comparative examinations were performed by the macroscopic view, histological view, scanning electron microscope (SEM), cartilage vitality assay and toluidine blue staining.


RESULTS
Macroscopic view showed that the larynxes perfused by sodium dodecyl sulphate (SDS) became transparent after 2 hours of perfusion, but the larynxes immersed by SDS over 16 hours still appeared pink-white. Histology and SEM indicated that compared with immersion group, perfusion group showed better acellular effect, more vintages and collagen fibers were retained, no intact cell or nuclei remained in acellular matrix and chondrocytes were still survival. The porosity was 85.39% +/- 3.16% in perfusion group and 34.72% +/- 4.51% in immersion group, showing significant difference (P < 0.01). The chondrocyte vitality rate of perfusion group (86.93% +/- 1.52%) was higher than that of immersion group (77.73% +/- 1.66%), showing significant difference (P < 0.01). Toluidine blue staining showed that the chondrocyte heterochromatin was stronger in perfusion group than that in immersion group.


CONCLUSIONS
Compared with immersion method, perfusion method is a better way to construct acellular larynx scaffold because it can achieve better acellular effect and retain chondrocyte vitality at the greatest extent in the acellular larynx scaffold.
Hou
Nan
N
Department of Otorlaryngology, Tangdu Hospital, the Fourth Military Medical University, Xi'an Shaanxi, 710038, P.R. China.
Cui
Pengcheng
P
Luo
Jiasheng
J
Zheng
Deyu
D
D000818:Animals; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D007831:Larynx, Artificial; D008297:Male; D011817:Rabbits; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds