成纤维细胞促进真皮替代物血管化的作用机制

MECHANISM OF THE FIBROBLAST INDUCING THE VASCULARIZATION OF DERMAL SUBSTITUTE

目的 探讨成纤维细胞促进真皮替代物移植后血管化的作用机制。 方法 将成纤维细胞种植于无细胞真皮表面培养 ,形成活性真皮替代物 ,采用 EL ISA法测定培养上清中白介素 - 8(IL- 8)和转化生长因子 - β1 (TGF- β1 )的含量 ,RT- PCR法检测成纤维细胞中酸、碱性成纤维细胞生长因子 (a FGF、b FGF) m RNA的表达量。将含成纤维细胞的真皮替代物植入 BAL B/ C小鼠全层皮肤缺损创面 ,采用原位杂交的方法观察成纤维细胞的转归。 结果 成纤维细胞种植于无细胞真皮表面生长良好 ,可形成单层细胞膜片 ,并分泌 IL - 8(192 .3± 15 .9) pg/ ml和TGF- β1 (1.10 5± 0 .0 5 1) pg/ ml,表达 a FGF、b FGF。植入创面后 3周 ,成纤维细胞仍成活并生长、增殖。 结论 成纤维细胞分泌多种与血管形成有关的活性肽 ,移植后继续生长、增殖 ,可能对真皮替代物的血管化具有重要作用。

Objective To investigate the possible mechanism of the fibroblasts inducing the vascularization of dermal substitute. Methods Fibroblasts were seeded on the surface of acellular dermal matrix and cultivated in vitro to construct the living dermal substitute. The release of interleukin 8 (IL 8) and transfonming growth factor β 1(TGF β 1) in culture supernatants were assayed by enzyme linked immunosorbent assay, the mRNA expression of acid fibroblast growth factor (aFGF) and basic fibroblast growth factor (bFGF) were detected by RT PCR. Then, the living substitute was sutured to full thickness excised wound on BALB/C mice, and the fate of fibroblast was observed by using in situ hybridization. Results Fibroblasts cultured on acellular dermal matrix proliferated and reached a single layer confluence. Fibroblasts could secret IL 8(192.3±15.9)pg/ml and TGF β 1(1.105±0.051)pg/ml. There was the mRNA exparession of aFGF and bFGF. Fibroblasts still survived and proliferated 3 weeks after grafting. Conclusion Peptides secreted by fibroblasts and its survival after grafting may be relative to the vascularization of the dermal substitute.