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器官培养中维持的小鼠胎儿肺切口伤口的再生愈合。

Regenerative healing of incisional wounds in murine fetal lungs maintained in organ culture.

作者信息

Blewett C J, Cilley R E, Ehrlich H P, Dillon P W, Blackburn J H, Krummel T M

机构信息

Division of Pediatric Surgery, Pennsylvania State University College of Medicine, Milton S. Hershey Medical Center, Hershey 17033, USA.

出版信息

J Pediatr Surg. 1995 Jul;30(7):945-8. doi: 10.1016/0022-3468(95)90318-6.

DOI:10.1016/0022-3468(95)90318-6
PMID:7472950
Abstract

Although fetal dermal repair is known to be fundamentally different from adult healing, the response to wounding in other organs is less well characterized. Scarless repair in mid-gestation dermis with a transition to adult-type healing at term has been shown in fetal organ culture. A lung explant culture system was used to investigate whether wound repair in the fetal lung shows characteristics similar to those found in fetal dermis. Lungs from 14-day and 18-day Cd-1 murine fetuses and 2-day-old newborns, (term = 20 days, n = 24) were wounded by linear incision and incubated at 37 degrees C, in a 21% O2, 5% CO2 environment, in BGJb supplemented with vitamin C and antibiotics. Medium was changed daily. Samples were fixed at 7 days and embedded in paraffin. Sections were stained with hematoxalyn-eosin and Masson Trichrome. Additional 14-day and 18-day samples were frozen in freon and immunohistochemical staining for TGF-beta performed. Other frozen tissues from each time point were homogenized and used to assay for endogenous TGF-beta levels by Western blot analysis. Histology showed reconstitution of tissue architecture across the wound in 14-day and 18-day specimens. In representative histological sections, intact bronchial architecture developed across the previous wound site. No cellular inflammatory response was observed, and collagen deposition was undetectable at the site of the wound by Trichrome staining. By 22 days the lung explants showed a much less ordered repair, including disorganized collagen deposition.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

尽管已知胎儿皮肤修复与成人愈合在根本上存在差异,但其他器官对创伤的反应却鲜为人知。胎儿器官培养显示,妊娠中期真皮无瘢痕修复,足月时转变为成人型愈合。采用肺外植体培养系统研究胎儿肺的伤口修复是否具有与胎儿真皮相似的特征。将14日龄和18日龄的Cd-1小鼠胎儿以及2日龄新生儿(足月为20天,n = 24)的肺进行线性切口创伤,并在补充了维生素C和抗生素的BGJb培养基中,于37℃、21% O₂、5% CO₂环境中培养。每天更换培养基。7天后固定样本并石蜡包埋。切片用苏木精-伊红和Masson三色染色。另外,将14日龄和18日龄的样本冷冻于氟利昂中,进行TGF-β的免疫组化染色。对每个时间点的其他冷冻组织进行匀浆,并用蛋白质免疫印迹分析测定内源性TGF-β水平。组织学显示,14日龄和18日龄样本的伤口处组织结构得以重建。在代表性组织学切片中,完整的支气管结构在先前伤口部位形成。未观察到细胞炎症反应,三色染色未在伤口部位检测到胶原沉积。到22天时,肺外植体的修复则不太有序,包括胶原沉积紊乱。(摘要截选至250词)

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