Bittinger F, Schepp C, Brochhausen C, Lehr H A, Otto M, Köhler H, Skarke C, Walgenbach S, Kirkpatrick C J
Department of Pathology, Johannes Gutenberg University, Mainz, Federal Republic of Germany.
J Surg Res. 1999 Mar;82(1):28-33. doi: 10.1006/jsre.1998.5449.
Intraabdominal adhesions are a common complication following laparotomy. Since the exact mechanisms involved in this processes are unknown we have analyzed in vitro the role of mesothelial cells in peritoneal healing.
Human mesothelial cells from omental tissue were cultivated for 2 weeks in a three-dimensional culture either on or in a collagen type I matrix. The effects of blood and collagen matrix were analyzed by exposing mesothelial cells to an overlying blood clot, simulating intraperitoneal bleeding, or a second collagen layer. The production of collagen types III and IV, fibronectin, and laminin was analyzed with immunohistochemical methods.
Mesothelial cells grown on a collagen matrix formed a monolayer of flat or cobblestone-like cells whereas those cultivated in a collagen matrix exhibited spindle-like morphology. Mesothelial cells failed to grow into an overlying collagen matrix, but did grow into a blood clot, emphasizing a potential role of blood clots in peritoneal adhesion formation. Independent of the culture systems mesothelial cells produced collagen type III, fibronectin, and laminin but not collagen type IV.
Our experiments demonstrate remodeling of peritoneal-like structures by mesothelial cells in a three-dimensional culture reflecting their putative role in the reepithelialization after serosal defects, and also in the formation of peritoneal adhesions.
腹腔粘连是剖腹手术后常见的并发症。由于该过程的确切机制尚不清楚,我们在体外分析了间皮细胞在腹膜愈合中的作用。
将来自网膜组织的人间皮细胞在三维培养中于I型胶原基质上或基质内培养2周。通过将间皮细胞暴露于覆盖的血凝块(模拟腹腔内出血)或第二层胶原层来分析血液和胶原基质的作用。采用免疫组织化学方法分析III型和IV型胶原、纤连蛋白和层粘连蛋白的产生。
在胶原基质上生长的间皮细胞形成单层扁平或鹅卵石样细胞,而在胶原基质内培养的间皮细胞呈现纺锤样形态。间皮细胞不能生长到覆盖的胶原基质中,但能生长到血凝块中,这强调了血凝块在腹膜粘连形成中的潜在作用。无论培养系统如何,间皮细胞都能产生III型胶原、纤连蛋白和层粘连蛋白,但不产生IV型胶原。
我们的实验证明了间皮细胞在三维培养中对类似腹膜结构的重塑,这反映了它们在浆膜缺损后再上皮化以及腹膜粘连形成中的假定作用。