Zünd G, Hoerstrup S P, Schoeberlein A, Lachat M, Uhlschmid G, Vogt P R, Turina M
Clinic for Cardiovascular Surgery, University Hospital, Zurich, Switzerland.
Eur J Cardiothorac Surg. 1998 Feb;13(2):160-4. doi: 10.1016/s1010-7940(97)00309-6.
In tissue engineering the material properties of synthetic compounds are chosen to enable delivery of dissociated cells onto a scaffold in a manner that will result in in vitro formation of a new functional tissue. The seeding of human fibroblasts followed by human endothelial cells on resorbable mesh is a precondition of a successful creation of human tissues such as vessels or cardiac valves.
Polymeric scaffolds (n = 18) composed of polyglycolic acid (PGA) with a fiber diameter of 12-15 microm and a polymer density of 70 mg/ml were used as square sheets of 1 x 1 x 0.3 cm. Fibroblasts (passage 7) harvested from human foreskin were seeded (3.4 x 10(6)) and cultured over a 3 week period on a PGA-mesh, followed by seeding of endothelial cells (passage 5, 2.8 x 10(6)) harvested from human ascending aorta. Thereafter the new tissue was stained for HE, van Gieson, Trichrom Masson, Factor VIII and CD 34 and proved by scanning electron microscopy.
Microscopic examination of the seeded mesh demonstrated that the human fibroblasts were attached to the polymeric fibers and had begun to spread out and divide. The scanning electron microscopic examination demonstrated a homogeneous scaffold resembling a solid sheet of tissue. The seeded endothelial cells formed a monolayer on the fibroblasts and no endothelial cell invasion or new formation of capillaris could be detected.
These results are a first step to demonstrate that seeding of human fibroblasts and endothelial cells on PGA-mesh might be a feasible model to construct human tissues such as vessels or cardiac valves.
在组织工程中,选择合成化合物的材料特性,以便能够以一种方式将解离的细胞递送到支架上,从而在体外形成新的功能组织。将人成纤维细胞接种到可吸收网片上,随后接种人内皮细胞,这是成功创建诸如血管或心脏瓣膜等人造组织的前提条件。
使用由聚乙醇酸(PGA)组成的聚合物支架(n = 18),其纤维直径为12 - 15微米,聚合物密度为70毫克/毫升,制成1×1×0.3厘米的方形薄片。从人包皮中获取的成纤维细胞(第7代)接种(3.4×10⁶个),并在PGA网片上培养3周,随后接种从人升主动脉获取的内皮细胞(第5代,2.8×10⁶个)。此后,对新组织进行苏木精 - 伊红(HE)、范吉森、三色马森、因子VIII和CD 34染色,并通过扫描电子显微镜进行验证。
对接种网片的显微镜检查表明,人成纤维细胞附着在聚合物纤维上,并已开始伸展和分裂。扫描电子显微镜检查显示出一个类似实体组织薄片的均匀支架。接种的内皮细胞在成纤维细胞上形成单层,未检测到内皮细胞侵袭或新毛细血管形成。
这些结果是证明在PGA网片上接种人成纤维细胞和内皮细胞可能是构建诸如血管或心脏瓣膜等人造组织的可行模型的第一步。