Drewa T, Sir J, Czajkowski R, Wozniak A
The Laboratory for Tissue Engineering, Medical Biology Department, N. Copernicus University, ul. Karlowicza 24, 85-092 Bydgoszcz, Torun, Poland.
Transplant Proc. 2006 Jan-Feb;38(1):133-5. doi: 10.1016/j.transproceed.2005.11.086.
Tissue-engineering methods using synthetic biodegradable scaffolds seeded with cells have potential to induce regeneration to a functional bladder wall. The aim of the study was to induce in vivo urothelial growth on implanted scaffolds previously seeded with stromal cells as compared with matrices implanted without cells for rat cystoplasty augmentation.
3T3 mouse fibroblasts were multiplied up to total of 10(8) cells. Cells were grown on Dulbecco's modified essential medium supplemented with 10% of fetal bovine serum and antibiotics in CO(2) chambers. Cells were seeded on biodegradable polyglycolic acid (PGA) scaffolds in eight rats: four bladders were augmented with cell-seeded grafts and the other four with acellular scaffolds. Rats were sacrificed after 4 months in preparation for hematoxylin and eosin staining.
One death in the acellular cystoplasty group was observed after 3 weeks. No epithelial layer was observed in the central part of the acellular graft. The cell-seeded grafts showed good visible multilayered epithelium with at least five layers of epithelial cells in the central part. The epithelium resembled rat native urothelium. The cell-seeded grafts showed a high degree of implanted 3T3 cells infiltration with good degradation of PGA fibers.
Our data indicated that urothelial proliferation on PGA grafts was intensified using a "feeder layer" of fibroblasts.
利用接种细胞的合成可生物降解支架的组织工程方法有潜力诱导功能性膀胱壁再生。本研究的目的是与未接种细胞的植入基质相比,诱导预先接种基质细胞的植入支架在体内产生尿路上皮生长,用于大鼠膀胱扩大成形术。
将3T3小鼠成纤维细胞增殖至总数达10⁸个细胞。细胞在补充有10%胎牛血清和抗生素的杜尔贝科改良伊格尔培养基中于二氧化碳培养箱中生长。将细胞接种在八只大鼠的可生物降解聚乙醇酸(PGA)支架上:四只膀胱用接种细胞的移植物进行扩大,另外四只用无细胞支架。4个月后处死大鼠,准备进行苏木精和伊红染色。
无细胞膀胱扩大成形术组在3周后观察到1只死亡。在无细胞移植物的中央部分未观察到上皮层。接种细胞的移植物在中央部分显示出良好的可见多层上皮,至少有五层上皮细胞。该上皮类似于大鼠天然尿路上皮。接种细胞的移植物显示出高度的植入3T3细胞浸润,PGA纤维降解良好。
我们的数据表明,使用成纤维细胞“饲养层”可增强PGA移植物上的尿路上皮增殖。