Schultheiss Dirk, Lorenz Ralf R, Meister Roland, Westphal Mareike, Gabouev Alexander I, Mertsching Heike, Biancosino Christian, Schlote Norbert, Wefer Jörg, Winkler Michael, Stief Christian G, Jonas Udo
Department of Urology and Pediatirc Urology and Tissue Engineering Network, Hannover Medical School, Germany.
Eur Urol. 2004 Jun;45(6):781-6. doi: 10.1016/j.eururo.2004.01.001.
The aim of the present study was to generate a tissue engineered type of mechanically stable graft suitable for surgical replacement of the tunica albuginea penis.
Porcine fibroblasts isolated from open fascia biopsies were seeded on decellularized collagen matrices and then cultivated in a bioreactor under continuous multiaxial stress for up to 21 days (n=12). Static cultures without mechanical stress served as controls. Cell proliferation, cell alignment, and de novo synthesis of extracellular matrix proteins (proteoglycans, procollagen I, elastin) in these grafts was evaluated by hematoxylin-eosin, pentachrome, and immuno-staining. Additionally, the enzymatic isolation of porcine fibroblasts from X4mm skin punch biopsies (n=8) was evaluated.
Mechanically strained cultures of fibroblasts showed a homogeneous multilayer matrix infiltration and a regular cell alignment in the direction of strain axis after 7 days, as well as a de novo production of extracellular matrix proteins compared to the static control. A large amount of viable fibroblasts was easily obtained from small skin punch biopsies.
This study shows that continuous multiaxial stimuli improve proliferation and extracellular matrix synthesis of mature fibroblasts reseeded on a biological matrix making this a feasible autologous tissue engineered graft for penile surgery. For the clinical setting fibroblasts harvested from small skin biopsies can be a comfortable cell source.
本研究的目的是生成一种组织工程化的机械稳定移植物，适用于阴茎白膜的手术替代。
从开放筋膜活检中分离出的猪成纤维细胞接种在脱细胞胶原基质上，然后在生物反应器中在持续多轴应力下培养长达21天（n = 12）。无机械应力的静态培养作为对照。通过苏木精 - 伊红染色、丽春红染色和免疫染色评估这些移植物中的细胞增殖、细胞排列以及细胞外基质蛋白（蛋白聚糖、原胶原I、弹性蛋白）的重新合成。此外，还评估了从4mm皮肤穿刺活检中酶法分离猪成纤维细胞（n = 8）的情况。
与静态对照相比，成纤维细胞的机械应变培养在7天后显示出均匀的多层基质浸润和沿应变轴方向的规则细胞排列，以及细胞外基质蛋白的重新合成。从小皮肤穿刺活检中很容易获得大量存活的成纤维细胞。
本研究表明，持续多轴刺激可改善接种在生物基质上的成熟成纤维细胞的增殖和细胞外基质合成，使其成为阴茎手术可行的自体组织工程移植物。对于临床应用，从小皮肤活检中获取的成纤维细胞可以是一种方便的细胞来源。
