Department of Materials Science and Engineering, Kroto Research Institute, Broad Lane, Sheffield, University of Sheffield, Sheffield, UK.
Eur Urol. 2011 Oct;60(4):856-61. doi: 10.1016/j.eururo.2011.07.045. Epub 2011 Jul 26.
We previously described the production and clinical outcomes of tissue-engineered buccal mucosa (TEBM) used to treat recurrent urethral strictures. In this study, two patients developed a recurrent stricture and there was also evidence of graft contraction.
Assess possible preclinical methods to reduce contraction of TEBM.
DESIGN, SETTING AND PARTICIPANTS: Using the model of TEBM in use clinically (ie, oral keratinocytes and fibroblasts cultured on de-epidermised acellular dermal scaffold), three methods of reducing TEBM contraction were investigated in vitro.
The techniques assessed were pretreatment of de-epidermised dermis (DED) with glutaraldehyde, culture with β-aminopropionitrile (β-APN; a lysyl oxidase inhibitor), and physical restraint of TEBM grafts during culture.
Contraction was assessed using serial digital image analysis. The cytotoxicity of the pharmacologic manipulations was assessed using monolayer cultures of oral mucosa cells.
Control TEBM lost a mean of 45.4% of its original surface area over 28 d of culture. Treating TEBM with glutaraldehyde, β-APN, or mechanical restraint during culture all significantly inhibited graft contraction. Glutaraldehyde treatment was most effective (only 5.5% loss of area with 0.1% glutaraldehyde), followed by mechanical restraint for at least 7 d (21.4% loss of area), and then β-APN (28.7% loss of area). None of the treatments had any significant effect on cell viability. This in vitro study identifies solutions for graft contracture to explore in the clinic.
Glutaraldehyde pretreatment and restraint of TEBM grafts during culture both reduce graft contraction.
我们之前描述了用于治疗复发性尿道狭窄的组织工程颊黏膜（TEBM）的生产和临床结果。在这项研究中，两名患者出现了复发性狭窄，并且有移植物收缩的证据。
评估减少 TEBM 收缩的可能临床前方法。
设计、设置和参与者：使用临床上使用的 TEBM 模型（即口腔角质形成细胞和纤维母细胞在去表皮化无细胞真皮支架上培养），在体外研究了三种减少 TEBM 收缩的方法。
评估的技术包括用戊二醛预处理去表皮真皮（DED）、用β-氨基丙腈（β-APN；赖氨酰氧化酶抑制剂）培养和在培养过程中物理限制 TEBM 移植物。
使用连续数字图像分析评估收缩。使用口腔黏膜细胞单层培养评估药物处理的细胞毒性。
对照 TEBM 在 28 天的培养过程中平均损失了其原始表面积的 45.4%。用戊二醛、β-APN 或机械限制处理 TEBM 在培养过程中均可显著抑制移植物收缩。戊二醛处理最有效（仅用 0.1%戊二醛处理，面积损失 5.5%），其次是机械限制至少 7 天（面积损失 21.4%），然后是β-APN（面积损失 28.7%）。这些处理都没有对细胞活力产生任何显著影响。这项体外研究确定了可在临床上探索的移植物挛缩解决方案。
戊二醛预处理和培养过程中 TEBM 移植物的限制都可以减少移植物收缩。
