Hepfer R Glenn, Brockbank Kelvin G M, Chen Zhen, Greene Elizabeth D, Campbell Lia H, Wright Gregory J, Linthurst-Jones Alyce, Yao Hai
Clemson-MUSC Joint Bioengineering Program, Department of Bioengineering, Clemson University, 173 Ashley Avenue MSC 508, Charleston, SC, 29425, USA.
Tissue Testing Technologies LLC, North Charleston, SC, USA.
Cell Tissue Bank. 2016 Sep;17(3):531-9. doi: 10.1007/s10561-016-9560-y. Epub 2016 Apr 29.
In regard to evaluating tissue banking methods used to preserve or otherwise treat (process) soft allograft tissue, current tests may not be sufficiently sensitive to detect potential damage inflicted before, during, and after processing. Using controlled parameters, we aim to examine the sensitivity of specific biomechanical, electrical, and biological tests in detecting mild damage to collagen. Fresh porcine pulmonary heart valves were treated with an enzyme, collagenase, and incubated using various times. Controls received no incubation. All valves were cryopreserved and stored at -135 °C until being rewarmed for evaluation using biomechanical, permeability, and cell viability tests. Statistically significant time dependent changes in leaflet ultimate stress, (p = 0.006), permeability (p = 0.01), and viability (p ≤ 0.02, four different days of culture) were found between heart valves subjected to 0-15 min of collagenase treatment (ANOVA). However, no statistical significance was found between the tensile modulus of treated and untreated valves (p = 0.07). Furthermore, the trends of decreasing and increasing ultimate stress and viability, respectively, were somewhat inconsistent across treatment times. These results suggest that permeability tests may offer a sensitive, quantitative assay to complement traditional biomechanical and viability tests in evaluating processing methods used for soft tissue allografts, or when making changes to current validated methods. Multiple test evaluation may also offer insight into the mechanism of potential tissue damage such as, as is the case here, reduced collagen content and increased tissue porosity.
关于评估用于保存或处理(加工)同种异体软组织的组织库方法,当前的测试可能不够灵敏,无法检测在加工前、加工过程中和加工后造成的潜在损伤。我们旨在通过控制参数,检验特定生物力学、电学和生物学测试在检测胶原蛋白轻度损伤方面的敏感性。用胶原酶对新鲜猪肺动脉瓣进行处理,并在不同时间进行孵育。对照组不进行孵育。所有瓣膜均进行冷冻保存,并在-135°C下储存,直至复温后使用生物力学、通透性和细胞活力测试进行评估。在接受0至15分钟胶原酶处理的心脏瓣膜之间,发现瓣叶极限应力(p = 0.006)、通透性(p = 0.01)和活力(p≤0.02,四天不同培养时间)存在统计学上显著的时间依赖性变化(方差分析)。然而,处理过的瓣膜和未处理的瓣膜的拉伸模量之间未发现统计学显著性(p = 0.07)。此外,极限应力和活力分别下降和上升的趋势在不同处理时间内有些不一致。这些结果表明,通透性测试可能提供一种灵敏的定量分析方法,以补充传统的生物力学和活力测试,用于评估同种异体软组织的加工方法,或在对当前经过验证的方法进行更改时。多种测试评估也可能有助于深入了解潜在组织损伤机制,如此处所示,即胶原蛋白含量降低和组织孔隙率增加。