Marsit Nagi, Dwejen Samira, Saad Ibrahim, Abdalla Sedigh, Shaab Arej, Salem Salma, Khanfas Enas, Hasan Anas, Mansur Mohamed, Abdul Sammad Mohamed
Tissue Banking Research Group, Biotechnology Research Center, Tweisha, Libyan Authority for Research, Science and Technology, P. O. Box 30313, Tripoli, Libya,
Cell Tissue Bank. 2014 Dec;15(4):603-11. doi: 10.1007/s10561-014-9433-1. Epub 2014 Mar 28.
Preparation of amniotic membrane (AM) by air drying method followed by radiation sterilization is simple and valuable approach; sterility and quality of the final AM product are depending on the quality management system at the tissue bank. Validation and substantiation of radiation sterilization dose (RSD) for tissue allografts is an essential step for the development and validation of the standard operating procedures (SOP). Application of SOP is perfectly relying on trained staff. Skills differences among personnel involved in AM preparation could have an effect on microbiological quality of the finished product and subsequently on the RSD required. AM were processed by four different couples of the tissue bank technicians. The AM grafts were randomly selected and subjected to bioburden test to validate and substantiate the 25 kGy RSD. Bioburden test for AM grafts were also useful to evaluate the skill of the tissue bank technicians and thus, to validate the current SOP for air dried AM. Moreover, the effect of placental source on bioburden counts on AM grafts was assessed. Substantiation of the 25 kGy RSD at a sterility assurance level of 10(-1), and sample item portion = 1, was carried out using Method VD max (25) of the International Organization for Standardization, document no. 11137-2 (ISO in Sterilization of healthcare products-radiation-part 2: establishing the sterilization dose, Method VDmax-substantiation of 25 kGy or 15 kGy as the sterilization dose, International Standard Organization, 2006). The results showed that there were no significant differences in the bioburdens of the four batches (α = 1 %), this means no significant differences in the skill of the four couples of the tissue bank technicians in terms of their ability to process AM according to the air dried AM SOP. The 25 kGy RSD was validated and substantiated as a valid sterilization dose for the AM prepared with the current established SOP at the Biotechnology Research Center experimental tissue bank. The donor's type of delivery, normal or caesarean, showed no significant effect on the levels of microbial counts on the tested AMs (α = 1 %).
采用空气干燥法制备羊膜（AM）并进行辐射灭菌是一种简单且有价值的方法；最终AM产品的无菌性和质量取决于组织库的质量管理体系。组织同种异体移植物辐射灭菌剂量（RSD）的验证和证实是制定和验证标准操作规程（SOP）的关键步骤。SOP的应用完全依赖于训练有素的工作人员。参与AM制备的人员技能差异可能会影响成品的微生物质量，进而影响所需的RSD。AM由组织库技术人员的四对不同组合进行处理。随机选择AM移植物并进行生物负荷测试，以验证和证实25 kGy的RSD。AM移植物的生物负荷测试也有助于评估组织库技术人员的技能，从而验证当前空气干燥AM的SOP。此外，还评估了胎盘来源对AM移植物生物负荷计数的影响。使用国际标准化组织文件编号为11137 - 2（《医疗保健产品灭菌 - 辐射 - 第2部分：确定灭菌剂量，方法VDmax - 证实25 kGy或15 kGy作为灭菌剂量》，国际标准化组织，2006年）中的方法VD max (25)，在无菌保证水平为10(-1)且样品项目部分 = 1的情况下，对25 kGy的RSD进行了证实。结果表明，四批产品的生物负荷无显著差异（α = 1%），这意味着组织库技术人员的四对组合在按照空气干燥AM SOP处理AM的能力方面技能无显著差异。25 kGy的RSD被验证并证实为生物技术研究中心实验组织库采用当前既定SOP制备的AM的有效灭菌剂量。供体的分娩方式（顺产或剖宫产）对测试的AM上的微生物计数水平无显著影响（α = 1%）。
