Quantifying the ultrastructure changes of air-dried and irradiated human amniotic membrane using atomic force microscopy: a preliminary study
- 32 Downloads
Air-dried and sterilized amnion has been widely used as a dressing to treat burn and partial thickness wounds. Sterilisation at the standard dose of 25 kGy was reported to cause changes in the morphological structure as observed under the scanning electron microscope. This study aimed to quantify the changes in the ultrastructure of the air-dried amnion after gamma-irradiated at several doses by using atomic force microscope. Human placentae were retrieved from mothers who had undergone cesarean elective surgery. Amnion separated from chorion was processed and air-dried for 16 h. It was cut into 10 × 10 mm, individually packed and exposed to gamma irradiation at 5, 15, 25 and 35 kGy. Changes in the ultrastructural images of the amnion were quantified in term of diameter of the epithelial cells, size of the intercellular gap and membrane surface roughness. The longest diameter of the amnion cells reduced significantly after radiation (p < 0.01) however the effect was not dose dependent. No significant changes in the shortest diameter after radiation, except at 35 kGy which decreased significantly when compared to 5 kGy (p < 0.01). The size of the irradiated air-dried amnion cells reduced in the same direction without affecting the gross ultrastructure. At 15 kGy the intercellular gap decreased significantly (p < 0.01) with Ra and Rq, values reflecting surface roughness, were significantly the highest (p < 0.01). Changes in the ultrastructure quantified by using atomic force microscope could complement results from other microscopic techniques.
KeywordsAir-dried amniotic membrane Atomic force microscopy Gamma irradiation Ultrastructure Epithelial cells
This study was supported by Bone Bank Internal Fund and University Malaya Research Grant (RG542-13HTM) and HIR-MoE Grant; UM.C/625/1/HIR/MOHE/CHAN/03-A000003-50001). The authors thanked Ms Nurhafizatul Nadia Hanafi and Mr Mohd Khalil Salleh for their technical assistance in the study.
Compliance with ethical standards
Conflict of interest
All authors declared no competing of interest in the study.
- AATB (2008) Standards for tissue banking, 12th edn. American Association of Tissue Banks, VirginiaGoogle Scholar
- Deocaris CC, Abad LB, Enriquez EP, Guzman ZMD, Aliganga AA, Tangonan MB, Tolentino MM, Ignacio LV, Deocaris CC (2005) Radiolytic damage to freeze-dried human amniotic membrane. Philipp J Sci 134(1):45–50Google Scholar
- Eskandarlou M, Azimi M, Rabiee S, Seif RMA (2016) The healing effect of amniotic membrane in burn patients. World J Plast Surg 5(1):39–44Google Scholar
- Ficicoglu C, Ozden S, Kavak Z, Okar I (1998) Effects of active labour on the ultrastructure of amnion epithelium. Med Sci Res 26(2):135–139Google Scholar
- Hilmy N, Siddik S, Gentur S, Gulardi W (1987) Physical and chemical properties of freeze-dried amnion chorion membranes sterilized by γ-Irradiation. Atom Indones 13(2):1–14Google Scholar
- Kamalia NZ, Suzina SABH, Yusof N (2014) Changes in biophysical properties of human amniotic membranes after different preservation methods and radiation sterilization. Regen Res 3(1):64–70Google Scholar
- Kanthan SR, Ganesan K, Azura M, Kamarul T (2010) The difference preparations of human amniotic membrane (HAM) as a potential cell carrier for condrocytes. Eur Cell Mater 20(2):46Google Scholar
- Loeffelbein DJ, Baumann C, Stoeckelhuber M, Hasler R, Mücke T, Steinsträßer L, Drecoll E, Wolff KD, Kesting MR (2012) Amniotic membrane as part of a skin substitute for full thickness wounds: an experimental evaluation in a porcine model. J Biomed Mater Res B Appl Biomater 100(5):1245–1256. https://doi.org/10.1002/jbm.b.32689 CrossRefGoogle Scholar
- Nguyen H, Cassady AI, Bennett MB, Gineyts E, Wu A, Morgan DA, Forwood MR (2013) Reducing the radiation sterilization dose improves mechanical and biological quality while retaining sterility assurance levels of bone allografts. Bone 57(1):194–200. https://doi.org/10.1016/j.bone.2013.07.036 CrossRefGoogle Scholar
- Yusof N, Hilmy N (2010) Physical and mechanical properties of radiation-sterilized amnion. In: Nather A, Yusof N, Hilmy N (eds) Allograft procurement, processing and transplantation—a comprehensive guide for tissue banks. World Scientific Publishing, Singapore, pp 155–168Google Scholar