The International Atomic Energy Agency
(IAEA) research on radiation sterilization in
tissue banking Oleg Belyakov
Radiation Biologist
Applied Radiation Biology and Radiotherapy Section
Division of Human Health
Department of Nuclear Sciences and Applications
IAEA is involved more
than 30 years in the use
of the ionising radiation
techniques for tissue
banking
Pedraza JM. The Use of the Ionizing Radiation Technique for Tissue Sterilization: The International Atomic Energy Agency (IAEA) Experience: Nova Science; 2011.
Mr Jorge Morales
Pedraza, Radiation
and Tissue Banking
Programme former
Manager in the
Technical Cooperation
department of IAEA
• The use of tissue allografts in surgical procedures has
increased tremendously over the last several years.
• Radiation has been used to sterilize tissue allografts on a
large scale.
• Tissue banks use radiation doses just based on empirical
and/or historical data, or use a fixed dose of 15 or 25 kGy for
all tissue allografts.
• In order to improve the knowledge on radiosterilisation of
tissue allografts studies were designed and conducted
under controlled and validated conditions.
• Five main categories of tissues were studied: bone,
demineralized bone, cartilage allograft, skin and amnion.
IAEA Coordinated Research Project (CRP)
E31006 “Safety and Optimization of Radiation
Sterilization in Tissue Banking: Studies on
Functional Properties of Irradiated Tissue Grafts”
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16 participating institutions
from 16 IAEA Members States
Five year CRP (2010-2015, to be formally closed in 2017)
Research Contracts – black, Research Agreements – green 4
AUL - GU AUL - VIFM GFR – UKC
UK - OTBVTB ARG - CNEA BRA - IPEN
CHI - CCHEN COL - SDS CUB - CEADEN
IND - TMH INS - AU MAL - USM
PER - IPEN POL - WUM SLR - UNB
URU - INDT
IAEA Project Officer – Oleg Belyakov, Alternate - Eduardo
Rosenblatt, started by Jan Wondergem
Overall Objectives
Main aims of the project:
• to find the optimal radiation dose and processing methods for several tissues providing allograft sterility (SAL 10-6) without compromising tissue biological or structural function for clinical use;
• to facilitate research and disseminate research results in IAEA Member States to study radiation-induced effects on tissue grafts processed and preserved by different methods and;
• to initiate collaboration between relevant laboratories, establishing of exchange programmes and joint applications for future additional funding.
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Specific research objectives
• Study interaction between processing methods and irradiation process.
• Establish dose response relationship and validation of relevant physical, chemical, biological and clinical end-points.
• Study effect of irradiation conditions (temperature, oxygen, water content, dose rate, radio-protectants, etc.) on biological properties of tissue allografts.
• To improve tissue banking processing and preservation protocols a number of questions have been addressed.
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• What type of testing methods should be applied for tissue
allografts?
• What is the optimum processing and preservation method to
preserve functionality of tissue grafts?
• What is the maximum tolerated irradiation dose for different levels
of damage in different tissue allografts (type of tissue, method of
processing)?
• Which processing combinations have the highest potential
reducing the radiation-induced tissue toxicity in specific tissues?
• Which processing combinations have the highest potential
preserving functionality of irradiated tissues?
• Is there relation of dose-rate on physical, chemical, biological
properties of tissue allografts?
Results: bone and
demineralized bone
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• Bone: adoption of sterilization doses below 15 kGy improves tissue
quality and surgical outcomes (left panel). Higher radiation
decreases several mechanical properties of bone rings (right
panel).
• Demineralized bone (DMB): sufficient osteoinduction was observed
after experiments using DBM treated at 15 kGy radiation dose.
Courtesy Artur Kamisnski, Warsaw, Poland
Courtesy of
Sven Scheffler,
Berlin,
Germany
Results: cartilage and tendons
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Studies demonstrated that Optical Coherence Tomography (OCT) is a
suitable non-destructive technique to evaluate the changes in costal
cartilage after sterilisation by ionizing radiation. Biomechanical testing of
tendons showed decreased failure load, stiffness and AP-laxity, in the E-
beam group compared to the fresh frozen allograft group and a significant
decreased failure load compared to autograft group (P = 0.024).
Comparison of failure load, stiffness and ap-laxity
values of Ebeam, fresh frozen allograft (FFA),
autograft (Auto) and native ACL (ACL) at 6 and 12
weeks, asterisk on ACL indicates significant
difference compared to all other groups. In the
Ebeam group, stiffness and failure load decreased
from 6 to 12 weeks and AP-laxity increased. Contrary
to this, failure load increased in fresh frozen allografts
(FFA), while stiffness and AP-laxity stayed nearly
constant. In autograft group, all the aforementioned
parameters increased.
Courtesy of
Sven Scheffler,
Berlin,
Germany
Results: skin and amnion
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Most sensitive tissue to structural changes induced by radiation proved to
be animal skin, least sensitive human skin. Irradiation dose up to 25 kGy
had minor impact on the ultrastructure of the irradiated skin and its
functionality (evaporation index). In contrast to skin, morphological changes
related to the radiation dose were seen at doses of 25 kGy and up.
Surface morphology of glycerol preserved
and gamma irradiated human amnion
membrane (HAM) at magnification ×2,000,
using high vacuum scanning electron
microscopy (HVSEM); (a) Non-irradiated, (b)
15 kGy, (c) 25 kGy, (d) 35 kGy (courtesy
Suzina S. Ab Hamid, Kelatan, Malaysia).
Research Outputs
• New data on efficacy of irradiated tissue allografts for bone, tendon, cartilage, skin and amniotic membrane, vascular grafts and heart valves were obtained.
• Methodology for development of quality control testing of different types of (irradiated) tissues was formulated.
• Significant research progress in radiation sterilization and tissue banking was achieved in MSs.
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Research Outcomes
• Provision of technical expertise in processing procedures of tissues subsequently radiation-sterilized.
• Knowledge and expertise in tissue allograft irradiation was improved.
• Quality of tissue grafts was improved.
• Significant progress has been made by the participating tissue banks/institutes leading to the production of safer allografts both with respect to its functionality and sterility.
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Selected publications Suzina Sheikh Ab Hamid, Nor Kamalia Zahari, Norimah Yusof , Asnah Hassan (2014).
“Scanning electron microscopic assessment on surface morphology of preserved human
amniotic membrane after gamma sterilisation.” Cell Tissue Bank, (15):15–24.
Anna Jastrzebska , Artur Kaminski, Ewelina Grazka, Joanna Marowska, Jaroslaw Sadlo,
Grzegorz Gut, Izabela Uhrynowska-Tyszkiewicz (2013). “Effect of gamma radiation and
accelerated electron beam on stable paramagnetic centers induction in bone mineral: influence
of dose, irradiation temperature and bone defatting.” Cell Tissue Bank, electronic version.
Artur Kaminski, Ewelina Grazka , Anna Jastrzebska, Joanna Marowska, Grzegorz Gut, Artur
Wojciechowski,Izabela Uhrynowska-Tyszkiewicz (2012). “Effect of accelerated electron beam
on mechanical properties of human cortical bone: influence of different processing methods.”
Cell Tissue Bank (13):375–386.
George Morales, Astrid Lobo Gajwala (2012). “A review of the International Atomic Energy
Agency (IAEA) International Standards of Tissue banking.” Cell Tissue bank, (13): 15-25.
Peter Myint, Jan Wondergem, Yaroslav Pynda, Glyn O. Phillips (2013). “An international
survey of tissue banking, a preliminary report.” Cell Tissue Bank, (14):159- 165.
Huynh Nguyen, Alan I. Cassdy, Michael B. Bennett, Andy Wu, David A.F. Morgan, Mark R.
Forwood (2013). “Reducing the radiation sterilization dose improves mechanical and biological
quality while retaining sterility assurance levels of bone allografts.” Bone, (57):194-200.
Schmidt T, Hoburg A, Broziat C, Smith MD, Gohs U, Pruss A, Scheffler S. Cell Tissue Bank
(2012). “Sterilization with electron beam irradiation influences the biomechanical properties
and the early remodeling of tendon allografts for reconstruction of the anterior cruciate
ligament (ACL).” Cell Tissue Bank, (13): 387-400. 12
Future direction
• Tissue engineering is revolutionising medicine by shifting the
focus from addressing symptoms to repair and regeneration.
• New IAEA CRP F23030-E31007 Instructive Surfaces and
Scaffolds for Tissue Engineering Using Radiation Technology
(2014-2021) was launched to reflect this change.
• The CRP is jointly conducted by the Radioisotope Products
and Radiation Technology Section (RPRT)/Division of
Physical and Chemical Sciences (NAPC) as well as the
Applied Radiation Biology and Radiotherapy Section
(ARBR)/Division of Human Health (NAHU).
• NAPC implements the part related to the development and
testing of the instructive surfaces and scaffolds, while NAHU
implements the biomedical application part related to the end
uses. 13
Success story
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• TC project PER6017 Improving
National Capabilities to Treat
Patients with Burns, Lesions and
Polytraumatized Conditions
Through the Application of
Radiation-Processed Cells,
Scaffolds and Tissues (2014-
2017) helped to transfer skin
regeneration technology to Peru.
• 13-year patient with 80 per cent
of skin severely burnt, usually
such medical condition leads to
death.
• Thanks to the technology of skin
cultivation, doctors were able to
grow from 2 cm2 of healthy
patient's skins 2 m2 of skin to
cover wounds.
• The patient recovered in 2 month.
IAEA News Story at:
https://www.iaea.org/newscenter/news/peruvi
an-boys-life-saved-thanks-to-radiation-
technology
I would like to acknowledge Chief
Scientific Investigators of Institutions,
contributed to CRP E31006: Paulina
Maria Estela Aguirre Herrera (CHI), Dr
Emma Castro Gamero (PER), Mark
Forwood (AUS), Clara Linda Guerrero
(COL), Marisa Herson (AUS), Eulogia
Kairiyama (ARG), Artur Marek
Kaminski (POL), Astrid Ann Lobo
Gajwala (IND), Jan Koller (SLO),
Menkher Manjas (INS), Monica
Beatriz Mathor and Wilson Aparecido
Calvo (BRA), Peter Myint (UK), Isabel
Maria Otero Abreu (CUB), María del
Carmen Salidas Farinella (URU),
Suzina Sheikh Ab. Hamid (MAL),Sven
Scheffler (GFR).
Participants of the 3rd RCM of CRP E31006,
IAEA Headquarters, VIC, Vienna, Austria, 3-5
September 2014