A State-of-the-art Review of Cytotoxicity Evaluation of Biomaterials
Fengsen MA1(),Yan YU1,Jie ZHANG1,Haibo CHEN2
1 Biologics and Biomaterials Laboratory, College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014 2 Zhejiang Institute of Medical Device Testing, Hangzhou 310018
Biomaterials are widely applied in medical field, and their biocompability evaluation before clinical application has become increasingly important. Cytotoxicity assay is an experimental method in vitro, which takes the advantages of simplicity, high speed, good repeatability and no ethical problems compared with the method in vivo. Accordingly, cytotoxicity assay has become a conventional examination for biomaterials before the clinical application, and played a significant role in biocompatibility evaluation. With the rapid development of modern cell biology and molecular biology technology, some new evaluation indicators and methods are urged to be included in the cytotoxicity evaluation standards at home and abroad. Simultaneously, the diversity of cytotoxicity assay methods highly depends on the development of new materials, the different purposes and application locations. This review summarized the methods of cytotoxicity evaluation, the cytotoxicity tests of the most commonly used materials and the problems in current cytotoxicity evaluation of biomaterials, which aims to pave the way for the researchers in relative fields.
Hussein K H, Park K M, Kang K S, et al.Biocompatibility evaluation of tissue-engineered decellularized scaffolds for biomedical application[J]. Materials Science and Engineering C, 2016,67:766.
[2]
Zhu M Q.Research on absorbable hemostatic gauze[D]. Hangzhou: Zhejiang University, 2014(in Chinese).
[2]
朱美琴. 可吸收止血纱布研究[D]. 杭州: 浙江大学, 2014.
[3]
奚廷斐. 医疗器械生物学评价[M]. 北京: 中国标准出版社, 2012:98.
[4]
Zange R, Kissel T.Comparative in vitro biocompatibility testing of polycyanoacrylates and poly (D, L-lactide-co-glycolide) using different mouse fibroblast (L929) biocompatibility test models[J]. European Journal of Pharmaceutics and Biopharmaceutics, 1997,44(2):149.
[5]
Lin H S, Wang C R, Wang Z J, et al.Progress in the studies of methods for cytobiocompatibility evaluation of biomaterials China Medical Device Information, 2011,17(9):10(in Chinese).
Lai F, Shen Z, Wen H, et al.A morphological identification cell cytotoxicity assay using cytoplasm-localized fluorescent probe (CLFP) to distinguish living and dead cells[J]. Biochemical and Biophysical Research Communications, 2017,482(2):257.
[7]
Uggeri J, Guizzardi S, Scandroglio R, et al.Adhesion of human osteoblasts to titanium: A morpho-functional analysis with confocal microscopy[J]. Micron, 2010,41(3):210.
[8]
Formentín P, Catalán ú, Alba M, et al.Effects of SiO2 micropillar arrays on endothelial cells’ morphology[J]. New Biotechnology, 2016,33(6):781.
[9]
Rocha V, Marques C, Figueiredo J L, et al.In vitro cytotoxicity evaluation of resveratrol-loaded nanoparticles: Focus on the challenges of in vitro methodologies[J]. Food & Chemical Toxicology, 2017,103:214.
[10]
WangH Y, Li R Y, Wang X H, et al. Application of neutral red staining in antifungal susceptibility test Chinese Journal of Laboratory Medicine, 1998, 21(1):43(in Chinese).
GuangS, Tian J, Wei G, et al. A modified fluorescein derivative with improved water-solubility for turn-on fluorescent determination of Hg2+ in aqueous and living cells[J]. Talanta, 2017,170:89.
[12]
VajrabhayaL, Sithisarn P, Wilairat P, et al. Comparison between Sulphorhodamine-B dye staining and 51Cr-release method in cytotoxicity assay of endodontic sealers[J]. Journal of Endodontics, 1997,23(6):355.
[13]
DengS H, Liu Y X. Research on optimal conditions for detecting LAK cell killing activity by 51 Crrelease assay Journal of Chengde Medical College, 1992,9(3):167(in Chinese).
FuH Y, Shao A L, Fang Y, et al. Study on key parameters of the in vitro cytotoxicity test in nanomaterials assessment Chinese Journal of Pharmaceutical Analysis, 2013,33(7):1096(in Chinese).
WangF H, Chen S X, Guo C Y, et al. Research on methodology for detecting cytotoxicity of NK cell by LDH release assay Chinese Journal of Immunology, 1990,6(2):115(in Chinese).
HexigB, Nakaoka R, Tsuchiya T. Safety evaluation of surgical materials by cytotoxicity testing[J]. Journal of Artificial Organs, 2008,11(4):204.
[17]
SangJ, Zhu S M, Kuang R. Comparison of MTT assay and relative growth rate method in vitro cytotoxicity test on pharmaceutical packaging material Chinese Journal of Modern Applied Pharmacy, 2015,32(5):571(in Chinese).
GaoS H, Wang H L, Wang S Y. Correlative study of proliferation of aorto smooth muscular cells by using flow cytometry and 3H-TdR incorporation in rats Journal of China Medical University, 2005,34(1):10(in Chinese).
TanY S, Xu Y X, Wei X H, et al. Comparative study of MTT colorimetric assay and 3H-TdR incorporation assay on the effects of MLC in swine Chinese Journal of Agricultural Biotechnology, 1997,5(3):274(in Chinese).
ChenW, Xing C, Hou F. Intra-laboratory study to determine the reproducibility of LLNA: BrdU-ELISA for the prediction of the skin sensitizing potential of chemicals[J]. Journal of Pharmacological and Toxicological Methods, 2016,82:26.
[21]
ZengB, Tang Y H, Wang A L. Cardiac cell proliferation assessed by EdU: A novel analysis of cardiac regeneration Medical Journal of Wuhan University, 2014,35(6):862(in Chinese).
LiaoN. Experimental study on the effect of labeling SD rat bone marrow mesenchymal stem cells with Edu[D]. Urumqi: Xinjiang Medical University, 2014(in Chinese).
23La?tovcka J, Rataj M, Bartuňková J. Assessment of lymphocyte proliferation for diagnostic purpose: Comparison of CFSE staining, Ki-67 expression and 3H-thymidine incorporation[J]. Human Immunology, 2016,77(12):1215.
[24]
OngT H D, Yu N, Meenashisundaram G K, et al. Insight into cytotoxicity of Mg nanocomposites using MTT assay technique[J]. Materials Science & Engineering C, 2017,78:647.
[25]
TanW D, Jin H, et al. Comparison of MTT with SRB assay in vitro anticancer drug screening Natural Product Research and Development, 1999,11(3):17(in Chinese).
GhoshA, Ali M A, Selvanesan L, et al. Structure-function characteristics of the biomaterials based on milk-derived proteins[J]. International Journal of Biological Macromolecules, 2010,46(4):404.
[27]
27T?lli M A, Ferreira M P A, Kinnunen S M, et al. In vivo biocompatibility of porous silicon biomaterials for drug delivery to the heart[J]. Biomaterials, 2014,35(29):8394.
[28]
28 LiK, Zhou R, Jia W W, et al. Zanthoxylum bungeanum essential oil induces apoptosis of HaCaT human keratinocytes[J]. Journal of Ethnopharmacology, 2016,186:351.
[29]
29 MaiL P, Zhong S L, Yang M, et al. Compare two different staining methods used in flow cytometric analysis of cell cycle Journal of Tropical Medicine, 2011,11(12):1363(in Chinese).
30 YangX F, He C E, Tang R H, et al. A comparative study on the measurement of neuronal cell apoptosis by Hoechst33342/PI double staining and TUNEL assay Carcinogenesis, Teratogenesis and Mutagenesis, 2014,26(3):180(in Chinese).
31 ZhangW, Liang Z H. Comparison between annexin V-FITC/PI and Hoechst33342/PI double stainings in the detection of apoptosis by flow cytometry Chinese Journal of Cellular and Molecular Immunology, 2014,30(11):1209(in Chinese).
32 TanX H, Zhang Y L, Jiang B, et al. Investigation of the influence factors in cell apoptosis by using flow cytometry with PI staining Journal of Southern Medical University, 2000,20(4):344(in Chinese).
33 LiuP, Ma Q Y, Song Y X, et al. Application of DAPI staining in detecting cell cycle of BHK-21 Shandong Journal of Animal Science and Veterinary Medicine, 2016,37(8):10(in Chinese).
34 ZhangC X, Liu J X, Dong X X, et al. The operating experience for TUNEL staining of apoptotic cell Laboratory Animal Science, 2014,31(4):46(in Chinese).
35 ZhaoT X, Jiang X, Wang X L, et al. Assay of apoptosis in human lung carcinoma cell under laser confocal microscope by FITC-Annexin V/PI Journal of Hubei College of Traditional Chinese, 2005,7(4):9(in Chinese).
36 MaoJ S, Cui Y L, Wang X H, et al. A preliminary study on chitosan and gelatin polyelectrolyte complex cytocompatibility by cell cycle and apoptosis analysis[J]. Biomaterials, 2004,25(18):3973.
[37]
37 CuiX, Liang T, Liu C, et al. Correlation of particle properties with cytotoxicity and cellular uptake of hydroxyapatite nanoparticles in human gastric cancer cells[J]. Materials Science and Engineering C, 2016,67:453.
[38]
38 RenD X. Development of the investigations on the bio-safety of nanomaterials[D]. Changchun: Northeast Normal University, 2010(in Chinese).
[38]
任德香. 纳米材料的生物安全性研究进展[D]. 长春: 东北师范大学, 2010.
[39]
39 ChenF, Liu Q, Wang L P, et al. Progress of the causes of cell toxicity of nanomaterials Orthopaedic Biomechanics Materials and Clinical Study, 2015,12(1):67(in Chinese).
40 ZhouZ J, Zhang J B, Zhu J L, et al. Research progress of cytotoxicity of nanomaterials Chinese Journal of Birth Health and Heredity, 2016,24(4):5(in Chinese).
42 ChaturvediA, Bajpai A K, Bajpai J, et al. Evaluation of poly (vinyl alcohol) based cryogel-zinc oxide nanocomposites for possible applications as wound dressing materials[J]. Materials Science and Engineering C, 2016,65:408.
[43]
43 SathishkumarM, Pavagadhi S, Mahadevan A, et al. Biosynjournal of gold nanoparticles and related cytotoxicity evaluation using A549 cells[J]. Ecotoxicology and Environmental Safety, 2015,114:232.
[44]
44 Kumb??ak ü, ?ava? T, ?ink?l?? N, et al. Evaluation of in vitro cytotoxicity and genotoxicity of copper-zinc alloy nanoparticles in human lung epithelial cells[J]. Food and Chemical Toxicology, 2014,73:105.
[45]
45 WangS G, Yu H T, Wickliffe J K. Limitation of the MTT and XTT assays for measuring cell viability due to superoxide formation induced by nano-scale TiO2[J]. Toxicology in Vitro, 2011,25:2147.
[46]
46Steinh?user K G, Sayre P G. Reliability of methods and data for regulatory assessment of nanomaterial risks[J]. NanoImpact, 2017,7:66.
[47]
47 ZhaoX Y. Advantages and disadvantages of the current in vitro cytotoxic test for dental restorative materials Chinese Journal of Dental Materials and Devices, 2009,18(4):173(in Chinese).
48 YuanY B, Zhang W Y. Progress in the studies of methods for testing cytotoxicity of dental biomaterial Journal of Biomedical Engineering, 2009,26(3):688(in Chinese).
49 DeSouza Costa C A, Hebling J, Scheffel D L S, et al. Methods to evaluate and strategies to improve the biocompatibility of dental materials and operative techniques[J]. Dental Materials, 2014,30(7):769.
[50]
50 IvanovaE P, Bazaka K, Crawford R J. 6-Cytotoxicity and biocompatibility of metallic biomaterials[M]∥New Functional Biomaterials for Medicine & Healthcare, Woodhead Publishing, 2014:148.
[51]
51 WillboldE, Weizbauer A, Loos A, et al. Magnesium alloys: A stony pathway from intensive research to clinical reality. Different test methods and approval-related considerations[J]. Journal of Biomedical Materials Research Part A, 2017,105(1):329.
[52]
52 WangJ, Witte F, Xi T, et al. Recommendation for modifying current cytotoxicity testing standards for biodegradable magnesium-based materials[J]. Acta Biomaterialia, 2015,21:237.
[53]
53 LiH, Peng Q, Li X, et al. Microstructures, mechanical and cytocompatibility of degradable Mg-Zn based orthopedic biomaterials[J]. Materials & Design, 2014,58:43.
[54]
54 MurniN S, Dambatta M S, Yeap S K, et al. Cytotoxicity evaluation of biodegradable Zn-3Mg alloy toward normal human osteoblast cells[J]. Materials Science and Engineering C, 2015,49:560.
[55]
55 FeyerabendF. 8-In vitro, analysis of magnesium corrosion in orthopaedic biomaterials[M]∥Peter Dubruel, Sandra van Vlierberghe, Biomaterials for Bone Regeneration, Woodhead Publishing, 2014:225.
[56]
56 DengL J, Fan H H, Li W D, et al. Progress in the study on biocompatibility of absorbable hemostatic materials Chinese Journal of Biomedical Engineering, 2016,35(2):241(in Chinese).
57 GranjaP L, De Jéso B, Bareille R, et al. Cellulose phosphates as biomaterials. In vitro biocompatibility studies[J]. Reactive and Functional Polymers, 2006,66(7):728.
[58]
58 GagoL A, Saed G M, Wang R X, et al. Effects of oxidized regenerated cellulose on the expression of extracellular matrix and transforming growth factor-β1 in human peritoneal fibroblasts and mesothelial cells[J]. American Journal of Obstetrics and Gynecology, 2003,189(6):1620.
[59]
59 YuT, Qu S F, Huang J, et al. Comparison of the relevant standards for cytotoxicity of medical devices in vitro and discussion of the related content International Journal of Laboratory Medicine, 2015,36(6):858(in Chinese).
ZhangM, Li J, Wang R, et al. Biological effect of disposable infusion sets in different extraction conditions Medical Equipment, 2016,29(9):56(in Chinese).
ISO10993-12-2007, Biological evaluation of medical devices-Part 12: Sample preparation and reference materials[S].
[63]
KuangH, Liu Y, Cao P, et al. Effect of different extracting vehicle on cytotoxicity evaluation for single use balloon catheter[J]. Journal of Biomedical Engineering Research, 2011(2):113(in Chinese).
LiuC L, Sun X J, Fang J Y, et al. Effect of different extracting vehicle on cytotoxicity evaluation for medical under pad China Medical Device Information, 2014,20(2):65(in Chinese).
ChenF Y, Cheng X R. In vitro studies on the cytotoxicity of three commercial denture adhesives Journal of Oral Science Research, 2015,31(5):457(in Chinese).
SangsanohP, Suwantong O, Neamnark A, et al. In vitro biocompatibility of electrospun and solvent-cast chitosan substrata towards Schwann, osteoblast, keratinocyte and fibroblast cells[J]. European Polymer Journal, 2010,46(3):428.
[67]
HowlingG I, Dettmar P W, Goddard P A, et al. The effect of chitin and chitosan on the proliferation of human skin fibroblasts and keratinocytes in vitro[J]. Biomaterials, 2001,22(22):2959.
[68]
WangY. Bioadaptability: An innovative concept for biomaterials[J]. Journal of Materials Science & Technology, 2016,32(9):801.
[69]
ZhangX, Jin R M. Progress of cell co-culture technique Chinese Journal of Pharmacology and Toxicology, 2011,25(3):330(in Chinese).
StioM, Martinesi M, Treves C, et al. Cultures and co-cultures of human blood mononuclear cells and endothelial cells for the biocompatibility assessment of surface modified AISI 316 Laustenitic stainless steel[J]. Materials Science and Engineering C, 2016,69:1081.
[71]
BalszuweitF, Menacher G, Bloemeke B, et al. Development of a co-culture of keratinocytes and immune cells for in vitro investigation of cutaneous sulfur mustard toxicity[J]. Chemico-biological Interactions, 2014,223:117.
[72]
JiangM M. The molecular mechanism of MSC and fibroblast proliferation/differentiation in wound healing[D]. Chongqing: Third Military Medical University, 2015(in Chinese).
NingR, Wang F, Lin L. Biomaterial-based microfluidics for cell culture and analysis[J]. TrAC Trends in Analytical Chemistry, 2016,80:255.
[74]
Monteiro-Riviere N A, Inman A O. Challenges for assessing carbon nanomaterial toxicity to the skin[J]. Carbon, 2006,44(6):1070.
[75]
JongW H D, Carraway J W, Geertsma R E. 7-In vivo, and in vitro, testing for the biological safety evaluation of biomaterials and medical devices[M]∥Biocompatibility & Performance of Medical Devices, Woodhead Publishing, 2012:120.
[76]
ChuP K. Surface engineering and modification of biomaterials[J]. Thin Solid Films, 2013,528:93.
[77]
ShiP, Jiang J H. The research progress of oral material cell toxicity evaluation method Chinese Journal of Aesthetic Medicine, 2014,23(14):1222(in Chinese).