Evaluation of Cell-detaching Effect of EDTA in Combination with Oxaliplatin for a Possible Application in HIPEC After Cytoreductive Surgery: A Preliminary in-vitro Study

Author(s): Justyna Schubert, Tanja Khosrawipour, Alessio Pigazzi, Joanna Kulas, Jacek Bania, Pawel Migdal, Mohamed Arafkas, Veria Khosrawipour*.

Journal Name: Current Pharmaceutical Design

Volume 25 , Issue 45 , 2019

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Abstract:

Background: Ethylenediaminetetraacetic acid (EDTA), a commonly used compound in laboratory medicine, is known for its membrane-destabilization capacity and cell-detaching effect. This preliminary study aims to assess the potential of EDTA in removing residual tumor cell clusters. Using an in-vitro model, this effect is then compared to the cytotoxic effect of oxaliplatin which is routinely administered during HIPEC procedures. The overall cell toxicity and cell detaching effects of EDTA are compared to those of Oxaliplatin and the additive effect is quantified.

Methods: HT-29 (ATCC® HTB-38™) cells were treated with A) EDTA only B) Oxaliplatin only and C) both agents using an in-vitro model. Cytotoxicity and cell detachment following EDTA application were measured via colorimetric MTS assay. Additionally, detached cell groups were visualized using light microscopy and further analyzed by means of electron microscopy.

Results: When solely applied, EDTA does not exhibit any cell toxicity nor does it add any toxicity to oxaliplatin. However, EDTA enhances the detachment of adherent colon carcinoma cells by removing up to 65% (p<0.05) of the total initial cell amount. In comparison, the sole application of highly concentrated oxaliplatin induced cell mortality by up to 66% (p<0.05). While detached cells showed no mortality after EDTA treatment, cell clusters exhibited a decreased amount of extracellular and adhesive matrix in-between cells. When combined, Oxaliplatin and EDTA display a significant additive effect with only 30% (mean p <0.01) of residual vitality detected in the initial well. EDTA and Oxaliplatin remove up to 81% (p <0.01) of adhesive HT-29 cells from the surface either by cytotoxic effects or cell detachment.

Conclusion: Our data support EDTA’s potential to remove microscopical tumor cell clusters from the peritoneum and possibly act as a supplementary agent in HIPEC procedures with chemotherapy. While adding EDTA to HIPEC procedures may significantly decrease the risk of PM recurrence, further in-vivo and clinical trials are required to evaluate this effect.

Keywords: EDTA, oxaliplatin, HIPEC, cytoreductive surgery, peritoneal metastases, cytotoxicity.

[1]
Desiderio J, Chao J, Melstrom L, et al. The 30-year experience-A meta-analysis of randomised and high-quality non-randomised studies of hyperthermic intraperitoneal chemotherapy in the treatment of gastric cancer. Review Eur J Cancer 2017; 79: 1-14.
[http://dx.doi.org/10.1016/j.ejca.2017.03.030] [PMID: 28456089]
[2]
Mo S, Cai G. Multidisciplinary treatment for colorectal peritoneal metastases: review of the literature. Gastroenterol Res Pract 2016; 2016(2): 1516259.
[http://dx.doi.org/10.1155/2016/1516259] [PMID: 28105045]
[3]
Piso P, Nedelcut SD, Rau B, et al. Morbidity and mortality following cytoreductive surgery and hyperthermic intraperitoneal chemotherapy: data from the DGAV StuDoQ registry with 2149 consecutive patients. Ann Surg Oncol 2019; 26(1): 148-54.
[http://dx.doi.org/10.1245/s10434-018-6992-6] [PMID: 30456672]
[4]
Hotouras A, Desai D, Bhan C, Murphy J, Lampe B, Sugarbaker PH. Heated IntraPEritoneal Chemotherapy (HIPEC) for patients with recurrent ovarian cancer: A systematic literature review. Int J Gynecol Cancer 2016; 26(4): 661-70.
[http://dx.doi.org/10.1097/IGC.0000000000000664] [PMID: 26844612]
[5]
Foster JM, Sleightholm R, Patel A, et al. Morbidity and mortality rates following cytoreductive surgery combined with hyperthermic intraperitoneal chemotherapy compared with other high-risk surgical oncology procedures. JAMA Netw Open 2019; 2(1): e186847.
[http://dx.doi.org/10.1001/jamanetworkopen.2018.6847] [PMID: 30646202]
[6]
Paredes AZ, Abdel-Misih S, Schmidt C, Dillhoff ME, Pawlik TM, Cloyd JM. Predictors of readmission after cytoreductive surgery and hyperthermic intraperitoneal chemotherapy. J Surg Res 2019; 234: 103-9.
[http://dx.doi.org/10.1016/j.jss.2018.09.022] [PMID: 30527460]
[7]
de Jong LAW, Elekonawo FMK, de Reuver PR, et al. Hyperthermic intraperitoneal chemotherapy with oxaliplatin for peritoneal carcinomatosis: a clinical pharmacological perspective on a surgical procedure. Br J Clin Pharmacol 2019; 85(1): 47-58.
[http://dx.doi.org/10.1111/bcp.13773] [PMID: 30255585]
[8]
Halkia E, Spiliotis J, Sugarbaker P. Diagnosis and management of peritoneal metastases from ovarian cancer. Gastroenterol Res Pract 2012; 2012: 541842.
[http://dx.doi.org/10.1155/2012/541842] [PMID: 22888339]
[9]
Khosrawipour T, Wu D, Bellendorf A, et al. Feasibility of single tumorspot treatment in peritoneal carcinomatosis via close range doxorubicin impaction in pressurized intra-peritoneal aerosol chemotherapy (PIPAC). J Clin Exp 2017; 6(3.)
[http://dx.doi.org/10.4172/2324-9110.1000187]
[10]
Khosrawipour V, Khosrawipour T, Falkenstein TA, et al. Evaluating the effect of Micropump© position, internal pressure and doxorubicin dosage on efficacy of Pressurized Intra-peritoneal Aerosol Chemotherapy (PIPAC) in an ex vivo model. Anticancer Res 2016; 36(9): 4595-600.
[http://dx.doi.org/10.21873/anticanres.11008] [PMID: 27630300]
[11]
Mikolajczyk A, Khosrawipour V, Kulas J, et al. Release of doxorubicin from its liposomal coating via high intensity ultrasound. Mol Clin Oncol 2019; 11(5): 483-7.
[http://dx.doi.org/10.3892/mco.2019.1917] [PMID: 31620279]
[12]
Khosrawipour V, Mikolajczyk A, Schubert J, Khosrawipour T. Pressurized Intra-peritoneal Aerosol Chemotherapy (PIPAC) via endoscopical microcatheter system. Anticancer Res 2018; 38(6): 3447-52.
[http://dx.doi.org/10.21873/anticanres.12613] [PMID: 29848695]
[13]
Mikolajczyk A, Khosrawipour V, Schubert J, et al. Effect of liposomal doxorubicin in Pressurized Intra-Peritoneal Aerosol Chemotherapy (PIPAC). J Cancer 2018; 9(23): 4301-5.
[http://dx.doi.org/10.7150/jca.26860] [PMID: 30519333]
[14]
Mikolajczyk A, Khosrawipour V, Schubert J, Chaudhry H, Pigazzi A, Khosrawipour T. Particle stability during Pressurized Intra-peritoneal Aerosol Chemotherapy (PIPAC). Anticancer Res 2018; 38(8): 4645-9.
[http://dx.doi.org/10.21873/anticanres.12769] [PMID: 30061231]
[15]
Khosrawipour V, Bellendorf A, Khosrawipour C, et al. Irradiation does not increase the penetration depth of doxorubicin in normal tissue after Pressurized Intra-peritoneal Aerosol Chemotherapy (PIPAC) in an ex vivo model. In Vivo 2016; 30(5): 593-7.
[PMID: 27566077]
[16]
Khosrawipour V, Giger-Pabst U, Khosrawipour T, et al. Effect of irradiation on tissue penetration depth of doxorubicin after Pressurized Intra-Peritoneal Aerosol Chemotherapy (PIPAC) in a novel ex-vivo model. J Cancer 2016; 7(8): 910-4.
[http://dx.doi.org/10.7150/jca.14714] [PMID: 27313780]
[17]
Khosrawipour V, Khosrawipour T, Hedayat-Pour Y, et al. Effect of whole-abdominal irradiation on penetration depth of doxorubicin in normal tissue after Pressurized Intraperitoneal Aerosol Chemotherapy (PIPAC) in a post-mortem swine model. Anticancer Res 2017; 37(4): 1677-80.
[http://dx.doi.org/10.21873/anticanres.11498] [PMID: 28373428]
[18]
Hoogstins CE, Weixler B, Boogerd LS, et al. In search for optimal targets for intraoperative fluorescence imaging of peritoneal metastasis from colorectal cancerBiomark Cancer 2017; 9.: 1179299X17728254
[http://dx.doi.org/10.1177/1179299X17728254]
[19]
Khosrawipour V, Diaz-Carballo D, Acikelli AH, et al. Cytotoxic effect of different treatment parameters in pressurized intraperitoneal aerosol chemotherapy (PIPAC) on the in vitro proliferation of human colonic cancer cells. World J Surg Oncol 2017; 15(1): 43.
[http://dx.doi.org/10.1186/s12957-017-1109-4] [PMID: 28183319]
[20]
Hendrix RJ, Damle A, Williams C, et al. Restrictive Intraoperative fluid therapy is associated with decreased morbidity and length of stay following hyperthermic intraperitoneal chemoperfusion. Ann Surg Oncol 2019; 26(2): 490-6.
[http://dx.doi.org/10.1245/s10434-018-07092-y] [PMID: 30515670]
[21]
Huo YR, Richards A, Liauw W, Morris DL. Hyperthermic Intraperitoneal Chemotherapy (HIPEC) and cytoreductive surgery (CRS) in ovarian cancer: a systematic review and meta-analysis. Eur J Surg Oncol 2015; 41(12): 1578-89.
[http://dx.doi.org/10.1016/j.ejso.2015.08.172] [PMID: 26453145]
[22]
Amiri M, Salavati-Niasari M, Akbari A. A magnetic CoFe2O4/SiO2 nanocomposite fabricated by the sol-gel method for electrocatalytic oxidation and determination of L-cysteine. Mikrochim Acta 2017; 184(3): 825-33.
[http://dx.doi.org/10.1007/s00604-016-2064-4]
[23]
Mortazavi-Derazkola S, Salavati-Niasari M, Khojasteh H, Amiri O, Ghoreishi SM. Green synthesis of magnetic Fe3O4/SiO2/HAp nanocomposite for atenolol delivery and in vivo toxicity study. J Clean Prod 2017; 168: 39-50.
[http://dx.doi.org/10.1016/j.jclepro.2017.08.235]
[24]
Mohandes F, Salavati-Niasari M. Particle size and shape modification of hydroxyapatite nanostructures synthesized via a complexing agent-assisted route. Mater Sci Eng C 2014; 40: 288-98.
[http://dx.doi.org/10.1016/j.msec.2014.04.008] [PMID: 24857496]
[25]
Amiri M, Salavati-Niasari M, Pardakhty A, Ahmadi M, Akbari A. Caffeine: a novel green precursor for synthesis of magnetic CoFe2O4 nanoparticles and pH-sensitive magnetic alginate beads for drug delivery. Mater Sci Eng C 2017; 76: 1085-93.
[http://dx.doi.org/10.1016/j.msec.2017.03.208] [PMID: 28482472]
[26]
Zinatloo-Ajabshir S, Mortazavi-Derazkola S, Salavati-Niasari M. Preparation, characterization and photocatalytic degradation of methyl violet pollutant of holmium oxide nanostructures prepared through a facile precipitation method. J Mol Liq 2017; 231: 306-13.
[http://dx.doi.org/10.1016/j.molliq.2017.02.002]
[27]
Mohandes F, Salavati-Niasari M, Fathi M, Fereshteh Z. Hydroxyapatite nanocrystals: simple preparation, characterization and formation mechanism. Mater Sci Eng C 2014; 45: 29-36.
[http://dx.doi.org/10.1016/j.msec.2014.08.058] [PMID: 25491798]
[28]
Mohandes F, Salavati-Niasari M. In vitro comparative study of pure hydroxyapatite nanorods and novel polyethylene glycol/graphene oxide/hydroxyapatite nanocomposite. Nanoparticle Res 2014; 16(9): 2604.
[http://dx.doi.org/10.1007/s11051-014-2604-y]
[29]
Goudarzi M, Mir N, Mousavi-Kamazani M, Bagheri S, Salavati-Niasari M. Biosynthesis and characterization of silver nanoparticles prepared from two novel natural precursors by facile thermal decomposition methods. Sci Rep 2016; 6: 32539.
[http://dx.doi.org/10.1038/srep32539] [PMID: 27581681]
[30]
Zinatloo-Ajabshir S, Mortazavi-Derazkola S, Salavati-Niasari M. Nd2O3-SiO2 nanocomposites: a simple sonochemical preparation, characterization and photocatalytic activity. Ultrason Sonochem 2018; 42: 171-82.
[http://dx.doi.org/10.1016/j.ultsonch.2017.11.026] [PMID: 29429658]
[31]
Auld DS. Use of chelating agents to inhibit enzymes. Methods Enzymol 1988; 158: 110-4.
[http://dx.doi.org/10.1016/0076-6879(88)58051-5] [PMID: 3374366]
[32]
Gandolfi MG, Taddei P, Pondrelli A, Zamparini F, Prati C, Spagnuolo G. Demineralization, collagen modification and remineralization degree of human dentin after EDTA and citric acid treatments. Materials (Basel) 2018; 12(1): 25.
[33]
Epand RF, Sarig H, Mor A, Epand RM. Cell-wall interactions and the selective bacteriostatic activity of a miniature oligo-acyl-lysyl. Biophys J 2009; 97(8): 2250-7.
[http://dx.doi.org/10.1016/j.bpj.2009.08.006] [PMID: 19843457]
[34]
Prachayasittikul V, Isarankura-Na-Ayudhya C, Tantimongcolwat T, Nantasenamat C, Galla HJ. EDTA-induced membrane fluidization and destabilization: biophysical studies on artificial lipid membranes. Acta Biochim Biophys Sin (Shanghai) 2007; 39(11): 901-13.
[http://dx.doi.org/10.1111/j.1745-7270.2007.00350.x] [PMID: 17989882]
[35]
Rosenblum MG, Verschraegen CF, Murray JL, et al. Phase I study of 90Y-labeled B72.3 intraperitoneal administration in patients with ovarian cancer: effect of dose and EDTA coadministration on pharmacokinetics and toxicity. Clin Cancer Res 1999; 5(5): 953-61.
[PMID: 10353726]
[36]
Mattheij JA, Kuipers MA, Swarts JJ, Verstijnen CP. Intraperitoneal infusion of EDTA in the rat blocks completely the prolactin rise in the plasma during suckling. Horm Res 1982; 16(4): 219-29.
[http://dx.doi.org/10.1159/000179505] [PMID: 6813213]
[37]
Schubert J, Khosrawipour V, Chaudhry H, et al. Comparing the cytotoxicity of taurolidine, mitomycin C, and oxaliplatin on the proliferation of in vitro colon carcinoma cells following pressurized intra-peritoneal aerosol chemotherapy (PIPAC). World J Surg Oncol 2019; 17(1): 93.
[http://dx.doi.org/10.1186/s12957-019-1633-5] [PMID: 31159819]
[38]
Ferrero ME. Rationale for the successful management of EDTA chelation therapy in human burden by toxic metals. BioMed Res Int 2016; 2016: 8274504.
[http://dx.doi.org/10.1155/2016/8274504] [PMID: 27896275]
[39]
Simkens GA, van Oudheusden TR, Braam HJ, et al. Treatment-related mortality after cytoreductive surgery and HIPEC in patients with colorectal peritoneal carcinomatosis is underestimated by conventional parameters. Ann Surg Oncol 2016; 23(1): 99-105.
[http://dx.doi.org/10.1245/s10434-015-4699-5] [PMID: 26148758]
[40]
Yang XJ, Huang CQ, Suo T, et al. Cytoreductive surgery and hyperthermic intraperitoneal chemotherapy improves survival of patients with peritoneal carcinomatosis from gastric cancer: final results of a phase III randomized clinical trial. Ann Surg Oncol 2011; 18(6): 1575-81.
[http://dx.doi.org/10.1245/s10434-011-1631-5] [PMID: 21431408]
[41]
Mohammadi Z, Shalavi S, Jafarzadeh H. Ethylenediaminetetraacetic acid in endodontics. Eur J Dent 2013; 7(Suppl. 1): S135-42.
[http://dx.doi.org/10.4103/1305-7456.119091]
[42]
Wang G1, Zabner J, Deering C, et al. Increasing epithelial junction permeability enhances gene transfer to airway epithelia In vivo. Am J Respir Cell Mol Biol 2000; 22: 129-38.
[http://dx.doi.org/10.1165/ajrcmb.22.2.3938]
[43]
Niessen CM, Leckband D, Yap AS. Tissue organization by cadherin adhesion molecules: dynamic molecular and cellular mechanisms of morphogenetic regulation. Physiol Rev 2011; 91(2): 691-731.
[http://dx.doi.org/10.1152/physrev.00004.2010] [PMID: 21527735]
[44]
Schmidt TP, Goetz C, Huemer M, Schneider G, Wessler S. Calcium binding protects E-cadherin from cleavage by helicobacter pylori HtrA. Gut Pathog 2016; 8(8): 29.
[http://dx.doi.org/10.1186/s13099-016-0112-6] [PMID: 27274359]
[45]
Le TL, Yap AS, Stow JL. Recycling of E-cadherin: a potential mechanism for regulating cadherin dynamics. J Cell Biol 1999; 146(1): 219-32.
[http://dx.doi.org/10.1083/jcb.146.1.219] [PMID: 10402472]
[46]
McNeill H, Ryan TA, Smith SJ, Nelson WJ. Spatial and temporal dissection of immediate and early events following cadherin-mediated epithelial cell adhesion. J Cell Biol 1993; 120(5): 1217-26.
[http://dx.doi.org/10.1083/jcb.120.5.1217] [PMID: 8436592]
[47]
Adams CL, Chen YT, Smith SJ, Nelson WJ. Mechanisms of epithelial cell-cell adhesion and cell compaction revealed by high-resolution tracking of E-cadherin-green fluorescent protein. J Cell Biol 1998; 142(4): 1105-19.
[http://dx.doi.org/10.1083/jcb.142.4.1105] [PMID: 9722621]
[48]
Myat MM, Chang S, Rodriguez-Boulan E, Aderem A. Identification of the basolateral targeting determinant of a peripheral membrane protein, MacMARCKS, in polarized cells. Curr Biol 1998; 8(12): 677-83.
[http://dx.doi.org/10.1016/S0960-9822(98)70273-8] [PMID: 9637918]
[49]
Kartenbeck J, Schmelz M, Franke WW, Geiger B. Endocytosis of junctional cadherins in bovine kidney epithelial (MDBK) cells cultured in low Ca2+ ion medium. J Cell Biol 1991; 113(4): 881-92.
[http://dx.doi.org/10.1083/jcb.113.4.881] [PMID: 2026652]
[50]
Kartenbeck J, Schmid E, Franke WW, Geiger B. Different modes of internalization of proteins associated with adhaerens junctions and desmosomes: experimental separation of lateral contacts induces endocytosis of desmosomal plaque material. EMBO J 1982; 1(6): 725-32.
[http://dx.doi.org/10.1002/j.1460-2075.1982.tb01237.x] [PMID: 6821357]
[51]
Duden R, Franke WW. Organization of desmosomal plaque proteins in cells growing at low calcium concentrations. J Cell Biol 1988; 107(3): 1049-63.
[http://dx.doi.org/10.1083/jcb.107.3.1049] [PMID: 2458360]
[52]
Miller JR, McClay DR. Characterization of the role of cadherin in regulating cell adhesion during sea urchin development. Dev Biol 1997; 192(2): 323-39.
[http://dx.doi.org/10.1006/dbio.1997.8740] [PMID: 9441671]


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VOLUME: 25
ISSUE: 45
Year: 2019
Page: [4813 - 4819]
Pages: 7
DOI: 10.2174/1381612825666191106153623
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