Cell and Tissue Banking

, Volume 19, Issue 4, pp 581–589 | Cite as

Antimicrobial efficiency and stability of two decontamination solutions

  • Ingrida Smeringaiova
  • Otakar Nyc
  • Peter Trosan
  • Jaroslav Spatenka
  • Jan Burkert
  • Jan Bednar
  • Katerina Jirsova


Two decontamination solutions, commercially produced BASE•128 and laboratory decontamination solution (LDS), with analogous content of antibiotic and antimycotic agents, were compared in their antimicrobial efficiency and stability (pH and osmolarity). Both solutions were compared immediately after thawing aliquots frozen for 1, 3 or 6 months. Agar well diffusion method was used to test their antimicrobial efficiency against five human pathogens: Staphylococcus aureus, Pseudomonas aeruginosa, Proteus mirabilis, Escherichia coli and Enterococcus faecalis. The difference in the inhibition of growth between the two decontamination solutions was mostly not statistically significant, with few exceptions. The most pronounced difference between the LDS and BASE•128 was observed in their decontamination efficacy against E. coli and E. faecalis, where the LDS showed to be more efficient than BASE•128. The osmolarity value of LDS decreased with cold-storage, the osmolarity values of the BASE•128 could not be measured as they were below the range of the osmometer. Slight changes were found in pH of the less stable LDS solution, whose pH increased from initial value 7.36 ± 0.07 to 7.72 ± 0.19 after 6 m-storage. We verified that BASE•128 and LDS are similarly efficient in elimination of possible placental bacterial contaminants and may be used for decontamination of various tissues.


Tissue decontamination Amniotic membrane decontamination Antimicrobial efficiency Decontamination solution 



This work was supported by European Regional Development Fund, Project BBMRI-CZ III: EF16_013/0001674. Institutional support was provided by Progres-Q26/LF1 and by SVV, Project 260367/2017.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest. The funding organizations had no role in the design or conduct of this research.


  1. Adds PJ, Hunt C, Hartley S (2001) Bacterial contamination of amniotic membrane. Br J Ophthalmol 85(2):228–230CrossRefPubMedPubMedCentralGoogle Scholar
  2. Aghayan HR, Goodarzi P, Baradaran-Rafii A, Larijani B, Moradabadi L, Rahim F, Arjmand B (2013) Bacterial contamination of amniotic membrane in a tissue bank from Iran. Cell Tissue Bank 14(3):401–406CrossRefPubMedGoogle Scholar
  3. Ashraf NN, Siyal NA, Sultan S, Adhi MI (2015) Comparison of efficacy of storage of amniotic membrane at-20 and-80 degrees centigrade. J Coll Phys Surg Pak 25(4):264–267Google Scholar
  4. Balouiri M, Sadiki M, Ibnsouda SK (2016) Methods for in vitro evaluating antimicrobial activity: a review. J Pharm Anal 6(2):71–79CrossRefPubMedGoogle Scholar
  5. Baorto EP (2017) Staphylococcus aureus infection. Medscape. Accessed 11 May 2018
  6. Binte Atique F, Ahmed KT, Asaduzzaman SM, Hasan KN (2013) Effects of gamma irradiation on bacterial microflora associated with human amniotic membrane. Biomed Res Int 2013:586561CrossRefPubMedPubMedCentralGoogle Scholar
  7. Burgos H, Sergeant RJ (1983) lyophilized human amniotic membranes used in reconstruction of the ear. J R Soc Med 76(5):433CrossRefPubMedPubMedCentralGoogle Scholar
  8. Clinical and Laboratory Standards Institute (2018) M100—performance standards for antimicrobial susceptibility testing, 28th edn. Clinical and Laboratory Standards Institute, WayneGoogle Scholar
  9. de Kraker ME, Wolkewitz M, Davey PG, Koller W, Berger J, Nagler J et al (2011) Clinical impact of antimicrobial resistance in European hospitals: excess mortality and length of hospital stay related to methicillin-resistant Staphylococcus aureus bloodstream infections. Antimicrob Agents Chemother 55(4):1598–1605CrossRefPubMedPubMedCentralGoogle Scholar
  10. Dua HS, Azuara-Blanco A (1999) Amniotic membrane transplantation. Br J Ophthalmol 83(6):748–752CrossRefPubMedPubMedCentralGoogle Scholar
  11. Duan-Arnold Y, Gyurdieva A, Johnson A, Jacobstein DA, Danilkovitch A (2015) Soluble factors released by endogenous viable cells enhance the antioxidant and chemoattractive activities of cryopreserved amniotic membrane. Adv Wound Care 4(6):329–338CrossRefGoogle Scholar
  12. Faron ML, Ledeboer NA, Buchan BW (2016) resistance mechanisms, epidemiology, and approaches to screening for vancomycin-resistant enterococcus in the health care setting. J Clin Microbiol 54(10):2436–2447CrossRefPubMedPubMedCentralGoogle Scholar
  13. Friedrich M (2017) Pseudomonas aeruginosa infections. Medscape. Accessed 10 May 2018
  14. Gannaway WL, Barry AL, Trelford JD (1984) Preparation of amniotic membranes for surgical use with antibiotic solutions. Surgery 95(5):580–585PubMedGoogle Scholar
  15. Gatto C, Giurgola L, D’Amato Tothova J (2013) A suitable and efficient procedure for the removal of decontaminating antibiotics from tissue allografts. Cell Tissue Bank 14(1):107–115CrossRefPubMedGoogle Scholar
  16. Gonzales G (2017) Proteus infections. Medscape. Accessed 10 May 2018
  17. Guiral E, Bosch J, Vila J, Soto SM (2011) Prevalence of Escherichia coli among samples collected from the genital tract in pregnant and nonpregnant women: relationship with virulence. FEMS Microbiol Lett 314(2):170–173CrossRefPubMedGoogle Scholar
  18. Hanada K, Shimazaki J, Shimmura S, Tsubota K (2001) Multilayered amniotic membrane transplantation for severe ulceration of the cornea and sclera. Am J Ophthalmol 131(3):324–331CrossRefPubMedGoogle Scholar
  19. Hennerbichler S, Reichl B, Pleiner D, Gabriel CH, Eibl J, Redl H (2007) The influence of various storage conditions on cell viability in amniotic membrane. Cell Tissue Bank 8(1):1–8CrossRefPubMedGoogle Scholar
  20. Herndon DN, Branski LK (2017) Contemporary methods allowing for safe and convenient use of amniotic membrane as a biologic wound dressing for burns. Ann Plast Surg 78(2 Suppl 1):S9–S10CrossRefPubMedGoogle Scholar
  21. Holder IA, Boyce ST (1994) Agar well diffusion assay testing of bacterial susceptibility to various antimicrobials in concentrations non-toxic for human cells in culture. Burns 20(5):426–429CrossRefPubMedGoogle Scholar
  22. Hopkinson A, McIntosh RS, Tighe PJ, James DK, Dua HS (2006) Amniotic membrane for ocular surface reconstruction: donor variations and the effect of handling on TGF-beta content. Investig Ophthalmol Vis Sci 47(10):4316–4322CrossRefGoogle Scholar
  23. Ilic D, Vicovac L, Nikolic M, Lazic Ilic E (2016) Human amniotic membrane grafts in therapy of chronic non-healing wounds. Br Med Bull 117(1):59–67CrossRefPubMedGoogle Scholar
  24. Jirsova K, Jones GLA (2017) Amniotic membrane in ophthalmology: properties, preparation, storage and indications for grafting—a review. Cell Tissue Bank 18(2):193–204CrossRefPubMedGoogle Scholar
  25. Keitel S (2017) Guide to the quality and safety of tissues and cells for human application, European Committee (Partial Agreement) on organ transplantation, European Directorate for the Quality of Medicines and HealthCare (EDQM), 3nd edn, Council of Europe, pp 197–201Google Scholar
  26. Khokhar S, Sharma N, Kumar H, Soni A (2001) Infection after use of nonpreserved human amniotic membrane for the reconstruction of the ocular surface. Cornea 20(7):773–774CrossRefPubMedGoogle Scholar
  27. Kim JC, Tseng SC (1995) Transplantation of preserved human amniotic membrane for surface reconstruction in severely damaged rabbit corneas. Cornea 14(5):473–484CrossRefPubMedGoogle Scholar
  28. Kim JS, Kim JC, Na BK, Jeong JM, Song CY (2000) Amniotic membrane patching promotes healing and inhibits proteinase activity on wound healing following acute corneal alkali burn. Exp Eye Res 70(3):329–337CrossRefPubMedGoogle Scholar
  29. Lam K, Bayer AS (1984) In vitro bactericidal synergy of gentamicin combined with penicillin G, vancomycin, or cefotaxime against group G streptococci. Antimicrob Agents Chemother 26(2):260–262CrossRefPubMedPubMedCentralGoogle Scholar
  30. Laurent R, Nallet A, Obert L, Nicod L, Gindraux F (2014) Storage and qualification of viable intact human amniotic graft and technology transfer to a tissue bank. Cell Tissue Bank 15(2):267–275CrossRefPubMedGoogle Scholar
  31. Lee SH, Tseng SC (1997) Amniotic membrane transplantation for persistent epithelial defects with ulceration. Am J Ophthalmol 123(3):303–312CrossRefPubMedGoogle Scholar
  32. Madappa T (2017) Escherichia coli (E. coli) infections medication. medscape. Accessed 10 Feb 2018
  33. Malhotra C, Jain AK (2014) Human amniotic membrane transplantation: different modalities of its use in ophthalmology. World J Transplant 4(2):111–121CrossRefPubMedPubMedCentralGoogle Scholar
  34. Maral T, Borman H, Arslan H, Demirhan B, Akinbingol G, Haberal M (1999) Effectiveness of human amnion preserved long-term in glycerol as a temporary biological dressing. Burns 25(7):625–635CrossRefPubMedGoogle Scholar
  35. Marangon FB, Alfonso EC, Miller D, Remonda NM, Muallem MS, Tseng SC (2004) Incidence of microbial infection after amniotic membrane transplantation. Cornea 23(3):264–269CrossRefPubMedGoogle Scholar
  36. Messmer EM (2001) Hypopyon after amniotic membrane transplantation. Ophthalmology 108(10):1714–1715CrossRefPubMedGoogle Scholar
  37. Mrázová H, Koller J, Kubišová K, Fujeríková G, Klincová E, Babal P (2016) Comparison of structural changes in skin and amnion tissue grafts for transplantation induced by gamma and electron beam irradiation for sterilization. Cell Tissue Bank 17(2):255–260CrossRefPubMedGoogle Scholar
  38. Niknejad H, Deihim T, Solati-Hashjin M, Peirovi H (2011) The effects of preservation procedures on amniotic membrane’s ability to serve as a substrate for cultivation of endothelial cells. Cryobiology 63(3):145–151CrossRefPubMedGoogle Scholar
  39. Paolin A, Cogliati E, Trojan D, Griffoni C, Grassetto A, Elbadawy HM, Ponzin D (2016) Amniotic membranes in ophthalmology: long term data on transplantation outcomes. Cell Tissue Bank 17(1):51–58CrossRefPubMedGoogle Scholar
  40. Perepelkin NMJ, Hayward K, Mokoena T, Bentley MJ, Ross-Rodriguez LU, Marquez-Curtis L, McGann LE, Holovati JL, Elliott JA (2016) Cryopreserved amniotic membrane as transplant allograft: viability and post-transplant outcome. Cell Tissue Bank 17(1):39–50CrossRefPubMedGoogle Scholar
  41. Rahman I, Said DG, Maharajan VS, Dua HS (2009) Amniotic membrane in ophthalmology: indications and limitations. Eye (Lond) 23(10):1954–1961CrossRefGoogle Scholar
  42. Rama P, Giannini R, Bruni A, Gatto C, Tiso R, Ponzin D (2001) Further evaluation of amniotic membrane banking for transplantation in ocular surface diseases. Cell Tissue Bank 2(3):155–163CrossRefPubMedGoogle Scholar
  43. Ramirez Estrada S, Borgatta B, Rello J (2016) Pseudomonas aeruginosa ventilator-associated pneumonia management. Infect Drug Resist 20(9):7–18Google Scholar
  44. Rautmann G, Daas A, Buchheit KH (2010) Collaborative study for the establishment of the second international standard for amphotericin B. Pharmeuropa Bio Sci Notes 1:1–13Google Scholar
  45. Riau AK, Beuerman RW, Lim LS, Mehta JS (2010) Preservation, sterilization and de-epithelialization of human amniotic membrane for use in ocular surface reconstruction. Biomaterials 31(2):216–225CrossRefPubMedGoogle Scholar
  46. Rodriguez-Ares MT, López-Valladares MJ, Touriño R, Vieites B, Gude F, Silva MT, Couceiro J (2009) Effects of lyophilization on human amniotic membrane. Acta Ophthalmol 87(4):396–403CrossRefPubMedGoogle Scholar
  47. Singh R, Gupta P, Kumar P, Kumar A, Chacharkar MP (2003) Properties of air dried radiation processed amniotic membranes under different storage conditions. Cell Tissue Bank 4(2–4):95–100CrossRefPubMedGoogle Scholar
  48. Singh R, Gupta P, Purohit S, Kumar P, Vaijapurkar SG, Chacharkar MP (2006) Radiation resistance of the microflora associated with amniotic membranes. World J Microbiol Biotechnol 22(1):23–27CrossRefGoogle Scholar
  49. Singh M, Sharma R, Gupta PK, Rana JK, Sharma M, Taneja N (2012) Comparative efficacy evaluation of disinfectants routinely used in hospital practice: India. Indian J Crit Care Med 16(3):123CrossRefPubMedPubMedCentralGoogle Scholar
  50. Smeringaiova I, Trosan P, Mrstinova MB, Matecha J, Burkert J, Bednar J, Jirsova K (2017) Comparison of impact of two decontamination solutions on the viability of the cells in human amnion. Cell Tissue Bank 18(3):413–423CrossRefPubMedGoogle Scholar
  51. Solomon A, Meller D, Prabhasawat P, John T, Espana EM, Steuhl KP, Tseng SC (2002) Amniotic membrane grafts for nontraumatic corneal perforations, descemetoceles, and deep ulcers. Ophthalmology 109(4):694–703CrossRefPubMedGoogle Scholar
  52. The European Committee on Antimicrobial Susceptibility Testing (EUCAST) (2018) Breakpoint tables for interpretation of MICs and zone diameters. Version 8.0. Accessed 5 Feb 2018
  53. Thomasen H, Pauklin M, Steuhl KP, Meller D (2009) Comparison of cryopreserved and air-dried human amniotic membrane for ophthalmologic applications. Graefes Arch Clin Exp Ophthalmol 247(12):1691–1700CrossRefPubMedGoogle Scholar
  54. Wolbank S, Hildner F, Redl H, van Griensven M, Gabriel C, Hennerbichler S (2009) Impact of human amniotic membrane preparation on release of angiogenic factors. J Tissue Eng Regen Med 3(8):651–654CrossRefPubMedGoogle Scholar
  55. World Health Organization (2018) WHO publishes list of bacteria for which new antibiotics are urgently needed. Accessed 11 May 2018
  56. Zidan SM, Eleowa SA, Nasef MA, Abd-Almoktader MA, Elbatawy AM, Borhamy AG, Aboliela MA, Ali AM, Algamal MR (2015) Maximizing the safety of glycerol preserved human amniotic membrane as a biological dressing. Burns 41(7):1498–1503CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Laboratory of the Biology and Pathology of the Eye, Department of Paediatrics and Adolescent Medicine, First Faculty of MedicineCharles University and General University HospitalPragueCzech Republic
  2. 2.Laboratory of the Biology and Pathology of the Eye, Institute of Biology and Medical Genetics, First Faculty of MedicineCharles University and General University HospitalPragueCzech Republic
  3. 3.Department of Clinical Microbiology, Second Faculty of MedicineCharles UniversityPragueCzech Republic
  4. 4.Department of Transplantation and Tissue BankMotol University HospitalPragueCzech Republic

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