Effects of biocidal solutions and wet tissue treatment on the biological properties of porcine aortic wall

Aim.To evaluate the effects of long-term wet treatment of porcine aortic wall fragments with biocidal solutions (glutaraldehyde, antibiotics, alcohol-glycerin mixture, complex alcohol solution (CAS)) on the severity of degradation of samples’ connective matrix and their long-term ability to accumulate calcium during subcutaneous implantation in laboratory rats (1 and 3 months).

Methods.Histological analysis of freshly collected material was performed after 50 days of treatment with various biocidal solutions and after subcutaneous implantation of samples in small laboratory animals at 1 and 3 months. Calcium was measured with the atomic absorption spectroscopy after 3 months of subcutaneous implantation.

Results.Histological analysis reported that connective matrix of samples treated with glutaraldehyde, antibiotics, and CAS was similar to the native one (with the exception of partial loss of cellular components in samples). After 1 month of implantation, a moderate degradation of connective tissue was found. The treatment with the alcohol-glycerin mixture was associated with the most severe degradation. After 3 months of implantation, glutaraldehyde-treated samples had the most preserved connective matrix, while alcohol-glycerin-treated samples demonstrated severe connective matrix injury. However, severe calcific deposits were found in samples treated with glutaraldehyde, whereas mild ones were detected in samples treated with alcohol-glycerin. Atomic absorption spectroscopy reported that calcific deposits in Cas- and alcohol-glycerin-treated samples were 2.3 and 1.8 times lower than those in glutaraldehyde-treated samples. The calcium content in samples treated with antibiotics did not differ significantly from those in glutaraldehyde-treated samples

Conclusion.The comprehensive analysis of the effects of various treatment media on the stability of connective matrix to subsequent degradation and its ability to accumulate calcium found that CAS was the most preferred medium for long-term wet treatment of xenoprosthetic vascular tissue.

1. Takkenberg J.J.M., Bogers A.J.J.C. Allografts for aortic valve and root replacement: Veni vidi vici? Expert Rev Cardiovasc Ther. 2004; 2(1):97-105.

2. Arabkhani B., Bekkers J.A., Andrinopoulou E.R., RoosHesselink J.W., Takkenberg J.J.M., Bogers A.J.J.C. Allografts in aortic position: Insights from a 27-year, single-center prospective study. J Thorac Cardiovasc Surg. 2016; 152(6): 1572-1579.

3. Demidov D.P., Astapov D.A., Bogachev-Prokophiev A.V., Zheleznev S.I. Quality of life after aortic valve replacement with biological prostheses in elderly patients. Patologiya krovoobrashcheniya i kardiokhirurgiya = Circulation Pathology and Cardiac Surgery. 2017; 21(3):40-47. (in Russian). doi: 10.21688/1681-3472-2017-3-40-47

4. Spiridonov S.V., Odincov V.O., Schetinko N.N., Mozgova E.A., Grinchuk I.I., Ostrovskiy Y.P. Aortic allografts in world cardiac surgery: Historical aspects of implementation in clinical practice and review of results of use. Medical Journal. 2015; 1:55-67. (in Russian).

5. Odarenko Y.N., Rutkovskaya N.V., Rogulina N.V., Stasev A.N., Kokorin S.G., Kagan E.S., Barbarash L.S. Analysis of 23-year experience epoxy treated xenoaortic bioprosthesisin surgery mitral heart disease. Research factors of recipients by positions of influence on the development of calcium degeneration. Complex Issues of Cardiovascular Diseases. 2015;(4):17-25. (in Russian).

6. Lisy M., Kalender G., Schenke-Layland K., Brockbank K.G.M., Biermann A., Stock U.A. Allograft Heart Valves: Current Aspects and Future Applications. Biopreserv Biobank. 2017; 15(2):148-157. doi: 10.1089/bio.2016.0070

7. Brockbank K.G.M., Lightfoot F.G., Song Y.C., Taylor M.J. Interstitial ice formation in cryopreserved homografts: A possible cause of tissue deterioration and calcification in vivo. J Heart Valve Dis. 2000; 9(2): 200-206.

8. Cebotari S., Tudorache I., Ciubotaru A., Boethig D., Sarikouch S., Goerler A., Lichtenberg A., Cheptanaru E., Barnaciuc S., Cazacu A., Maliga O., Repin O., Maniuc L., Breymann T., Haverich A. Use of fresh decellularized allografts for pulmonary valve replacement may reduce the reoperation rate in children and young adults: Early report. Circulation.2011;124(11SUPPL.1):115-124. doi: 10.1161/ CIRCULATIONAHA.110.012161

9. Barratt-Boyes B.G., Roche A.H.G., Subramanyan R., Pemberton J.R., Whitlock R.M. Long-term follow-up of patients with the antibiotic-sterilized aortic homograft valve inserted freehand in the aortic position. Circulation. 1987; 75(4):768-777.

10. Gatto C., Giurgola L., D’Amato Tothova J. A suitable and efficient procedure for the removal of decontaminating antibiotics from tissue allografts. Cell Tissue Bank. 2013; 14(1):107-115. doi: 10.1007/s10561-012-9305-5

11. Vasilyeva M.B., Krasilnikova A.A., Kuznetsova E.V., Lunina M.V., Samoylova L.M., Rusakova Y.L., Chepeleva E.V., Dokuchaeva A.A., Sergeevichev D.S. Alternative biocidal solutions for storage of allogeneic vascular grafts used for the replacement of cardiovascular elements. Patologiya krovoobrashcheniya i kardiokhirurgiya = Circulation Pathology and Cardiac Surgery. 2018;22(4):95-102. (in Russian). doi: 10.21688/1681-3472-2018-4-95-102

12. Cheung D.T., Weber P.A., Grobe A.C., Shomura Y., Choo S.J., Luo H.H., Marchion D.C., Duran C.M. A new method for the preservation of aortic valve homografts. J Heart Valve Dis. 2001;10(6):728-34.

13. Zhuravleva I.Y. Biocidal composition for aseptic storage of prosthetic material from animal tissue. RF patent for invention RU 2580621C1. Bull. No. 10 (10.04.2016).

14. Zhuravleva I.Y., Vasilieva M.B., Timchenko T.P., Kuznetsova E.V., Polienko Y.F., Nichay N.R., Grigoryev I.A., Bogachev-Prokofiev A.V. Сalcification of elastin-containing xenogenic biomaterials: influence of conservants and bisphosphonates The Siberian Scientific Medical Journal. 2017; 37(6):28-37. (in Russian).

15. Zhuravleva I.Y, Nichay N.R, Kulyabin Y.Y, Timchenko T.P, Korobeinikov A.A, Polienko Y.F, Shatskaya S.S, Kuznetsova E.V, Voitov A.V, Bogachev-Prokophiev A.V, Karaskov A.M. In search of the best xenogeneic material for a paediatric conduit: an experimental study. Interact Cardiovasc Thorac Surg. 2018; 26(5):738-744. . doi: 10.1093/icvts/ivx445

16. Gallyamov M. O., Chaschin I. S., Khokhlova M. A., Grigorev T. E., Bakuleva N. P., Lyutova I. G., et al. Collagen tissue treated with chitosan solutions in carbonic acid for improved biological prosthetic heart valves. MaterSciEng. 2014; 37:127–40. doi: 10.1016/j.msec.2014.01.017

17. Eliaz N (Ed). Degradation of Implant Materials. New York: Springer Science+Business Media; 2012.521р.

18. Levy R J, Schoen F J, Levy J T, Nelson A C, Howard S L, Oshry L J. Biologic determinants of dystrophic calcification and osteocalcin deposition in glutaraldehyde-preserved porcine aortic valve leaflets implanted subcutaneously in rats. AmJPathol 1983; 113: 143–55.

19. Wollmann L.C., Suss P.H., Kraft L., Stadler Ribeiro V., Noronha L., da Costa F.D.A., Tuon F.F. Histological and biomechanical characteristics of human decellularized allograft heart valves after eighteen months of storage in saline solution. Biopreserv Biobank. 2020; doi: 10.1089/bio.2019.0106. In press.

20. Tedder M.E., Liao J., Weed B., Stabler C., Zhang H., Simionescu A., Simionescu D.T. Stabilized collagen scaffolds for heart valve tissue engineering. Tissue Eng Part A. 2009; 15:1257–1268.