Vol. 29 No. 3 (2025): Patologiya krovoobrashcheniya i kardiokhirurgiya
EXPERIMENTAL STUDIES

The Optimization of decellularization and biocompatibility assessment of laser-perforated vascular matrices

Elena Chepeleva
Meshalkin National Medical Research Center, Ministry of Health of Russian Federation, Novosibirsk, Russian Federation
Anna Dokuchaeva
Meshalkin National Medical Research Center, Ministry of Health of Russian Federation, Novosibirsk, Russian Federation
Sergey Vladimirov
Meshalkin National Medical Research Center, Ministry of Health of Russian Federation, Novosibirsk, Russian Federation
Elena Kuznetsova
Meshalkin National Medical Research Center, Ministry of Health of Russian Federation, Novosibirsk, Russian Federation
David Sergeevichev
Meshalkin National Medical Research Center, Ministry of Health of Russian Federation, Novosibirsk, Russian Federation

Published 2025-12-18

How to Cite

Chepeleva, E., Dokuchaeva, A., Vladimirov, S., Kuznetsova, E., & Sergeevichev, D. (2025). The Optimization of decellularization and biocompatibility assessment of laser-perforated vascular matrices. Patologiya Krovoobrashcheniya I Kardiokhirurgiya, 29(3), 46–57. https://doi.org/10.21688/1681-3472-2025-3-46-57

Abstract

Background: To enable the adaptation of biological materials intended for the prosthetics of various parts of the cardiovascular system to the physiological conditions of the body, research aimed at a comparative analysis of decellularization protocols for creating an optimal scaffold, as well as improving its properties, in particular through surface modification, is of current importance.

Objective: The study aims to optimize the protocol on decellularization of the porcine pulmonary trunk wall by comparative analysis of detergent compositions followed by assessement of the effect of laser microperforation on the biocompatibility and cellular repopulation of the decellularized matrix in vitro and in vivo.

Methods: The first stage of the work was aimed at selecting the incubation time for porcine pulmonary trunk fragments in solutions with various concentrations of detergents, followed by tensometric tests to determine the optimal combination of conditions. Further, the decellularized material was perforated using laser processing. Some of the resulting samples were repopulated with fibroblast cells for 7 days, while other samples were subcutaneously implanted into the paravertebral region of rats for 30 days. Histological studies of the biomaterials were conducted at each stage.

Results: The optimal decellularization protocol for the porcine pulmonary trunk wall was determined to be its treatment with a 0.5 % sodium dodecyl sulfate solution in combination with 0.5 % sodium deoxycholate, which enables to produce an acellular matrix with preserved ultrastructure within 16 hours. Culturing mouse fibroblasts on the surface of laser-treated samples proved the sustention of cell viability. The investigation of the cellular composition of explants retrieved 30 days after the experimental surgery showed that laser perforation preserves the material’s biocompatibility.

Conclusion: The conducted research resulted in the development and validation of an effective decellularization protocol for obtaining an acellular pulmonary trunk wall matrix with preserved extracellular structure. Moreover, laser perforation of the matrices does not compromise their biocompatibility and enhances their integrative potential by expanding the contact surface area.

References

  1. Waqanivavalagi S.W.F.R., Bhat S., Ground M.B., Milsom P.F., Cornish J. Clinical performance of decellularized heart valves versus standard tissue conduits: a systematic review and meta-analysis. J Cardiothorac Surg. 2020;15(1):260. PMID: 32948234; PMCID: PMC7501674. https://doi.org/10.1186/s13019-020-01292-y
  2. Arabkhani B., Bekkers J.A., Andrinopoulou E.R., Roos-Hesselink J.W., Takkenberg J.J., Bogers A.J. Allografts in aortic posi- tion: Insights from a 27-year, single-center prospective study. J Thorac Cardiovasc Surg. 2016;152(6):1572-1579.e3. PMID: 27842683. https://doi.org/10.1016/j.jtcvs.2016.08.013
  3. Демидов Д.П., Астапов Д.А., Богачев-Прокофьев А.В., Железнев С.И. Оценка качества жизни после протезирования аортального клапана биологическими протезами у паци- ентов пожилого возраста. Патология кровообращения и кардиохирургия. 2017;21(3):40-47. https://doi.org/10.21688/1681-3472-2017-3-40-47 Demidov D.P., Astaspov D.A., Bogachev-Prokophiev A.V., Zheleznev S.I. Quality of life after aortic valve replacement with biological prostheses in elderly patients. Patologiya krovoobrash- cheniya i kardiokhirurgiya = Circulation Pathology and Cardiac Surgery. 2017;21(3):40-47. (In Russ.) https://doi.org/10.21688/1681-3472-2017-3-40-47
  4. Hirji S.A., Kolkailah A.A., Ramirez-Del Val F., Lee J., McGurk S., Pelletier M., Singh S., Mallidi H.R., Aranki S., Shekar P., Kaneko T. Mechanical Versus Bioprosthetic Aortic Valve Re- placement in Patients Aged 50 Years and Younger. Ann Thorac Surg. 2018;106(4):1113-1120. PMID: 29966596. https://doi.org/10.1016/j.athoracsur.2018.05.073
  5. David T. How to Decide Between a Bioprosthetic and Mechani- cal Valve. Can J Cardiol. 2021;37(7):1121-1123. PMID: 33002585. https://doi.org/10.1016/j.cjca.2020.09.011
  6. Horke A., Tudorache I., Laufer G., Andreas M., Pomar J.L., Pereda D., Quintana E., Sitges M., Meyns B., Rega F., Hazekamp M., Hübler M., Schmiady M., Pepper J., Rosendahl U., Lichtenberg A., Akhyari P., Jashari R., Boethig D., Bobylev D., Avsar M., Cebotari S., Haverich A., Sarikouch S. Early results from a prospective, sin- gle-arm European trial on decellularized allografts for aortic valve replacement: the ARISE study and ARISE Registry data. Eur J Cardiothorac Surg. 2020;58(5):1045-1053. PMID: 32386409; PMCID: PMC7577293. https://doi.org/10.1093/ejcts/ezaa100
  7. Спиридонов С.В., Одинцов В.О., Щетинко Н.Н., Мозгова Е.А., Гринчук И.И., Островский Ю.П. Аортальные аллографты в мировой кардиохирургии: исторические аспекты внедрения в клиническую практику и обзор результатов использования. Медицинский журнал. 2015;1:55-67. Spirydonau S.V., Adzintsou V.O., Shchatsinka N.N., Mazgova E.A., Hrynchuk I.I., Ostrovsky Yu.P. Aortic allografts in the world cardiac surgery: historical aspects of clinical implementation, and re- view of implantation outcomes. Meditsinskii zhurnal. 2015;1:55-67. (In Russ.)
  8. Fioretta E.S., von Boehmer L., Motta S.E., Lintas V., Hoerstrup S.P., Emmert M.Y. Cardiovascular tissue engineering: From basic sci- ence to clinical application. Exp Gerontol. 2019;117:1-12. PMID: 29604404. https://doi.org/10.1016/j.exger.2018.03.022
  9. VeDepo M.C., Detamore M.S., Hopkins R.A., Converse G.L. Recellularization of decellularized heart valves: Prog- ress toward the tissue-engineered heart valve. J Tissue Eng. 2017;8:2041731417726327. PMID: 28890780; PMCID: PMC5574480. https://doi.org/10.1177/2041731417726327
  10. 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(11 Suppl):S115-23. PMID: 21911800. https://doi.org/10.1161/circulationaha.110.012161
  11. Argento G., Simonet M., Oomens C.W., Baaijens F.P. Multi-scale mechanical characterization of scaffolds for heart valve tissue engineering. J Biomech. 2012;45(16):2893-8. PMID: 22999107. https://doi.org/10.1016/j.jbiomech.2012.07.037
  12. Converse G.L., Armstrong M., Quinn R.W., Buse E.E., Cromwell M.L., Moriarty S.J., Lofland G.K., Hilbert S.L., Hopkins R.A. Effects of cryopreservation, decellularization and novel extra- cellular matrix conditioning on the quasi-static and time-depen- dent properties of the pulmonary valve leaflet. Acta Biomater. 2012;8(7):2722-9. PMID: 22484150. https://doi.org/10.1016/j.actbio.2012.03.047
  13. Tefft B.J., Choe J.A., Young M.D., Hennessy R.S., Morse D.W., Bouchard J.A., Hedberg H.J., Consiglio J.F., Dragomir-Daescu D., Simari R.D., Lerman A. Cardiac Valve Bioreactor for Physiological Conditioning and Hydrodynamic Performance Assessment. Car- diovasc Eng Technol. 2019;10(1):80-94. PMID: 30311149; PMCID: PMC6541400. https://doi.org/10.1007/s13239-018-00382-2
  14. Converse G.L., Buse E.E., Neill K.R., McFall C.R., Lewis H.N., VeDe- po M.C., Quinn R.W., Hopkins R.A. Design and efficacy of a sin- gle-use bioreactor for heart valve tissue engineering. J Biomed Mater Res B Appl Biomater. 2017;105(2):249-259. PMID: 26469196. https://doi.org/10.1002/jbm.b.33552
  15. Koenig F., Lee J.S., Akra B., Hollweck T., Wintermantel E., Hagl C., Thierfelder N. Is Transcatheter Aortic Valve Implantation of Living Tissue-En- gineered Valves Feasible? An In Vitro Evaluation Utilizing a Decellular- ized and Reseeded Biohybrid Valve. Artif Organs. 2016;40(8):727-37. PMID: 27187768. https://doi.org/10.1111/aor.12683
  16. Parvin Nejad S., Blaser M.C., Santerre J.P., Caldarone C.A., Simmons C.A. Biomechanical conditioning of tissue engineered heart valves: Too much of a good thing? Adv Drug Deliv Rev. 2016;96:161-75. PMID: 26555371. https://doi.org/10.1016/j.addr.2015.11.003
  17. Berry J.L., Steen J.A., Koudy Williams J., Jordan J.E., Atala A., Yoo J.J. Bioreactors for development of tissue engineered heart valves. Ann Biomed Eng. 2010;38(11):3272-9. PMID: 20820920. https://doi.org/10.1007/s10439-010-0148-6
  18. VeDepo M.C., Buse E.E., Paul A., Converse G.L., Hopkins R.A. Non-physiologic Bioreactor Processing Conditions for Heart Valve Tissue Engineering. Cardiovasc Eng Technol. 2019;10(4):628- 637. PMID: 31650518. https://doi.org/10.1007/s13239-019-00438-x
  19. Mendibil U., Ruiz-Hernandez R., Retegi-Carrion S., Garcia-Ur- quia N., Olalde-Graells B., Abarrategi A. Tissue-Specific Decel- lularization Methods: Rationale and Strategies to Achieve Re- generative Compounds. Int J Mol Sci. 2020;21(15):5447. PMID: 32751654; PMCID: PMC7432 490.https://doi.org/10.3390/ijms21155447
  20. Roosens A., Asadian M., De Geyter N., Somers P., Cornelissen R. Complete Static Repopulation of Decellularized Porcine Tissues for Heart Valve Engineering: An in vitro Study. Cells Tissues Or- gans. 2017;204(5-6):270-282. PMID: 29131080. https://doi.org/10.1159/000480660
  21. Theodoridis K., Tudorache I., Calistru A., Cebotari S., Meyer T., Sarikouch S., Bara C., Brehm R., Haverich A., Hilfiker A. Successful matrix guided tissue regeneration of decellularized pulmonary heart valve allografts in elderly sheep. Biomaterials. 2015;52:221-8. PMID: 25818428. https://doi.org/10.1016/j.biomaterials.2015.02.023
  22. Badria A.F., Koutsoukos P.G., Mavrilas D. Decellularized tissue-engineered heart valves calcification: what do animal and clinical studies tell us? J Mater Sci Mater Med. 2020;31(12):132. PMID: 33278023; PMCID: PMC7719105. https://doi.org/10.1007/s10856-020-06462-x
  23. Bergmeister H., Boeck P., Kasimir M.T., Fleck T., Fitzal F., Husinsky W., Mittlboeck M., Stoehr H.G., Losert U., Wolner E., Grabenwoeger M. Effect of laser perforation on the remodeling of acellular matrix grafts. J Biomed Mater Res B Appl Biomater. 2005;74(1):495-503. PMID: 15912530. https://doi.org/10.1002/jbm.b.30228
  24. Васильева М.Б., Кузнецова Е.В., Русакова Я.Л., Чепелева Е.В., Сергеевичев Д.С., Журавлева И.Ю. Механические свойства нативной и децеллюляризованной стенки аорты после длительного хранения в биоцидных растворах. Вестник транcплантологии и искусственных органов. 2021;23(4):86-94. https://doi.org/10.15825/1995-1191-2021-4-86-94 Vasilyeva M.B., Kuznetsova E.V., Rusakova Ya.L., Chepeleva E.V., Sergeevichev D.S., Juravleva I.Yu. Mechanical properties of na- tive and decellularized aortic wall after long-term storage in bio- cide solutions. Russian Journal of Transplantology and Artificial Organs. 2021;23(4):86-94. (In Russ.) https://doi.org/10.15825/1995-1191-2021-4-86-94
  25. Tzavellas N.P., Simos Y.V., Tsamis K.I., Markopoulos G.S., Lekkas P., Peschos D., Lakkas L. Decellularized scaffolds and heart valve treatment: present techniques, long-standing hurdles and the challenging future. Biomater Adv. 2025;177:214367. PMID: 40505382. https://doi.org/10.1016/j.bioadv.2025.214367
  26. Mangani S., Vetoulas M., Mineschou K., Spanopoulos K., Vivanco M.D., Piperigkou Z., Karamanos N.K. Design and Applications of Extracellular Matrix Scaffolds in Tissue Engineering and Regeneration. Cells. 2025;14(14):1076. PMID: 40710329; PMCID: PMC12293650. https://doi.org/10.3390/cells14141076
  27. Rabbani M., Zakian N., Alimoradi N. Contribution of Physical Methods in Decellularization of Animal Tissues. J Med Signals Sens. 2021;11(1):1–11. PMID: 34026585. PMCID: PMC8043117. https://doi.org/10.4103/jmss.jmss_2_20
  28. Васильева М.Б., Сергеевичев Д.С., Юношев А.С., Ларионов П.М., Новрузов Р.Б., Караськов А.М. Морфофункциональная оценка ферментативного и детергентного способов децеллюляризации сердечных аллографтов. Патология кровообращения и кардиохирургия. 2012;(16)2:77-80. https://doi.org/10.21688/1681-3472-2012-2-77-80 Vasilyeva M.B., Sergeevichev D.S., Yunoshev A.S., Larionov P.M., Novruzov R.B., Karaskov A.M. Morphofunctional assessment ofenzymatic and detergent methods of decellularization of cardi- ac allografts. Patologiya krovoobrashcheniya i kardiokhirurgiya = Circulation Pathology and Cardiac Surgery. 2012;(16)2:77–80. (In Russ.) https://doi.org/10.21688/1681-3472-2012-2-77-80
  29. Liao J., Joyce E.M., Sacks M.S. Effects of decellularization on the me- chanical and structural properties of the porcine aortic valve leaflet. Biomaterials. 2008;29(8):1065-1074. PMID: 8096223; PMCID: PMC2253688. https://doi.org/10.1016/j.biomaterials.2007.11.007
  30. Сергеевичев Д.С., Сергеевичева В.В., Субботовская А.И., Подхватилина Н.А., Васильев В.Ю., Русакова Я.Л. Токсическое влияние детергентов на мезенхимальные стромальные клетки человека при заселении графтов. Патология кровообращения и кардиохирургия. 2013;17(2):67–71. https://doi.org/10.21688/1681-3472-2013-2-67-71 Sergeevichev D. S., Sergeevicheva V. V., Subbotovskaya A. I., Podkhvatilina N. A., Vasiliev V. Yu., Rusakova Ya. L. Toxic effect of detergents on human mesenchymal stromal cells during graft colonization. Patologiya krovoobrashcheniya i kardiokhirurgiya = Circulation Pathology and Cardiac Surgery. 2013;17(2):67-71. (In Russ.) https://doi.org/10.21688/1681-3472-2013-2-67-71
  31. Grabenwöger M., Fitzal F., Sider J., Csekö C., Bergmeister H., Schima H., Husinsky W., Böck P., Wolner E. Endothelialization of biosynthetic vascular prostheses after laser perforation. Ann Thorac Surg. 1998;66(6 Suppl):S110-4. PMID: 9930428. https://doi.org/10.1016/s0003-4975(98)00982-5
  32. Anderson J.M. Biological responses to materials. Annu Rev Mater Res. 2001;31(1):81-110. https://doi.org/10.1146/annurev.matsci.31.1.81
  33. Иванов А.В., Липатов В.А., Лазаренко С.В., Жердев Н.Н., Северинов Д.А. Исследование реакции тканей лабораторных животных на имплантацию новых образцов сосудистых эндопротезов. Патология кровообращения и кардиохирургия. 2016;20(2):87-94. https://doi.org/10.21688/1681-3472-2016-2-87-94 Ivanov A.V., Lipatov V.A., Lazarenko S.V., Zherdev N.N., Severinov D.A. Studying the response of tissues of laboratory animals to implantation of vascular endoprostheses. Patologiya krovoobrashcheniya i kardiokhirurgiya = Circulation Pathology and Cardiac Surgery. 2016;20(2):87-94. (In Russ.) https://doi.org/10.21688/1681-3472-2016-2-87-94
  34. Мелконян К.И., Накохов Р.З., Русинова Т.В., Козмай Я.А., Быков И.М., Редько А.Н., Алексеенко С.Н. Показатели цитокинового профиля в сыворотке крови крыс при подкожной имплантации децеллюляризированного матрикса пищевода. Гены и клетки. 2020;15(4):46-56. https://doi.org/10.23868/202012008 Melkonyan K.I., Nakokhov R.Z., Rusinova T.V., Kozmai Y.A., Bykov I.M., Redko A.N., Alekseenko S.N. Serum cytokine profile indicators after subcutaneous implantation of the decellularized esophagus matrix in rats. Genes & Cells. 2020;15(4):46-56. (In Russ.) https://doi.org/10.23868/202012008
  35. Adam K., Günay H., Vaske B., Flohr M., Staufenbiel I. The granulation tissue preservation technique in regenerative periodontal surgery-a randomized controlled clinical trial. Clin Exp Dent Res. 2022;8(1):9-19. PMID: 35018724; PMCID: PMC8874108. https://doi.org/10.1002/cre2.532