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BMC biotechnology2017; 17(1); 13; doi: 10.1186/s12896-017-0329-6

Automated freeze-thaw cycles for decellularization of tendon tissue – a pilot study.

Abstract: Decellularization of tendon tissue plays a pivotal role in current tissue engineering approaches for in vitro research as well as for translation of graft-based tendon restoration into clinics. Automation of essential decellularization steps like freeze-thawing is crucial for the development of more standardized decellularization protocols and commercial graft production under good manufacturing practice (GMP) conditions in the future. In this study, a liquid nitrogen-based controlled rate freezer was utilized for automation of repeated freeze-thawing for decellularization of equine superficial digital flexor tendons. Additional tendon specimens underwent manually performed freeze-thaw cycles based on an established procedure. Tendon decellularization was completed by using non-ionic detergent treatment (Triton X-100). Effectiveness of decellularization was assessed by residual nuclei count and calculation of DNA content. Cytocompatibility was evaluated by culturing allogeneic adipose tissue-derived mesenchymal stromal cells on the tendon scaffolds. There were no significant differences in decellularization effectiveness between samples decellularized by the automated freeze-thaw procedure and samples that underwent manual freeze-thaw cycles. Further, we inferred no significant differences in the effectiveness of decellularization between two different cooling and heating rates applied in the automated freeze-thaw process. Both the automated protocols and the manually performed protocol resulted in roughly 2% residual nuclei and 13% residual DNA content. Successful cell culture was achieved with samples decellularized by automated freeze-thawing as well as with tendon samples decellularized by manually performed freeze-thaw cycles. Automated freeze-thaw cycles performed by using a liquid nitrogen-based controlled rate freezer were as effective as previously described manual freeze-thaw procedures for decellularization of equine superficial digital flexor tendons. The automation of this key procedure in decellularization of large tendon samples is an important step towards the processing of large sample quantities under standardized conditions. Furthermore, with a view to the production of commercially available tendon graft-based materials for application in human and veterinary medicine, the automation of key procedural steps is highly required to develop manufacturing processes under GMP conditions.
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Publication Date: 2017-02-14 PubMed ID: 28193263PubMed Central: PMC5307874DOI: 10.1186/s12896-017-0329-6Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research explores the use of automated freeze-thaw cycles, using a liquid nitrogen based controlled-rate freezer, to decellularize equine tendon tissue. The results indicated that automated processes were as effective as established manual methods, showing promise for standardization and commercial production of graft-based tendon restoration under good manufacturing practices.

Tissue Decellularization and Its Importance

  • The decellularization of tendon tissue plays a central role in tissue engineering, both for in-vitro studies and in methods using graft-based techniques for tendon restoration.
  • The study is focused on how this decellularization process can be automated, in a push for greater standardization, consistency, and efficiency in the process.

Methodology

  • The research team used a liquid nitrogen-based controlled rate freezer to automate the repeatability of the freeze-thaw cycle that is integral to tendon decellularization.
  • For comparison, manual freeze-thaw cycles were also performed using previous procedural methods.
  • The final stage of the tendon decellularization involved the use of a non-ionic detergent treatment.
  • The team evaluated the effectiveness of its decellularization technique by calculating residual nuclei and DNA content.
  • It further studied the cytocompatibility of the treated tissue by culturing mesenchymal stromal cells derived from allogeneic adipose tissue on the tendon scaffolds.

Findings

  • The results showed no significant variation between tendons decellularized using the automated freeze-thaw cycles and those decellularized via manual methods.
  • Tendons treated using automated and non-automated methods showed roughly 2% residual nuclei and 13% residual DNA content, which is a successful result for the desired decellularization.
  • Additionally, successful cell culture was feasible with samples processed by both automated and manual freeze-thaw cycles.

Significance and Future Directions

  • This study is an important advancement in creating a standardized, consistent, and efficient method of tendon decellularization.
  • The method proved promising for bringing commercialized graft-based tendon restoration closer to adherence with Good Manufacturing Practices (GMP), which is crucial for quality control in producing clinically-approved materials.
  • Looking forward, automating key procedural steps—like the freeze-thaw cycles used here—will be integral to developing manufacturing processes that meet GMP standards; this study makes a significant contribution to that goal.

Cite This Article

APA
Roth SP, Glauche SM, Plenge A, Erbe I, Heller S, Burk J. (2017). Automated freeze-thaw cycles for decellularization of tendon tissue – a pilot study. BMC Biotechnol, 17(1), 13. https://doi.org/10.1186/s12896-017-0329-6

Publication

BMC biotechnology
ISSN: 1472-6750
NlmUniqueID: 101088663
Country: England
Language: English
Volume: 17
Issue: 1
Pages: 13
PII: 13

Researcher Affiliations

Roth, Susanne Pauline
  • Large Animal Clinic for Surgery, University of Leipzig, An den Tierkliniken 21, Leipzig, 04103, Germany. susanne.roth@uni-leipzig.de.
  • Saxonian Incubator for Clinical Translation, University of Leipzig, Philipp-Rosenthal-Strasse 55, Leipzig, 04103, Germany. susanne.roth@uni-leipzig.de.
Glauche, Sina Marie
  • Saxonian Incubator for Clinical Translation, University of Leipzig, Philipp-Rosenthal-Strasse 55, Leipzig, 04103, Germany.
Plenge, Amelie
  • Tierklinik Kaufungen, Pfingstweide 2, Kaufungen, 34260, Germany.
Erbe, Ina
  • Large Animal Clinic for Surgery, University of Leipzig, An den Tierkliniken 21, Leipzig, 04103, Germany.
Heller, Sandra
  • Department of Pathology and Laboratory Medicine, Tulane University, New Orleans, USA.
Burk, Janina
  • Large Animal Clinic for Surgery, University of Leipzig, An den Tierkliniken 21, Leipzig, 04103, Germany.
  • Saxonian Incubator for Clinical Translation, University of Leipzig, Philipp-Rosenthal-Strasse 55, Leipzig, 04103, Germany.
  • Institute of Veterinary Physiology, University of Leipzig, An den Tierkliniken 7, Leipzig, 04103, Germany.

MeSH Terms

  • Animals
  • Cell Separation / instrumentation
  • Cells, Cultured
  • Equipment Design
  • Equipment Failure Analysis
  • Extracellular Matrix / chemistry
  • Freezing
  • Horses
  • Mesenchymal Stem Cell Transplantation / instrumentation
  • Mesenchymal Stem Cells / cytology
  • Pilot Projects
  • Robotics / instrumentation
  • Tendons / chemistry
  • Tendons / cytology
  • Tissue Engineering / instrumentation
  • Tissue Scaffolds

References

This article includes 43 references
  1. Cheng CW, Solorio LD, Alsberg E. Decellularized tissue and cell-derived extracellular matrices as scaffolds for orthopaedic tissue engineering.. Biotechnol Adv 2014 Mar-Apr;32(2):462-84.
    doi: 10.1016/j.biotechadv.2013.12.012pmc: PMC3959761pubmed: 24417915google scholar: lookup
  2. Yang G, Rothrauff BB, Tuan RS. Tendon and ligament regeneration and repair: clinical relevance and developmental paradigm.. Birth Defects Res C Embryo Today 2013 Sep;99(3):203-222.
    doi: 10.1002/bdrc.21041pmc: PMC4041869pubmed: 24078497google scholar: lookup
  3. Verdiyeva G, Koshy K, Glibbery N, Mann H, Seifalian AM. Tendon Reconstruction with Tissue Engineering Approach--A Review.. J Biomed Nanotechnol 2015 Sep;11(9):1495-523.
    doi: 10.1166/jbn.2015.2121pubmed: 26485923google scholar: lookup
  4. Siegel L, Vandenakker-Albanese C, Siegel D. Anterior cruciate ligament injuries: anatomy, physiology, biomechanics, and management.. Clin J Sport Med 2012 Jul;22(4):349-55.
    doi: 10.1097/JSM.0b013e3182580cd0pubmed: 22695402google scholar: lookup
  5. Lovati AB, Bottagisio M, Moretti M. Decellularized and Engineered Tendons as Biological Substitutes: A Critical Review.. Stem Cells Int 2016;2016:7276150.
    doi: 10.1155/2016/7276150pmc: PMC4736572pubmed: 26880985google scholar: lookup
  6. Youngstrom DW, Barrett JG. Engineering Tendon: Scaffolds, Bioreactors, and Models of Regeneration.. Stem Cells Int 2016;2016:3919030.
    doi: 10.1155/2016/3919030pmc: PMC4709784pubmed: 26839559google scholar: lookup
  7. Crowe CS, Chattopadhyay A, McGoldrick R, Chiou G, Pham H, Chang J. Characteristics of Reconstituted Lyophilized Tendon Hydrogel: An Injectable Scaffold for Tendon Regeneration.. Plast Reconstr Surg 2016 Mar;137(3):843-851.
    doi: 10.1097/01.prs.0000480012.41411.7cpubmed: 26910664google scholar: lookup
  8. Schulze-Tanzil G, Al-Sadi O, Ertel W, Lohan A. Decellularized tendon extracellular matrix-a valuable approach for tendon reconstruction?. Cells 2012 Nov 5;1(4):1010-28.
    doi: 10.3390/cells1041010pmc: PMC3901141pubmed: 24710540google scholar: lookup
  9. Gilbert TW. Strategies for tissue and organ decellularization.. J Cell Biochem 2012 Jul;113(7):2217-22.
    doi: 10.1002/jcb.24130pubmed: 22415903google scholar: lookup
  10. Gilbert TW, Sellaro TL, Badylak SF. Decellularization of tissues and organs.. Biomaterials 2006 Jul;27(19):3675-83.
    pubmed: 16519932doi: 10.1016/j.biomaterials.2006.02.014google scholar: lookup
  11. Burk J, Erbe I, Berner D, Kacza J, Kasper C, Pfeiffer B, Winter K, Brehm W. Freeze-thaw cycles enhance decellularization of large tendons.. Tissue Eng Part C Methods 2014 Apr;20(4):276-84.
    doi: 10.1089/ten.tec.2012.0760pmc: PMC3968887pubmed: 23879725google scholar: lookup
  12. Azuma C, Tohyama H, Nakamura H, Kanaya F, Yasuda K. Antibody neutralization of TGF-beta enhances the deterioration of collagen fascicles in a tissue-cultured tendon matrix with ex vivo fibroblast infiltration.. J Biomech 2007;40(10):2184-90.
    doi: 10.1016/j.jbiomech.2006.10.023pubmed: 17462658google scholar: lookup
  13. Omae H, Zhao C, Sun YL, An KN, Amadio PC. Multilayer tendon slices seeded with bone marrow stromal cells: a novel composite for tendon engineering.. J Orthop Res 2009 Jul;27(7):937-42.
    doi: 10.1002/jor.20823pmc: PMC5175470pubmed: 19105224google scholar: lookup
  14. Stewart AA, Barrett JG, Byron CR, Yates AC, Durgam SS, Evans RB, Stewart MC. Comparison of equine tendon-, muscle-, and bone marrow-derived cells cultured on tendon matrix.. Am J Vet Res 2009 Jun;70(6):750-7.
    doi: 10.2460/ajvr.70.6.750pubmed: 19496665google scholar: lookup
  15. Crapo PM, Gilbert TW, Badylak SF. An overview of tissue and whole organ decellularization processes.. Biomaterials 2011 Apr;32(12):3233-43.
    doi: 10.1016/j.biomaterials.2011.01.057pmc: PMC3084613pubmed: 21296410google scholar: lookup
  16. Pridgen BC, Woon CY, Kim M, Thorfinn J, Lindsey D, Pham H, Chang J. Flexor tendon tissue engineering: acellularization of human flexor tendons with preservation of biomechanical properties and biocompatibility.. Tissue Eng Part C Methods 2011 Aug;17(8):819-28.
    doi: 10.1089/ten.tec.2010.0457pubmed: 21548795google scholar: lookup
  17. Vavken P, Joshi S, Murray MM. TRITON-X is most effective among three decellularization agents for ACL tissue engineering.. J Orthop Res 2009 Dec;27(12):1612-8.
    doi: 10.1002/jor.20932pmc: PMC2824567pubmed: 19504590google scholar: lookup
  18. Whitlock PW, Smith TL, Poehling GG, Shilt JS, Van Dyke M. A naturally derived, cytocompatible, and architecturally optimized scaffold for tendon and ligament regeneration.. Biomaterials 2007 Oct;28(29):4321-9.
    doi: 10.1016/j.biomaterials.2007.05.029pubmed: 17610948google scholar: lookup
  19. Harrison RD, Gratzer PF. Effect of extraction protocols and epidermal growth factor on the cellular repopulation of decellularized anterior cruciate ligament allografts.. J Biomed Mater Res A 2005 Dec 15;75(4):841-54.
    doi: 10.1002/jbm.a.30486pubmed: 16123978google scholar: lookup
  20. Woods T, Gratzer PF. Effectiveness of three extraction techniques in the development of a decellularized bone-anterior cruciate ligament-bone graft.. Biomaterials 2005 Dec;26(35):7339-49.
    doi: 10.1016/j.biomaterials.2005.05.066pubmed: 16023194google scholar: lookup
  21. Cartmell JS, Dunn MG. Effect of chemical treatments on tendon cellularity and mechanical properties.. J Biomed Mater Res 2000 Jan;49(1):134-40.
    doi: 10.1002/(SICI)1097-4636(200001)49:1<134::AID-JBM17>3.0.CO;2-Dpubmed: 10559756google scholar: lookup
  22. Deeken CR, White AK, Bachman SL, Ramshaw BJ, Cleveland DS, Loy TS, Grant SA. Method of preparing a decellularized porcine tendon using tributyl phosphate.. J Biomed Mater Res B Appl Biomater 2011 Feb;96(2):199-206.
    doi: 10.1002/jbm.b.31753pubmed: 21210498google scholar: lookup
  23. Youngstrom DW, Barrett JG, Jose RR, Kaplan DL. Functional characterization of detergent-decellularized equine tendon extracellular matrix for tissue engineering applications.. PLoS One 2013;8(5):e64151.
    doi: 10.1371/journal.pone.0064151pmc: PMC3664617pubmed: 23724028google scholar: lookup
  24. Bottagisio M, Pellegata AF, Boschetti F, Ferroni M, Moretti M, Lovati AB. A new strategy for the decellularisation of large equine tendons as biocompatible tendon substitutes.. Eur Cell Mater 2016 Jul 8;32:58-73.
    doi: 10.22203/eCM.v032a04pubmed: 27386840google scholar: lookup
  25. Ning LJ, Zhang Y, Chen XH, Luo JC, Li XQ, Yang ZM, Qin TW. Preparation and characterization of decellularized tendon slices for tendon tissue engineering.. J Biomed Mater Res A 2012 Jun;100(6):1448-56.
    doi: 10.1002/jbm.a.34083pubmed: 22378703google scholar: lookup
  26. Ning LJ, Zhang YJ, Zhang Y, Qing Q, Jiang YL, Yang JL, Luo JC, Qin TW. The utilization of decellularized tendon slices to provide an inductive microenvironment for the proliferation and tenogenic differentiation of stem cells.. Biomaterials 2015 Jun;52:539-50.
    doi: 10.1016/j.biomaterials.2015.02.061pubmed: 25818459google scholar: lookup
  27. Durgam SS, Stewart AA, Pondenis HC, Gutierrez-Nibeyro SM, Evans RB, Stewart MC. Comparison of equine tendon- and bone marrow-derived cells cultured on tendon matrix with or without insulin-like growth factor-I supplementation.. Am J Vet Res 2012 Jan;73(1):153-61.
    doi: 10.2460/ajvr.73.1.153pubmed: 22204302google scholar: lookup
  28. Massie I, Selden C, Hodgson H, Fuller B, Gibbons S, Morris GJ. GMP cryopreservation of large volumes of cells for regenerative medicine: active control of the freezing process.. Tissue Eng Part C Methods 2014 Sep;20(9):693-702.
    doi: 10.1089/ten.tec.2013.0571pmc: PMC4152792pubmed: 24410575google scholar: lookup
  29. Devireddy RV, Neidert MR, Bischof JC, Tranquillo RT. Cryopreservation of collagen-based tissue equivalents. I. Effect of freezing in the absence of cryoprotective agents.. Tissue Eng 2003 Dec;9(6):1089-100.
    doi: 10.1089/10763270360728008pubmed: 14670097google scholar: lookup
  30. Gage AA, Baust J. Mechanisms of tissue injury in cryosurgery.. Cryobiology 1998 Nov;37(3):171-86.
    doi: 10.1006/cryo.1998.2115pubmed: 9787063google scholar: lookup
  31. Yiu WK, Basco MT, Aruny JE, Cheng SW, Sumpio BE. Cryosurgery: A review.. Int J Angiol 2007 Spring;16(1):1-6.
    doi: 10.1055/s-0031-1278235pmc: PMC2732998pubmed: 22477240google scholar: lookup
  32. Suto K, Urabe K, Naruse K, Uchida K, Matsuura T, Mikuni-Takagaki Y, Suto M, Nemoto N, Kamiya K, Itoman M. Repeated freeze-thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons.. Cell Tissue Bank 2012 Mar;13(1):71-80.
    doi: 10.1007/s10561-010-9234-0pmc: PMC3286509pubmed: 21116722google scholar: lookup
  33. Güngörmüş C, Kolankaya D, Aydin E. Histopathological and biomechanical evaluation of tenocyte seeded allografts on rat Achilles tendon regeneration.. Biomaterials 2015 May;51:108-118.
    doi: 10.1016/j.biomaterials.2015.01.077pubmed: 25771002google scholar: lookup
  34. Whitlock PW, Seyler TM, Parks GD, Ornelles DA, Smith TL, Van Dyke ME, Poehling GG. A novel process for optimizing musculoskeletal allograft tissue to improve safety, ultrastructural properties, and cell infiltration.. J Bone Joint Surg Am 2012 Aug 15;94(16):1458-67.
    doi: 10.2106/JBJS.K.01397pubmed: 22786867google scholar: lookup
  35. Liu W, Chen B, Deng D, Xu F, Cui L, Cao Y. Repair of tendon defect with dermal fibroblast engineered tendon in a porcine model.. Tissue Eng 2006 Apr;12(4):775-88.
    doi: 10.1089/ten.2006.12.775pubmed: 16674291google scholar: lookup
  36. Yin Z, Chen X, Zhu T, Hu JJ, Song HX, Shen WL, Jiang LY, Heng BC, Ji JF, Ouyang HW. The effect of decellularized matrices on human tendon stem/progenitor cell differentiation and tendon repair.. Acta Biomater 2013 Dec;9(12):9317-29.
    doi: 10.1016/j.actbio.2013.07.022pubmed: 23896565google scholar: lookup
  37. Ingram JH, Korossis S, Howling G, Fisher J, Ingham E. The use of ultrasonication to aid recellularization of acellular natural tissue scaffolds for use in anterior cruciate ligament reconstruction.. Tissue Eng 2007 Jul;13(7):1561-72.
    doi: 10.1089/ten.2006.0362pubmed: 17518726google scholar: lookup
  38. Martinello T, Bronzini I, Volpin A, Vindigni V, Maccatrozzo L, Caporale G, Bassetto F, Patruno M. Successful recellularization of human tendon scaffolds using adipose-derived mesenchymal stem cells and collagen gel.. J Tissue Eng Regen Med 2014 Aug;8(8):612-9.
    doi: 10.1002/term.1557pubmed: 22711488google scholar: lookup
  39. Woon CYL, Pridgen BC, Kraus A, Bari S, Pham H, Chang J. Optimization of human tendon tissue engineering: peracetic acid oxidation for enhanced reseeding of acellularized intrasynovial tendon.. Plast Reconstr Surg 2011 Mar;127(3):1107-1117.
    doi: 10.1097/PRS.0b013e318205f298pubmed: 21364414google scholar: lookup
  40. Issa RI, Engebretson B, Rustom L, McFetridge PS, Sikavitsas VI. The effect of cell seeding density on the cellular and mechanical properties of a mechanostimulated tissue-engineered tendon.. Tissue Eng Part A 2011 Jun;17(11-12):1479-87.
    doi: 10.1089/ten.tea.2010.0484pubmed: 21275843google scholar: lookup
  41. Thorpe CT, Clegg PD, Birch HL. A review of tendon injury: why is the equine superficial digital flexor tendon most at risk?. Equine Vet J 2010 Mar;42(2):174-80.
    doi: 10.2746/042516409X480395pubmed: 20156256google scholar: lookup
  42. Dowling BA, Dart AJ. Mechanical and functional properties of the equine superficial digital flexor tendon.. Vet J 2005 Sep;170(2):184-92.
    doi: 10.1016/j.tvjl.2004.03.021pubmed: 16129339google scholar: lookup
  43. Patterson-Kane JC, Rich T. Achilles tendon injuries in elite athletes: lessons in pathophysiology from their equine counterparts.. ILAR J 2014;55(1):86-99.
    doi: 10.1093/ilar/ilu004pubmed: 24936032google scholar: lookup

Citations

This article has been cited 34 times.
  1. Jin Y, Sun Q, Ma R, Li R, Qiao R, Li J, Wang L, Hu Y. The trend of allogeneic tendon decellularization: literature review. Cell Tissue Bank 2023 Jun 24;.
    doi: 10.1007/s10561-023-10097-xpubmed: 37355504google scholar: lookup
  2. Luo P, Huang R, Wu Y, Liu X, Shan Z, Gong L, Deng S, Liu H, Fang J, Wu S, Wu X, Liu Q, Chen Z, Yeung KWK, Qiao W, Chen S, Chen Z. Tailoring the multiscale mechanics of tunable decellularized extracellular matrix (dECM) for wound healing through immunomodulation. Bioact Mater 2023 Oct;28:95-111.
    doi: 10.1016/j.bioactmat.2023.05.011pubmed: 37250862google scholar: lookup
  3. Pantoja BTDS, Carvalho RC, Miglino MA, Carreira ACO. The Canine Pancreatic Extracellular Matrix in Diabetes Mellitus and Pancreatitis: Its Essential Role and Therapeutic Perspective. Animals (Basel) 2023 Feb 15;13(4).
    doi: 10.3390/ani13040684pubmed: 36830471google scholar: lookup
  4. Shang Y, Wang G, Zhen Y, Liu N, Nie F, Zhao Z, Li H, An Y. Application of decellularization-recellularization technique in plastic and reconstructive surgery. Chin Med J (Engl) 2023 Sep 5;136(17):2017-2027.
    doi: 10.1097/CM9.0000000000002085pubmed: 36752783google scholar: lookup
  5. Kuşoğlu A, Yangın K, Özkan SN, Sarıca S, Örnek D, Solcan N, Karaoğlu İC, Kızılel S, Bulutay P, Fırat P, Erus S, Tanju S, Dilege Ş, Öztürk E. Different Decellularization Methods in Bovine Lung Tissue Reveals Distinct Biochemical Composition, Stiffness, and Viscoelasticity in Reconstituted Hydrogels. ACS Appl Bio Mater 2023 Feb 20;6(2):793-805.
    doi: 10.1021/acsabm.2c00968pubmed: 36728815google scholar: lookup
  6. Wang B, Qinglai T, Yang Q, Li M, Zeng S, Yang X, Xiao Z, Tong X, Lei L, Li S. Functional acellular matrix for tissue repair. Mater Today Bio 2023 Feb;18:100530.
    doi: 10.1016/j.mtbio.2022.100530pubmed: 36601535google scholar: lookup
  7. Santos AL, Silva CGD, Barreto LSS, Tamaoki MJS, Almeida FG, Faloppa F. Automated Assessment of Cell Infiltration and Removal in Decellularized Scaffolds - Experimental Study in Rabbits. Rev Bras Ortop (Sao Paulo) 2022 Dec;57(6):992-1000.
    doi: 10.1055/s-0041-1739174pubmed: 36540747google scholar: lookup
  8. Marvin JC, Mochida A, Paredes J, Vaughn B, Andarawis-Puri N. Detergent-Free Decellularization Preserves the Mechanical and Biological Integrity of Murine Tendon. Tissue Eng Part C Methods 2022 Dec;28(12):646-655.
    doi: 10.1089/ten.TEC.2022.0135pubmed: 36326204google scholar: lookup
  9. Zhang Q, Hu Y, Long X, Hu L, Wu Y, Wu J, Shi X, Xie R, Bi Y, Yu F, Li P, Yang Y. Preparation and Application of Decellularized ECM-Based Biological Scaffolds for Articular Cartilage Repair: A Review. Front Bioeng Biotechnol 2022;10:908082.
    doi: 10.3389/fbioe.2022.908082pubmed: 35845417google scholar: lookup
  10. Moffat D, Ye K, Jin S. Decellularization for the retention of tissue niches. J Tissue Eng 2022 Jan-Dec;13:20417314221101151.
    doi: 10.1177/20417314221101151pubmed: 35620656google scholar: lookup
  11. Neishabouri A, Soltani Khaboushan A, Daghigh F, Kajbafzadeh AM, Majidi Zolbin M. Decellularization in Tissue Engineering and Regenerative Medicine: Evaluation, Modification, and Application Methods. Front Bioeng Biotechnol 2022;10:805299.
    doi: 10.3389/fbioe.2022.805299pubmed: 35547166google scholar: lookup
  12. Baranovskii D, Demner J, Nürnberger S, Lyundup A, Redl H, Hilpert M, Pigeot S, Krasheninnikov M, Krasilnikova O, Klabukov I, Parshin V, Martin I, Lardinois D, Barbero A. Engineering of Tracheal Grafts Based on Recellularization of Laser-Engraved Human Airway Cartilage Substrates. Cartilage 2022 Jan-Mar;13(1):19476035221075951.
    doi: 10.1177/19476035221075951pubmed: 35189712google scholar: lookup
  13. Zemmyo D, Yamamoto M, Miyata S. Efficient Decellularization by Application of Moderate High Hydrostatic Pressure with Supercooling Pretreatment. Micromachines (Basel) 2021 Nov 30;12(12).
    doi: 10.3390/mi12121486pubmed: 34945339google scholar: lookup
  14. Zhang X, Chen X, Hong H, Hu R, Liu J, Liu C. Decellularized extracellular matrix scaffolds: Recent trends and emerging strategies in tissue engineering. Bioact Mater 2022 Apr;10:15-31.
    doi: 10.1016/j.bioactmat.2021.09.014pubmed: 34901526google scholar: lookup
  15. Melzer M, Schubert S, Müller SF, Geyer J, Hagen A, Niebert S, Burk J. Rho/ROCK Inhibition Promotes TGF-β3-Induced Tenogenic Differentiation in Mesenchymal Stromal Cells. Stem Cells Int 2021;2021:8284690.
    doi: 10.1155/2021/8284690pubmed: 34659420google scholar: lookup
  16. Rabbani M, Zakian N, Alimoradi N. Contribution of Physical Methods in Decellularization of Animal Tissues. J Med Signals Sens 2021 Jan-Mar;11(1):1-11.
    doi: 10.4103/jmss.JMSS_2_20pubmed: 34026585google scholar: lookup
  17. Wei F, Liu S, Chen M, Tian G, Zha K, Yang Z, Jiang S, Li M, Sui X, Chen Z, Guo Q. Host Response to Biomaterials for Cartilage Tissue Engineering: Key to Remodeling. Front Bioeng Biotechnol 2021;9:664592.
    doi: 10.3389/fbioe.2021.664592pubmed: 34017827google scholar: lookup
  18. Heidarzadeh M, Rahbarghazi R, Saberianpour S, Delkhosh A, Amini H, Sokullu E, Hassanpour M. Distinct chemical composition and enzymatic treatment induced human endothelial cells survival in acellular ovine aortae. BMC Res Notes 2021 Apr 7;14(1):126.
    doi: 10.1186/s13104-021-05538-3pubmed: 33827673google scholar: lookup
  19. Schubert S, Brandt L, Burk J. A 3D Dynamic In Vitro Model of Inflammatory Tendon Disease. Methods Mol Biol 2021;2269:167-174.
    doi: 10.1007/978-1-0716-1225-5_12pubmed: 33687679google scholar: lookup
  20. Zemmyo D, Yamamoto M, Miyata S. Fundamental Study of Decellularization Method Using Cyclic Application of High Hydrostatic Pressure. Micromachines (Basel) 2020 Nov 15;11(11).
    doi: 10.3390/mi11111008pubmed: 33203164google scholar: lookup
  21. Mendibil U, Ruiz-Hernandez R, Retegi-Carrion S, Garcia-Urquia N, Olalde-Graells B, Abarrategi A. Tissue-Specific Decellularization Methods: Rationale and Strategies to Achieve Regenerative Compounds. Int J Mol Sci 2020 Jul 30;21(15).
    doi: 10.3390/ijms21155447pubmed: 32751654google scholar: lookup
  22. Dzobo K, Motaung KSCM, Adesida A. Recent Trends in Decellularized Extracellular Matrix Bioinks for 3D Printing: An Updated Review. Int J Mol Sci 2019 Sep 18;20(18).
    doi: 10.3390/ijms20184628pubmed: 31540457google scholar: lookup
  23. Jung SM, Chung GY, Shin HS. Artificial controlled model of blood circulation system for adhesive evaluation. Sci Rep 2017 Dec 1;7(1):16720.
    doi: 10.1038/s41598-017-16814-3pubmed: 29196674google scholar: lookup
  24. Egnot ML, Knight KM, Meyn LA, Shetty NA, Abramowitch SD, Moalli PA. Decellularization method and tissue region impact the structural properties and composition of porcine vaginal extracellular matrix. Acta Biomater 2026 Jan;210:206-220.
    doi: 10.1016/j.actbio.2025.12.025pubmed: 41397485google scholar: lookup
  25. Baranovskii D, Smirnova A, Yakimova A, Kisel A, Koryakin S, Atiakshin D, Ignatyuk M, Potievskiy M, Saburov V, Budnik S, Sulina Y, Stepanenko VN, Churyukin R, Akhmedov B, Shegay P, Kaprin AD, Klabukov I. Tissue-Cultured Chondrocytes Survive After Irradiation in 1300 Gy Dose. Biomedicines 2025 Sep 4;13(9).
    doi: 10.3390/biomedicines13092153pubmed: 41007716google scholar: lookup
  26. Zhang Y, Yu W. Recent advances in bionic scaffolds for cartilage tissue engineering. Front Bioeng Biotechnol 2025;13:1625550.
    doi: 10.3389/fbioe.2025.1625550pubmed: 40718699google scholar: lookup
  27. Li J, Wen M, Zhang S, Du L, Fan X, Liang H, Wang H, Sun J, Ding Y, Ge L, Ma J, Zhang J. Research Progress on the Preparation and Application of Decellularized Tendons. Curr Issues Mol Biol 2025 Apr 6;47(4).
    doi: 10.3390/cimb47040251pubmed: 40699650google scholar: lookup
  28. Shestakova VA, Klabukov ID, Kolobaev IV, Rao L, Atiakshin DA, Ignatyuk MA, Krasheninnikov ME, Ahmedov BG, Ivanov SA, Shegay PV, Kaprin AD, Baranovskii DS. Pathologically altered articular cartilage attracts intense chondrocyte invasion into the extracellular matrix: in vitro pilot study. Knee Surg Relat Res 2024 Dec 3;36(1):42.
    doi: 10.1186/s43019-024-00249-ypubmed: 39627845google scholar: lookup
  29. Kisel AA, Stepanov VA, Isaeva EV, Demyashkin GA, Isaev EI, Smirnova EI, Yatsenko EM, Afonin GV, Ivanov SA, Atiakshin DA, Shegay PV, Kaprin AD, Klabukov ID, Baranovskii DS. Cartilage Laser Engraving for Fast-Track Tissue Engineering of Auricular Grafts. Int J Mol Sci 2024 Oct 27;25(21).
    doi: 10.3390/ijms252111538pubmed: 39519090google scholar: lookup
  30. Hussein KH, Ahmadzada B, Correa JC, Sultan A, Wilken S, Amiot B, Nyberg SL. Liver tissue engineering using decellularized scaffolds: Current progress, challenges, and opportunities. Bioact Mater 2024 Oct;40:280-305.
    doi: 10.1016/j.bioactmat.2024.06.001pubmed: 38973992google scholar: lookup
  31. Lan X, Luo M, Li M, Mu L, Li G, Chen G, He Z, Xiao J. Swim bladder-derived biomaterials: structures, compositions, properties, modifications, and biomedical applications. J Nanobiotechnology 2024 Apr 17;22(1):186.
    doi: 10.1186/s12951-024-02449-wpubmed: 38632585google scholar: lookup
  32. Gadre M, Kasturi M, Agarwal P, Vasanthan KS. Decellularization and Their Significance for Tissue Regeneration in the Era of 3D Bioprinting. ACS Omega 2024 Feb 20;9(7):7375-7392.
    doi: 10.1021/acsomega.3c08930pubmed: 38405516google scholar: lookup
  33. Noro J, Vilaça-Faria H, Reis RL, Pirraco RP. Extracellular matrix-derived materials for tissue engineering and regenerative medicine: A journey from isolation to characterization and application. Bioact Mater 2024 Apr;34:494-519.
    doi: 10.1016/j.bioactmat.2024.01.004pubmed: 38298755google scholar: lookup
  34. Data K, Kulus M, Ziemak H, Chwarzyński M, Piotrowska-Kempisty H, Bukowska D, Antosik P, Mozdziak P, Kempisty B. Decellularization of Dense Regular Connective Tissue-Cellular and Molecular Modification with Applications in Regenerative Medicine. Cells 2023 Sep 16;12(18).
    doi: 10.3390/cells12182293pubmed: 37759515google scholar: lookup
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