Decellularized Scaffolds for Ligament/Tendon Regeneration: Challenges, Opportunities and Future Directions
DOI:
https://doi.org/10.65795/pfv27w47Keywords:
tendon, ligament, injuries, decellularized, regeneration, extracellular matrixAbstract
Decellularized scaffolds have gained significant attention in tissue engineering, particularly for tendon and ligament regeneration, due to their ability to retain native extracellular matrix (ECM) architecture and biochemical cues. These decellularized scaffolds offer a biocompatible and bioactive framework that closely mimics the native tissue niche, promoting cellular infiltration, proliferation, and differentiation. Unlike synthetic materials, decellularized tissues can better support functional healing while minimizing immune responses. Tendon and ligament injuries pose a unique challenge due to the dense collagenous structure, limited vascularization, and poor intrinsic healing capacity of these tissues. While autografts and allografts are commonly used, limitations such as donor site morbidity, limited availability, and risk of immune rejection pose treats. Decellularized scaffolds offer a compelling solution by providing mechanical support while promoting natural tissue regeneration. Yet, developing these scaffolds remains challenging, as differences in tissue size, ECM composition, and porosity make it difficult to apply a universal decellularization approach. The major decellularization hurdle is to remove cellular components effectively without compromising the scaffold’s structural integrity or biological activity. In addition, achieving immune tolerance and enhancing vascular growth are essential for proper integration and long-lasting performance of the graft. This review highlights the latest advancements in decellularized scaffolds for tendon and ligament repair, discussing the various physical, chemical, and enzymatic methods used for decellularization. It also examines current limitations and strategies being explored to overcome them, such as combining scaffolds with growth factors, stem cells, or pro-angiogenic agents. Beyond musculoskeletal repair, the potential of decellularized scaffolds is expanding into broader applications in tissue and organ engineering. Future research should focus on standardizing decellularization protocols, improving vascular integration, and translating laboratory success into clinically viable products that can enhance patient outcomes in regenerative medicine.
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Jung H-J, Fisher MB, Woo SL-Y. Role of biomechanics in the understanding of normal, injured, and healing ligaments and tendons. BMC Sports Science, Medicine and Rehabilitation 2009 1:1 2009;1:1–17. https://doi.org/10.1186/1758-2555-1-9.
Moshiri A. Tendon and Ligament Tissue Engineering, Healing and Regenerative Medicine. J Sports Med Doping Stud 2013;03. https://doi.org/10.4172/2161-0673.1000126.
Lim WL, Liau LL, Ng MH, Chowdhury SR, Law JX. Current Progress in Tendon and Ligament Tissue Engineering. Tissue Eng Regen Med 2019;16:549–71. https://doi.org/10.1007/S13770-019-00196-W/FIGURES/9.
Wu F, Nerlich M, Docheva D. Tendon injuries: Basic science and new repair proposals. EFORT Open Rev 2017;2:332–42. https://doi.org/10.1302/2058-5241.2.160075.
Yelin E, Weinstein S, King T. The burden of musculoskeletal diseases in the United States. Semin Arthritis Rheum 2016;46:259–60. https://doi.org/10.1016/j.semarthrit.2016.07.013.
John R, Dhillon MS, Syam K, Prabhakar S, Behera P, Singh H. Epidemiological profile of sports-related knee injuries in northern India: An observational study at a tertiary care centre. J Clin Orthop Trauma 2016;7:207–11. https://doi.org/10.1016/J.JCOT.2016.02.003.
Hootman JM, Dick R, Agel J. Epidemiology of Collegiate Injuries for 15 Sports: Summary and Recommendations for Injury Prevention Initiatives. J Athl Train 2007;42:311.
Blaudez F, Ivanovski S, Hamlet S, Vaquette C. An overview of decellularisation techniques of native tissues and tissue engineered products for bone, ligament and tendon regeneration. Methods 2020;171:28–40. https://doi.org/10.1016/j.ymeth.2019.08.002.
Leong NL, Kator JL, Clemens TL, James A, Enamoto-Iwamoto M, Jiang J. Tendon and Ligament Healing and Current Approaches to Tendon and Ligament Regeneration. J Orthop Res 2019;38:7. https://doi.org/10.1002/JOR.24475.
Zhao X, Zhou Y, Li J, Zhang C, Wang J. Opportunities and challenges of hydrogel microspheres for tendon–bone healing after anterior cruciate ligament reconstruction. J Biomed Mater Res B Appl Biomater 2022;110:289–301. https://doi.org/10.1002/JBM.B.34925.
Anjum S, Li T, Saeed M, Ao Q. Exploring polysaccharide and protein-enriched decellularized matrix scaffolds for tendon and ligament repair: A review. Int J Biol Macromol 2024;254. https://doi.org/10.1016/J.IJBIOMAC.2023.127891.
K N, CA V, Joseph J, U A, John A, Abraham A. Mesenchymal Stem Cells Seeded Decellularized Tendon Scaffold for Tissue Engineering. Curr Stem Cell Res Ther 2020;16:155–64. https://doi.org/10.2174/1574888X15666200723123901.
Lim WL, Liau LL, Ng MH, Chowdhury SR, Law JX. Current Progress in Tendon and Ligament Tissue Engineering. Tissue Eng Regen Med 2019;16:549–71. https://doi.org/10.1007/S13770-019-00196-W.
Elmashhady HH, Kraemer BA, Patel KH, Sell SA, Garg K. Decellularized extracellular matrices for tissue engineering applications. Electrospinning 2017;1. https://doi.org/10.1515/ESP-2017-0005.
van Hengel EVA, van der Laan LJW, de Jonge J, Verstegen MMA. Towards Safety and Regulation Criteria for Clinical Applications of Decellularized Organ-Derived Matrices. Bioengineering 2025;12. https://doi.org/10.3390/BIOENGINEERING12020136.
Taylor DA, Sampaio LC, Ferdous Z, Gobin AS, Taite LJ. Decellularized matrices in regenerative medicine. Acta Biomater 2018;74:74–89. https://doi.org/10.1016/j.actbio.2018.04.044.
Amiel D, Frank C, Harwood F, Fronek J, Akeson W. Tendons and ligaments: A morphological and biochemical comparison. Journal of Orthopaedic Research 1983;1:257–65. https://doi.org/10.1002/JOR.1100010305.
Subramanian A, Schilling TF. Tendon development and musculoskeletal assembly: emerging roles for the extracellular matrix. Development 2015;142:4191. https://doi.org/10.1242/DEV.114777.
Tai CC, Hui J, Williams A. Ligaments and Tendons. Basic Orthopaedic Sciences 2018:123–35. https://doi.org/10.1201/9781315117294-8.
Kirkendall DT, Garrett WE. Function and biomechanics of tendons. Scand J Med Sci Sports 1997;7:62–6. https://doi.org/10.1111/J.1600-0838.1997.TB00120.X.
Holmes DF, Graham HK, Kadler KE. Collagen fibrils forming in developing tendon show an early and abrupt limitation in diameter at the growing tips. J Mol Biol 1998;283:1049–58. https://doi.org/10.1006/jmbi.1998.2153.
Smith KD, Clegg PD, Innes JF, Comerford EJ. Elastin content is high in the canine cruciate ligament and is associated with degeneration. Vet J 2014;199:169. https://doi.org/10.1016/J.TVJL.2013.11.002.
Svärd A, Hammerman M, Eliasson P. Elastin levels are higher in healing tendons than in intact tendons and influence tissue compliance. FASEB Journal 2020;34:13409–18. https://doi.org/10.1096/FJ.202001255R.
Tozer S, Duprez D. Tendon and ligament: Development, repair and disease. Birth Defects Res C Embryo Today 2005;75:226–36. https://doi.org/10.1002/BDRC.20049.
Özer Kaya D. Architecture of tendon and ligament and their adaptation to pathological conditions. Comparative Kinesiology of the Human Body: Normal and Pathological Conditions 2020:115–47. https://doi.org/10.1016/B978-0-12-812162-7.00009-6.
Chamberlain CS, Crowley EM, Kobayashi H, Eliceiri KW, Vanderby R. Quantification of Collagen Organization and Extracellular Matrix Factors within the Healing Ligament. Microscopy and Microanalysis 2011;17:779. https://doi.org/10.1017/S1431927611011925.
Franchi M, Trirè A, Quaranta M, Orsini E, Ottani V. Collagen structure of tendon relates to function. ScientificWorldJournal 2007;7:404–20. https://doi.org/10.1100/TSW.2007.92.
Hauser RA, Blakemore PJ, Wang J, Steilen D. Structural Basis of Joint Instability as Cause for Chronic Musculoskeletal Pain and Its Successful Treatment with Regenerative Injection Therapy (Prolotherapy). AACE Clin Case Rep 2021;7:1. https://doi.org/10.2174/1876386301407010009.
O’Brien M. Structure and metabolism of tendons. Scand J Med Sci Sports 1997;7:55–61. https://doi.org/10.1111/J.1600-0838.1997.TB00119.X.
Franchi M, Raspanti M, Dell’Orbo C, Quaranta M, De Pasquale V, Ottani V, et al. Different crimp patterns in collagen fibrils relate to the subfibrillar arrangement. Connect Tissue Res 2008;49:85–91. https://doi.org/10.1080/03008200801913635.
Thorpe CT, Screen HRC. Tendon structure and composition. Adv Exp Med Biol 2016;920:3–10. https://doi.org/10.1007/978-3-319-33943-6_1.
Wu F, Nerlich M, Docheva D. Tendon injuries: Basic science and new repair proposals. EFORT Open Rev 2017;2:332–42. https://doi.org/10.1302/2058-5241.2.160075.
LaBella CR, Hennrikus W, Hewett TE, Brenner JS, Brooks A, Demorest RA, et al. Anterior cruciate ligament injuries: Diagnosis, treatment, and prevention. Pediatrics 2014;133. https://doi.org/10.1542/PEDS.2014-0623.
Beck NA, Lawrence JTR, Nordin JD, DeFor TA, Tompkins M. ACL tears in school-aged children and adolescents over 20 years. Pediatrics 2017;139. https://doi.org/10.1542/PEDS.2016-1877.
Hauser RA. Ligament Injury and Healing: A Review of Current Clinical Diagnostics and Therapeutics. Open Rehabil J 2013;6:1–20. https://doi.org/10.2174/1874943701306010001.
Siegel L, Vandenakker-Albanese C, Siegel D. Anterior cruciate ligament injuries: Anatomy, physiology, biomechanics, and management. Clinical Journal of Sport Medicine 2012;22:349–55. https://doi.org/10.1097/JSM.0B013E3182580CD0.
Thomopoulos S, Parks WC, Rifkin DB, Derwin KA. Mechanisms of tendon injury and repair. Journal of Orthopaedic Research 2015;33:832–9. https://doi.org/10.1002/JOR.22806.
Rhode BA, Rhode WS. Occupational Risk Factors for Shoulder Tendon Disorders 2015 Update n.d. https://doi.org/10.15406/mojor.2015.03.00104.
Kjær M, Petersen J, Dünweber MR, Andersen JL, Engebretsen L, Magnusson SP. Dilemma in the Treatment of Sports Injuries in Athletes: Tendon Overuse, Muscle Strain, and Tendon Rupture. Scand J Med Sci Sports 2025;35:e70026. https://doi.org/10.1111/SMS.70026;WGROUP:STRING:PUBLICATION.
Leong HT, Fu SC, He X, Oh JH, Yamamoto N, Yung SHP. Risk factors for rotator cuff tendinopathy: A systematic review and meta-analysis. J Rehabil Med 2019;51:627–37. https://doi.org/10.2340/16501977-2598.
Thomopoulos S, Parks WC, Rifkin DB, Derwin KA. Mechanisms of tendon injury and repair. Journal of Orthopaedic Research 2015;33:832–9. https://doi.org/10.1002/JOR.22806.
Hauser RA, Woldin BA. Joint Instability as the Cause of Chronic Musculoskeletal Pain and Its Successful Treatment with Prolotherapy. Anatomy, Posture, Prevalence, Pain, Treatment and Interventions of Musculoskeletal Disorders 2018. https://doi.org/10.5772/INTECHOPEN.74384.
Capotosto S, Nazemi AK, Komatsu DE, Penna J. Prolotherapy in the Treatment of Sports-Related Tendinopathies: A Systematic Review of Randomized Controlled Trials. Orthop J Sports Med 2024;12. https://doi.org/10.1177/23259671241275087/SUPPL_FILE/SJ-PDF-1-OJS-10.1177_23259671241275087.PDF.
Duchman KR, Lemmex DB, Patel SH, Ledbetter L, Garrigues GE, Riboh JC. The Effect of Non-Steroidal Anti-Inflammatory Drugs on Tendon-to-Bone Healing: A Systematic Review with Subgroup Meta-Analysis. Iowa Orthop J 2019;39:107.
Ghosh N, Kolade OO, Shontz E, Rosenthal Y, Zuckerman JD, Bosco JA, et al. Nonsteroidal Anti-Inflammatory Drugs (NSAIDs) and Their Effect on Musculoskeletal Soft-Tissue Healing: A Scoping Review. JBJS Rev 2019;7. https://doi.org/10.2106/JBJS.RVW.19.00055.
Elder CL, Dahners LE, Weinhold PS. A Cyclooxygenase-2 Inhibitor Impairs Ligament Healing in the Rat. Am J Sports Med 2001;29:801–5. https://doi.org/10.1177/03635465010290062101.
Meheux CJ, Jack RA, McCulloch PC, Lintner DM, Harris JD. Surgical Management of Simultaneous Anterior Cruciate Ligament and Patellar Tendon Ruptures: A Systematic Review. Journal of Knee Surgery 2018;31:875–83. https://doi.org/10.1055/S-0037-1615814/ID/JR170135OA-32/BIB.
Lim WL, Liau LL, Ng MH, Chowdhury SR, Law JX. Current Progress in Tendon and Ligament Tissue Engineering. Tissue Eng Regen Med 2019;16:549–71. https://doi.org/10.1007/S13770-019-00196-W/FIGURES/9.
Sepúlveda F, Sánchez L, Amy E, Micheo W. Anterior cruciate ligament injury: Return to play, function and long-term considerations. Curr Sports Med Rep 2017;16:172–8. https://doi.org/10.1249/JSR.0000000000000356.
Mooney RA, Sampson ER, Lerea J, Rosier RN, Zuscik MJ. High-fat diet accelerates progression of osteoarthritis after meniscal/ligamentous injury. Arthritis Res Ther 2011;13. https://doi.org/10.1186/AR3529.
Messina OD, Vidal Wilman M, Vidal Neira LF. Nutrition, osteoarthritis and cartilage metabolism. Aging Clin Exp Res 2019;31:807–13. https://doi.org/10.1007/S40520-019-01191-W.
Tarantino D, Mottola R, Sirico F, Corrado B, Ruosi C, Saggini R, et al. Exploring the impact of vitamin D on tendon health: A comprehensive review. J Basic Clin Physiol Pharmacol 2024;35:143–52. https://doi.org/10.1515/JBCPP-2024-0061/MACHINEREADABLECITATION/RIS.
Rabago D, Nourani B. Prolotherapy for Osteoarthritis and Tendinopathy: a Descriptive Review. Curr Rheumatol Rep 2017;19:1–8. https://doi.org/10.1007/S11926-017-0659-3/METRICS.
Hauser RA, Blakemore PJ, Wang J, Steilen D. Structural Basis of Joint Instability as Cause for Chronic Musculoskeletal Pain and Its Successful Treatment with Regenerative Injection Therapy (Prolotherapy). Open Pain J 2014;7:09–22. https://doi.org/10.2174/1876386301407010009.
Dhammi IK, Rehan-Ul-Haq, Kumar S. Graft choices for anterior cruciate ligament reconstruction. Indian J Orthop 2015;49:127. https://doi.org/10.4103/0019-5413.152393.
Macaulay AA, Perfetti DC, Levine WN. Anterior Cruciate Ligament Graft Choices. Sports Health 2012;4:63. https://doi.org/10.1177/1941738111409890.
Pujji O, Keswani N, Collier N, Black M, Doos L. Evaluating the functional results and complications of autograft vs allograft use for reconstruction of the anterior cruciate ligament: A systematic review. Orthop Rev (Pavia) 2017;9:32–7. https://doi.org/10.4081/OR.2017.6833.
Smolle MA, Fischerauer SF, Zötsch S, Kiegerl A V., Sadoghi P, Gruber G, et al. Long-term outcomes of surgery using the Ligament Advanced Reinforcement System as treatment for anterior cruciate ligament tears. Bone and Joint Journal 2022;104:242–8. https://doi.org/10.1302/0301-620X.104B2.BJJ-2021-0798.R2/LETTERTOEDITOR.
Ratcliffe A, Butler DL, Dyment NA, Cagle PJ, Proctor CS, Ratcliffe SS, et al. Scaffolds for Tendon and Ligament Repair and Regeneration. Ann Biomed Eng 2015;43:819–31. https://doi.org/10.1007/S10439-015-1263-1/FIGURES/5.
Jayasree A, Kottappally Thankappan S, Ramachandran R, Sundaram MN, Chen CH, Mony U, et al. Bioengineered Braided Micro-Nano (Multiscale) Fibrous Scaffolds for Tendon Reconstruction. ACS Biomater Sci Eng 2019;5:1476–86. https://doi.org/10.1021/ACSBIOMATERIALS.8B01328/ASSET/IMAGES/MEDIUM/AB-2018-013286_0009.GIF.
Rothrauff BB, Lauro BB, Yang G, Debski RE, Musahl V, Tuan RS. Braided and Stacked Electrospun Nanofibrous Scaffolds for Tendon and Ligament Tissue Engineering. Tissue Eng Part A 2017;23:378–89. https://doi.org/10.1089/TEN.TEA.2016.0319.
Yao S, Liang Z, Lee YW, Yung PSH, Lui PPY. Bioactive Decellularized Tendon-Derived Stem Cell Sheet for Promoting Graft Healing After Anterior Cruciate Ligament Reconstruction. American Journal of Sports Medicine 2023;51:66–80. https://doi.org/10.1177/03635465221135770/SUPPL_FILE/SJ-PDF-1-AJS-10.1177_03635465221135770.PDF.
Anjum S, Li T, Saeed M, Ao Q. Exploring polysaccharide and protein-enriched decellularized matrix scaffolds for tendon and ligament repair: A review. Int J Biol Macromol 2024;254:127891. https://doi.org/10.1016/J.IJBIOMAC.2023.127891.
Wang S, Wang Y, Song L, Chen J, Ma Y, Chen Y, et al. Decellularized tendon as a prospective scaffold for tendon repair. Materials Science and Engineering C 2017;77:1290–301. https://doi.org/10.1016/j.msec.2017.03.279.
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. https://doi.org/10.3389/FBIOE.2022.805299/XML.
Keane TJ, Swinehart IT, Badylak SF. Methods of tissue decellularization used for preparation of biologic scaffolds and in vivo relevance. Methods 2015;84:25–34. https://doi.org/10.1016/J.YMETH.2015.03.005.
Rabbani M, Zakian N, Alimoradi N. Contribution of Physical Methods in Decellularization of Animal Tissues. J Med Signals Sens 2021;11:1–11. https://doi.org/10.4103/JMSS.JMSS_2_20.
Miranda CMFC, Leonel LCPC, Cañada RR, Maria DA, Miglino MA, Del Sol M, et al. Effects of chemical and physical methods on decellularization of murine skeletal muscles. An Acad Bras Cienc 2021;93:e20190942. https://doi.org/10.1590/0001-3765202120190942.
Roth SP, Glauche SM, Plenge A, Erbe I, Heller S, Burk J. Automated freeze-thaw cycles for decellularization of tendon tissue - a pilot study. BMC Biotechnol 2017;17:1–10. https://doi.org/10.1186/S12896-017-0329-6/FIGURES/5.
Szynkaruk M, Kemp SWP, Wood MD, Gordon T, Borschel GH. Experimental and clinical evidence for use of decellularized nerve allografts in peripheral nerve gap reconstruction. Tissue Eng Part B Rev 2013;19:83–96. https://doi.org/10.1089/TEN.TEB.2012.0275;ISSUE:ISSUE:10.1089/TEB.2013.19.ISSUE-1;WGROUP:STRING:PUBLICATION.
Kishida A. Preparation of the materials for regenerative medicine from biological tissues/organs by using high hydrostatic pressure decellularization method. Review of High Pressure Science and Technology/Koatsuryoku No Kagaku To Gijutsu 2020;30:36–46. https://doi.org/10.4131/JSHPREVIEW.30.36.
Mahara A, Morimoto N, Sakuma T, Fujisato T, Yamaoka T. Complete cell killing by applying high hydrostatic pressure for acellular vascular graft preparation. Biomed Res Int 2014;2014. https://doi.org/10.1155/2014/379607.
Zager Y, Kain D, Landa N, Leor J, Maor E. Optimization of Irreversible Electroporation Protocols for In-vivo Myocardial Decellularization. PLoS One 2016;11:e0165475. https://doi.org/10.1371/JOURNAL.PONE.0165475.
Sano MB, Neal RE, Garcia PA, Gerber D, Robertson J, Davalos R V. Towards the creation of decellularized organ constructs using irreversible electroporation and active mechanical perfusion. Biomed Eng Online 2010;9:1–16. https://doi.org/10.1186/1475-925X-9-83/FIGURES/6.
Sherifi I, Bachy M, Laumonier T, Petite H, Hannouche D. Use of supercritical carbon dioxide technology for fabricating a tissue engineering scaffold for anterior cruciate ligament repair. Sci Rep 2020;10:1–12. https://doi.org/10.1038/S41598-020-70994-Z;SUBJMETA=54,61,631,990;KWRD=BIOMATERIALS,BIOMEDICAL+MATERIALS,BIOTECHNOLOGY.
Huang C-C. Newly Designed Decellularized Scaffolds for Scaffold-based Gene Therapy from Elastic Cartilages via Supercritical Carbon Dioxide Fluid and Alkaline/ Protease Treatments. Curr Gene Ther 2021;22:162–7. https://doi.org/10.2174/1566523219666210618151843/CITE/REFWORKS.
de Wit RJJ, van Dis DJ, Bertrand ME, Tiemessen D, Siddiqi S, Oosterwijk E, et al. Scaffold-based tissue engineering: Supercritical carbon dioxide as an alternative method for decellularization and sterilization of dense materials. Acta Biomater 2023;155:323–32. https://doi.org/10.1016/J.ACTBIO.2022.11.028.
Akbari Zahmati AH, Alipoor R, Rezaei Shahmirzadi A, Khori V, Abolhasani MM. Chemical Decellularization Methods and Its Effects on Extracellular Matrix. Internal Medicine and Medical Investigation Journal 2017;2:76. https://doi.org/10.24200/IMMINV.V2I3.63.
Yamanaka H, Morimoto N, Yamaoka T. Decellularization of submillimeter-diameter vascular scaffolds using peracetic acid. Journal of Artificial Organs 2020;23:156–62. https://doi.org/10.1007/S10047-019-01152-0/METRICS.
Lin HJ, Wang W, Huang YY, Liao WT, Lin TY, Lin SY, et al. Decellularized Lymph Node Scaffolding as a Carrier for Dendritic Cells to Induce Anti-Tumor Immunity. Pharmaceutics 2019, Vol 11, Page 553 2019;11:553. https://doi.org/10.3390/PHARMACEUTICS11110553.
Sengyoku H, Tsuchiya T, Obata T, Doi R, Hashimoto Y, Ishii M, et al. Sodium hydroxide based non-detergent decellularizing solution for rat lung. Organogenesis 2018;14:94–106. https://doi.org/10.1080/15476278.2018.1462432;JOURNAL:JOURNAL:KOGG20;PAGE:STRING:ARTICLE/CHAPTER.
Emami A, Izadi E. Decellularizing Bone Tissue: Various Protocols. Regenerative Engineering and Translational Medicine 2024 2024:1–13. https://doi.org/10.1007/S40883-024-00375-9.
Moffat D, Ye K, Jin S. Decellularization for the retention of tissue niches. J Tissue Eng 2022;13. https://doi.org/10.1177/20417314221101151/ASSET/53194D1C-B190-4223-8469-0DDAA666DD82/ASSETS/IMAGES/LARGE/10.1177_20417314221101151-FIG2.JPG.
Roosens A, Somers P, De Somer F, Carriel V, Van Nooten G, Cornelissen R. Impact of Detergent-Based Decellularization Methods on Porcine Tissues for Heart Valve Engineering. Ann Biomed Eng 2016;44:2827–39. https://doi.org/10.1007/S10439-016-1555-0/FIGURES/6.
Gupta SK, Mishra NC, Dhasmana A. Decellularization Methods for Scaffold Fabrication. Methods in Molecular Biology 2017;1577:1–10. https://doi.org/10.1007/7651_2017_34.
Roth SP, Erbe I, Burk J. Decellularization of Large Tendon Specimens: Combination of Manually Performed Freeze-Thaw Cycles and Detergent Treatment. Methods in Molecular Biology 2017;1577:227–37. https://doi.org/10.1007/7651_2017_49.
Nellinger S, Mrsic I, Keller S, Heine S, Southan A, Bach M, et al. Cell-derived and enzyme-based decellularized extracellular matrix exhibit compositional and structural differences that are relevant for its use as a biomaterial. Biotechnol Bioeng 2022;119:1142–56. https://doi.org/10.1002/BIT.28047;WEBSITE:WEBSITE:ANALYTICALSCIENCEJOURNALS;WGROUP:STRING:PUBLICATION.
Shin YH, Park SY, Kim JK. Comparison of systematically combined detergent and nuclease-based decellularization methods for acellular nerve graft: An ex vivo characterization and in vivo evaluation. J Tissue Eng Regen Med 2019;13:1241–52. https://doi.org/10.1002/TERM.2874;JOURNAL:JOURNAL:19327005;PAGE:STRING:ARTICLE/CHAPTER.
Gilbert TW, Sellaro TL, Badylak SF. Decellularization of tissues and organs. Biomaterials 2006;27:3675–83. https://doi.org/10.1016/J.BIOMATERIALS.2006.02.014.
Giraldo-Gomez DM, Leon-Mancilla B, Del Prado-Audelo ML, Sotres-Vega A, Villalba-Caloca J, Garciadiego-Cazares D, et al. Trypsin as enhancement in cyclical tracheal decellularization: Morphological and biophysical characterization. Materials Science and Engineering: C 2016;59:930–7. https://doi.org/10.1016/J.MSEC.2015.10.094.
Lin CH, Kao YC, Ma H, Tsay RY. An investigation on the correlation between the mechanical property change and the alterations in composition and microstructure of a porcine vascular tissue underwent trypsin-based decellularization treatment. J Mech Behav Biomed Mater 2018;86:199–207. https://doi.org/10.1016/j.jmbbm.2018.06.029.
Liu YW, Huang CC, Wang YY, Xu J, Wang GD, Bai XP. Biological evaluations of decellularized extracellular matrix collagen microparticles prepared based on plant enzymes and aqueous two-phase method. Regen Biomater 2021;8. https://doi.org/10.1093/RB/RBAB002.
Fuetterer L. OPTIMIZATION AND BIOLOGICAL CHARACTERIZATION OF DECELLULARIZED ADIPOSE TISSUE SCAFFOLDS FOR SOFT TISSUE RECONSTRUCTION n.d.
Kuljanin M, Brown CFC, Raleigh MJ, Lajoie GA, Flynn LE. Collagenase treatment enhances proteomic coverage of low-abundance proteins in decellularized matrix bioscaffolds. Biomaterials 2017;144:130–43. https://doi.org/10.1016/J.BIOMATERIALS.2017.08.012.
Godehardt AW, Ramm R, Gulich B, Tönjes RR, Hilfiker A. Decellularized pig pulmonary heart valves—Depletion of nucleic acids measured by proviral PERV pol. Xenotransplantation 2020;27:e12565. https://doi.org/10.1111/XEN.12565;WGROUP:STRING:PUBLICATION.
Schmitz TC, Van Doeselaar M, Tryfonidou MA, Ito K. Detergent-Free Decellularization of Notochordal Cell-Derived Matrix Yields a Regenerative, Injectable, and Swellable Biomaterial. ACS Biomater Sci Eng 2022;8:3912–23. https://doi.org/10.1021/ACSBIOMATERIALS.2C00790/ASSET/IMAGES/LARGE/AB2C00790_0009.JPEG.
Wu Z, Zhou Y, Li N, Huang M, Duan H, Ge J, et al. The use of phospholipase A2 to prepare acellular porcine corneal stroma as a tissue engineering scaffold. Biomaterials 2009;30:3513–22. https://doi.org/10.1016/J.BIOMATERIALS.2009.03.003.
Xie S, Zhou Y, Tang Y, Chen C, Li S, Zhao C, et al. ¬Book-shaped decellularized tendon matrix scaffold combined with bone marrow mesenchymal stem cells-sheets for repair of achilles tendon defect in rabbit. Journal of Orthopaedic Research 2019;37:887–97. https://doi.org/10.1002/JOR.24255.
Suzuki M, Kimura T, Yoshida Y, Kobayashi M, Hashimoto Y, Takahashi H, et al. In Vitro Tissue Reconstruction Using Decellularized Pericardium Cultured with Cells for Ligament Regeneration. Polymers 2022, Vol 14, Page 2351 2022;14:2351. https://doi.org/10.3390/POLYM14122351.
Whitlock PW, Seyler TM, Parks GD, Ornelles DA, Smith TL, Van Dyke ME, et al. A novel process for optimizing musculoskeletal allograft tissue to improve safety, ultrastructural properties, and cell infiltration. Journal of Bone and Joint Surgery 2012;94:1458–67. https://doi.org/10.2106/JBJS.K.01397.
Seyler TM, Bracey DN, Plate JF, Lively MO, Mannava S, Smith TL, et al. The Development of a Xenograft-Derived Scaffold for Tendon and Ligament Reconstruction Using a Decellularization and Oxidation Protocol. Arthroscopy: The Journal of Arthroscopic & Related Surgery 2017;33:374–86. https://doi.org/10.1016/J.ARTHRO.2016.07.016.
Zhou S yuan, Yuan B, Huang W mao, Chen X sheng, Jia L shun. Aponeurosis discission, a low-detergent method for tissue-engineered acellular ligament scaffolds. J Mater Sci Mater Med 2022;33:1–10. https://doi.org/10.1007/S10856-022-06661-8/FIGURES/11.
Kim BS, Kim EJ, Choi JS, Jeong JH, Jo CH, Cho YW. Human collagen-based multilayer scaffolds for tendon-to-bone interface tissue engineering. J Biomed Mater Res A 2014;102:4044–54. https://doi.org/10.1002/JBM.A.35057.
Musson DS, Naot D, Chhana A, Matthews BG, McIntosh JD, Lin STC, et al. In Vitro Evaluation of a Novel Non-Mulberry Silk Scaffold for Use in Tendon Regeneration. Https://HomeLiebertpubCom/Tea 2015;21:1539–51. https://doi.org/10.1089/TEN.TEA.2014.0128.
Zhao T, Qi Y, Xiao S, Ran J, Wang J, Ghamor-Amegavi EP, et al. Integration of mesenchymal stem cell sheet and bFGF-loaded fibrin gel in knitted PLGA scaffolds favorable for tendon repair. J Mater Chem B 2019;7:2201–11. https://doi.org/10.1039/C8TB02759E.
Uyanik O, Pekkoc-Uyanik KC, Findik S, Avci A, Altuntas Z. Prevention of peritendinous adhesions with electrospun poly (lactic acid-co-glycolic acid) (PLGA) bioabsorbable nanofiber: An experimental study. Colloids Surf B Biointerfaces 2022;209:112181. https://doi.org/10.1016/J.COLSURFB.2021.112181.
Sheng D, Li J, Ai C, Feng S, Ying T, Liu X, et al. Electrospun PCL/Gel-aligned scaffolds enhance the biomechanical strength in tendon repair. J Mater Chem B 2019;7:4801–10. https://doi.org/10.1039/C9TB00837C.
Zhang X, Song W, Han K, Fang Z, Cho E, Huangfu X, et al. Three-Dimensional Bioprinting of a Structure-, Composition-, and Mechanics-Graded Biomimetic Scaffold Coated with Specific Decellularized Extracellular Matrix to Improve the Tendon-to-Bone Healing. ACS Appl Mater Interfaces 2023;15:28964–80. https://doi.org/10.1021/ACSAMI.3C03793/SUPPL_FILE/AM3C03793_SI_001.PDF.
Huang L, Chen L, Chen H, Wang M, Jin L, Zhou S, et al. Biomimetic Scaffolds for Tendon Tissue Regeneration. Biomimetics 2023, Vol 8, Page 246 2023;8:246. https://doi.org/10.3390/BIOMIMETICS8020246.
Costa-Almeida R, Calejo I, Gomes ME. Mesenchymal Stem Cells Empowering Tendon Regenerative Therapies. International Journal of Molecular Sciences 2019, Vol 20, Page 3002 2019;20:3002. https://doi.org/10.3390/IJMS20123002.
Lu J, Chen H, Lyu K, Jiang L, Chen Y, Long L, et al. The Functions and Mechanisms of Tendon Stem/Progenitor Cells in Tendon Healing. Stem Cells Int 2023;2023:1258024. https://doi.org/10.1155/2023/1258024.
Chen B, Li B, Qi YJ, Ni QB, Pan ZQ, Wang H, et al. Enhancement of tendon-to-bone healing after anterior cruciate ligament reconstruction using bone marrow-derived mesenchymal stem cells genetically modified with bFGF/BMP2. Scientific Reports 2016 6:1 2016;6:1–9. https://doi.org/10.1038/srep25940.
Hoffmann A, Gross G. Tendon and ligament engineering in the adult organism: Mesenchymal stem cells and gene-therapeutic approaches. Int Orthop 2007;31:791–7. https://doi.org/10.1007/S00264-007-0395-9/METRICS.
Yu H, Cheng J, Shi W, Ren B, Zhao F, Shi Y, et al. Bone marrow mesenchymal stem cell-derived exosomes promote tendon regeneration by facilitating the proliferation and migration of endogenous tendon stem/progenitor cells. Acta Biomater 2020;106:328–41. https://doi.org/10.1016/J.ACTBIO.2020.01.051.
Müller SA, Majewski M, Heisterbach P, Jaquiéry C, Dürselen L, Todorov A, et al. IMPROVED TENDON HEALING USING bFGF, BMP-12 AND TGFβ 1 IN A RAT MODEL. Eur Cell Mater 2018;35:318–34. https://doi.org/10.22203/eCM.v035a22.
Yonemitsu R, Tokunaga T, Shukunami C, Ideo K, Arimura H, Karasugi T, et al. Fibroblast Growth Factor 2 Enhances Tendon-to-Bone Healing in a Rat Rotator Cuff Repair of Chronic Tears. American Journal of Sports Medicine 2019;47:1701–12. https://doi.org/10.1177/0363546519836959/SUPPL_FILE/DS_10.1177_0363546519836959.PDF.
Lai F, Wang J, Tang H, Huang P, Liu J, He G, et al. VEGF promotes tendon regeneration of aged rats by inhibiting adipogenic differentiation of tendon stem/progenitor cells and promoting vascularization. The FASEB Journal 2022;36:e22433. https://doi.org/10.1096/FJ.202200213R.
Xu K, Al-ani MK, Sun Y, Xu W, Pan L, Song Y, et al. Platelet-rich plasma activates tendon-derived stem cells to promote regeneration of Achilles tendon rupture in rats. J Tissue Eng Regen Med 2017;11:1173–84. https://doi.org/10.1002/TERM.2020.
Chalidis B, Givissis P, Papadopoulos P, Pitsilos C. Molecular and Biologic Effects of Platelet-Rich Plasma (PRP) in Ligament and Tendon Healing and Regeneration: A Systematic Review. International Journal of Molecular Sciences 2023, Vol 24, Page 2744 2023;24:2744. https://doi.org/10.3390/IJMS24032744.
Song F, Jiang D, Wang T, Wang Y, Chen F, Xu G, et al. Mechanical Loading Improves Tendon-Bone Healing in a Rabbit Anterior Cruciate Ligament Reconstruction Model by Promoting Proliferation and Matrix Formation of Mesenchymal Stem Cells and Tendon Cells. Cellular Physiology and Biochemistry 2017;41:875–89. https://doi.org/10.1159/000460005.
Seeliger C, Falldorf K, Sachtleben J, Griensven M Van. Low-frequency pulsed electromagnetic fields significantly improve time of closure and proliferation of human tendon fibroblasts. Eur J Med Res 2014;19:1–7. https://doi.org/10.1186/2047-783X-19-37/FIGURES/3.
Lai WC, Iglesias BC, Mark BJ, Wang D. Low-Intensity Pulsed Ultrasound Augments Tendon, Ligament, and Bone–Soft Tissue Healing in Preclinical Animal Models: A Systematic Review. Arthroscopy: The Journal of Arthroscopic & Related Surgery 2021;37:2318-2333.e3. https://doi.org/10.1016/J.ARTHRO.2021.02.019.
Cole BJ, Easley JT, Nyska A, Rousselle S. Anterior cruciate ligament reconstruction in a translational model in sheep using biointegrative mineral fiber reinforced screws. Sci Rep 2024;14:1–10. https://doi.org/10.1038/S41598-024-75914-Z;SUBJMETA=301,308,61,631,639,692;KWRD=BIOTECHNOLOGY,MATERIALS+SCIENCE,MEDICAL+RESEARCH.
Nelson AL, O’Hara KM, Nolte PC, Fukase N, Murata Y, Nolte AK, et al. Engineered Decellularized Tendon Matrix Putty Preserves Native Tendon Bioactivity to Promote Cell Proliferation and Enthesis Repair. J Tissue Eng Regen Med 2023;2023. https://doi.org/10.1155/2023/4665795.
Xie S, Zhou Y, Tang Y, Chen C, Li S, Zhao C, et al. ¬Book-shaped decellularized tendon matrix scaffold combined with bone marrow mesenchymal stem cells-sheets for repair of achilles tendon defect in rabbit. Journal of Orthopaedic Research 2019;37:887–97. https://doi.org/10.1002/JOR.24255.
Anjum S, Li T, Saeed M, Ao Q. Exploring polysaccharide and protein-enriched decellularized matrix scaffolds for tendon and ligament repair: A review. Int J Biol Macromol 2024;254:127891. https://doi.org/10.1016/J.IJBIOMAC.2023.127891.
Lovati AB, Bottagisio M, Moretti M. Decellularized and Engineered Tendons as Biological Substitutes: A Critical Review. Stem Cells Int 2016;2016:7276150. https://doi.org/10.1155/2016/7276150.
Zhong S, Lan Y, Liu J, Seng Tam M, Hou Z, Zheng Q, et al. Advances focusing on the application of decellularization methods in tendon-bone healing. J Adv Res 2025;67:361–72. https://doi.org/10.1016/j.jare.2024.01.020.
Zhao J, Zhang D, Lan Q, Zhong G, Liu Y, Holwell N, et al. Tendon Decellularized Matrix Modified Fibrous Scaffolds with Porous and Crimped Microstructure for Tendon Regeneration. ACS Appl Bio Mater 2024;7:4747–59. https://doi.org/10.1021/ACSABM.4C00565.
Codispoti G, Carniato M, Brogini S, Romanelli A, Martini L, Giavaresi G, et al. Decellularized biological matrices for the repair of rotator cuff lesions: a systematic review of preclinical in vivo studies. Front Bioeng Biotechnol 2024;12. https://doi.org/10.3389/FBIOE.2024.1345343/PDF.
Song W, Guo Y, Liu W, Yao Y, Zhang X, Cai Z, et al. Circadian Rhythm-Regulated ADSC-Derived sEVs and a Triphasic Microneedle Delivery System to Enhance Tendon-to-Bone Healing. Advanced Materials 2024;36. https://doi.org/10.1002/ADMA.202408255.
He W, Jiang C, Zhou P, Hu X, Gu XP, Zhang SO. Role of tendon-derived stem cells in tendon and ligament repair: focus on tissue engineer. Front Bioeng Biotechnol 2024;12:1357696. https://doi.org/10.3389/FBIOE.2024.1357696/XML.
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Accepted 17-02-2026
Published 24-02-2026
