Collagen Biosynthesis to Engineered Biomaterials: Molecular Design, Synthetic Strategy, and Biomedical Application

Abstract

Collagen, a principal component of the extracellular matrix, provides mechanical strength and stability to tissues and organs through its structural organization. Its biocompatibility has established it as a crucial material in biomedical applications such as drug delivery systems, cell culture matrices, and tissue engineering scaffolds. However, the use of animal-derived collagen carries risks of pathogen transmission, which has driven research towards developing synthetic collagen alternatives. Advances in AI-assisted protein engineering are accelerating the design of synthetic collagens and their applications in biomaterials. This review examines collagen’s structural characteristics, biosynthesis strategies, biological activities as well as AI-assisting engineering.

Keywords

Collagen / Extracellular matrix / Biomaterial / AI-assisting engineering / Synthetic biology

Acknowledgments

We also would like to thank the iBiofoundary and Core Facility of Institute for Intelligent Bio/Chem Manufacturing, and ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University.

Author Contributions

Conceptualization, Y.Y. and X.Z.; Software, Z.W. and C.Z.; Writing—Original Draft Preparation, X.Z.; Writing—Review & Editing, X.Z., Y.Y. and Q.Z., X.L., B.L.; Visualization, X.Z.; Supervision, Y.Y.; Project Administration, Y.Y.; Funding Acquisition, Y.Y.

Ethics Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The datasets generated during the current study are available from the corresponding author on reasonable request.

Funding

This research was supported by Zhejiang Provincial Natural Science Foundation of China under Grant (No. LR25C100001), ‘Pioneer’ and ‘Leading Goose’ R&D Program of Zhejiang Province (No. 2025C01100, No. 2025C01102), the National Natural Science Foundation of China General Program (No.32471372).

Declaration of Competing Interest

The authors declare that they have no competing interests.

References

[1] Granito RN, Custódio MR, Rennó ACM. Natural Marine Sponges for Bone Tissue Engineering: The State of Art and Future Perspectives. J. Biomed. Mater. Res. B Appl. Biomater. 2017, 105, 1717-1727. doi:10.1002/jbm.b.33706.

[2] Viguet-Carrin S, Garnero P, Delmas PD. The Role of Collagen in Bone Strength. Osteoporos. Int. 2006, 17, 319-336. doi:10.1007/s00198-005-2035-9.

[3] Ricard-Blum S.The Collagen Family. Cold Spring Harb. Perspect. Biol. 2011, 3, a004978. doi:10.1101/cshperspect.a004978.

[4] Shoulders MD, Raines RT. Collagen Structure and Stability. Annu. Rev. Biochem. 2009, 78, 929-958. doi:10.1146/annurev.biochem.77.032207.120833.

[5] Fallas JA, O’Leary LER, Hartgerink JD. Heterotrimers and Synthetic Collagen Mimics: Self-Assembly of Homotrimers, Higher Order Structures. Chem. Soc. Rev. 2010, 39, 3510. doi:10.1039/b919455j.[GoogleScholar]

[6] Kannus P. Structure of the Tendon Connective Tissue. Scand. J. Med. Sci. Sports 2000, 10, 312-320. doi:10.1034/j.1600-0838.2000.010006312.x.[GoogleScholar]

[7] Hu X, Wu Z, Zhang Z, Yao H, Wang D-A. Type II Collagen Scaffolds for Tissue Engineering. Commun. Mater. 2024, 5, 149. doi:10.1038/s43246-024-00598-x.[GoogleScholar]

[8] Kuivaniemi H, Tromp G. Type III Collagen (COL3A1): Gene and Protein Structure, Tissue Distribution, and Associated Diseases. Gene 2019, 707, 151-171. doi:10.1016/j.gene.2019.05.003.

[9] Brown KL, Cummings CF, Vanacore RM, Hudson BG. Building Collagen IV Smart Scaffolds on the Outside of Cells. Protein Sci. 2017, 26, 2151-2161. doi:10.1002/pro.3283.

[10] Malak TM, Ockleford CD, Bell SC, Dalgleish R, Bright N, Macvicar J. Confocal Immunofluorescence Localization of Collagen Types I, III, IV, V and VI and Their Ultrastructural Organization in Term Human Fetal Membranes. Placenta 1993, 14, 385-406. doi:10.1016/S0143-4004(05)80460-6.

[11] Di Martino A, Cescon M, D’Agostino C, Schilardi F, Sabatelli P, Merlini L, et al. Collagen VI in the Musculoskeletal System. Int. J. Mol. Sci. 2023, 24, 5095. doi:10.3390/ijms24065095.

[12] Chung HJ, Uitto J. Type VII Collagen: The Anchoring Fibril Protein at Fault in Dystrophic Epidermolysis Bullosa. Dermatol. Clin. 2010, 28, 93-105. doi:10.1016/j.det.2009.10.011.

[13] Li Q, Tintut Y, Demer LL, Vazquez-Padron RI, Bendeck MP, Hsu JJ.Collagen VIII in Vascular Diseases. Matrix Biol. 2024, 133, 64-76. doi:10.1016/j.matbio.2024.08.006.

[14] Carlsen S, Nandakumar KS, Holmdahl R. Type IX Collagen Deficiency Enhances the Binding of Cartilage-Specific Antibodies and Arthritis Severity. Arthritis Res. 2006, 8, R102.

[15] Shen G. The Role of Type X Collagen in Facilitating and Regulating Endochondral Ossification of Articular Cartilage. Orthod. Craniofac. Res. 2005, 8, 11-17. doi:10.1111/j.1601-6343.2004.00308.x.[GoogleScholar]

[16] Sun M, Luo EY, Adams SM, Adams T, Ye Y, Shetye SS, et al. Collagen XI Regulates the Acquisition of Collagen Fibril Structure, Organization and Functional Properties in Tendon. Matrix Biol. 2020, 94, 77-94. doi:10.1016/j.matbio.2020.09.001.

[17] Papanicolaou M, Parker AL, Yam M, Filipe EC, Wu SZ, Chitty JL, et al. Temporal Profiling of the Breast Tumour Microenvironment Reveals Collagen XII as a Driver of Metastasis. Nat. Commun. 2022, 13, 4587. doi:10.1038/s41467-022-32255-7.

[18] Heikkinen A, Tu H, Pihlajaniemi T. Collagen XIII: A Type II Transmembrane Protein with Relevance to Musculoskeletal Tissues, Microvessels and Inflammation. Int. J. Biochem. Cell Biol. 2012, 44, 714-717. doi:10.1016/j.biocel.2012.01.024.

[19] Tao G, Levay AK, Peacock JD, Huk DJ, Both SN, Purcell NH, et al. Collagen XIV Is Important for Growth and Structural Integrity of the Myocardium. J. Mol. Cell. Cardiol. 2012, 53, 626-638. doi:10.1016/j.yjmcc.2012.08.002.

[20] Bretaud S, Guillon E, Karppinen S-M, Pihlajaniemi T, Ruggiero F. Collagen XV, a Multifaceted Multiplexin Present across Tissues and Species. Matrix Biol. Plus 2020, 6-7, 100023. doi:10.1016/j.mbplus.2020.100023.

[21] Grässel S, Bauer RJ. Collagen XVI in Health and Disease. Matrix Biol. 2013, 32, 64-73. doi:10.1016/j.matbio.2012.11.001.

[22] Natsuga K, Watanabe M, Nishie W, Shimizu H. Life before and beyond Blistering: The Role of Collagen XVII in Epidermal Physiology. Exp. Dermatol. 2019, 28, 1135-1141. doi:10.1111/exd.13550.

[23] Aikio M, Elamaa H, Vicente D, Izzi V, Kaur I, Seppinen L, et al. Specific Collagen XVIII Isoforms Promote Adipose Tissue Accrual via Mechanisms Determining Adipocyte Number and Affect Fat Deposition. Proc. Natl. Acad. Sci. USA 2014, 111, E3043-E3052. doi:10.1073/pnas.1405879111.

[24] Sadri G, Fischer AG, Brittian KR, Elliott E, Nystoriak MA, Uchida S, et al. Collagen Type XIX Regulates Cardiac Extracellular Matrix Structure and Ventricular Function. Matrix Biol. 2022, 109, 49-69. doi:10.1016/j.matbio.2022.03.007.

[25] Thorlacius-Ussing J, Jensen C, Madsen EA, Nissen NI, Manon-Jensen T, Chen IM, et al. Type XX Collagen Is Elevated in Circulation of Patients with Solid Tumors. Int. J. Mol. Sci. 2022, 23, 4144. doi:10.3390/ijms23084144.

[26] Chou M-Y, Li H-C. Genomic Organization and Characterization of the Human Type XXI Collagen (COL21A1) Gene. Genomics 2002, 79, 395-401. doi:10.1006/geno.2002.6712.

[27] Koch M, Schulze J, Hansen U, Ashwodt T, Keene DR, Brunken WJ, et al. Collagen XXII. A Novel Marker of Tissue Junctions, J. Biol. Chem. 2004, 279, 22514-22521. doi:10.1074/jbc.M400536200.

[28] Kehlet SN, Karsdal MA.Type XXIII Collagen. In Biochemistry of Collagens, Laminins and Elastin; Elsevier: Amsterdam, The Netherlands, 2016; pp 139-141. doi:10.1016/B978-0-12-809847-9.00023-4.

[29] Koch M, Laub F, Zhou P, Hahn RA, Tanaka S, Burgeson RE, et al. Collagen XXIV, a Vertebrate Fibrillar Collagen with Structural Features of Invertebrate Collagens. J. Biol. Chem. 2003, 278, 43236-43244. doi:10.1074/jbc.M302112200.

[30] Gonçalves TJM, Boutillon F, Lefebvre S, Goffin V, Iwatsubo T, Wakabayashi T, et al. Collagen XXV Promotes Myoblast Fusion during Myogenic Differentiation and Muscle Formation. Sci. Rep. 2019, 9, 5878. doi:10.1038/s41598-019-42296-6.

[31] Sato K, Yomogida K, Wada T, Yorihuzi T, Nishimune Y, Hosokawa N, et al. Type XXVI Collagen, a New Member of the Collagen Family, Is Specifically Expressed in the Testis and Ovary. J. Biol. Chem. 2002, 277, 37678-37684. doi:10.1074/jbc.M205347200.

[32] Hjorten R, Hansen U, Underwood RA, Telfer HE, Fernandes RJ, Krakow D, et al. Type XXVII Collagen at the Transition of Cartilage to Bone during Skeletogenesis. Bone 2007, 41, 535-542. doi:10.1016/j.bone.2007.06.024.

[33] Exposito J-Y, Valcourt U, Cluzel C, Lethias C.The Fibrillar Collagen Family. Int. J. Mol. Sci. 2010, 11, 407-426. doi:10.3390/ijms11020407.

[34] Grimal S, Puech S, Wagener R, Ventéo S, Carroll P, Fichard-Carroll A. Collagen XXVIII Is a Distinctive Component of the Peripheral Nervous System Nodes of Ranvier and Surrounds Nonmyelinating Glial Cells. Glia 2010, 58, 1977-1987. doi:10.1002/glia.21066.

[35] Bella J, Eaton M, Brodsky B, Berman HM. Crystal and Molecular Structure of a Collagen-Like Peptide at 1.9 Å Resolution. Science 1994, 266, 75-81. doi:10.1126/science.7695699.

[36] Berisio R, Vitagliano L, Mazzarella L, Zagari A. Crystal Structure of the Collagen Triple Helix Model [(Pro-Pro-Gly)₁₀]₃. Protein Sci. 2002, 11, 262-270. doi:10.1110/ps.32602.

[37] Fan D, Takawale A, Lee J, Kassiri Z. Cardiac Fibroblasts, Fibrosis and Extracellular Matrix Remodeling in Heart Disease. Fibrogenesis Tissue Repair 2012, 5, 15. doi:10.1186/1755-1536-5-15.

[38] Liu Y, Ramanath HS, Wang D-A. Tendon Tissue Engineering Using Scaffold Enhancing Strategies. Trends Biotechnol. 2008, 26, 201-209. doi:10.1016/j.tibtech.2008.01.003.

[39] Tuckwell DS, Ayad S, Grant ME, Takigawa M, Humphries MJ. Conformation Dependence of Integrin-Type II Collagen Binding: Inability of Collagen Peptides to Support Α2β1 Binding, and Mediation of Adhesion to Denatured Collagen by a Novel Α5β1-Fibronectin Bridge. J. Cell Sci. 1994, 107, 993-1005. doi:10.1242/jcs.107.4.993.

[40] Franzke C-W, Bruckner P, Bruckner-Tuderman L. Collagenous Transmembrane Proteins: Recent Insights into Biology and Pathology*. J. Biol. Chem. 2005, 280, 4005-4008. doi:10.1074/jbc.R400034200.

[41] Mouw JK, Ou G, Weaver VM. Extracellular Matrix Assembly: A Multiscale Deconstruction. Nat. Rev. Mol. Cell Biol. 2014, 15, 771-785. doi:10.1038/nrm3902.

[42] Myllyharju J. Intracellular Post-Translational Modifications of Collagens. In Collagen; Brinckmann J, Notbohm H, Müller PK,Eds.; Topics in Current Chemistry; Springer: Berlin/Heidelberg, Germany, 2005; Volume 247, pp. 115-147. doi:10.1007/b103821.

[43] Persikov AV, Ramshaw JAM, Kirkpatrick A, Brodsky B. Electrostatic Interactions Involving Lysine Make Major Contributions to Collagen Triple-Helix Stability. Biochemistry 2005, 44, 1414-1422. doi:10.1021/bi048216r.[GoogleScholar]

[44] Brodsky B, Persikov AV. Molecular Structure of the Collagen Triple Helix. In Advances in Protein Chemistry; Elsevier: Amsterdam, The Netherlands, 2005; Volume 70, pp. 301-339. doi:10.1016/S0065-3233(05)70009-7.

[45] Boudko SP, Engel J, Bächinger HP. The Crucial Role of Trimerization Domains in Collagen Folding. Int. J. Biochem. Cell Biol. 2012, 44, 21-32. doi:10.1016/j.biocel.2011.09.009.

[46] McLaughlin SH, Bulleid NJ. Molecular recognition in procollagen chain assembly. Matrix Biol. 1998, 16, 369-377. doi:10.1016/s0945-053x(98)90010-5.

[47] Bourhis J-M, Mariano N, Zhao Y, Harlos K, Exposito J-Y, Jones EY, et al. S. Structural Basis of Fibrillar Collagen Trimerization and Related Genetic Disorders. Nat. Struct. Mol. Biol. 2012, 19, 1031-1036. doi:10.1038/nsmb.2389.

[48] Birk DE, Trelstad RL. Extracellular Compartments in Tendon Morphogenesis: Collagen Fibril, Bundle, and Macroaggregate Formation. J. Cell Biol. 1986, 103, 231-240.

[49] Canty EG, Lu Y, Meadows RS, Shaw MK, Holmes DF, Kadler KE. Coalignment of Plasma Membrane Channels and Protrusions (Fibripositors) Specifies the Parallelism of Tendon. J. Cell Biol. 2004, 165, 553-563. doi:10.1083/jcb.200312071.

[50] Bruckner P. Suprastructures of Extracellular Matrices: Paradigms of Functions Controlled by Aggregates Rather than Molecules. Cell Tissue Res. 2010, 339, 7-18. doi:10.1007/s00441-009-0864-0.

[51] Wenstrup RJ, Florer JB, Brunskill EW, Bell SM, Chervoneva I, Birk DE. Type V Collagen Controls the Initiation of Collagen Fibril Assembly. J. Biol. Chem. 2004, 279, 53331-53337. doi:10.1074/jbc.M409622200.

[52] Eyre DR, Weis MA, Wu J-J.Advances in Collagen Cross-Link Analysis. Methods 2008, 45, 65-74. doi:10.1016/j.ymeth.2008.01.002.

[53] Lucero HA, Kagan HM. Lysyl Oxidase: An Oxidative Enzyme and Effector of Cell Function. Cell. Mol. Life Sci. 2006, 63, 2304-2316. doi:10.1007/s00018-006-6149-9.

[54] Orgel JP, Wess TJ, Miller A. The in Situ Conformation and Axial Location of the Intermolecular Cross-Linked Non-Helical Telopeptides of Type I Collagen. Structure 2000, 8, 137-142. doi:10.1016/S0969-2126(00)00089-7.[GoogleScholar]

[55] O’Leary LER, Fallas JA, Bakota EL, Kang MK, Hartgerink JD. Multi-Hierarchical Self-Assembly of a Collagen Mimetic Peptide from Triple Helix to Nanofibre and Hydrogel. Nat. Chem. 2011, 3, 821-828. doi:10.1038/nchem.1123.

[56] Knupp C, Squire JM.Molecular Packing in Network-Forming Collagens. Sci. World J. 2003, 3, 558-577. doi:10.1100/tsw.2003.40.

[57] Fidler AL, Boudko SP, Rokas A, Hudson BG. The Triple Helix of Collagens—an Ancient Protein Structure That Enabled Animal Multicellularity and Tissue Evolution. J. Cell Sci. 2018, 131, jcs203950. doi:10.1242/jcs.203950.

[58] Kadler KE, Hojima Y, Prockop DJ. Assembly of Collagen Fibrils de Novo by Cleavage of the Type I pC-Collagen with Procollagen C-Proteinase. Assay of Critical Concentration Demonstrates That Collagen Self-Assembly Is a Classical Example of an Entropy-Driven Process. J. Biol. Chem. 1987, 262, 15696-15701. doi:10.1016/S0021-9258(18)47783-6.

[59] Cummings CF, Pedchenko V, Brown KL, Colon S, Rafi M, Jones-Paris C, et al. Extracellular Chloride Signals Collagen IV Network Assembly during Basement Membrane Formation. J. Cell Biol. 2016, 213, 479-494. doi:10.1083/jcb.201510065.

[60] Bella J. A First Census of Collagen Interruptions: Collagen’s Own Stutters and Stammers. J. Struct. Biol. 2014, 186, 438-450. doi:10.1016/j.jsb.2014.03.022.

[61] Yurchenco PD, Ruben GC. Basement Membrane Structure in Situ: Evidence for Lateral Associations in the Type IV Collagen Network. J. Cell Biol. 1987, 105, 2559-2568. doi:10.1083/jcb.105.6.2559.

[62] Elango J, Hou C, Bao B, Wang S, Maté Sánchez De Val JE, Wenhui W. The Molecular Interaction of Collagen with Cell Receptors for Biological Function. Polymers 2022, 14, 876. doi:10.3390/polym14050876.

[63] Hu K, Hu M, Xiao Y, Cui Y, Yan J, Yang G, et al. Preparation Recombination Human-like Collagen/Fibroin Scaffold and Promoting the Cell Compatibility with Osteoblasts. J. Biomed. Mater. Res. A 2021, 109, 346-353. doi:10.1002/jbm.a.37027.

[64] Lowell CA, Mayadas TN. Overview: Studying Integrins In Vivo. In Integrin and Cell Adhesion Molecules; Shimaoka M, Ed.; Methods in Molecular Biology; Humana Press: Totowa, NJ, USA, 2011; Volume 757, pp. 369-397. doi:10.1007/978-1-61779-166-6_22.

[65] Velling T, Kusche-Gullberg M, Sejersen T, Gullberg D. cDNA Cloning and Chromosomal Localization of Human Α11 Integrin. J. Biol. Chem. 1999, 274, 25735-25742. doi:10.1074/jbc.274.36.25735.

[66] Briesewitz R, Epstein MR, Marcantonio EE. Expression of Native and Truncated Forms of the Human Integrin Alpha 1 Subunit. J. Biol. Chem. 1993, 268, 2989-2996. doi:10.1016/S0021-9258(18)53871-0.

[67] Iwamoto DV, Calderwood DA. Regulation of Integrin-Mediated Adhesions. Curr. Opin. Cell Biol. 2015, 36, 41-47. doi:10.1016/j.ceb.2015.06.009.

[68] Shrivastava A, Radziejewski C, Campbell E, Kovac L, McGlynn M, Ryan TE, et al. An Orphan Receptor Tyrosine Kinase Family Whose Members Serve as Nonintegrin Collagen Receptors. Mol. Cell 1997, 1, 25-34. doi:10.1016/S1097-2765(00)80004-0.

[69] Löffek S, Hurskainen T, Jackow J, Sigloch FC, Schilling O, Tasanen K, et al. Transmembrane Collagen XVII Modulates Integrin Dependent Keratinocyte Migration via PI3K/Rac1 Signaling. PLoS ONE 2014, 9, e87263. doi:10.1371/journal.pone.0087263.

[70] Yang S, Kim J, Ryu J-H, Oh H, Chun C-H, Kim BJ, et al. Hypoxia-Inducible Factor-2α Is a Catabolic Regulator of Osteoarthritic Cartilage Destruction. Nat. Med. 2010, 16, 687-693. doi:10.1038/nm.2153.

[71] Gonzalez DM, Medici D. Signaling Mechanisms of the Epithelial-Mesenchymal Transition. Sci. Signal. 2014, 7, re8. doi:10.1126/scisignal.2005189.

[72] Ozaki I, Hamajima H, Matsuhashi S, Mizuta T. Regulation of TGF-?1-Induced Pro-Apoptotic Signaling by Growth Factor Receptors and Extracellular Matrix Receptor Integrins in the Liver. Front. Physiol. 2011, 2, 78. doi:10.3389/fphys.2011.00078.

[73] Madamanchi A, Santoro SA, Zutter MM. α2β1 Integrin. Adv Exp Med Biol. 2014, 819, 41-60. doi:10.1007/978-94-017-9153-3_3.

[74] Calderwood DA, Tuckwell DS, Eble J, Kühn K, Humphries MJ. The Integrin Α1 A-Domain Is a Ligand Binding Site for Collagens and Laminin. J. Biol. Chem. 1997, 272, 12311-12317. doi:10.1074/jbc.272.19.12311.

[75] Nykvist P, Tu H, Ivaska J, Käpylä J, Pihlajaniemi T, Heino J. Distinct Recognition of Collagen Subtypes by Α1β1 and α2β1Integrins. J. Biol. Chem. 2000, 275, 8255-8261. doi:10.1074/jbc.275.11.8255.

[76] Kern A, Eble J, Golbik R, Kühn K. Interaction of Type IV Collagen with the Isolated Integrins Α1β1 and Α2β1. Eur. J. Biochem. 1993, 215, 151-159. doi:10.1111/j.1432-1033.1993.tb18017.x.[GoogleScholar]

[77] Jokinen J, Dadu E, Nykvist P, Käpylä J, White DJ, Ivaska J, et al. Integrin-Mediated Cell Adhesion to Type I Collagen Fibrils. J. Biol. Chem. 2004, 279, 31956-31963. doi:10.1074/jbc.M401409200.

[78] Käpylä J, Jäälinoja J, Tulla M, Ylöstalo J, Nissinen L, Viitasalo T, et al. The Fibril-Associated Collagen IX Provides a Novel Mechanism for Cell Adhesion to Cartilaginous Matrix. J. Biol. Chem. 2004, 279, 51677-51687. doi:10.1074/jbc.M409412200.

[79] Koivunen J, Tu H, Kemppainen A, Anbazhagan P, Finnilä MA, Saarakkala S, et al. Integrin Α11β1 Is a Receptor for Collagen XIII. Cell Tissue Res. 2021, 383, 1135-1153. doi:10.1007/s00441-020-03300-y.[GoogleScholar]

[80] Zhang W-M, Käpylä J, Puranen JS, Knight CG, Tiger C-F, Pentikäinen OT, et al. Α11β1 Integrin Recognizes the GFOGER Sequence in Interstitial Collagens. J. Biol. Chem. 2003, 278, 7270-7277. doi:10.1074/jbc.M210313200.

[81] Orgel JPRO, Madhurapantula RS. A Structural Prospective for Collagen Receptors Such as DDR and Their Binding of the Collagen Fibril. Biochim. Biophys. Acta BBA—Mol. Cell Res. 2019, 1866, 118478. doi:10.1016/j.bbamcr.2019.04.008.

[82] Jung SM, Takemura Y, Imamura Y, Hayashi T, Adachi E, Moroi M. Collagen-Type Specificity of Glycoprotein VI as a Determinant of Platelet Adhesion. Platelets 2008, 19, 32-42. doi:10.1080/09537100701609027.

[83] Schultz HS, Guo L, Keller P, Fleetwood AJ, Sun M, Guo W, et al. OSCAR-collagen Signaling in Monocytes Plays a Proinflammatory Role and May Contribute to the Pathogenesis of Rheumatoid Arthritis. Eur. J. Immunol. 2016, 46, 952-963. doi:10.1002/eji.201545986.

[84] Son M. Understanding the Contextual Functions of C1q and LAIR-1 and Their Applications. Exp. Mol. Med. 2022, 54, 567-572. doi:10.1038/s12276-022-00774-4.

[85] Steffensen B, Xu X, Martin PA, Zardeneta G. Human Fibronectin and MMP-2 Collagen Binding Domains Compete for Collagen Binding Sites and Modify Cellular Activation of MMP-2. Matrix Biol. 2002, 21, 399-414. doi:10.1016/S0945-053X(02)00032-X.

[86] Sano K, Asanuma-Date K, Arisaka F, Hattori S, Ogawa H. Changes in Glycosylation of Vitronectin Modulate Multimerization and Collagen Binding during Liver Regeneration. Glycobiology 2007, 17, 784-794. doi:10.1093/glycob/cwm031.

[87] Melander MC, Jürgensen HJ, Madsen DH, Engelholm LH, Behrendt N. The Collagen Receptor uPARAP/Endo180 in Tissue Degradation and Cancer (Review). Int. J. Oncol. 2015, 47, 1177-1188. doi:10.3892/ijo.2015.3120.

[88] Chattopadhyay S, Raines RT.Collagen-based Biomaterials for Wound Healing. Biopolymers 2014, 101, 821-833. doi:10.1002/bip.22486.

[89] Huang X, Lv J, Li Y, Mao S, Li Z, Jing Z, et al. Programmable C-to-U RNA Editing Using the Human APOBEC 3A Deaminase. EMBO J. 2020, 39, e104741. doi:10.15252/embj.2020104741.

[90] Sorushanova A, Delgado LM, Wu Z, Shologu N, Kshirsagar A, Raghunath R, et al. The Collagen Suprafamily: From Biosynthesis to Advanced Biomaterial Development. Adv. Mater. 2019, 31, 1801651. doi:10.1002/adma.201801651.

[91] Parenteau-Bareil R, Gauvin R, Berthod F. Collagen-Based Biomaterials for Tissue Engineering Applications. Materials 2010, 3, 1863-1887. doi:10.3390/ma3031863.

[92] Silvipriya K, Kumar K, Bhat A, Kumar B, John A, Lakshmanan P. Collagen: Animal Sources and Biomedical Application. J. Appl. Pharm. Sci. 2015, 123-127. doi:10.7324/JAPS.2015.50322.

[93] Fan J, Zhuang Y, Li B. Effects of Collagen and Collagen Hydrolysate from Jellyfish Umbrella on Histological and Immunity Changes of Mice Photoaging. Nutrients 2013, 5, 223-233. doi:10.3390/nu5010223.

[94] Gelse K. Collagens—Structure, Function, and Biosynthesis. Adv. Drug Deliv. Rev. 2003, 55, 1531-1546. doi:10.1016/j.addr.2003.08.002.

[95] Lima CA, Campos JF, Filho JLL, Converti A, Da Cunha MGC, Porto ALF. Antimicrobial and Radical Scavenging Properties of Bovine Collagen Hydrolysates Produced by Penicillium Aurantiogriseum URM 4622 Collagenase. J. Food Sci. Technol. 2015, 52, 4459-4466. doi:10.1007/s13197-014-1463-y.[GoogleScholar]

[96] Zhang Y, Olsen K, Grossi A, Otte J. Effect of Pretreatment on Enzymatic Hydrolysis of Bovine Collagen and Formation of ACE-Inhibitory Peptides. Food Chem. 2013, 141, 2343-2354. doi:10.1016/j.foodchem.2013.05.058.

[97] Choi D, Min S-G, Jo Y-J. Functionality of Porcine Skin Hydrolysates Produced by Hydrothermal Processing for Liposomal Delivery System. J. Food Biochem. 2018, 42, e12464. doi:10.1111/jfbc.12464.

[98] Dandagi GL, Byahatti SM. An Insight into the Swine-Influenza A (H1N1) Virus Infection. Lung India 2011, 28, 34-38.

[99] León-López A, Morales-Peñaloza A, Martínez-Juárez VM, Vargas-Torres A, Zeugolis DI, Aguirre-Álvarez G. Hydrolyzed Collagen—Sources and Applications. Molecules 2019, 24, 4031. doi:10.3390/molecules24224031.

[100] Gómez-Guillén MC, Giménez B, López-Caballero ME, Montero MP. Functional and Bioactive Properties of Collagen and Gelatin from Alternative Sources: A Review. Food Hydrocoll. 2011, 25, 1813-1827. doi:10.1016/j.foodhyd.2011.02.007.

[101] Skierka E, Sadowska M. The Influence of Different Acids and Pepsin on the Extractability of Collagen from the Skin of Baltic Cod (Gadus Morhua). Food Chem. 2007, 105, 1302-1306. doi:10.1016/j.foodchem.2007.04.030.

[102] Ketnawa S, Benjakul S, Martínez-Alvarez O, Rawdkuen S. Fish Skin Gelatin Hydrolysates Produced by Visceral Peptidase and Bovine Trypsin: Bioactivity and Stability. Food Chem. 2017, 215, 383-390. doi:10.1016/j.foodchem.2016.07.145.

[103] Hong H, Chaplot S, Chalamaiah M, Roy BC, Bruce HL, Wu J. Removing Cross-Linked Telopeptides Enhances the Production of Low-Molecular-Weight Collagen Peptides from Spent Hens. J. Agric. Food Chem. 2017, 65, 7491-7499. doi:10.1021/acs.jafc.7b02319.

[104] Hulgan SAH, Hartgerink JD. Recent Advances in Collagen Mimetic Peptide Structure and Design. Biomacromolecules 2022, 23, 1475-1489. doi:10.1021/acs.biomac.2c00028.

[105] Mukherjee S, Varshashankari V, Feba A, Ayyadurai N, Balamurugan K, Shanmugam G. Hyperstable and Fibril-Forming Collagen-Mimetic Peptides in Shortest Triple Helices: Empowering the Capping by π-Systems. Biomacromolecules 2025, 26, 2171-2185. doi:10.1021/acs.biomac.4c01455.

[106] Bourgot I, Primac I, Louis T, Noël A, Maquoi E. Reciprocal Interplay Between Fibrillar Collagens and Collagen-Binding Integrins: Implications in Cancer Progression and Metastasis. Front. Oncol. 2020, 10, 1488. doi:10.3389/fonc.2020.01488.

[107] Mhanna R, Öztürk E, Vallmajo-Martin Q, Millan C, Müller M, Zenobi-Wong M. GFOGER-Modified MMP-Sensitive Polyethylene Glycol Hydrogels Induce Chondrogenic Differentiation of Human Mesenchymal Stem Cells. Tissue Eng. Part A 2014, 20, 1165-1174. doi:10.1089/ten.tea.2013.0519.

[108] Zhao C, Xiao Y, Ling S, Pei Y, Ren J. Structure of Collagen. Methods Mol. Biol. 2021, 2347, 17-25. doi:10.1007/978-1-0716-1574-4_2.

[109] Zhang C, Fan D, Shang L, Ma X, Luo Y, Xue W, et al. Optimization of Fermentation Process for Human-like Collagen Production of Recombinant Escherichia Coli Using Response Surface Methodology. Chin. J. Chem. Eng. 2010, 18, 137-142. doi:10.1016/s1004-9541(08)60334-1.

[110] Tang Y, Yang X, Hang B, Li J, Huang L, Huang F, et al. Efficient Production of Hydroxylated Human-Like Collagen Via the Coexpression of Three Key Genes in Escherichia Coli Origami (DE3). Appl. Biochem. Biotechnol. 2016, 178, 1458-1470. doi:10.1007/s12010-015-1959-6.

[111] Nokelainen M, Tu H, Vuorela A, Notbohm H, Kivirikko KI, Myllyharju J. High-level Production of Human Type I Collagen in the Yeast Pichia Pastoris. Yeast 2001, 18, 797-806. doi:10.1002/yea.730.

[112] Werkmeister JA, Ramshaw JAM. Recombinant Protein Scaffolds for Tissue Engineering. Biomed. Mater. 2012, 7, 012002. doi:10.1088/1748-6041/7/1/012002.

[113] Rutschmann C, Baumann S, Cabalzar J, Luther KB, Hennet T. Recombinant Expression of Hydroxylated Human Collagen in Escherichia coli. Appl. Microbiol. Biotechnol. 2014, 98, 4445-4455. doi:10.1007/s00253-013-5447-z.[GoogleScholar]

[114] Ruggiero F, Exposito J-Y, Bournat P, Gruber V, Perret S, Comte J, et al. Triple Helix Assembly and Processing of Human Collagen Produced in Transgenic Tobacco Plants. FEBS Lett. 2000, 469, 132-136. doi:10.1016/S0014-5793(00)01259-X.

[115] Xu X, Gan Q, Clough RC, Pappu KM, Howard JA, Baez JA, et al. Hydroxylation of Recombinant Human Collagen Type I Alpha 1 in Transgenic Maize Co-Expressed with a Recombinant Human Prolyl 4-Hydroxylase. BMC Biotechnol. 2011, 11, 69. doi:10.1186/1472-6750-11-69.

[116] Vuorela A. Assembly of Human Prolyl 4-Hydroxylase and Type III Collagen in the Yeast Pichia Pastoris: Formation of a Stable Enzyme Tetramer Requires Coexpression with Collagen and Assembly of a Stable Collagen Requires Coexpression with Prolyl 4-Hydroxylase. EMBO J. 1997, 16, 6702-6712. doi:10.1093/emboj/16.22.6702.

[117] Myllyharju’ J, Nokelainen M, Vuorela A. Expression of Recombinant Human Type 1-111 Collagens in the Yeast Pichia Pastoris. Biochem. Soc. Trans. 2000, 28.

[118] Toman PD, Chisholm G, McMullin H, Giere LM, Olsen DR, Kovach RJ, et al. Production of Recombinant Human Type I Procollagen Trimers Using a Four-Gene Expression System in the Yeast Saccharomyces Cerevisiae. J. Biol. Chem. 2000, 275, 23303-23309. doi:10.1074/jbc.M002284200.

[119] Liu X, Yang Y, Zhang W, Sun Y, Peng F, Jeffrey L, et al. Expression of Recombinant Protein Using Corynebacterium Glutamicum: Progress, Challenges and Applications. Crit. Rev. Biotechnol. 2016, 36, 652-664. doi:10.3109/07388551.2015.1004519.

[120] Li X, Xue W, Zhu C, Fan D, Liu Y, Ma X. Novel Hydrogels Based on Carboxyl Pullulan and Collagen Crosslinking with 1, 4-Butanediol Diglycidylether for Use as a Dermal Filler: Initial in Vitro and in Vivo Investigations. Mater. Sci. Eng. C 2015, 57, 189-196. doi:10.1016/j.msec.2015.07.059.

[121] Kumar V, Kumar N, Gangwar AK, Singh H, Singh R. Comparative Histologic and Immunologic Evaluation of 1,4-Butanediol Diglycidyl Ether Crosslinked versus Noncrosslinked Acellular Swim Bladder Matrix for Healing of Full-Thickness Skin Wounds in Rabbits. J. Surg. Res. 2015, 197, 436-446. doi:10.1016/j.jss.2015.04.080.

[122] Pustlauk W, Paul B, Gelinsky M, Bernhardt A. Jellyfish Collagen and Alginate: Combined Marine Materials for Superior Chondrogenesis of hMSC. Mater. Sci. Eng. C 2016, 64, 190-198. doi:10.1016/j.msec.2016.03.081.

[123] Unsöld C, Pappano WN, Imamura Y, Steiglitz BM, Greenspan DS. Biosynthetic Processing of the Pro-Α1(V)2Pro-Α2(V) Collagen Heterotrimer by Bone Morphogenetic Protein-1 and Furin-like Proprotein Convertases. J. Biol. Chem. 2002, 277, 5596-5602. doi:10.1074/jbc.M110003200.

[124] Zhou H, Li W, Pan L, Zhu T, Zhou T, Xiao E, et al. Human Extracellular Matrix (ECM)-like Collagen and Its Bioactivity. Regen. Biomater. 2024, 11, rbae008. doi:10.1093/rb/rbae008.

[125] Jia Y, Han Y, Zhang Y, Li L, Zhang B, Yan X. Multifunctional Type Lll Recombinant Human Collagen Incorporated Sodium Alginate Hydrogel with Sustained Release of Extra Cellular Vehicles for Wound Healing Multimodal Therapy in Diabetic Mice. Regen. Ther. 2024, 27, 329-341. doi:10.1016/j.reth.2024.03.010.

[126] Zhuang Z-M, Wang Y, Chen L, Wu Z-R, Zhang T, Bei H-P, et al. Triple-Molded, Reinforced Arrowhead Microneedle Patch of Dual Human-Derived Matrix for Integrated Management of Diabetic Wounds. Biomaterials 2026, 324, 123520. doi:10.1016/j.biomaterials.2025.123520.

[127] Wang Q, An Q, Wang Y, Yang J, Zhang X, Jiang S, et al. Development and Mechanistic Investigation of Recombinant Type III Humanized Collagen Gel for Mid-Facial Soft Tissue Repair. Collagen Leather 2025, 7, 14. doi:10.1186/s42825-025-00196-8.

[128] Munyemana JC, He H, Fu C, Fan Y, Sun X, Xiao J. Recombinant Collagen-Templated Biomineralized Synthesis of Biocompatible pH-Responsive Porous Calcium Carbonate Nanospheres. ACS Omega 2023, 8, 30879-30887. doi:10.1021/acsomega.3c01467.

[129] Zhang H, Wang J, Wei J, Fu X, Ma J, Chen J. Molecular Engineering of Recombinant Protein Hydrogels: Programmable Design and Biomedical Applications. Gels 2025, 11, 579. doi:10.3390/gels11080579.

[130] Vitale M, Ligorio C, Smith IP, Richardson SM, Hoyland JA, Bella J. Incorporation of Natural and Recombinant Collagen Proteins within Fmoc-Based Self-Assembling Peptide Hydrogels. Gels 2022, 8, 254. doi:10.3390/gels8050254.

[131] Gu J, He Y, He C, Zhang Q, Huang Q, Bai S, et al. Advances in the Structures, Mechanisms and Targeting of Molecular Chaperones. Signal Transduct. Target. Ther. 2025, 10, 84. doi:10.1038/s41392-025-02166-2.

[132] Masuda R, Phyu Thant KP, Kawahara K, Oki H, Kadonosono T, Kobayashi Y, et al. A Yeast Two-Hybrid System to Obtain Triple-Helical Ligands from Combinatorial Random Peptide Libraries. J. Biol. Chem. 2024, 300, 107794. doi:10.1016/j.jbc.2024.107794.

[133] Ruggiero F, Koch M.Making Recombinant Extracellular Matrix Proteins. Methods 2008, 45, 75-85. doi:10.1016/j.ymeth.2008.01.003.

[134] Li H-C, Huang C-C, Chen S-F, Chou M-Y. Assembly of Homotrimeric Type XXI Minicollagen by Coexpression of Prolyl 4-Hydroxylase in Stably Transfected Drosophila Melanogaster S 2 Cells. Biochem. Biophys. Res. Commun. 2005, 336, 375-385. doi:10.1016/j.bbrc.2005.08.018.

[135] Shi C, Wang C, Lu J, Zhong B, Tang J. Protein Sequence and Structure Co-Design with Equivariant Translation. arXiv 2023, arXiv:2210.08761.

[136] Evans R, O’Neill M, Pritzel A, Antropova N, Senior A, Green T, et al.Protein Complex Prediction with AlphaFold-Multimer. BioRxiv 2021. doi:10.1101/2021.10.04.463034.

[137] Elder S, Clune J, Walker J, Gloth P. Suitability of EGCG as a Means of Stabilizing a Porcine Osteochondral Xenograft. J. Funct. Biomater. 2017, 8, 43. doi:10.3390/jfb8040043.

[138] Pawelec KM, Confalonieri D, Ehlicke F, Van Boxtel HA, Walles H, Kluijtmans SGJM. Osteogenesis and Mineralization of Mesenchymal Stem Cells in Collagen Type I-based Recombinant Peptide Scaffolds. J. Biomed. Mater. Res. A 2017, 105, 1856-1866. doi:10.1002/jbm.a.36049.

[139] Lee JC, Pereira CT, Ren X, Huang W, Bischoff D, Weisgerber DW, et al. Optimizing Collagen Scaffolds for Bone Engineering: Effects of Cross-Linking and Mineral Content on Structural Contraction and Osteogenesis. J. Craniofac. Surg. 2015, 26, 1992-1996. doi:10.1097/SCS.0000000000001918.

[140] Berndt M, Li Y, Seyedhassantehrani N, Yao L. Fabrication and Characterization of Microspheres Encapsulating Astrocytes for Neural Regeneration. ACS Biomater. Sci. Eng. 2017, 3, 1313-1321. doi:10.1021/acsbiomaterials.6b00229.

[141] Han L, Zhang Z, Wang B, Wen Z. Construction and Biocompatibility of a Thin Type I/II Collagen Composite Scaffold. Cell Tissue Bank. 2018, 19, 47-59. doi:10.1007/s10561-017-9653-2.

[142] Wang X, Ma B, Chang J. Preparation of Decellularized Vascular Matrix by Co-Crosslinking of Procyanidins and Glutaraldehyde. Biomed. Mater. Eng. 2015, 26, 19-30. doi:10.3233/BME-151548.

[143] Wang X, Xu K, Ma L, Sun R, Wang K, Wang R, et al. Diffusion Model Assisted Designing Self-Assembling Collagen Mimetic Peptides as Biocompatible Materials. Brief. Bioinform. 2024, 26, bbae622. doi:10.1093/bib/bbae622.

[144] Khare E, Yu C-H, Gonzalez Obeso C, Milazzo M, Kaplan DL, Buehler MJ. Discovering Design Principles of Collagen Molecular Stability Using a Genetic Algorithm, Deep Learning, and Experimental Validation. Proc. Natl. Acad. Sci. USA 2022, 119, e2209524119. doi:10.1073/pnas.2209524119.

[145] Hu J, Li J, Jiang J, Wang L, Roth J, McGuinness KN, et al. Design of Synthetic Collagens That Assemble into Supramolecular Banded Fibers as a Functional Biomaterial Testbed. Nat. Commun. 2022, 13, 6761. doi:10.1038/s41467-022-34127-6.

[146] Köster S, Evans HM, Wong JY, Pfohl T. An In Situ Study of Collagen Self-Assembly Processes. Biomacromolecules 2008, 9, 199-207. doi:10.1021/bm700973t.[GoogleScholar]

[147] Hafner AE, Gyori NG, Bench CA, Davis LK, Šarić A. Modeling Fibrillogenesis of Collagen-Mimetic Molecules. Biophys. J. 2020, 119, 1791-1799. doi:10.1016/j.bpj.2020.09.013.

[148] Li Z, Du T, Ruan C, Niu X. Bioinspired Mineralized Collagen Scaffolds for Bone Tissue Engineering. Bioact. Mater. 2021, 6, 1491-1511. doi:10.1016/j.bioactmat.2020.11.004.

[149] Shen H, Fallas JA, Lynch E, Sheffler W, Parry B, Jannetty N, et al. De Novo Design of Self-Assembling Helical Protein Filaments. Science 2018, 362, 705-709. doi:10.1126/science.aau3775.

[150] LeCun Y, Bengio Y, Hinton G. Deep Learning. Nature 2015, 521, 436-444. doi:10.1038/nature14539.

[151] Srinivasan A, Gupta A, Narayanamurthy V. A Comprehensive Review of AI-Based Collagen Valorization: Recent Trends, Innovations in Extraction, and Applications. Green Anal. Chem. 2025, 12, 100212. doi:10.1016/j.greeac.2025.100212.

[152] Li W, Peng J, Yao D, Rao B, Xia Y, Wang Q, et al. The Structural Basis for the Collagen Processing by Human P3H1/CRTAP/PPIB Ternary Complex. Nat. Commun. 2024, 15, 7844. doi:10.1038/s41467-024-52321-6.

[153] Qian Y, Xu C, Xiong W, Jiang N, Zheng Y, He X, et al. Dual Cross-Linked Organic-Inorganic Hybrid Hydrogels Accelerate Diabetic Skin Wound Healing. Chem. Eng. J. 2021, 417, 129335. doi:10.1016/j.cej.2021.129335.

[154] Yao T, Chen H, Wang R, Rivero R, Wang F, Kessels L, et al. Thiol-Ene Conjugation of a VEGF Peptide to Electrospun Scaffolds for Potential Applications in Angiogenesis. Bioact. Mater. 2023, 20, 306-317. doi:10.1016/j.bioactmat.2022.05.029.

[155] Conde J, Oliva N, Atilano M, Song HS, Artzi N. Self-Assembled RNA-Triple-Helix Hydrogel Scaffold for microRNA Modulation in the Tumour Microenvironment. Nat. Mater. 2016, 15, 353-363. doi:10.1038/nmat4497.

[156] Kubyshkin V. Stabilization of the Triple Helix in Collagen Mimicking Peptides. Org. Biomol. Chem. 2019, 17, 8031-8047. doi:10.1039/C9OB01646E.[GoogleScholar]

[157] Lyu C, Kong W, Liu Z, Wang S, Zhao P, Liang K, et al. Advanced Glycation End-Products as Mediators of the Aberrant Crosslinking of Extracellular Matrix in Scarred Liver Tissue. Nat. Biomed. Eng. 2023, 7, 1437-1454. doi:10.1038/s41551-023-01019-z.[GoogleScholar]

[158] Wang J, Wu Q, Wang Y, Xiang L, Feng J, Zhou Z, et al. Collagenase-Loaded pH-Sensitive Nanocarriers Efficiently Remodeled Tumor Stroma Matrixes and Improved the Enrichment of Nanomedicines. Nanoscale 2021, 13, 9402-9414. doi:10.1039/D1NR00950H.[GoogleScholar]

[159] Le DHT, Ibrahimova V, Van Den Wildenberg SAH, Wu H, Fonseca A, Torres T, et al. Light-Responsive Elastin-Like Peptide-Based Targeted Nanoparticles for Enhanced Spheroid Penetration. Angew. Chem. Int. Ed. 2023, 62, e202300511. doi:10.1002/anie.202300511.

[160] Li Y, Xia Y, Liu X, Wang J, Sun Y, Huang J, et al. Rational Design of Bioengineered Recombinant Collagen-like Protein Enhances GelMA Hydrogel for Diabetic Wound Healing. Int. J. Biol. Macromol. 2024, 280, 136012. doi:10.1016/j.ijbiomac.2024.136012.

[161] Sun TL, Luo F, Kurokawa T, Karobi SN, Nakajima T, Gong JP. Molecular Structure of Self-Healing Polyampholyte Hydrogels Analyzed from Tensile Behaviors. Soft Matter 2015, 11, 9355-9366. doi:10.1039/C5SM01423A.[GoogleScholar]

[162] Yu HC, Zheng SY, Fang L, Ying Z, Du M, Wang J, et al. Reversibly Transforming a Highly Swollen Polyelectrolyte Hydrogel to an Extremely Tough One and Its Application as a Tubular Grasper. Adv. Mater. 2020, 32, 2005171. doi:10.1002/adma.202005171.

[163] Meyers MA, McKittrick J, Chen P-Y. Structural Biological Materials: Critical Mechanics-Materials Connections. Science 2013, 339, 773-779. doi:10.1126/science.1220854.

[164] Su R, Ma C, Han B, Zhang H, Liu K.Proteins for Hyperelastic Materials. Small 2025, 21, 2406388. doi:10.1002/smll.202406388.

[165] Mozdzen LC, Rodgers R, Banks JM, Bailey RC, Harley BAC. Increasing the Strength and Bioactivity of Collagen Scaffolds Using Customizable Arrays of 3D-Printed Polymer Fibers. Acta Biomater. 2016, 33, 25-33. doi:10.1016/j.actbio.2016.02.004.

[166] Hadidi P, Cissell DD, Hu JC, Athanasiou KA. Temporal Development of Near-Native Functional Properties and Correlations with qMRI in Self-Assembling Fibrocartilage Treated with Exogenous Lysyl Oxidase Homolog 2. Acta Biomater. 2017, 64, 29-40. doi:10.1016/j.actbio.2017.09.035.

[167] Priyadarshani P, Li Y, Yang S, Yao L. Injectable Hydrogel Provides Growth-permissive Environment for Human Nucleus Pulposus Cells. J. Biomed. Mater. Res. A 2016, 104, 419-426. doi:10.1002/jbm.a.35580.

[168] Zhou X, Wang J, Fang W, Tao Y, Zhao T, Xia K, et al. Genipin Cross-Linked Type II Collagen/Chondroitin Sulfate Composite Hydrogel-like Cell Delivery System Induces Differentiation of Adipose-Derived Stem Cells and Regenerates Degenerated Nucleus Pulposus. Acta Biomater. 2018, 71, 496-509. doi:10.1016/j.actbio.2018.03.019.

[169] Bryś M, Urbańska K, Olas B. Novel Findings Regarding the Bioactivity of the Natural Blue Pigment Genipin in Human Diseases. Int. J. Mol. Sci. 2022, 23, 902.

[170] Vardar E, Larsson HM, Allazetta S, Engelhardt EM, Pinnagoda K, Vythilingam G, et al. Microfluidic Production of Bioactive Fibrin Micro-Beads Embedded in Crosslinked Collagen Used as an Injectable Bulking Agent for Urinary Incontinence Treatment. Acta Biomater. 2018, 67, 156-166. doi:10.1016/j.actbio.2017.11.034.

[171] Quinlan E, Thompson EM, Matsiko A, O’Brien FJ, López-Noriega A. Functionalization of a Collagen-Hydroxyapatite Scaffold with Osteostatin to Facilitate Enhanced Bone Regeneration. Adv. Healthc. Mater. 2015, 4, 2649-2656. doi:10.1002/adhm.201500439.

[172] Liu T, Dan W, Dan N, Liu X, Liu X, Peng X. A Novel Grapheme Oxide-Modified Collagen-Chitosan Bio-Film for Controlled Growth Factor Release in Wound Healing Applications. Mater. Sci. Eng. C 2017, 77, 202-211. doi:10.1016/j.msec.2017.03.256.

[173] Baird CW, Myers PO, Piekarski B, Borisuk M, Majeed A, Emani SM, et al. Photo-Oxidized Bovine Pericardium in Congenital Cardiac Surgery: Single-Centre Experience. Interact. Cardiovasc. Thorac. Surg. 2016, 24, 240-244. doi:10.1093/icvts/ivw315.

[174] Rosellini E, Zhang YS, Migliori B, Barbani N, Lazzeri L, Shin SR, et al. Protein/Polysaccharide-based Scaffolds Mimicking Native Extracellular Matrix for Cardiac Tissue Engineering Applications. J. Biomed. Mater. Res. A 2018, 106, 769-781. doi:10.1002/jbm.a.36272.

[175] Chen C, Zhao M, Zhang R, Lu G, Zhao C, Fu F, et al. Collagen/Heparin Sulfate Scaffolds Fabricated by a 3D Bioprinter Improved Mechanical Properties and Neurological Function after Spinal Cord Injury in Rats. J. Biomed. Mater. Res. A 2017, 105, 1324-1332. doi:10.1002/jbm.a.36011.

[176] Siriwardane ML, DeRosa K, Collins G, Pfister BJ. Controlled Formation of Cross-Linked Collagen Fibers for Neural Tissue Engineering Applications. Biofabrication 2014, 6, 015012. doi:10.1088/1758-5082/6/1/015012.

[177] Li Y, He Q, Hu X, Liu Y, Cheng X, Li X, et al. Improved Performance of Collagen Scaffolds Crosslinked by Traut’s Reagent and Sulfo-SMCC. J. Biomater. Sci. Polym. Ed. 2017, 28, 629-647. doi:10.1080/09205063.2017.1291296.

[178] Bell A, Kofron M, Nistor V. Multiphoton Crosslinking for Biocompatible 3D Printing of Type I Collagen. Biofabrication 2015, 7, 035007. doi:10.1088/1758-5090/7/3/035007.

[179] Fadlallah A, Zhu H, Arafat S, Kochevar I, Melki S, Ciolino JB. Corneal Resistance to Keratolysis After Collagen Crosslinking with Rose Bengal and Green Light. Investig. Opthalmology Vis. Sci. 2016, 57, 6610. doi:10.1167/iovs.15-18764.[GoogleScholar]

[180] Moreira MA, Souza NO, Sousa RS, Freitas DQ, Lemos MV, De Paula DM, et al. Efficacy of New Natural Biomodification Agents from Anacardiaceae Extracts on Dentin Collagen Cross-Linking. Dent. Mater. 2017, 33, 1103-1109. doi:10.1016/j.dental.2017.07.003.

[181] Lim E, Lim M-J, Min K-S, Kwon Y-S, Hwang Y-C, Yu M-K, et al. Effects of Epicatechin, a Crosslinking Agent, on Human Dental Pulp Cells Cultured in Collagen Scaffolds. J. Appl. Oral Sci. 2016, 24, 76-84. doi:10.1590/1678-775720150383.