Horseradish peroxidase (HRP)-catalyzed hydrogels are considered to be an important platform for tissue engineering applications. In this study, we investigated the chondrogenic capacity of phenolated (1.2%) alginate-(0.5%) collagen hydrogel on human amniotic mesenchymal stem cells after 21 days. Using NMR, FTIR analyses, and SEM imaging, we studied the phenolation and structure of alginate-collagen hydrogel. For physicochemical evaluations, gelation time, mechanical properties, swelling, and degradation rate were assessed. The survival rate was monitored using the MTT assay and DAPI staining. Western blotting was performed to measure the chondrogenic differentiation of cells. NMR showed successful phenolation of the alginate-collagen hydrogel. FTIR exhibited the interaction between the functional groups of collagen with phenolated alginate. SEM showed the existence of collagen microfibrils in the alginate-collagen hydrogel. Compared to phenolated alginate, the addition of collagen increased hydrogel elasticity by 10%. Both swelling rate and biodegradability were reduced in the presence of collagen. We noted an increased survival rate in phenolated alginate-collagen compared to the control cells (p < 0.05). Western blotting revealed the increase of chondrocyte-associated proteins such as SOX9 and COL2A1 in phenolated-alginate-collagen hydrogels after 21 days. These data showed that phenolated alginate-collagen hydrogel is an appropriate 3 D substrate to induce chondrogenic capacity of human mesenchymal stem cells.
RimYANamYJuJH.Application of cord blood and cord blood-derived induced pluripotent stem cells for cartilage regeneration. Cell Transplant2019; 28: 529–537.
2.
LevingstoneTJMatsikoADicksonGR, et al. A biomimetic multi-layered collagen-based scaffold for osteochondral repair. Acta Biomater2014; 10: 1996–2004.
3.
LiuJWangXLuG, et al. Bionic cartilage acellular matrix microspheres as a scaffold for engineering cartilage. J Mater Chem B2019; 7: 640–650.
4.
ArmientoAStoddartMAliniM, et al. Biomaterials for articular cartilage tissue engineering: learning from biology. Acta Biomater2018; 65: 1–20.
5.
VinatierCGuicheuxJ.Cartilage tissue engineering: from biomaterials and stem cells to osteoarthritis treatments. Ann Phys Rehabil Med2016; 59: 139–144.
6.
SaghatiSNasrabadiHTKhoshfetratAB, et al. Tissue engineering strategies to increase osteochondral regeneration of stem cells; a close look at different modalities. Stem Cell Rev Rep. Epub ahead of print February 2021. DOI: 10.1007/s12015-021-10130-0.
7.
DengCChangJWuC.Bioactive scaffolds for osteochondral regeneration. J Orthop Translat2019; 17: 15–25.
8.
Fuentes-MeraLCamachoAEngelE, et al. Therapeutic potential of articular cartilage regeneration using tissue engineering based on multiphase designs. In: Nikolopoulos DD, Safos GK and Dimitrios K (eds) Cartilage Tissue Engineering and Regeneration Techniques. London: IntechOpen, 2019. Epub ahead of print 27 March 2019. DOI: 10.5772/intechopen.84697.
9.
Martínez-MorenoDJiménezGGálvez-MartínP, et al. Cartilage biomechanics: a key factor for osteoarthritis regenerative medicine. Biochim Biophys Acta Mol Basis Dis2019; 1865: 1067–1075.
10.
VinodEKachrooURebekahG, et al. In vitro chondrogenic differentiation of human articular cartilage derived chondroprogenitors using pulsed electromagnetic field. J Clin Orthop Trauma2021; 14: 22–28.
11.
PereiraDRReisRLOliveiraJM.Layered scaffolds for osteochondral tissue engineering. Adv Exp Med Biol2018; 1058: 193–218.
12.
ChungJHNaficySYueZ, et al. Bio-ink properties and printability for extrusion printing living cells. Biomater Sci2013; 1: 763–773.
13.
Ricard-BlumS.The collagen family. Cold Spring Harb Perspect Biol2011; 3: a004978.
14.
CondelloVFilardoGMadonnaV, et al. Use of a biomimetic scaffold for the treatment of osteochondral lesions in early osteoarthritis. BioMed Res Int2018; 2018: 7937089.
15.
YangXLuZWuH, et al. Collagen-alginate as bioink for three-dimensional (3D) cell printing based cartilage tissue engineering. Mater Sci Eng C Mater Biol Appl2018; 83: 195–201.
16.
OmidiSPirhayatiMKakanejadifardA.Co-delivery of doxorubicin and curcumin by a pH-sensitive, injectable, and in situ hydrogel composed of chitosan, graphene, and cellulose nanowhisker. Carbohydr Polym2020; 231: 115745.
17.
EichhornSJYoungRJDaviesGR.Modeling crystal and molecular deformation in regenerated cellulose fibers. Biomacromolecules2005; 6: 507–513.
18.
BrownRMJrandSaxenaIM.Cellulose biosynthesis: a model for understanding the assembly of biopolymers. Plant Physiol Biochem2000; 38: 57–67.
19.
DemirtaşTTIrmakGGümüşderelioğluM.A bioprintable form of chitosan hydrogel for bone tissue engineering. Biofabrication2017; 9: 035003.
20.
ZhouHYJiangLJCaoPP, et al. Glycerophosphate-based chitosan thermosensitive hydrogels and their biomedical applications. Carbohydr Polym2015; 117: 524–536.
21.
HaoTWenNCaoJ-K, et al. The support of matrix accumulation and the promotion of sheep articular cartilage defects repair in vivo by chitosan hydrogels. Osteoarthr Cartil2010; 18: 257–265.
22.
VickersNJ.Animal communication: when I’m calling you, will you answer too?Curr Biol2017; 27: R713–R715.
23.
NevesSCGomesDBSousaA, et al. Biofunctionalized pectin hydrogels as 3D cellular microenvironments. J Mater Chem B2015; 3: 2096–2108.
24.
SakaiSKawakamiK.Both ionically and enzymatically crosslinkable alginate–tyramine conjugate as materials for cell encapsulation. J Biomed Mater Res A2008; 85: 345–351.
25.
AminiHHashemzadehSHeidarzadehM, et al. Cytoprotective and cytofunctional effect of polyanionic polysaccharide alginate and gelatin microspheres on rat cardiac cells. Int J Biol Macromol2020; 161: 969–976.
26.
DalyACCritchleySERencsokEM, et al. A comparison of different bioinks for 3D bioprinting of fibrocartilage and hyaline cartilage. Biofabrication2016; 8: 045002.
27.
KellerLWagnerQSchwintéP, et al. Double compartmented and hybrid implant outfitted with well-organized 3D stem cells for osteochondral regenerative nanomedicine. Nanomedicine (Lond)2015; 10: 2833–2845.
28.
AugstADKongHJMooneyDJ.Alginate hydrogels as biomaterials. Macromol Biosci2006; 6: 623–633.
29.
LansdownAPayneM.An evaluation of the local reaction and biodegradation of calcium sodium alginate (Kaltostat) following subcutaneous implantation in the rat. J R Coll Surg Edinb1994; 39: 284–288.
30.
DhootNOTobiasCAFischerI, et al. Peptide‐modified alginate surfaces as a growth permissive substrate for neurite outgrowth. J Biomed Mater Res A2004; 71: 191–200.
31.
MosahebiAWibergMTerenghiG.Addition of fibronectin to alginate matrix improves peripheral nerve regeneration in tissue-engineered conduits. Tissue Eng2003; 9: 209–218.
32.
SakaiSHashimotoIOgushiY, et al. Peroxidase-catalyzed cell encapsulation in subsieve-size capsules of alginate with phenol moieties in water-immiscible fluid dissolving H2O2. Biomacromolecules2007; 8: 2622–2626.
33.
SakaiSAshidaTOginoS, et al. Horseradish peroxidase-mediated encapsulation of mammalian cells in hydrogel particles by dropping. J Microencapsul2014; 31: 100–104.
34.
SmidsrødOSkjaG.Alginate as immobilization matrix for cells. Trends Biotechnol1990; 8: 71–78.
35.
FurunoKWangJSuzukiK, et al. Gelatin-based electrospun fibers insolubilized by horseradish peroxidase-catalyzed cross-linking for biomedical applications. ACS Omega2020; 5: 21254–21259.
36.
GantumurESakaiSNakahataM, et al. Cytocompatible enzymatic hydrogelation mediated by glucose and cysteine residues. ACS Macro Lett2017; 6: 485–488.
37.
SakaiSNakahataM.Horseradish peroxidase catalyzed hydrogelation for biomedical, biopharmaceutical, and biofabrication applications. Chem Asian J2017; 12: 3098–3109.
38.
SakaiSUedaKGantumurE, et al. Drop‐on‐drop multimaterial 3D bioprinting realized by peroxidase‐mediated cross‐linking. Macromol Rapid Commun2018; 39: 1700534.
39.
TeixeiraLSMFeijenJvan BlitterswijkCA, et al. Enzyme-catalyzed crosslinkable hydrogels: emerging strategies for tissue engineering. Biomaterials2012; 33: 1281–1290.
40.
LeeFBaeKHKurisawaM.Injectable hydrogel systems crosslinked by horseradish peroxidase. Biomed Mater2015; 11: 014101.
41.
ChirilaTVSuzukiSPapollaC.A comparative investigation of Bombyx mori silk fibroin hydrogels generated by chemical and enzymatic cross‐linking. Biotechnol Appl Biochem2017; 64: 771–781.
42.
HosoyaKOhtsukiCKawaiT, et al. A novel covalently crosslinked gel of alginate and silane with the ability to form bone‐like apatite. J Biomed Mater Res A2004; 71: 596–601.
43.
RowleyJAMadlambayanGMooneyDJ.Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials1999; 20: 45–53.
44.
Bernkop-SchnürchAKastCERichterMF.Improvement in the mucoadhesive properties of alginate by the covalent attachment of cysteine. J Controlled Rel2001; 71: 277–285.
45.
AhmadianMKhoshfetratABKhatamiN, et al. Influence of gelatin and collagen incorporation on peroxidase-mediated injectable pectin-based hydrogel and bioactivity of fibroblasts. J Biomater Appl2020; 885328220977601.
46.
MousaviSKhoshfetratABKhatamiN, et al. Comparative study of collagen and gelatin in chitosan-based hydrogels for effective wound dressing: physical properties and fibroblastic cell behavior. Biochem Biophys Res Commun2019; 518: 625–631.
47.
KhatamiNKhoshfetratABKhaksarM, et al. Collagen-alginate-nano-silica microspheres improved the osteogenic potential of human osteoblast-like MG-63 cells. J Cell Biochem2019; 120: 15069–15082.
48.
MishraMTiwariSGomesAV.Protein purification and analysis: next generation Western blotting techniques. Expert Rev Proteomics2017; 14: 1037–1053.
49.
KhoshfetratABKhanmohammadiMSakaiS, et al. Enzymatically-gellable galactosylated chitosan: hydrogel characteristics and hepatic cell behavior. Int J Biol Macromol2016; 92: 892–899.
50.
SmejkalovaDSpacciniRFontaineB, et al. Binding of phenol and differently halogenated phenols to dissolved humic matter as measured by NMR spectroscopy. Environ Sci Technol2009; 43: 5377–5382.
51.
FertahMBelfkiraADahmaneEm, et al. Extraction and characterization of sodium alginate from Moroccan Laminaria digitata brown seaweed. Arab J Chem2017; 10: S3707–S3714.
52.
NaderiNGriffinMMalinsE, et al. Slow chlorine releasing compounds: a viable sterilisation method for bioabsorbable nanocomposite biomaterials. J Biomater Appl2016; 30: 1114–1124.
53.
WasyłeczkoMSikorskaWChwojnowskiA.Review of synthetic and hybrid scaffolds incartilage tissue engineering. Membranes2020; 10: 348.
54.
KruegerSAchillesSZimmermannJ, et al. Re-differentiation capacity of human chondrocytes in vitro following electrical stimulation with capacitively coupled fields. J Clin Med2019; 8: 1771.
PrasadhSWongRCW.Unraveling the mechanical strength of biomaterials used as a bone scaffold in oral and maxillofacial defects. Oral Sci Int2018; 15: 48–55.
57.
HuJKurokawaTNakajimaT, et al. High fracture efficiency and stress concentration phenomenon for microgel-reinforced hydrogels based on double-network principle. Macromolecules2012; 45: 9445–9451.
58.
LuTWangZTangJ, et al. A pseudo-elasticity theory to model the strain-softening behavior of tough hydrogels. J Mech Phys Solids2020; 137: 103832.
59.
DrexlerJWPowellHM.Dehydrothermal crosslinking of electrospun collagen. Tissue Eng Part C Methods2011; 17: 9–17.
60.
ZitnayJLLiYQinZ, et al. Molecular level detection and localization of mechanical damage in collagen enabled by collagen hybridizing peptides. Nat Commun2017; 8: 14913.
61.
ChangS-WBuehlerMJ.Molecular biomechanics of collagen molecules. Mater Today2014; 17: 70–76.
62.
ChangS-WShefelbineSJBuehlerMJ.Structural and mechanical differences between collagen homo-and heterotrimers: relevance for the molecular origin of brittle bone disease. Biophys J2012; 102: 640–648.
63.
ChangS-WFlynnBPRubertiJW, et al. Molecular mechanism of force induced stabilization of collagen against enzymatic breakdown. Biomaterials2012; 33: 3852–3859.
64.
Davidovich-PinhasMBianco-PeledH.A quantitative analysis of alginate swelling. Carbohydr Polym2010; 79: 1020–1027.
65.
ParkHGuoXTemenoffJS, et al. Effect of swelling ratio of injectable hydrogel composites on chondrogenic differentiation of encapsulated rabbit marrow mesenchymal stem cells in vitro. Biomacromolecules2009; 10: 541–546.
66.
MorshedlooFKhoshfetratABKazemiD, et al. Gelatin improves peroxidase‐mediated alginate hydrogel characteristics as a potential injectable hydrogel for soft tissue engineering applications. J Biomed Mater Res2020; 108: 2950–2960.
67.
HoshinoK-iNakajimaTMatsudaT, et al. Network elasticity of a model hydrogel as a function of swelling ratio: from shrinking to extreme swelling states. Soft Matter2018; 14: 9693–9701.
68.
NematiSRezabakhshAKhoshfetratAB, et al. Alginate-gelatin encapsulation of human endothelial cells promoted angiogenesis in in vivo and in vitro milieu. Biotechnol Bioeng2017; 114: 2920–2930.
69.
IbusukiSHalbesmaGJRandolphMA, et al. Photochemically cross-linked collagen gels as three-dimensional scaffolds for tissue engineering. Tissue Eng2007; 13: 1995–2001.
70.
CohenBPHooperRCPuetzerJL, et al. Long-term morphological and microarchitectural stability of tissue-engineered, patient-specific auricles in vivo. Tissue Eng Part A2016; 22: 461–468.
71.
PuetzerJLKooEBonassarLJ.Induction of fiber alignment and mechanical anisotropy in tissue engineered menisci with mechanical anchoring. J Biomech2015; 48: 1436–1443.
72.
RheeSPuetzerJLMasonBN, et al. 3D bioprinting of spatially heterogeneous collagen constructs for cartilage tissue engineering. ACS Biomater Sci Eng2016; 2: 1800–1805.
73.
HeinoJ.The collagen family members as cell adhesion proteins. Bioessays2007; 29: 1001–1010.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
