The main goal of the study was to produce chitosan-collagen hydrogel composite scaffolds consisting of 3D printed poly(lactic acid) (PLA) strut and nanofibrous cellulose for meniscus cartilage tissue engineering. For this purpose, first PLA strut containing microchannels was incorporated into cellulose nanofibers and then they were embedded into chitosan-collagen matrix to obtain micro- and nano-sized topographical features for better cellular activities as well as mechanical properties. All the hydrogel composite scaffolds produced by using three different concentrations of genipin (0.1, 0.3, and 0.5%) had an interconnected microporous structure with a swelling ratio of about 400% and water content values between 77 and 83% which is similar to native cartilage extracellular matrix. The compressive strength of all the hydrogel composite scaffolds was found to be similar (∼32 kPa) and suitable for cartilage tissue engineering applications. Besides, the hydrogel composite scaffold comprising 0.3% (w/v) genipin had the highest tan δ value (0.044) at a frequency of 1 Hz which is around the walking frequency of a person. According to the in vitro analysis, this hydrogel composite scaffold did not show any cytotoxic effect on the rabbit mesenchymal stem cells and enabled cells to attach, proliferate and also migrate through the inner area of the scaffold. In conclusion, the produced hydrogel composite scaffold holds great promise for meniscus tissue engineering.
EslahiNSimchiAMehrjooM, et al.Hybrid cross-linked hydrogels based on fibrous protein/block copolymers and layered silicate nanoparticles: tunable thermosensitivity, biodegradability and mechanical durability. RSC Adv2016; 6(67): 62944–62957.
2.
SharmaCGautamSDindaAK, et al.Cartilage tissue engineering: current scenario and challenges. Adv Mater Lett2011; 2: 90–99.
3.
Van Der BrachtHVerdonkRVerbruggenA, et al.Cell-based meniscus tissue engineering. In: Topics in tissue engineering. Biomaterials and Tissue Engineering GroupBTE), 2007, Oulu University, Finland: ch2_1-ch2_13.
4.
De MoriAPena FernandezMBlunnG, et al.3D printing and electrospinning of composite hydrogels for cartilage and bone tissue engineering. Polymers2018; 10(3): 285.
5.
LiuMZengXMaC, et al.Injectable hydrogels for cartilage and bone tissue engineering. Bone Res2017; 5: 17014.
6.
AinolaMTomaszewskiWOstrowskaB, et al.A bioactive hybrid three-dimensional tissue-engineering construct for cartilage repair. J Biomater Appl2016; 30(6): 873–885.
7.
GugjooMAmarpalGTSharmaG, et al.Cartilage tissue engineering: role of mesenchymal stem cells along with growth factors & scaffolds. Indian J Med Res2016; 144(3): 339–347.
8.
CoburnJMGibsonMMonagleS, et al.Bioinspired nanofibers support chondrogenesis for articular cartilage repair. Proc Natl Acad Sci USA2012; 109(25): 10012–10017.
9.
SeolYJParkJYJeongW, et al.Development of hybrid scaffolds using ceramic and hydrogel for articular cartilage tissue regeneration. J Biomed Mater Res A2015; 103(4): 1404–1413.
10.
IwasakiNKasaharaYYamaneS, et al.Chitosan-based hyaluronic acid hybrid polymer fibers as a scaffold biomaterial for cartilage tissue engineering. Polymers2010; 3(1): 100–113.
11.
HolzwarthJMMaPX. Biomimetic nanofibrous scaffolds for bone tissue engineering. Biomaterials2011; 32(36): 9622–9629.
12.
LiWJLaurencinCTCatersonEJ, et al.Electrospun nanofibrous structure: a novel scaffold for tissue engineering. J Biomed Mater2002; 60(4): 613–621.
LinkeRDUlmerMImhoffAB. Replacement of the meniscus with a collagen implant (CMI). Oper Orthop Traumatol2006; 18(5–6): 453–462.
17.
YodmuangSMcNamaraSLNoverAB, et al.Silk microfiber-reinforced silk hydrogel composites for functional cartilage tissue repair. Acta Biomater2015; 11: 27–36.
18.
VaraprasadKRaghavendraGMJayaramuduT, et al.A mini review on hydrogels classification and recent developments in miscellaneous applications. Mater Sci Eng C2017; 79: 958–971.
19.
DimidaSBarcaACancelliN, et al.Effects of genipin concentration on cross-linked chitosan scaffolds for bone tissue engineering: structural characterization and evidence of biocompatibility features. Int J Polym Sci2017; 2017: 1–8.
20.
YangJZhangYSYueK, et al.Cell-laden hydrogels for osteochondral and cartilage tissue engineering. Acta Biomater2017; 57: 1–25.
21.
MirahmadiFTafazzoli-ShadpourMShokrgozarMA, et al.Enhanced mechanical properties of thermosensitive chitosan hydrogel by silk fibers for cartilage tissue engineering. Mater Sci Eng: C2013; 33(8): 4786–4794.
22.
KangH-WLeeSJKoIK, et al.A 3D bioprinting system to produce human-scale tissue constructs with structural integrity. Nat Biotechnol2016; 34(3): 312–319.
23.
JangJ-HCastanoOKimH-W. Electrospun materials as potential platforms for bone tissue engineering. Adv Drug Delivery Rev2009; 61(12): 1065–1083.
24.
KostakovaESepsMPokornyP, et al.Study of polycaprolactone wet electrospinning process. Exp Polym Lett2014; 8(8): 554–564.
25.
TaskinMBXuRGregersenH, et al.3D polydopamine functionalized coiled microfibrous scaffolds enhance human mesenchymal stem cells colonization and mild myofibroblastic differentiation. ACS Appl Mater Interfaces2016; 8(25): 15864–15873.
26.
ZhangMDingCChenL, et al.The preparation of cellulose/collagen composite films using 1-ethyl-3-methylimidazolium acetate as a solvent. BioResources2014; 9(1): 756–771.
27.
PrasanthRNageswaranSThakurVK, et al.Electrospinning of cellulose: process and applications. In: ThakurVK (ed) Nanocellulose polymer nanocomposites: fundamentals and applications. Hoboken, NJ, USA: John Wiley & Sons, 2014, pp. 311–340.
28.
KaraAKocturkSBiliciG, et al.Development of biological meniscus scaffold: decellularization method and recellularization with meniscal cell population derived from mesenchymal stem cells. J Biomater Appl2021; 35(9): 1192–1207.
29.
ChiengBWAzowaINYunusW, et al.Effects of graphene nanopletelets on poly (lactic acid)/poly (ethylene glycol) polymer nanocomposites. Adv Mater Res2014; 1024: 136–139.
30.
AliSKhatriZOhKW, et al.Zein/cellulose acetate hybrid nanofibers: electrospinning and characterization. Macromol Res2014; 22(9): 971–977.
31.
RodríguezKRenneckarSGatenholmP. Biomimetic calcium phosphate crystal mineralization on electrospun cellulose-based scaffolds. ACS Appl Mater Interfaces2011; 3(3): 681–689.
32.
SungHWLiangILChenCN, et al.Stability of a biological tissue fixed with a naturally occurring crosslinking agent (genipin)The Japanese Society for Biomaterials. J Biomed Mater Res2001; 55(4): 538–546.
33.
WeigelTSchinkelGLendleinA. Design and preparation of polymeric scaffolds for tissue engineering. Expert Rev Med Devices2006; 3(6): 835–851.
34.
RaghunathJRolloJSalesK, et al.Biomaterials and scaffold design: key to tissue‐engineering cartilage. Biotechnol Appl Biochem2007; 46(2): 73–84.
35.
YanJMiaoYTanH, et al.Injectable alginate/hydroxyapatite gel scaffold combined with gelatin microspheres for drug delivery and bone tissue engineering. Mater Sci Eng C2016; 63: 274–284.
36.
ZhangYWangQSYanK, et al.Preparation, characterization, and evaluation of genipin crosslinked chitosan/gelatin three‐dimensional scaffolds for liver tissue engineering applications. J Biomed Mater Res Part A2016; 104(8): 1863–1870.
37.
KumarGVSuC-HVelusamyP.Ciprofloxacin loaded genipin cross-linked chitosan/heparin nanoparticles for drug delivery application. Mater Lett2016; 180: 119–122.
38.
SundararaghavanHGMonteiroGALapinNA, et al.Genipin‐induced changes in collagen gels: correlation of mechanical properties to fluorescence. J Biomed Mater Res A2008; 87(2): 308–320.
39.
LiQWangXLouX, et al.Genipin-crosslinked electrospun chitosan nanofibers: determination of crosslinking conditions and evaluation of cytocompatibility. Carbohydr Polym2015; 130: 166–174.
40.
Sophia FoxAJBediARodeoSA. The basic science of articular cartilage: structure, composition, and function. Sports Health2009; 1(6): 461–468.
41.
GilarskaALewandowska-LancuckaJHorakW, et al.Collagen/chitosan/hyaluronic acid–based injectable hydrogels for tissue engineering applications–design, physicochemical and biological characterization. Colloids Surf B: Biointerfaces2018; 170: 152–162.
42.
ChenKZhangDYangX, et al.Research on viscoelastic behavior and mechanism of hydrogel grafted with UHMWPE. Soft Mater2016; 14(4): 244–252.
43.
YangZWuYLiC, et al.Improved mesenchymal stem cells attachment and in vitro cartilage tissue formation on chitosan-modified poly (L-lactide-co-epsilon-caprolactone) scaffold. Tissue Eng A2011; 18(3–4): 242–251.
44.
ByjuAGKulkarniAGundiahN. Mechanics of gelatin and elastin based hydrogels as tissue engineered constructs. In: Proceedings of the 13th International Conference on Fracture, Beijing, China, 16-21 June, 2013, pp. 4406–4415.
45.
LawlessBMSadeghiHTempleDK, et al.Viscoelasticity of articular cartilage: Analysing the effect of induced stress and the restraint of bone in a dynamic environment. J Mech Behav Biomed Mater2017; 75: 293–301.
46.
ReisRLCohnD (eds) Polymer based systems on tissue engineering, replacement and regenerationSpringer Science & Business Media, Germany, 2012, Vol. 86.
47.
LiuYChan-ParkMB. Hydrogel based on interpenetrating polymer networks of dextran and gelatin for vascular tissue engineering. Biomaterials2009; 30(2): 196–207.
48.
ColuccinoLPeresCGottardiR, et al.Anisotropy in the viscoelastic response of knee meniscus cartilage. J Appl Biomater Funct Mater2017; 15(1): 77–83.
49.
TempleDKCederlundAALawlessBM, et al.Viscoelastic properties of human and bovine articular cartilage: a comparison of frequency-dependent trends. BMC Musculoskeletal Disord2016; 17(1): 419.
50.
YanLPWangYJRenL, et al.Genipin‐cross‐linked collagen/chitosan biomimetic scaffolds for articular cartilage tissue engineering applications. J Biomed Mater Res A2010; 95(2): 465–475.
51.
PereiraHCaridadeSFriasA, et al.Biomechanical and cellular segmental characterization of human meniscus: building the basis for Tissue Engineering therapies. Osteoarthritis Cartilage2014; 22(9): 1271–1281.
52.
SlapnikJBobovnikRMeslM, et al.Modified polylactide filaments for 3D printing with improved mechanical properties. Contemp Mater2016; 7(2): 142–150.
53.
JohnstoneBHMillerHMBeckMR, et al.Identification and characterization of a large source of primary mesenchymal stem cells tightly adhered to bone surfaces of human vertebral body marrow cavities. Cytotherapy2020; 22(11): 617–628.
54.
FrancisSLDi BellaCWallaceGG, et al.Cartilage tissue engineering using stem cells and bioprinting technology—barriers to clinical translation. Front Surg2018; 5: 70.
55.
NavaMMDraghiLGiordanoC, et al.The effect of scaffold pore size in cartilage tissue engineering. J Appl Biomater Funct Mater2016; 14(3): e223–e229.
56.
DinescuSGalateanuBAlbuM, et al.Biocompatibility assessment of novel collagen-sericin scaffolds improved with hyaluronic acid and chondroitin sulfate for cartilage regeneration. Biomed Res Int2013; 2013: 598056.
57.
KumarPNagarajanAUchilPD. Analysis of cell viability by the lactate dehydrogenase assay. Cold Spring Harbor Protoc2018; 2018(6): 465–469.
58.
VýbornýKVallovaJKociZ, et al.Genipin and EDC crosslinking of extracellular matrix hydrogel derived from human umbilical cord for neural tissue repair. Sci Rep2019; 9(1): 10674.
59.
KarabayUHüsemoğluRBEğrilmezMY, et al.3D printed polylactic acid scaffold For dermal tissue engineering application: the fibroblast proliferation in vitro. J Med Innov Technol2019; 1(2): 51–56.
60.
GeyikÖGNalbantBHüsemoğluRB, et al.Investigation of surface adhesion of MCF-7 cells in 3D printed PET and PLA tissue Scaffold models. J Med Innov Technol2019; 1(2): 45–50.
61.
NelsonCMChenCS. Cell-cell signaling by direct contact increases cell proliferation via a PI3K-dependent signal. FEBS Lett2002; 514(2–3): 238–242.
62.
NgI. C.PawijitPTanJ.HYu, Anatomy and physiology for biomaterials research and development. Encyclopedia of Biomedical Engineering, Elsevier, Amsterdam, Holland, 2019, vol. 1–3.
63.
MartínezABlancoMDavidenkoN, et al.Tailoring chitosan/collagen scaffolds for tissue engineering: effect of composition and different crosslinking agents on scaffold properties. Carbohydr Polym2015; 132: 606–619.
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.
