Abstract
Background
Recently biomaterials utilized for designing scaffolds in tissue engineering are not cost-effective and eco-friendly. As a result, we design and develop biocompatible and bioactive hydrogels for osteo-tissue regeneration based on the natural polysaccharide chitosan. Three distinct hydrogel components were used for this.
Methods
Hydrogels networks were created using chitosan 2% (CTS 2%), carboxymethyl chitosan 2% (CMC 2%), and 50:50 mixtures of CTS and CMC (CTS/CMC 50:50). Furthermore, scanning electron microscopy (SEM), Fourier transforms infrared spectroscopy (FTIR), degradation, and swelling behavior of design hydrogels were studied. Also, the cytocompatibility and osteo-differentiation potency were examined by encapsulating mesenchymal stem cells derived from adipose tissue (AMSCs) on the designed hydrogels.
Results
According to the findings, our results showed an acceptable pore structure, functional groups, and degradation rate of the designed hydrogels for in vitro evaluation. In addition, employing CMC instead of CTS or adding 50% CMC to the hydrogel component could improve the hydrogel's osteo-bioactivity without the use of external osteogenic differentiation agents.
Conclusion
The CMC-containing hydrogel not only caused early osteogenesis but also accelerated differentiation to the maturity phase of osteoblasts.
Similar content being viewed by others
Data availability
They are available on the request.
References:
Fares MM, Sani ES, Lara RP, Oliveira RB, Khademhosseini A, Annabi N (2018) Interpenetrating network gelatin methacryloyl (GelMA) and pectin-g-PCL hydrogels with tunable properties for tissue engineering. Biomater Sci 6(11):2938–2950
Chang S, Liu Z, Wang X (2022) Advances of stimulus-responsive hydrogels for bone defects repair in tissue engineering. Gels 8:389
Ghalei S, Handa H (2021) Nitric oxide-releasing gelatin methacryloyl/silk fibroin interpenetrating polymer network hydrogels for tissue engineering applications. ACS Biomater Sci Eng 8(1):273–283
Zou Z, Wang L, Zhou Z, Sun Q, Liu D, Chen Y, Zou X et al (2021) Simultaneous incorporation of PTH (1–34) and nano-hydroxyapatite into chitosan/alginate hydrogels for efficient bone regeneration. Bioactive Mater 6(6):1839–1851
Nie J, Pei B, Wang Z, Hu Q (2019) Construction of ordered structure in polysaccharide hydrogel: a review. Carbohydr Polym 205:225–235
Nie H, Liu M, Zhan F, Guo M (2004) Factors on the preparation of carboxymethylcellulose hydrogel and its degradation behavior in soil. Carbohydr Polym 58(2):185–189
Dutta SD, Hexiu J, Patel DK, Ganguly K, Lim KT (2021) 3D-printed bioactive and biodegradable hydrogel scaffolds of alginate/gelatin/cellulose nanocrystals for tissue engineering. Int J Biol Macromol 167:644–658
Sharifi F et al (2018) Polycaprolactone/carboxymethyl chitosan nanofibrous scaffolds for bone tissue engineering application. Int J Biol Macromol 115:243–248
Tang Y, Du Y, Li Y, Wang X, Hu X (2009) A thermosensitive chitosan/poly (vinyl alcohol) hydrogel containing hydroxyapatite for protein delivery. J Biomed Mater Res Part A 91(4):953–963
Anitha A, Sowmya S, Kumar PS, Deepthi S, Chennazhi KP, Ehrlich H, Jayakumar R et al (2014) Chitin and chitosan in selected biomedical applications. Progr Polym Sci 39(9):1644–1667
Qi L, Xu Z, Jiang X, Hu C, Zou X (2004) Preparation and antibacterial activity of chitosan nanoparticles. Carbohydr Res 339(16):2693–2700
No HK, Park NY, Lee SH, Meyers SP (2002) Antibacterial activity of chitosans and chitosan oligomers with different molecular weights. Int J Food Microbiol 74(1–2):65–72
Li Q, Dunn ET, Grandmaison EW, Goosen MF (2020) Applications and properties of chitosan. In: Goosen MFA (ed) Applications of chitin and chitosan. Boca Raton, CRC Press, pp 3–29
Tsai GJ, Su WH (1999) Antibacterial activity of shrimp chitosan against Escherichia coli. J Food Prot 62(3):239–243
Hunt JA, Chen R, van Veen T, Bryan N (2014) Hydrogels for tissue engineering and regenerative medicine. J Mater Chem B 2(33):5319–5338
Ji X, Yang W, Wang T, Mao C, Guo L, Xiao J, He N (2013) Coaxially electrospun core/shell structured poly (l-lactide) acid/chitosan nanofibers for potential drug carrier in tissue engineering. J Biomed Nanotechnol 9(10):1672–1678
Wu T, Huang J, Jiang Y, Hu Y, Ye X, Liu D, Chen J (2018) Formation of hydrogels based on chitosan/alginate for the delivery of lysozyme and their antibacterial activity. Food Chem 240:361–369
Zhong QK, Wu ZY, Qin YQ, Hu Z, Li SD, Yang ZM, Li PW (2019) Preparation and properties of carboxymethyl chitosan/alginate/tranexamic acid composite films. Membranes 9(1):11
Alemi PS et al (2019) Synergistic effect of pressure cold atmospheric plasma and carboxymethyl chitosan to mesenchymal stem cell differentiation on PCL/CMC nanofibers for cartilage tissue engineering. Polym Adv Technol 30:1356
Wach RA, Mitomo H, Yoshii F, Kumo T (2001) Hydrogel of biodegradation cellulose derivatives. II. Effect of some factors on radation-induced crosslinking of CMC. J Appl Polym Sci 81:3000–3017
Tao F, Cheng Y, Tao H, Jin L, Wan Z, Dai F, Deng H et al (2020) Carboxymethyl chitosan/sodium alginate-based micron-fibers fabricated by emulsion electrospinning for periosteal tissue engineering. Mater Design 194:108849
Sharifi F, Atyabi SM, Irani S, Bakhshi H (2020) Bone morphogenic protein-2 immobilization by cold atmospheric plasma to enhance the osteoinductivity of carboxymethyl chitosan-based nanofibers. Carbohydr Polym 231:115681
Arab-Ahmadi S, Irani S, Bakhshi H, Atyabi F, Ghalandari B (2021) Immobilization of carboxymethyl chitosan/laponite on polycaprolactone nanofibers as osteoinductive bone scaffolds. Polym Adv Technol 32(2):755–765
Arab-Ahmadi S, Irani S, Bakhshi H, Atyabi F, Ghalandari B (2021) Immobilization of cobalt-loaded laponite/carboxymethyl chitosan on polycaprolactone nanofiber for improving osteogenesis and angiogenesis activities. Polym Adv Technol 32:4362
Gupta D, Tator CH, Shoichet MS (2006) Fast-gelling injectable blend of hyaluronan and methylcellulose for intrathecal, localized delivery to the injured spinal cord. Biomaterials 27(11):2370–2379
Anderson HC (1995) Molecular biology of matrix vesicles. Clin Orthop Relat Res 314:266–280
Zhong C, Chu CC (2012) Biomimetic mineralization of acid polysaccharide-based hydrogels: towards porous 3-dimensional bone-like biocomposites. J Mater Chem B 22(13):6080–6087
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Res 29(9):45
Mohammadnezhad J, Bakhshi H et al (2016) Preparation and evaluation of chitosan-coated eggshell particles as copper(II) biosorbent. Desalin Water Treatment 57:1693–1704
Ray M, Anis A, Banthia AK (2010) Development and characterization of chitosan based polymeric hydrogel membranes. Des Monomers Polym 13:193–206
Hassani F, Ebrahimi B, Moini A, Ghiaseddin A, Bazrafkan M, Hassanzadeh G, Valojerdi MR (2020) Chitosan hydrogel supports integrity of ovarian follicles during in vitro culture: a preliminary of a novel biomaterial for three dimensional culture of ovarian follicles. Cell J (Yakhteh) 21(4):193
Yar M, Gigliobianco G, Shahzadi L, Dew L, Siddiqi SA, Khan AF, MacNeil S et al (2016) Production of chitosan PVA PCL hydrogels to bind heparin and induce angiogenesis. Int J Polym Mater Polym Biomater 65(9):466–476
Sharifi F, Irani S, Azadegan G, Pezeshki-Modaress M, Zandi M, Saeed M (2020) Co-electrospun gelatin-chondroitin sulfate/polycaprolactone nanofibrous scaffolds for cartilage tissue engineering. Bioactive Carbohydr Dietary Fibre 22:100215
Rusdianto Budiraharjo KGN (2012) Hydroxyapatite-coated carboxymethyl chitosan scaffolds for promoting osteoblast and stem cell differentiation. J Colloid Interface Sci 366:224–232
Upadhyaya L, Agarwal V, Tewari RP (2014) The im plications of recent advances in carboxym eth yl chitosan based targeted drug delivery an d tissu e engin eering applicat ions. J Controlled Release 186:54–87
Neuss S et al (2008) Assessment of stem cell/biomaterial combinations for stem cell-based tissue engineering. Biomaterials 29(3):302–313
Siddiqui N, Jabbari E (2015) Osteogenic differentiation of human mesenchymal stem cells in freeze-gelled chitosan/nano β-tricalcium phosphate porous scaffolds crosslinked with genipin. Mater Sci Eng C 54:76–83
Zaharia A, Muşat V, Anghel EM, Atkinson I, Mocioiu OC, Buşilă M, Pleşcan VG (2017) Biomimetic chitosan-hydroxyapatite hybrid biocoatings for enamel remineralization. Ceram Int 43:11390–11402
Salama A (2018) Chitosan based hydrogel assisted spongelike calcium phosphate mineralization for in-vitro BSA release. Int J Biol Macromol 108:471–476
Liang H, Sheng F, Zhou B, Pei Y, Li B, Li J (2017) Phosphoprotein/chitosan electrospun nanofibrous scaffold for biomineralization. Int J Biol Macromol 102:218–224
Douglas TE, Skwarczynska A, Modrzejewska Z, Balcaen L, Schaubroeck D, Lycke S, Leeuwenburgh SC (2013) Acceleration of gelation and promotion of mineralization of chitosan hydrogels by alkaline phosphatase. Int J Biol Macromol 56:122–132
Li N, Zhou L, Xie W, Zeng D, Cai D, Wang H, Li L et al (2019) Alkaline phosphatase enzyme-induced biomineralization of chitosan scaffolds with enhanced osteogenesis for bone tissue engineering. Chem Eng J 371:618–630
Orafa Z, Irani S, Zamanian A, Bakhshi H, Nikukar H, Ghalandari B (2021) Evaluation of biocompatibility of PLA scaffold coated with laponite on human bone marrow mesenchymal stem cells. J Anim Biol 13(4):101–117
Fu C, Yang X, Tan S, Song L (2017) Enhancing cell proliferation and osteogenic differentiation of MC3T3-E1 pre-osteoblasts by BMP-2 delivery in graphene oxide-incorporated PLGA/HA biodegradable microcarriers. Sci Rep 7(1):1–13
Kim JA, Yun HS, Choi YA, Kim JE, Choi SY, Kwon TG, Park EK (2018) Magnesium phosphate ceramics incorporating a novel indene compound promote osteoblast differentiation in vitro and bone regeneration in vivo. Biomaterials 157:51–61
Roberts S, Narisawa S, Harmey D, Millán JL, Farquharson C (2007) Functional involvement of PHOSPHO1 in matrix vesicle–mediated skeletal mineralization. J Bone Miner Res 22(4):617–627
Rader BA (2017) Alkaline phosphatase, an unconventional immune protein. Front Immunol. https://doi.org/10.3389/fimmu.2017.00897
Favarin BZ, Andrade MAR, Bolean M, Simão AMS, Ramos AP, Hoylaerts MF, Millán JL, Ciancaglini P (2017) Effect of the presence of cholesterol in the interfacial microenvironment on the modulation of the alkaline phosphatase activity during in vitro mineralization. Colloids Surf B 155:466–476
Khorasani MT, Joorabloo A, Moghaddam A, Shamsi H, MansooriMoghadam Z (2018) Incorporation of ZnO nanoparticles into heparinised polyvinyl alcohol/chitosan hydrogels for wound dressing application. Int J Biol Macromol 114:1203–1215
Domingos M, Gloria A, Gristina R, Ambrosio L, Bártolo PJ, Favia P, Uovo F, Gloria A, Gristina R, Ambrosio L, Bártolo PJ, Favia P (2013) Improved osteoblast cell affinity on plasma-modified 3-D extruded PCL scaffolds. Acta Biomater 9(4):5997–6005
Marolt D, Vunjak-Novakovic G (2010) Bone tissue engineering with human stem cells. Stem Cell Res Therapy. 1(2):10
Moghadam FH, Dehghan M, Eslami G, Nadri H, Moradi A, Vahedian-Ardakani H, Barzegar K (2014) Differentiation of bone marrow mesenchymal stem cells into chondrocytes after short term culture in alkaline medium. Int J Hematol-Oncol Stem Cell Res 8(4):12
Birmingham E, Niebur G, McHugh P (2012) Osteogenic differentiation of mesenchymal stem cells is regulated by osteocyte and osteoblast cells in a simplified bone niche. Eur Cell Mater 23:13–27
Shi Z, Neoh K, Kang E, Poh CK, Wang W (2009) Surface functionalization of titanium with carboxymethyl chitosan and immobilized bone morphogenetic protein-2 for enhanced osseointegration. Biomacromol 10:1603–1611
Baratta JL, Ngo A, Lopez B, Kasabwalla N, Longmuir KJ, Robertson RT (2009) Cellular organization of normal mouse liver: a histological, quantitative immunocytochemical, and fine structural analysis. Histochem Cell Biol 131(6):713–726
Anne Neumann AC (2013) BMP2-loaded nanoporous silica nanoparticles promote osteogenic differentiation of human mesenchymal stem cells. RSC Adv 3:24222–24230
Gamblin A-L et al (2014) Bone tissue formation with human mesenchymal stem cells and biphasic calcium phosphate ceramics: the local implication of osteoclasts and macrophages. Biomaterials 35(36):9660–9667
Kilmer CE, Battistoni CM, Cox A, Breur GJ, Panitch A, Liu JC (2020) Collagen type I and II blend hydrogel with autologous mesenchymal stem cells as a scaffold for articular cartilage defect repair. ACS Biomater Sci Eng 6(6):3464–3476
Funding
This research received no external funding.
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare no competing interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Sharifi, F., Hasani, M., Atyabi, S.M. et al. Mesenchymal stem cells encapsulation in chitosan and carboxymethyl chitosan hydrogels to enhance osteo-differentiation. Mol Biol Rep 49, 12063–12075 (2022). https://doi.org/10.1007/s11033-022-08013-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-022-08013-9