Abstract
Our previous study showed that thermosensitive chitosan/corn starch/β-glycerol phosphate (C/S/β-GP) hydrogel was an effective carrier for chondrocytes and their transforming factor, TGF-β1. In the present study, MSCs were grown in C/S/β-GP hydrogels as an effective tool for chondrocyte-like cell differentiation. The MSCs-encapsulated hydrogel was prepared by blending chitosan solution (1.70% w/v in 0.1 M HCl) with pregelatinized corn starch solution (1.70% w/v). The total final concentration of the blended polymers was 1.53% w/v, and the weight ratio of chitosan to corn starch was 4 to 1. The TGF-β1 (final concentration of 25 ng/mL) and 5 × 105 MSCs were added to 500 μL chitosan/starch solution. Finally, β-GP (60% w/v) was added to obtain 6.0% w/v final concentration. The C/S/β-GP hydrogel changed from a liquid at room temperature to a gel at 37 ± 2°C. It converted the fibroblast-like MSCs into spheroid cells. In hydrogels containing TGF-β1, these cells further differentiated into chondrocyte-like cells. This was shown by their expressions of type II collagen and aggrecan mRNA. Type I collagen mRNA was initially expressed but this disappeared by 6 weeks in culture suggesting a complete chondrocyte differentiation by that time. Type II collagen protein production was detected by immunohistochemistry and immunofluorescence, and successively increased after 4–6 weeks in culture. Neither the mRNA nor the collagen expression could be detected in the absence of TGF-β1. The data indicate that MSCs would be an appropriate chondrocyte precursor in conjunction with our hydrogel loading TGF-β1 which is able to sustain chondrocyte function.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
R Cancedda, B Dozin, P Giannoni, et al., Tissue engineering and cell therapy of cartilage and bone, Matrix Biol, 22, 81, (2003).
D Magne, C Vinatier, M Julien, et al., Mesenchymal stem cell therapy to rebuild cartilage, Trends Mol Med, 11, 519 (2005).
WJ Li, R Tuli, C Okafor, et al., A three-dimensional nanofibrous scaffold for cartilage tissue engineering using human mesenchymal stem cells, Biomaterials, 26, 599 (2005).
JM Dang, DD Sun, Y Shin-Ya, et al., Temperature-responsive hydroxybutyl chitosan for the culture of mesenchymal stem cells and intervertebral disk cells, Biomaterials, 27, 406 (2006).
RL Mauck, X Yuan, RS Tuan, Chondrogenic differentiation and functional maturation of bovine mesenchymal stem cells in longterm agarose culture, Osteoarthr Cartilage, 14, 179 (2006).
AI Caplan, Adult mesenchymal stem cells for tissue engineering versus regenerative medicine, J Cell Physiol, 213, 341 (2007).
RF Pereira, KW Halford, MD O’Hara, et al., Cultured adherent cells from marrow can serve as long-lasting precursor cells for bone, cartilage, and lung in irradiated mice, Proc Natl Acad Sci USA, 92, 4857 (1995).
EM Horwitz, DJ Prockop, LA Fitzpatrick, et al., Transplantability and therapeutic effects of bone marrow-derived mesenchymal cells in children with osteogenesis imperfect, Nat Med, 5, 309 (1999).
MF Pittenger, AM Mackay, SC Beck, et al., Multilineage potential of adult human mesenchymal stem cells, Science, 284, 143 (1999).
SE Kim, JH Park, YW Cho, et al., Porous chitosan scaffold containing microspheres loaded with transforming growth factor-β1: Implications for cartilage tissue engineering, J Control Release, 91, 365 (2003).
M-N Kang, H-H Yoon, Y-K Seo, et al., Human umbilical cordderived mesenchymal stem cells differentiate into ligament-like cells with mechanical stimulation in various media, Tissue Eng Regen Med, 9, 185 (2012).
SH Lee, H Shin, Matrices and scaffolds for delivery of bioactive molecules in bone and cartilage tissue engineering, Adv Drug Deliver Rev, 59, 339 (2007).
TA Holland, JKV Tessmar, Y Tabata, et al., Transforming growth factor-β1 release from oligo(poly(ethylene glycol) fumarate) hydrogels in conditions that model the cartilage wound healing environment, J Control Release, 94, 101 (2004).
WJ Li, R Tuli, C Okafor, et al., A three-dimensional nanofibrous scaffold for cartilage tissue engineering using human mesenchymal stem cells, Biomaterials, 26, 599 (2005).
LY Sung, WH Lo, HY Chiu, et al., Modulation of chondrocyte phenotype via baculovirus-mediated growth factor expression, Biomaterials, 28, 3437 (2007).
GM Williams, KJ Dills, CR Flores, et al., Differential regulation of immature articular cartilage compressive moduli and Poisson’s ratios by in vitro stimulation with IGF-1 and TGFbeta1, J Biomech, 43, 2501 (2010).
J Ngoenkam, A Faikrua, S Yasothornsrikul, et al., Potential of an injectable chitosan/starch/β-glycerol phosphate hydrogel for sustaining normal chondrocyte function, Int J Pharm, 391, 115 (2010).
A Faikrua, S Wittaya-Areekul, B Oonkhanond, et al., In vivo chondrocyte and transforming growth factor-β1 delivery using the thermosensitive chitosan/starch/β-glycerol phosphate hydrogel, J Biomater Appl, 28, 175 (2013).
A Faikrua, R Jeenapongsa, M Sila-asna, et al., Properties of âglycerol phosphate/collagen/chitosan blend scaffolds for application in skin tissue engineering, ScienceAsia, 35, 247 (2009).
MA Lee, L Cai, N Hübner, et al., Tissue and development specific expression of multiple alternatively spliced transcripts of rat neuronal nitric oxide synthase, J Clin Invest, 100, 1507 (1997).
A Gartland, J Mechler, A Mason-Savas, et al., In vitro chondrocyte differentiation using costochondral chondrocytes as a source of primary rat chondrocyte cultures: An improved isolation and cryopreservation method, Bone, 37, 530 (2005).
F Barry, RE Boynton, B Liu, et al., Chondrogenic differentiation of mesenchymal stem cells from bone marrow: Differentiationdependent gene expression of matrix components, Exp Cell Res, 268, 189 (2001).
AD Murdoch, LM Grady, MP Ablett, et al., Chondrogenic differentiation of human bone marrow stem cells in transwell cultures: Generation of scaffold-free cartilage, Stem Cells, 25, 2786 (2007).
M Endres, N Wenda, H Woehlecke, et al., Microencapsulation and chondrogenic differentiation of human mesenchymal progenitor cells from subchondral bone marrow in Ca-alginate for cell injection, Acta Biomater, 6, 436 (2010).
HJ Kim, KK Kim, IL Kyu Park, et al., Hybrid scaffolds composed of hyaluronic acid and collagen for cartilage regeneration, Tissue Eng Regen Med, 9, 57 (2012).
TJ Kang, JE Yeom, HJ Lee, et al., Growth kinetics of human mesenchymal stem cells from bone marrow and umbrilical cord blood, Acta Haematol, 112, 230 (2004).
SP Bruder, N Jaiswal, SE Haynesworth, Growth kinetics, selfrenewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation, J Cell Biochem, 64, 278 (1997).
KM Park, SY Lee, YK Joung, et al., Thermosensitive chitosanpluronic hydrogel as an injectable cell delivery carrier for cartilage regeneration, Acta Biomater, 5, 1956 (2009).
AD Murdoch, LM Grady, MP Ablett, et al., Chondrogenic differentiation of human bone marrow stem cells in transwell cultures: Generation of scaffold-free cartilage, Stem Cells, 25, 2786 (2007).
JL Moreau, HH Xu, Mesenchymal stem cell proliferation and differentiation on injectable calcium phosphate-chitosan composite scaffold, Biomaterials, 30, 2675 (2009).
G Karsenty, HM Kronenberg, C Settembre, Genetic control of bone formation, Annu Rev Cell Dev Biol, 25, 629 (2009).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Faikrua, A., Wittaya-areekul, S., Oonkhanond, B. et al. A thermosensitive chitosan/corn starch/β-glycerol phosphate hydrogel containing TGF-β1 promotes differentiation of MSCs into chondrocyte-like cells. Tissue Eng Regen Med 11, 355–361 (2014). https://doi.org/10.1007/s13770-014-0030-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13770-014-0030-y