Temperature-responsive hydroxybutyl chitosan for the culture of mesenchymal stem cells and intervertebral disk cells

Jiyoung M. Dang, Daniel D N Sun, Yoshitsune Shin-Ya, Ann N. Sieber, John P. Kostuik, Kam W. Leong

Research output: Contribution to journalArticle

Abstract

Temperature-responsive polymers are attractive candidates for applications related to injectable delivery of biologically active therapeutics, such as stem cells. In this study, we evaluate the potential of thermosensitive hydroxybutyl chitosan (HBC) as a biomaterial for the culture of human mesenchymal stem cells (hMSC) and cells derived from the intervertebral disk, with the eventual goal of using the HBC polymer as an injectable matrix/cell therapeutic. Conjugation of hydroxybutyl groups to chitosan renders the polymer water soluble and thermally responsive. Below its lower critical solution temperature, a solution of HBC can be maintained indefinitely in its solvated state. Upon exposure to a 37°C environment, within 60 s, a 3.8 wt% HBC solution rapidly forms a gel that can be maneuvered with forceps. Upon cooling, the gel once again is able to revert to its solvated state. The gel exhibits a dramatic increase in both G′ and G″ with increasing temperature, signifying a temperature-dependent enhancement of gel mechanical properties. Although a solid structure upon gelation, due to its physical nature of polymer interaction and gel formation, the gel exhibits a fluid-like viscoelastic behavior when exposed to shear stresses of up to 10% strain, with both G′ and G″ approaching zero with increasing shear stress. Formulations of HBC gels presented in this study have gelation temperatures ranging from 13.0 to 34.6°C and water contents of 67-95%. Minimal cytotoxicity in MSC and disk cell cultures was observed with these polymers up to a concentration of 5 wt%. Detection of metabolic activity, genetic analysis of synthesized mRNA, and histological staining of MSC and disk cell cultures in these gels collectively indicate cell proliferation without a loss in metabolic activity and extracellular matrix production. This study suggests the potential of HBC gel as an injectable carrier for future applications of delivering therapeutics to encourage a biologically relevant reconstruction of the degenerated disk.

Original languageEnglish (US)
Pages (from-to)406-418
Number of pages13
JournalBiomaterials
Volume27
Issue number3
DOIs
StatePublished - Jan 2006

Fingerprint

Intervertebral Disc
Chitosan
Stem cells
Mesenchymal Stromal Cells
Cell culture
Gels
Temperature
Polymers
Gelation
Injections
Shear stress
Cell Culture Techniques
Water
Cell proliferation
Biocompatible Materials
Cytotoxicity
Surgical Instruments
Biomaterials
Water content
Extracellular Matrix

Keywords

  • Chitin/chitosan
  • Hydrogel
  • Intervertebral disk
  • Mesenchymal stem cells
  • Thermally responsive material

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering

Cite this

Temperature-responsive hydroxybutyl chitosan for the culture of mesenchymal stem cells and intervertebral disk cells. / Dang, Jiyoung M.; Sun, Daniel D N; Shin-Ya, Yoshitsune; Sieber, Ann N.; Kostuik, John P.; Leong, Kam W.

In: Biomaterials, Vol. 27, No. 3, 01.2006, p. 406-418.

Research output: Contribution to journalArticle

Dang, Jiyoung M. ; Sun, Daniel D N ; Shin-Ya, Yoshitsune ; Sieber, Ann N. ; Kostuik, John P. ; Leong, Kam W. / Temperature-responsive hydroxybutyl chitosan for the culture of mesenchymal stem cells and intervertebral disk cells. In: Biomaterials. 2006 ; Vol. 27, No. 3. pp. 406-418.
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