Characterisation of hyaluronic acid methylcellulose hydrogels for 3D bioprinting.

Hydrogels containing hyaluronic acid (HA) and methylcellulose (MC) have shown promising results for three dimensional (3D) bioprinting applications. However, several parameters influence the applicability bioprinting and there is scarce data in the literature characterising HAMC. We assessed eight concentrations of HAMC for printability, swelling and stability over time, rheological and structural behaviour, and viability of mesenchymal stem cells. We show that HAMC blends behave as viscous solutions at 4°C and have faster gelation times at higher temperatures, typically gelling upon reaching 37°C. We found the storage, loss and compressive moduli to be dependent on HAMC concentration and incubation time at 37°C, and show the compressive modulus to be strain-rate dependent. Swelling and stability was influenced by time, more so than pH environment. We demonstrated that mesenchymal stem cell viability was above 75% in bioprinted structures and cells remain viable for at least one week after 3D bioprinting. The mechanical properties of HAMC are highly tuneable and we show that higher concentrations of HAMC are particularly suited to cell-encapsulated 3D bioprinting applications that require scaffold structure and delivery of cells.

[1]  Song Li,et al.  Anisotropic mechanosensing by mesenchymal stem cells , 2006, Proceedings of the National Academy of Sciences.

[2]  H. Shivakumar,et al.  Investigation of Swelling Behavior and Mechanical Properties of a pH-Sensitive Superporous Hydrogel Composite , 2012, Iranian journal of pharmaceutical research : IJPR.

[3]  M. Maiuri,et al.  Effect of hyaluronic acid on the thermogelation and biocompatibility of its blends with methyl cellulose. , 2014, Carbohydrate polymers.

[4]  T. Wong,et al.  Hyaluronan Regulates Cell Behavior: A Potential Niche Matrix for Stem Cells , 2012, Biochemistry research international.

[5]  Charles Tator,et al.  Intrathecal delivery of a polymeric nanocomposite hydrogel after spinal cord injury. , 2010, Biomaterials.

[6]  Cindi M Morshead,et al.  Controlled epi-cortical delivery of epidermal growth factor for the stimulation of endogenous neural stem cell proliferation in stroke-injured brain. , 2011, Biomaterials.

[7]  Cindi M Morshead,et al.  A hydrogel composite system for sustained epi-cortical delivery of Cyclosporin A to the brain for treatment of stroke. , 2013, Journal of controlled release : official journal of the Controlled Release Society.

[8]  M. Shoichet,et al.  An injectable drug delivery platform for sustained combination therapy. , 2009, Journal of controlled release : official journal of the Controlled Release Society.

[9]  Charles Tator,et al.  Characterization of hyaluronan-methylcellulose hydrogels for cell delivery to the injured spinal cord. , 2013, Journal of biomedical materials research. Part A.

[10]  F. Huss,et al.  Transplantation of cultured human keratinocytes in single cell suspension: a comparative in vitro study of different application techniques. , 2008, Burns : journal of the International Society for Burn Injuries.

[11]  Charles H Tator,et al.  Fast-gelling injectable blend of hyaluronan and methylcellulose for intrathecal, localized delivery to the injured spinal cord. , 2006, Biomaterials.

[12]  Sharon Gerecht,et al.  Dextran hydrogel scaffolds enhance angiogenic responses and promote complete skin regeneration during burn wound healing , 2011, Proceedings of the National Academy of Sciences.

[13]  D. van der Kooy,et al.  A Hyaluronan-Based Injectable Hydrogel Improves the Survival and Integration of Stem Cell Progeny following Transplantation , 2015, Stem cell reports.

[14]  Anup Tuladhar,et al.  Circumventing the blood-brain barrier: Local delivery of cyclosporin A stimulates stem cells in stroke-injured rat brain. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[15]  B. Duan,et al.  3D bioprinting of heterogeneous aortic valve conduits with alginate/gelatin hydrogels. , 2013, Journal of biomedical materials research. Part A.

[16]  P. Janmey,et al.  Tissue Cells Feel and Respond to the Stiffness of Their Substrate , 2005, Science.

[17]  Barry J. Doyle,et al.  Parameter optimization for 3D bioprinting of hydrogels , 2017 .

[18]  Subbu Venkatraman,et al.  Printing cell-laden gelatin constructs by free-form fabrication and enzymatic protein crosslinking , 2015, Biomedical microdevices.

[19]  G. Gethin,et al.  The significance of surface pH in chronic wounds , 2007 .

[20]  Anthony Atala,et al.  Evaluation of hydrogels for bio-printing applications. , 2013, Journal of biomedical materials research. Part A.

[21]  Ali Khademhosseini,et al.  3D biofabrication strategies for tissue engineering and regenerative medicine. , 2014, Annual review of biomedical engineering.

[22]  James C. Weaver,et al.  Hydrogels with tunable stress relaxation regulate stem cell fate and activity , 2015, Nature materials.

[23]  D. Beebe,et al.  Equilibrium swelling and kinetics of pH-responsive hydrogels: models, experiments, and simulations , 2002 .

[24]  M. Shoichet,et al.  Accelerated release of a sparingly soluble drug from an injectable hyaluronan-methylcellulose hydrogel. , 2009, Journal of controlled release : official journal of the Controlled Release Society.

[25]  Jennifer S. Park,et al.  The effect of matrix stiffness on the differentiation of mesenchymal stem cells in response to TGF-β. , 2011, Biomaterials.

[26]  R. Wells The role of matrix stiffness in regulating cell behavior , 2008, Hepatology.

[27]  J. Hunt,et al.  The effects of pH on wound healing, biofilms, and antimicrobial efficacy , 2014, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[28]  D. van der Kooy,et al.  A hydrogel-based stem cell delivery system to treat retinal degenerative diseases. , 2010, Biomaterials.

[29]  Joachim P Spatz,et al.  Stem cell migration and mechanotransduction on linear stiffness gradient hydrogels , 2017, Proceedings of the National Academy of Sciences.

[30]  Z. Upton,et al.  Optimized delivery of skin keratinocytes by aerosolization and suspension in fibrin tissue adhesive , 2006, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[31]  Jason A. Burdick,et al.  Hyaluronic Acid Hydrogels for Biomedical Applications , 2011, Advanced materials.

[32]  L. Mullins Softening of Rubber by Deformation , 1969 .