In situ-forming chitosan/nano-hydroxyapatite/collagen gel for the delivery of bone marrow mesenchymal stem cells

Abstract A biomimetic and thermosensitive gel scaffold was prepared from chitosan (CS), nano-hydroxyapatite (HA) and collagen (Col). The changes of storage modulus, loss modulus, pH and conductivity of the CS/HA/Col system as a function of temperature confirmed the liquid-to-gel phase transition process occurred under physiological conditions. The formed gel acted as a biocompatible substrate for the proliferation of rat bone marrow stem cells (rBMSCs) in vitro. rBMSCs suspension was co-injected with CS/HA/Col solution into Wistar rats. The injected rBMSCs survived in the CS/HA/Col gel for 28 days in vivo. The CS/HA/Col/rBMSCs gels induced less inflammatory reaction in the host tissue than the CS/HA/Col gels. Our results suggest that the CS/HA/Col system can be used to load rBMSCs in vitro homogeneously. The CS/HA/Col system can be injected into body in a minimally invasive manner and provides a biocompatible environment for rBMSCs survival in vivo.

[1]  Fuzhai Cui,et al.  Chemical characteristics and cytocompatibility of collagen-based scaffold reinforced by chitin fibers for bone tissue engineering. , 2006, Journal of biomedical materials research. Part B, Applied biomaterials.

[2]  Krishnendu Roy,et al.  Biomaterials for stem cell differentiation. , 2008, Advanced drug delivery reviews.

[3]  C. Kennedy,et al.  Molecular interactions in collagen and chitosan blends. , 2004, Biomaterials.

[4]  P. Carreau,et al.  Physical gelation of chitosan in the presence of beta-glycerophosphate: the effect of temperature. , 2005, Biomacromolecules.

[5]  S. Bryant,et al.  Crosslinking density influences the morphology of chondrocytes photoencapsulated in PEG hydrogels during the application of compressive strain , 2004, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[6]  Q. Feng,et al.  Biomimetic properties of an injectable chitosan/nano-hydroxyapatite/collagen composite , 2011 .

[7]  James M. Anderson,et al.  Biological Responses to Materials , 2001 .

[8]  Dong Wang,et al.  Rheological characterisation of thermogelling chitosan/glycerol-phosphate solutions , 2001 .

[9]  S Tamai,et al.  Immediate bone forming capability of prefabricated osteogenic hydroxyapatite. , 1996, Journal of biomedical materials research.

[10]  J. Kennedy,et al.  Injectable hydrogels based on chitosan derivative/polyethylene glycol dimethacrylate/N,N-dimethylacrylamide as bone tissue engineering matrix , 2010 .

[11]  D. Mooney,et al.  Hydrogels for tissue engineering: scaffold design variables and applications. , 2003, Biomaterials.

[12]  J. Y. Lee,et al.  The osteogenic differentiation of rat muscle-derived stem cells in vivo within in situ-forming chitosan scaffolds. , 2008, Biomaterials.

[13]  Michael D Buschmann,et al.  Heat-induced transfer of protons from chitosan to glycerol phosphate produces chitosan precipitation and gelation. , 2008, Biomacromolecules.

[14]  V. Goldberg,et al.  Heterotopic osteogenesis in porous ceramics induced by marrow cells , 1989, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[15]  Qingling Feng,et al.  Porous poly-L-lactic acid scaffold reinforced by chitin fibers , 2005 .

[16]  C. Carlo-Stella,et al.  Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. , 2002, Blood.

[17]  Steve Weiner,et al.  THE MATERIAL BONE: Structure-Mechanical Function Relations , 1998 .

[18]  X. Mo,et al.  Intermolecular interactions in electrospun collagen–chitosan complex nanofibers , 2008 .

[19]  Kristi S Anseth,et al.  Mechanical Properties of Cellularly Responsive Hydrogels and Their Experimental Determination , 2010, Advanced materials.

[20]  J. H. Kim,et al.  In vivo osteogenic differentiation of human adipose-derived stem cells in an injectable in situ-forming gel scaffold. , 2009, Tissue engineering. Part A.

[21]  R. Muzzarelli.  Chitins and chitosans for the repair of wounded skin, nerve, cartilage and bone , 2009 .

[22]  Kevin McIntosh,et al.  Mesenchymal stem cells suppress lymphocyte proliferation in vitro and prolong skin graft survival in vivo. , 2002, Experimental hematology.

[23]  Jing Deng,et al.  Biocompatibility of a chitosan-based injectable thermosensitive hydrogel and its effects on dog periodontal tissue regeneration , 2010 .

[24]  F. Djouad,et al.  Immunosuppressive effect of mesenchymal stem cells favors tumor growth in allogeneic animals. , 2003, Blood.

[25]  H. Lee,et al.  Chitosan Gel as an In Situ–Forming Scaffold for Rat Bone Marrow Mesenchymal Stem Cells In Vivo , 2008 .

[26]  Xiguang Chen,et al.  Preparation and characteristics of novel porous hydrogel films based on chitosan and glycerophosphate , 2009 .

[27]  C. Bowman,et al.  Mechanical properties of hydrogels and their experimental determination. , 1996, Biomaterials.

[28]  R. Wright,et al.  The Fourth Pacific Rim International Conference on Advanced Materials and Processing (PRICM4) , 2001 .

[29]  Samuel I Stupp,et al.  Biomimetic systems for hydroxyapatite mineralization inspired by bone and enamel. , 2008, Chemical reviews.

[30]  K. Shakesheff,et al.  Injectable scaffolds for tissue regeneration , 2004 .

[31]  A. Malmström,et al.  Proposed protective mechanism of the pancreas in the rat , 2010, Journal of Inflammation.

[32]  P. Carreau,et al.  Study of the phase transition of Poly(N, N-diethylacrylamide) in water by rheology and dynamic light scattering , 2003 .

[33]  M. Machluf,et al.  Alginate–chitosan complex coacervation for cell encapsulation: Effect on mechanical properties and on long‐term viability , 2006, Biopolymers.

[34]  P. Carreau,et al.  Effect of urea on solution behavior and heat-induced gelationof chitosan-β-glycerophosphate , 2006 .

[35]  H. Ohgushi,et al.  Osteogenic Phenotype Expression of Allogeneic Rat Marrow Cells in Porous Hydroxyapatite Ceramics , 1999, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[36]  F. Cui,et al.  Hierarchical Self-Assembly of Nano-Fibrils in Mineralized Collagen , 2003 .