Chitosan/hyaluronic acid porous scaffold for bone tissue engineering
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1
Université de Reims Champagne Ardenne, Odontology, France
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2
Université de Lorraine, CNRS 7563, France
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3
Université de Cergy Pontoise, EA 1391, France
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4
Université de Reims Champagne Ardenne, Pharmacy, France
Introduction: Strategies for bone tissue engineering and regeneration rely on bioactive scaffolds to mimic the natural extracellular matrix and act as templates onto which cells attach, proliferate, and differentiate. Scaffolds need to be biocompatible, biodegradable during tissue regeneration process, structurally close to bone, and with some degree of mechanical strength. In the present work, we developed a porous three-dimensional (3D) hybrid chitosan (CHI)/hyaluronic acid (HA) scaffold suitable for tissue engineering applications. Cell colonization was studied using Wharton’s Jelly stem cells (WJ-SCs) which are considered as primitive stem cells much more proliferative, immunosuppressive, and even therapeutically active than adult stem cells.
Experimental Methods: CHI was dissolved in 0.1 M acetic acid, neutralized by the addition of β-glycerolphosphate (β-GP) and then supplemented by HA. The final CHI/HA ratio was fixed at 3/1. Further crosslink was obtained by adding Genipin solution into CHI/HA mixture. Freeze-drying approach was used to form 3D scaffolds. Porosity, pore size, water uptake ability as well as elastic modulus were first investigated. Cellular compatibility and proliferation were followed through DNA quantification assay and cytoskeleton labelling.
Results and Discussion: Formation of 3D porous scaffold was demonstrated by scanning electron microscopy and confocal microscopy. Adding β-glycerophosphate to the mixture was required for exhibiting interconnected pores. Through natural derived cross-linking reagents with low toxicity, genipin, CHI/HA scaffold exhibited a higher elastic modulus and more stable structure compared to uncrosslinked scaffold. DNA assay showed an increase in cell number, signature of cytotoxic agents free scaffold. Confocal imaging demonstrated the presence of cells inner the CHI/HA scaffold reflecting scaffold colonization by WJ-SCs. However, cells grow into the scaffold kept a round morphology in opposite to the plastic cell culture, where cells preserved their fibroblastic shape. A possible reason is that the 3D scaffold provides a geometrical distribution of binding sites to cells rather than planer conventional substrate.
Conclusion: In this work, a novel cell-colonized scaffold exhibiting highly porous and interconnected pore was developed. The original and simple thermogelling followed by freez-drying approach ensure an easy transfer to orthobiological industries. Nevertheless, understanding the mechanism and process of cell colonization in these 3D porous scaffolds are essential for further physiological tissue model applications.
Keywords:
Cell Adhesion,
Biocompatibility,
3D scaffold,
Bone repair
Conference:
10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.
Presentation Type:
Poster
Topic:
Biomaterials in constructing tissue substitutes
Citation:
Jing
J,
Rammal
H,
Dubus
M,
Rahouadj
R,
Pauthe
E,
Velard
F,
Braux
J,
Gangloff
SC,
Siad
L and
Kerdjoudj
H
(2016). Chitosan/hyaluronic acid porous scaffold for bone tissue engineering.
Front. Bioeng. Biotechnol.
Conference Abstract:
10th World Biomaterials Congress.
doi: 10.3389/conf.FBIOE.2016.01.01918
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Received:
27 Mar 2016;
Published Online:
30 Mar 2016.
*
Correspondence:
Dr. Jing Jing, Université de Reims Champagne Ardenne, Odontology, Reims, France, jing.jing@univ-reims.fr
Dr. Hassan Rammal, Université de Reims Champagne Ardenne, Odontology, Reims, France, hassan.rammal@univ-reims.fr
Dr. Rachid Rahouadj, Université de Lorraine, CNRS 7563, Nancy, France, rachid.rahouadj@univ-lorraine.fr
Dr. Emmanuel Pauthe, Université de Cergy Pontoise, EA 1391, Paris, France, emmanuel.pauthe@u-cergy.fr
Dr. Frederic Velard, Université de Reims Champagne Ardenne, Odontology, Reims, France, frederic.velard@univ-reims.fr
Dr. Julien Braux, Université de Reims Champagne Ardenne, Odontology, Reims, France, julien.braux@univ-reims.fr
Dr. Sophie C Gangloff, Université de Reims Champagne Ardenne, Pharmacy, Reims, France, sophie.gangloff@univ-reims.fr
Dr. Larbi Siad, Université de Reims Champagne Ardenne, Pharmacy, Reims, France, larbi.siad@univ-reims.fr
Dr. Halima Kerdjoudj, Université de Reims Champagne Ardenne, Odontology, Reims, France, Email1