X-ray diffraction enhanced imaging as a novel method to visualize low-density scaffolds in soft tissue engineering.

Scaffold visualization is challenging yet essential to the success of various tissue engineering applications. The aim of this study was to explore the potential of X-ray diffraction enhanced imaging (DEI) as a novel method for the visualization of low density engineered scaffolds in soft tissue. Imaging of the scaffolds made from poly(L-lactide) (PLLA) and chitosan was conducted using synchrotron radiation-based radiography, in-line phase-contrast imaging (in-line PCI), and DEI techniques as well as laboratory-based radiography. Scaffolds were visualized in air, water, and rat muscle tissue. Compared with the images from X-ray radiography and in-line PCI techniques, DEI images more clearly show the structure of the low density scaffold in air and have enhanced image contrast. DEI was the only technique able to visualize scaffolds embedded in unstained muscle tissue; this method could also define the microstructure of muscle tissue in the boundary areas. At a photon energy of 20 KeV, DEI had the capacity to image PLLA/chitosan scaffolds in soft tissue with a sample thickness of up to 4 cm. The DEI technique can be applied at high X-ray energies, thus facilitating lower in vivo radiation doses to tissues during imaging as compared to conventional radiography.

[1]  J. Mano,et al.  Bioactive poly(L-lactic acid)-chitosan hybrid scaffolds , 2008 .

[2]  Zhong Zhong,et al.  Design and implementation of a compact low-dose diffraction enhanced medical imaging system. , 2009, Academic radiology.

[3]  H. Riesemeier,et al.  Going beyond histology. Synchrotron micro-computed tomography as a methodology for biological tissue characterization: from tissue morphology to individual cells , 2009, Journal of The Royal Society Interface.

[4]  S. Wilkins,et al.  Phase-contrast imaging using polychromatic hard X-rays , 1996, Nature.

[5]  D. Chapman,et al.  A Brief Review of Visualization Techniques for Nerve Tissue Engineering Applications , 2010 .

[6]  David L. Kaplan,et al.  Non-Invasive Time-Lapsed Monitoring and Quantification of Engineered Bone-Like Tissue , 2007, Annals of Biomedical Engineering.

[7]  N. Turner,et al.  A novel hyaluronan-based biomaterial (Hyaff-11) as a scaffold for endothelial cells in tissue engineered vascular grafts. , 2004, Biomaterials.

[8]  P. Cloetens,et al.  Quantitative comparison between two phase contrast techniques: diffraction enhanced imaging and phase propagation imaging , 2006, Physics in medicine and biology.

[9]  E. Pisano,et al.  Characterization of diffraction-enhanced imaging contrast in breast cancer , 2009, Physics in medicine and biology.

[10]  X. B. Chen,et al.  Effects of laminin blended with chitosan on axon guidance on patterned substrates , 2010, Biofabrication.

[11]  Wei Sun,et al.  Recent development on computer aided tissue engineering - a review , 2002, Comput. Methods Programs Biomed..

[12]  Bruno De Man,et al.  An outlook on x-ray CT research and development. , 2008, Medical physics.

[13]  Judith M Curran,et al.  The differentiation of bone marrow mesenchymal stem cells into chondrocyte-like cells on poly-L-lactic acid (PLLA) scaffolds. , 2006, Biomaterials.

[14]  Gao,et al.  X-ray image contrast from a simple phase object. , 1995, Physical review letters.

[15]  Investigation of biomedical inner microstructures with hard X-ray phase-contrast imaging , 2007 .

[16]  E. Pisano,et al.  Diffraction enhanced x-ray imaging. , 1997, Physics in medicine and biology.

[17]  Warren S. Grundfest,et al.  Bioengineering and Imaging Research Opportunities Workshop V: A Summary , 2008, Annals of Biomedical Engineering.

[18]  K. Shakesheff,et al.  The influence of dispersant concentration on the pore morphology of hydroxyapatite ceramics for bone tissue engineering. , 2005, Biomaterials.

[19]  Françoise Peyrin,et al.  Quantification of the degree of mineralization of bone in three dimensions using synchrotron radiation microtomography. , 2002, Medical physics.

[20]  Charles Tator,et al.  Extramedullary chitosan channels promote survival of transplanted neural stem and progenitor cells and create a tissue bridge after complete spinal cord transection. , 2008, Tissue engineering. Part A.

[21]  D W Holdsworth,et al.  Fundamental image quality limits for microcomputed tomography in small animals. , 2003, Medical physics.

[22]  Chiara Renghini,et al.  Micro-CT studies on 3-D bioactive glass-ceramic scaffolds for bone regeneration. , 2009, Acta biomaterialia.

[23]  D R Dance,et al.  X-ray refraction effects: application to the imaging of biological tissues. , 2003, The British journal of radiology.

[24]  Donglin Zeng,et al.  Radiologist evaluation of an X-ray tube-based diffraction-enhanced imaging prototype using full-thickness breast specimens. , 2009, Academic radiology.

[25]  Francoise Peyrin,et al.  X-ray synchrotron radiation pseudo-holotomography as a new imaging technique to investigate angio- and microvasculogenesis with no usage of contrast agents. , 2009, Tissue engineering. Part C, Methods.

[26]  Alberto Bravin,et al.  High-resolution CT by diffraction-enhanced x-ray imaging: mapping of breast tissue samples and comparison with their histo-pathology , 2007, Physics in medicine and biology.

[27]  Charles Tator,et al.  Delayed implantation of intramedullary chitosan channels containing nerve grafts promotes extensive axonal regeneration after spinal cord injury. , 2008, Neurosurgery.

[28]  Highly sensitive detection of the soft tissues based on refraction contrast by in-plane diffraction-enhanced imaging CT , 2008 .

[29]  R. Guldberg,et al.  Imaging Techniques for Biomaterials Characterization , 2007 .

[30]  D. Bezuidenhout,et al.  Rapid three-dimensional quantification of VEGF-induced scaffold neovascularisation by microcomputed tomography. , 2009, Biomaterials.

[31]  Ingo Heschel,et al.  In vitro cell alignment obtained with a Schwann cell enriched microstructured nerve guide with longitudinal guidance channels. , 2009, Biomaterials.

[32]  Kun Hu,et al.  Three-dimensional fibroin/collagen scaffolds derived from aqueous solution and the use for HepG2 culture , 2005 .

[33]  William Thomlinson,et al.  Implementation of diffraction-enhanced imaging experiments: at the NSLS and APS , 2000 .

[34]  I. Robinson.  Elements of Modern X-ray Physics , 2002 .

[35]  Ralph Müller,et al.  Nondestructive micro-computed tomography for biological imaging and quantification of scaffold-bone interaction in vivo. , 2007, Biomaterials.

[36]  Hongtae Kim,et al.  High resolution X‐ray phase contrast synchrotron imaging of normal and ligation damaged rat sciatic nerves , 2008, Microscopy research and technique.

[37]  Dietmar W Hutmacher,et al.  Assessment of bone ingrowth into porous biomaterials using MICRO-CT. , 2007, Biomaterials.

[38]  Yuanwei Chen,et al.  Effect of strontium ions on the growth of ROS17/2.8 cells on porous calcium polyphosphate scaffolds. , 2006, Biomaterials.