Imaging human bone marrow stem cell morphogenesis in polyglycolic acid scaffold by multiphoton microscopy.

The noninvasive imaging of tissue engineering constructs is vital for understanding the physiological changes in construct formation and the design of improved products for therapeutic purposes. In this work, we use the combination of multiphoton autofluorescence and second harmonic generation (SHG) microscopy to image the physiological changes to the engineered constructs of human mesenchymal stem cells seeded in a polyglycolic acid (PGA) scaffold under induction by chondrogenic transforming growth factor-beta3. Without histological procedures, we found that multiphoton autofluorescence is useful for imaging the PGA scaffold and stem cells while SHG is useful for following the progress of extracellular matrix (ECM) formation. We found that the initial ECM formation tends to align along the PGA scaffold orientation and progressive induction alters the scaffold conformation, indicating that biomechanical forces or the chemical environment generated by chondrogenesis is sufficient for scaffold reorganization. Our results suggest that in the future this approach may be used for real-time monitoring of the physiological processes associated with tissue engineering.

[1]  B R Masters,et al.  Two-photon excitation fluorescence microscopy. , 2000, Annual review of biomedical engineering.

[2]  L. Griffith,et al.  Functional behavior of primary rat liver cells in a three-dimensional perfused microarray bioreactor. , 2002, Tissue engineering.

[3]  Wei Wu,et al.  Two-photon laser scanning microscopy of epithelial cell-modulated collagen density in engineered human lung tissue. , 2001, Tissue engineering.

[4]  U A Stock,et al.  Tissue engineering: current state and prospects. , 2001, Annual review of medicine.

[5]  M. L. Marquette,et al.  A novel in vitro three-dimensional skeletal muscle model , 2007, In Vitro Cellular & Developmental Biology - Animal.

[6]  C. Dong,et al.  Imaging tissue engineering scaffolds using multiphoton microscopy , 2008, Microscopy research and technique.

[7]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[8]  W. Denk,et al.  Two-photon laser scanning fluorescence microscopy. , 1990, Science.

[9]  L. Freed.  Tissue culture bioreactors ; chondrogenesis as a model system , 1997 .

[10]  Jeffrey A Hubbell,et al.  Analysis of progenitor cell-scaffold combinations by in vivo non-invasive photonic imaging. , 2007, Biomaterials.

[11]  R. Tuan,et al.  A three-dimensional nanofibrous scaffold for cartilage tissue engineering using human mesenchymal stem cells. , 2005, Biomaterials.

[12]  E. Caterson,et al.  Three-dimensional cartilage formation by bone marrow-derived cells seeded in polylactide/alginate amalgam. , 2001, Journal of biomedical materials research.

[13]  Ching-Chuan Jiang,et al.  Multipotential Mesenchymal Stem Cells from Femoral Bone Marrow Near the Site of Osteonecrosis , 2003, Stem cells.

[14]  W. Webb,et al.  Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[15]  B. Tromberg,et al.  Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[16]  S. Bruder,et al.  Mesenchymal stem cells: building blocks for molecular medicine in the 21st century. , 2001, Trends in molecular medicine.