A bioprosthetic ovary created using 3D printed microporous scaffolds restores ovarian function in sterilized mice
Monica M. Laronda, Alexandra L. Rutz, Shuo Xiao, Kelly A. Whelan, Francesca E. Duncan, Eric W. Roth, Teresa K. Woodruff & Ramille N. Shah
Nature Communications 8, Article number: 15261 (2017)
doi: 10.1038/ncomms15261
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Biomaterials Preclinical research Translational research
Received: 09 February 2017 Accepted: 14 March 2017
Published online:16 May 2017
Source/Fonte: Technology Networks
Abstract
Emerging additive manufacturing techniques enable investigation of the effects of pore geometry on cell behavior and function. Here, we 3D print microporous hydrogel scaffolds to test how varying pore geometry, accomplished by manipulating the advancing angle between printed layers, affects the survival of ovarian follicles. 30° and 60° scaffolds provide corners that surround follicles on multiple sides while 90° scaffolds have an open porosity that limits follicle–scaffold interaction. As the amount of scaffold interaction increases, follicle spreading is limited and survival increases. Follicle-seeded scaffolds become highly vascularized and ovarian function is fully restored when implanted in surgically sterilized mice. Moreover, pups are born through natural mating and thrive through maternal lactation. These findings present an in vivo functional ovarian implant designed with 3D printing, and indicate that scaffold pore architecture is a critical variable in additively manufactured scaffold design for functional tissue engineering.
Acknowledgements
M.M.L. and A.L.R. contributed equally to this work. The authors would like to thank Keisha Barreto (NU) of the Reproductive Science Histology Core, Center for Reproductive Science, Lindsay Reustle (KUMC) for her technical contribution on HMGB1 immunohistochemistry and Dr. Constadina Arvanitis (NU) of the Center of Advanced Microscopy. The authors would also like to thank Prof. Wesley Burghardt for use of his lab’s rheometer and his advisement on rheological data. This work was supported by the Watkins Chair of Obstetrics and Gynecology (TKW), the National Institutes of Health National Center for Translational Research in Reproduction and Infertility (NCTRI) Center for Reproductive Health After Disease (P50HD076188, T.K.W., M.M.L., Pilot FED), the UH3TR001207 (NCATS, NICHD, NIEHS, OWHR, NIH Common Fund, TKW), NIH 1K01DK099454-01 (R.N.S.), the Burroughs Wellcome Fund Career Award at the Scientific Interface (M.M.L.), and the NSF Graduate Research Fellowship Program (A.L.R., DGE-1324585). The University of Virginia Center for Research in Reproduction Ligand Assay and Analysis Core is supported by the Eunice Kennedy Shriver NICHD/NIH (NCTRI) Grant P50-HD28934. Imaging work was performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. This work made use of the EPIC facility of the NUANCE Center at Northwestern University, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF NNCI-1542205); the MRSEC program (NSF DMR-1121262) at the Materials Research Center; the International Institute for Nanotechnology (IIN); the Keck Foundation; and the State of Illinois, through the IIN.
Author information
Author notes
Monica M. Laronda & Alexandra L. Rutz
These authors contributed equally to this work.
Affiliations
Division of Reproductive Biology in Medicine, Department of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611, USA
Monica M. Laronda, Shuo Xiao, Kelly A. Whelan, Francesca E. Duncan & Teresa K. Woodruff
Center for Reproductive Science, Northwestern University, Chicago, Illinois 60611, USA
Monica M. Laronda, Shuo Xiao, Kelly A. Whelan, Francesca E. Duncan & Teresa K. Woodruff
Oncofertility Consortium, Northwestern University, Chicago, Illinois 60611, USA
Monica M. Laronda, Shuo Xiao, Kelly A. Whelan, Francesca E. Duncan & Teresa K. Woodruff
Simpson Querrey Institute for BioNanotechnology, Northwestern University, Chicago, Illinois 60611, USA
Alexandra L. Rutz & Ramille N. Shah
Department of Biomedical Engineering, Northwestern University, Evanston, Illinois 60208, USA
Alexandra L. Rutz & Ramille N. Shah
Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, Kansas 66160, USA
Francesca E. Duncan
Northwestern University Atomic and Nanoscale Characterization Experimental Center, Northwestern University, Evanston, Illinois 60208, USA
Eric W. Roth
Department of Materials Science and Engineering, Northwestern University, Evanston, Illinois 60208, USA
Ramille N. Shah
Department of Surgery, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611, USA
Ramille N. Shah
Contributions
M.M.L., A.L.R., T.K.W. and R.N.S. wrote this manuscript. T.K.W. envisioned the bioprosthetic ovary and A.L.R. and R.N.S. designed the 3D printing scaffolds. A.L.R. conceptualized and developed the ink and performed printing and material analyses. T.K.W., M.M.L. and K.A.W. designed the in vitro and in vivo ovarian follicle experiments. M.M.L. and K.A.W. performed all mouse experiments, including follicle culture and surgeries, and histological analysis of follicle culture and surgical tissue sections. F.E.D. determined the appropriate stroma cell marker for the ovary and analysed those histological samples. S.X. performed the MII egg staining. A.L.R. designed 3D analyses of scaffold–follicle interactions and performed immunostaining and confocal imaging of follicles seeded within 3D printed scaffolds. E.W.R. performed SEM imaging. M.M.L., A.L.R., S.X., T.K.W. and R.N.S. contributed to experimental design and interpretation.
Competing interests
M.M.L., A.L.R., R.N.S. and T.K.W. have filed an international Patent Application #PCT/US16/15398 titled in 2016. The remaining authors declare no competing financial interests.
Corresponding author
Correspondence to Ramille N. Shah.
