Nanofiber-reinforced decellularized amniotic membrane improves limbal stem cell transplantation in a rabbit model of corneal epithelial defect

Zhengbing Zhou; Da Long; Chih-Chien Hsu; Huanhuan Liu; Long Chen; Benjamin Slavin; Hui Lin; Xiaowei Li; Juyu Tang; Samuel Yiu; Sami Tuffaha; Hai-Quan Mao

doi:10.1016/j.actbio.2019.08.027

Nanofiber-reinforced decellularized amniotic membrane improves limbal stem cell transplantation in a rabbit model of corneal epithelial defect

Acta Biomater. 2019 Oct 1:97:310-320. doi: 10.1016/j.actbio.2019.08.027. Epub 2019 Aug 19.

Authors

Zhengbing Zhou¹, Da Long², Chih-Chien Hsu³, Huanhuan Liu⁴, Long Chen⁵, Benjamin Slavin⁶, Hui Lin⁷, Xiaowei Li⁸, Juyu Tang⁹, Samuel Yiu⁷, Sami Tuffaha¹⁰, Hai-Quan Mao¹¹

Affiliations

¹ Department of Hand and Microsurgery, Xiangya Hospital of Central South University, Changsha, Hunan Province 410008, PR China; Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA.
² Department of Ophthalmology, The First Hospital of Hunan University of Chinese Traditional Medicine, Changsha, Hunan Province 410007, PR China; Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
³ Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Department of Ophthalmology, Taipei Veterans General Hospital, National Yang-Ming University, No. 201, Shipai Rd. Sec. 2, Taipei, Taiwan.
⁴ Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
⁵ Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
⁶ Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
⁷ Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
⁸ Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Mary and Dick Holland Regenerative Medicine Program, Department of Neurological Sciences, University of Nebraska Medical Center, Omaha, NE 68198, USA.
⁹ Department of Hand and Microsurgery, Xiangya Hospital of Central South University, Changsha, Hunan Province 410008, PR China.
¹⁰ Department of Plastic and Reconstructive Surgery, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: stuffah1@jhmi.edu.
¹¹ Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA. Electronic address: hmao@jhu.edu.

PMID: 31437637
DOI: 10.1016/j.actbio.2019.08.027

Abstract

Human amniotic membrane (AM) offers unique advantages as a matrix to support the transplantation of limbal stem cells (LSCs) due to its inherent pro-regenerative and anti-inflammatory properties. However, the widespread use of AM in clinical treatments of ocular surface disorders is limited by its weak mechanical strength and fast degradation, and high cost associated with preserving freshly isolated AM. Here we constructed a composite membrane consisting of an electrospun bioabsorbable poly(ε-caprolactone) (PCL) nanofiber mesh to significantly improve the ultimate tensile strength, toughness, and suture retention strength by 4-10-fold in comparison with decellularized AM sheet. The composite membrane showed extended stability and conferred longer-lasting coverage on wounded cornea surface compared with dAM. The composite membrane maintained the pro-regenerative and immunomodulatory properties of dAM, promoted LSC survival, retention, and organization, improved re-epithelialization of the defect area, and reduced inflammation and neovascularization. This study demonstrates the translational potential of our composite membrane for stem cell-based treatment of ocular surface damage. STATEMENT OF SIGNIFICANCE: Human decellularized amniotic membrane (dAM) has been widely shown as a biodegradable and bioactive matrix for regenerative tissue repair. However, the weak mechanical property has limited its widespread use in the clinic. Here we constructed a composite membrane using a layer of electrospun poly(ε-caprolactone) (PCL) nanofiber mesh to reinforce the dAM sheet through covalent interfacial bonding, while retaining the unique bioactivity of dAM. In a rabbit model of limbal stem cell (LSC) deficiency induced by alkaline burn, we demonstrated the superior property of this PCL-dAM composite membrane for repairing damaged cornea through promoting LSC transplantation, improving re-epithelialization, and reducing inflammation and neovascularization. This new composite membrane offers great translational potential in supporting stem cell-based treatment of ocular surface damage.

Keywords: Amniotic membrane; Composite membrane; Electrospinning; Limbal stem cell deficiency; Nanofibers; Transplantation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amnion / chemistry*
Animals
Epithelium, Corneal* / injuries
Epithelium, Corneal* / metabolism
Epithelium, Corneal* / pathology
Extracellular Matrix / chemistry
Humans
Limbus Corneae / metabolism*
Limbus Corneae / pathology
Nanofibers / chemistry*
Rabbits
Stem Cell Transplantation*
Stem Cells / metabolism*
Stem Cells / pathology