Designing and fabrication of a bacterial nanocellulose film induced with corneal mimicking topography as a substrate for corneal wound cell therapy
Seyed Hashem Daryabari1 , Seyedeh Mona Mousavi Esfahani2 *, Sajad Mansourian3 , Mazaher Gholipourmalekabadi4
- Baghiatollah University of Medical Sciences, Tehran, Islamic Republic of Iran
- Department of Medical Biotechnology, Cellular and Molecular Research Center, Faculty of Allied Medicine, Tehran University of Medical Sciences, Tehran, Iran
- Department of Plastic and Reconstructive Surgery, Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran
- Department of Medical Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran
Abstract: In 1997, it was proposed that a small amount of limbal tissue containing limbal epithelial stem
cells (LESC) could be cultured, expanded, and then transplanted into patients with LESC
deficiencies. However, challenges in limbal epithelial stem cell culture, cell sheet preparation,
and delivery to corneal defects remain. Histological examinations have shown that the
surface and area where LESC proliferate and differentiate into epithelial cells are unique. The
limbal epithelial ring has a distinct topography, whereas differentiated epithelial cells are located
on a flat surface
Methods: In this study, we prepare bacterial cellulose from Acetobacter xylinum bacteria by culturing them
in optimized media. We induce a cornea-mimicking topography through lithography or other
techniques, which is then printed onto the bacterial cellulose following an optimized protocol.
We perform various morphological and biomechanical evaluations of the topography-induced
bacterial cellulose (Topo-BC) using tests such as scanning electron microscopy (SEM), tensile
and suture retention tests, swelling assessments, atomic force microscopy (AFM), biodegradation
analysis, and observations of morphological changes during biodegradation and transparency.
Results: We then examine LESC behaviors on Topo-BC in vitro to determine its potential as a carrier for
maintaining stemness, promoting proliferation, and facilitating differentiation of LESCs. For this
purpose, MTT assays are conducted at 1, 3, and 7 days of culture on standard plates, bacterial
cellulose (BC), and Topo-BC. LESC adhesion and distribution after 3 and 7 days of culture on
the substrates are assessed using SEM and DAPI staining. The effects of topography on the
expression of stemness, limbal, and epithelial genes are evaluated through reverse transcription
polymerase chain reaction (RT-PCR).Furthermore, Topo-BC seeded with LESC is applied to corneal defects in rabbits that have
experienced corneal failure due to chemical burns (HCl models), in order to histologically and
molecularly assess the healing of corneal wounds.
Conclusion: At the conclusion of this study, we anticipate
that our topographical bacterial nanofibrous cellulose will provide a suitable substrate for LESC
culture, sheet formation, and effective LESC delivery in vivo. This construct may be used
independently or seeded with LESCs as a stem cell delivery carrier for addressing various
corneal defects.
