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Establishment of a bi-layered tissue engineered conjunctiva using a 3D-printed melt electrowritten poly-(ε-caprolactone) scaffold.
Xie, Jiajun; Gao, Qi; Del Prado, Zelmira Nuñez; Venkateswaran, Nandini; Mousa, Hazem M; Salero, Enrique; Ye, Juan; De Juan-Pardo, Elena M; Sabater, Alfonso L; Perez, Victor L.
Afiliación
  • Xie J; Department of Ophthalmology, Ocular Surface Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
  • Gao Q; Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
  • Del Prado ZN; Department of Ophthalmology, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
  • Venkateswaran N; Department of Ophthalmology, Ocular Surface Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
  • Mousa HM; Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
  • Salero E; Department of Ophthalmology, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
  • Ye J; Department of Ophthalmology, Ocular Surface Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
  • De Juan-Pardo EM; Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
  • Sabater AL; Department of Ophthalmology, Ocular Surface Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
  • Perez VL; Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA, USA.
Int Ophthalmol ; 43(1): 215-232, 2023 Jan.
Article en En | MEDLINE | ID: mdl-35932420
ABSTRACT

PURPOSE:

To utilize melt electrowriting (MEW) technology using poly-(ε-caprolactone) (PCL) coupled with a 2-step co-culturing strategy for the development of a conjunctival bi-layer synthetic construct.

METHODS:

Melt electrowritten scaffolds using PCL were fabricated using an in-house-built MEW printer. Human conjunctival stromal cells (CjSCs) and epithelial cells (CjECs) were isolated from donor tissue. A 2-step co-culture method was done by first seeding the CjSCs and culturing for 4 weeks to establish a stromal layer, followed by CjECs and co-culturing for 2 more weeks. Cultured cells were each characterized by morphology and marker expression on immunofluorescence and qPCR. The produced construct was assessed for cellular proliferation using viability assays. The bi-layer morphology was assessed using scanning electron microscopy (SEM), confocal microscopy, and immunofluorescence imaging. The expression of extracellular matrix components and TGF-b was evaluated using qPCR.

RESULTS:

CjSCs were spindle-shaped and vimentin + while CjECs were polygonal and CK13 + . CjSCs showed consistent proliferation and optimal adherence with the scaffold at the 4-week culture mark. A 2-layered construct consisting of a CjSC-composed stromal layer and a CjEC-composed epithelial layer was appreciated on confocal microscopy, SEM, and immunofluorescence. CjSCs secreted collagens (types I, V, VI) but at differing amounts from natural tissue while TGF-b production was comparable.

CONCLUSION:

The 3D-printed melt electrowritten PCL scaffold paired with the 2-step co-culturing conditions of the scaffold allowed for the first approximation of a bi-layered stromal and epithelial reconstruction of the conjunctiva that can potentially improve the therapeutic arsenal in ocular surface reconstruction.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Poliésteres / Andamios del Tejido Límite: Humans Idioma: En Revista: Int Ophthalmol Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Poliésteres / Andamios del Tejido Límite: Humans Idioma: En Revista: Int Ophthalmol Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos