Journal List > J Korean Ophthalmol Soc > v.49(7) > 1008024

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Wook-Yang, Young Lee, Gwang Park, and Il Yang: The Role of Endothelial Progenital Cells and Fibrin on Vascularization and Stability in Orbital Implant
Original Article

J Korean Ophthalmol Soc 2008;49(7):1135-1145.

Published online: 7 September 2008

DOI: https://doi.org/10.3341/jkos.2008.49.7.1135

The Role of Endothelial Progenital Cells and Fibrin on Vascularization and Stability in Orbital Implant

Jae Wook-Yang, M.D., Ph.D.1, , Ho Young Lee, M.D.1, Sae Gwang Park, M.D., Ph.D.2, Young Il Yang, M.D., Ph.D.3

Department of Ophthalmology, College of Medicine, Inje University, Pusan, Kora

Department of Microbiology, College of Medicine, Inje University, Pusan, Kora

Department of Pathology, College of Medicine, Inje University, Pusan, Kora

Address reprint requests to Jae-Wook Yang, M.D., Ph.D. Department of Ophthalmology, College of Medicine, Inje University of Medicine #633-165 Kekum-dong, Pusanjin-gu, Pusan 614-735, Korea Tel: 82-51-890-6016, Fax: 82-51-890-6329, E-mail: jwsyhyo@yahoo.co.kr

Received 10 April 2008       Accepted 20 November 7

Copyright © 2015 Korean Ophthalmological Society

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

The effects of endothelial progenitor cells (EPCs) and fibrin on fibrovascular growth into porous polyethylene orbital implants (Medpor® sheet) were investigated using stem cells.

Methods

EPCs were separated from human adipose fat tissue for culture. Fluorescence-activated cell sorting (FACS) was used to identify the phenotype and to analyze the purity of EPCs cultivated from human adipose tissue. Processed Medpor® sheets were inserted in each quadrant of the subcutaneous fat layer under the dorsal surface of 20 anesthetized athymic nude mice, using sterile methods. Medpor® sheets processed with endothelial progenitor cells and fibrin were inserted into the two top quadrants, a Medpor® sheet processed with fibrin was inserted in the lower right quadrant, and an unprocessed Medpor® sheet was inserted in the lower left quadrant of each mouse. The mice were sacrificed on the seventh day. The adhesiveness and blood vessel formation were quantified by weight and the number of blood cells within the Medpor® sheets. Hematoxylin and eosin (H&E) and toluidine blue stains were used to analyze fibrovascular and cell growth within the Medpor® sheets.

Results

The sheets processed with EPCs and fibrin were heavier and contained more white and red blood cells (p<0.001) than the other sheets. The sheets processed with fibrin alone were heavier (p<0.01) and contained more blood cells (p<0.001) than the unprocessed sheets. The degree of vessel formation and tissue adhesiveness was greatest in the group of Medpor® sheets processed with EPCs and fibrin. The sheets processed with fibrin only had greater tissue adhesiveness and fibrovascular proliferation than the unprocessed Medpor® sheets.

Conclusions

Endothelial progenitor cells and fibrin applied to Medpor® sheets improve fibrovascular proliferation and tissue adhesiveness. When both are applied together, a synergistic effect is seen.

Keywords: Endothelial progenital cells, Fibrin, Fibrovascularization, Medpor® sheet

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Figure 1.
Thin Medpor® sheets (Porex Surgical Inc., CA, U.S.A.) were cut to 8 mm in diameter and were used after sterilization.
jkos-49-1135f1.tif
Figure 2.
Adipose stem cells (ASCs) and endothelial progenitor cells (EPCs). (A) Light microscope image of cultured ASCs; (B) Nile red O staining of ASCs(DAPI nuclear counter staining); (C) Phase contrast microscope image of cultured EPCs.
jkos-49-1135f2.tif
Figure 3.
Flow cytometry analysis of isolated and cultured EPCs.
jkos-49-1135f3.tif
Figure 4.
Confocal microscope image of EPCs. (A) FITC-labeled von-Willebrand factor staining; (B) PI staining; (C) overlapping (merge) image of A and B; (D) Phase contrast image of EPCs in 3-dimensional matrix culture; (E) FITC-labeled CD31; (F) PI & CD31 staining.
jkos-49-1135f4.tif
Figure 5.
Medpor® sheet and postimplanted mouse. (A) Medpor® sheet treated with EPCs and fibrin; (B) postoperated mouse 1) RUQ (Right upper quadrant) & LUQ(Left upper quadrant) were implanted with Medpor® sheet treated with EPCs and fibrin. 2) LLQ (Left lower quadrant) was with untreated Medpor® sheet. 3) RLQ (Right lower quadrant) was with Medpor® sheet treated with fibrin.
jkos-49-1135f5.tif
Figure 6.
Gross examination of Medpor® sheet; (A) Medpor® sheet treated with EPCs and fibrin; (B) Medpor® sheet treated with fibrin; (C) Untreated Medpor® sheet.
jkos-49-1135f6.tif
Figure 7.
Weight of Medpor® sheet. * p<0.001, ** p<0.01.
jkos-49-1135f7.tif
Figure 8.
Implanted Medpor® sheet and it's toluidine blue staining; (A) Gross finding of implanted Medpor® sheet; (B) Medpor® sheet treated with EPCs and fibrin; (C) Medpor® sheet treated with fibrin; (D) Untreated Medpor® sheet.
jkos-49-1135f8.tif
Figure 9.
H&E staining of external wall of implanted Medpor®; (A) Medpor® sheet treated with EPCs and fibrin; (B) Medpor® sheet treated with fibrin; (C) Untreated Medpor® sheet.
jkos-49-1135f9.tif
Figure 10.
H&E staining of internal spaces of implanted Medpor®. (A) Medpor® sheet treated with EPCs and fibrin; (B) Medpor® sheet treated with fibrin; (C) Untreated Medpor® sheet.
jkos-49-1135f10.tif
Table 1.
Quantitation of blood cells in Medpor®(mean±standard deviation) sheet
WBC RBC
(x103 cells/µl) (x106 cells/µl)
EPCs+Fibrin† 0.23±0.02* 0.1±0.01
Fibrin‡ 0.13±0.01* 0.1±0.01
Medpor® only§ 0.02±0.01 0.0±0.01

* p<0.001;

‡ Medpor® sheet treated with fibrin; sheet.

† Medpor® sheet treated with EPCs and fibrin;

§ Untreated Medpor®

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