April 2011
Volume 52, Issue 14
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ARVO Annual Meeting Abstract  |   April 2011
Stem Cell Donor Limbus: Preoperative Assessment and Wound Healing Evaluation by in vivo Confocal Microscopy
Author Affiliations & Notes
  • Beatriz E. Ramirez
    Ocular Surface Group-IOBA, University of Valladolid, Valladolid, Spain
  • Dario A. Victoria
    Ocular Surface Group-IOBA, University of Valladolid, Valladolid, Spain
  • Denise Hileeto
    Ocular Surface Group-IOBA, University of Valladolid, Valladolid, Spain
  • M. Eugenia Mateo
    Ocular Surface Group-IOBA, University of Valladolid, Valladolid, Spain
  • Jose M. Herreras
    Ocular Surface Group-IOBA, University of Valladolid, Valladolid, Spain
  • Margarita Calonge
    Ocular Surface Group-IOBA, University of Valladolid, Valladolid, Spain
  • Footnotes
    Commercial Relationships  Beatriz E. Ramirez, None; Dario A. Victoria, None; Denise Hileeto, None; M. Eugenia Mateo, None; Jose M. Herreras, None; Margarita Calonge, None
  • Footnotes
    Support  CIBER-BBN, Carlos III Health Institute, Spain. Junta de Castilla y León SAN 1178/200. D. Victoria and B. Ramirez have scholarships from the Carolina Foundation, Ministry of Foreign Affairs, Spain.
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 5145. doi:
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      Beatriz E. Ramirez, Dario A. Victoria, Denise Hileeto, M. Eugenia Mateo, Jose M. Herreras, Margarita Calonge; Stem Cell Donor Limbus: Preoperative Assessment and Wound Healing Evaluation by in vivo Confocal Microscopy. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5145.

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Abstract

Purpose: : To analyze the preoperative features of the corneoescleral limbus by slit lamp examination (SLE) and in vivo confocal microscopy (CM) to identify the limbal stem cell (LSC) rich regions and to follow the wound healing process after harvesting limbal tissue for cultured LSC transplantation.

Methods: : Five patients with unilateral LSC deficiency underwent evaluation for autologous cultured LSC transplantation. SLE and CM at 12, 3, 6, and 9 o’clock limbal meridians identified the most appropriate site for biopsy to harvest the LSC. After biopsy, similar evaluations were done at 24 hours, 1 week, 1, 3, and 6 months to assess the healing process and the possible limbal restoration.

Results: : At initial assessment, SLE showed visible palisades of Vogt (POV) in only 3 patients, just at 12 and 6 o’clock meridians. By contrast, CM detected the POV in all 5 patients at 12 and 6 o’clock meridians; additionally, at 9 o’clock in 3 patients, and in 2 patients in all 4 meridians. POV were consistently better defined at 12 o’clock meridian, therefore, were selected for limbal biopsy. Mean depth of POV, was 80.4+19.8µm. Limbal biopsy (mean depth 136.8+19.12µm) extended significantly beyond the depth of the POV, harvesting LSC niche completely. The outgrowth rate of cultured biopsy-derived limbal explants was similar in all patients, regardless of the presence/absence of POV by SLE. At 1-week postsurgery, SLE showed complete re-epithelialization and peripheral scarring in all donor sites. In addition to these findings, CM detected scarring at the bottom of the donor limbus. Vascularization was observed at 1 month by CM and at 2 months by SLE. In months 3 and 6, CM showed fibrovascular tissue advancing from periphery and bottom to the center of the donor site.

Conclusions: : In vivo CM allowed to accurately identifying limbal features which cannot be visualized by routine clinical evaluation methods, therefore making it a useful technique to identify the appropriate biopsy site for autologous cultured LSC transplantation. Additionally, it allows confirmation of an adequate depth of the limbal biopsies, assuring complete removal of the LSC niche. There was no restoration of the donor limbal structures after 6 months of follow up.

Keywords: comparative anatomy • cornea: clinical science • wound healing 
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