Skip to main content
SpringerLink
Log in

Langzeitergebnisse zur autologen Transplantation von ex vivo kultiviertem Limbusepithel bei limbaler Stammzellinsuffizienz

Long-term results of autologous transplantation of limbal epithelium cultivated ex vivo for limbal stem cell deficiency

  • Originalien
  • Published:
Der Ophthalmologe Aims and scope Submit manuscript

Zusammenfassung

Hintergrund

In dieser Studie sollen die Langzeitergebnisse der Ex-vivo-Expansion von autologem Limbusepithel zur Oberflächenrekonstruktion bei limbaler Stammzellinsuffizienz (LSZI) untersucht werden.

Patienten und Methoden

Bei 61 Augen von 57 Patienten (m:w 46:11) mit einer LSZI erfolgte eine autologe Transplantation von kultiviertem Limbusepithel. Folgende Ätiologien einer LSZI wurden behandelt: Verätzungen (n = 32), Verbrennungen (n = 2), Mitomycin C (MMC)/INF-α-induziert bei Zustand nach Tumorexzision (n = 9), rezidivierende, bzw. primäre, große Pterygien (n = 12), schwere infektiöse Keratitiden (n = 3), Zustand nach Perforationstrauma, Epidermolysis bullosa, kontaktlinsenassoziierte Keratopathie (jeweils n = 1). Nur Augen mit einem Nachbeobachtungszeitraum von mindestens 12 Monaten wurden in die Studie eingeschlossen.

Ergebnisse

Der Nachbeobachtungszeitraum betrug im Mittel 50,8 ± 32,7 Monate (Intervall: 12,5 bis 142 Monate). Ein stabiles okuläres Oberflächenepithel wurde bei 46 Augen (75,4 %) erzielt. Die Sehschärfe stieg bei 40 Augen (65,6 %) an, war stabil bei 12 Augen (19,7 %) und verschlechterte sich bei 9 Augen (14,8 %). Der durchschnittliche logMAR-Visus verbesserte sich signifikant (p < 0,0001) von präoperativ 1,4 ± 0,91 zu postoperativ 0,8 ± 0,67.

Diskussion

Transplantation von ex vivo expandiertem Limbusepithel führt zur Stabilisierung des kornealen Oberflächenepithels und zu einem signifikanten Visusgewinn. Autologe Verfahren haben auch nach Langzeitbeobachtung eine sehr gute Prognose.

Abstract

Objective

This study reports the long-term clinical outcome of autologous limbal epithelial cells cultivated ex vivo on intact amniotic membranes (AM) for ocular surface reconstruction in limbal stem cell deficiency (LSCD).

Patients and methods

A total of 61 eyes from 57 patients (46 males and 11 females) with LSCD were treated by transplantation of autologous limbal epithelial cells cultivated on intact AM. The etiology of the LSCD was chemical and thermal burns (n = 34), recurrent or primary large-sized pterygium (n = 12), mitomycin C and tumor excision-induced LSCD (n = 9), severe infectious keratitis (n = 3), perforating injury, epidermolysis bullosa and contact lens-associated keratopathy (each n = 1). Only eyes with a follow-up time of at least 12 months were included in the analysis. The main outcome end points were restoration of ocular surface integrity and improvement of visual acuity (VA).

Results

The mean follow-up time was 50.8 ± 32.7 months. An entirely stable corneal surface was reconstructed in 46 (75.4 %) eyes. Visual acuity significantly increased in 40 (65.6 %) eyes, was stable in 12 (19.7 %) eyes and decreased in 9 eyes (14.8 %). The mean visual acuity significantly increased (p < 0.0001) from 1.4 ± 0.91 LogMAR preoperatively to 0.8 ± 0.67 LogMAR postoperatively.

Conclusion

Transplantation of limbal epithelium cultivated ex vivo on intact AM leads to restoration of a stable corneal surface and resulted in a significant increase of visual acuity in most cases of LSCD. Autologous transplantation of cultivated limbal epithelium showed an excellent prognosis and outcome after long-term follow-up.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Abb. 1
Abb. 2
Abb. 3

Literatur

  1. Akle C, McColl I, Dean M, Adinolfi M, Brown S, Fensom AH, Marsh J, Welsh K (1985) Transplantation of amniotic epithelial membranes in patients with mucopolysaccharidoses. Exp Clin Immunogenet 2:43–48

    CAS  PubMed  Google Scholar 

  2. Akle CA, Adinolfi M, Welsh KI, Leibowitz S, McColl I (1981) Immunogenicity of human amniotic epithelial cells after transplantation into volunteers. Lancet 2:1003–1005

    Article  CAS  PubMed  Google Scholar 

  3. Baharvand H, Heidari M, Ebrahimi M, Valadbeigi T, Salekdeh GH (2007) Proteomic analysis of epithelium-denuded human amniotic membrane as a limbal stem cell niche. Mol Vis 13:1711–1721

    CAS  PubMed  Google Scholar 

  4. Balasubramanian S, Jasty S, Sitalakshmi G, Madhavan HN, Krishnakumar S (2008) Influence of feeder layer on the expression of stem cell markers in cultured limbal corneal epithelial cells. Indian J Med Res 128:616–622

    CAS  PubMed  Google Scholar 

  5. Basti S, Mathur U (1999) Unusual intermediate-term outcome in three cases of limbal autograft transplantation. Ophthalmology 106:958–963

    Article  CAS  PubMed  Google Scholar 

  6. Basu S, Ali H, Sangwan VS (2012) Clinical outcomes of repeat autologous cultivated limbal epithelial transplantation for ocular surface burns. Am J Ophthalmol 153:643–650, 650 e641–642

    Article  PubMed  Google Scholar 

  7. Dua HS, Azuara-Blanco A (2000) Autologous limbal transplantation in patients with unilateral corneal stem cell deficiency. Br J Ophthalmol 84:273–278

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Dua HS, Forrester JV (1990) The corneoscleral limbus in human corneal epithelial wound healing. Am J Ophthalmol 110:646–656

    Article  CAS  PubMed  Google Scholar 

  9. Egbert PR, Lauber S, Maurice DM (1977) A simple conjunctival biopsy. Am J Ophthal 84:798–801

    Article  CAS  PubMed  Google Scholar 

  10. Feng Y, Borrelli M, Reichl S, Schrader S, Geerling G (2014) Review of alternative carrier materials for ocular surface reconstruction. Current eye research 39:541–552

  11. Fernandes M, Sangwan VS, Rao SK, Basti S, Sridhar MS, Bansal AK, Dua HS (2004) Limbal stem cell transplantation. Indian J Ophthalmol 52:5–22

    PubMed  Google Scholar 

  12. Galindo EE, Theiss C, Pauklin M, Wettey FR, Steuhl KP, Meller D (2009) Correlation of cell cycle kinetics with p63 expression in human limbal epithelial cells expanded on intact human amniotic membrane. Ophthalmic Res 41:83–90

    Article  PubMed  Google Scholar 

  13. Grueterich M (2012) Klinik der Limbusstammzellinsuffizienz. Ophthalmologe 109:850–856

    Article  CAS  PubMed  Google Scholar 

  14. Grueterich M, Espana E, Tseng SC (2002) Connexin 43 expression and proliferation of human limbal epithelium on intact and denuded amniotic membrane. Invest Ophthalmol Vis Sci 43:63–71

    PubMed  Google Scholar 

  15. Grueterich M, Espana EM, Tseng SC (2003) Modulation of keratin and connexin expression in limbal epithelium expanded on denuded amniotic membrane with and without a 3T3 fibroblast feeder layer. Invest Ophthalmol Vis Sci 44:4230–4236

    Article  PubMed  Google Scholar 

  16. Ijiri S, Kobayashi A, Sugiyama K, Tseng SC (2007) Evaluation of visual acuity and color vision in normal human eyes with a sutureless temporary amniotic membrane patch. Am J Ophthalmol 144:938–942

    Article  PubMed  Google Scholar 

  17. Kenyon KR, Tseng SC (1989) Limbal autograft transplantation for ocular surface disorders. Ophthalmology 96:709–722 (discussion 722–703)

    Article  CAS  PubMed  Google Scholar 

  18. Kim JC, Tseng SC (1995) The effects on inhibition of corneal neovascularization after human amniotic membrane transplantation in severely damaged rabbit corneas. Korean J Ophthalmol 9:32–46

    Article  CAS  PubMed  Google Scholar 

  19. Kinoshita S, Koizumi N, Nakamura T (2004) Transplantable cultivated mucosal epithelial sheet for ocular surface reconstruction. Exp Eye Res 78:483–491

    Article  CAS  PubMed  Google Scholar 

  20. Koizumi N, Fullwood NJ, Bairaktaris G, Inatomi T, Kinoshita S, Quantock AJ (2000) Cultivation of corneal epithelial cells on intact and denuded human amniotic membrane. Invest Ophthalmol Vis Sci 41:2506–2513

    CAS  PubMed  Google Scholar 

  21. Koizumi N, Inatomi T, Suzuki T, Sotozono C, Kinoshita S (2001) Cultivated corneal epithelial stem cell transplantation in ocular surface disorders. Ophthalmology 108:1569–1574

    Article  CAS  PubMed  Google Scholar 

  22. Koizumi N, Rigby H, Fullwood NJ, Kawasaki S, Tanioka H, Koizumi K, Kociok N, Joussen AM, Kinoshita S (2007) Comparison of intact and denuded amniotic membrane as a substrate for cell-suspension culture of human limbal epithelial cells. Graefes Arch Clin Exp Ophthalmol 245:123–134

    Article  PubMed  Google Scholar 

  23. Kruse FE (1994) Stem cells and corneal epithelial regeneration. Eye (Lond) 8(Pt 2):170–183

    Article  Google Scholar 

  24. Kruse FE, Cursiefen C (2008) Surgery of the cornea: corneal, limbal stem cell and amniotic membrane transplantation. Dev Ophthalmol 41:159–170

    Article  CAS  PubMed  Google Scholar 

  25. Lange C, Feltgen N, Junker B, Schulze-Bonsel K, Bach M (2009) Resolving the clinical acuity categories „hand motion“ and „counting fingers“ using the Freiburg Visual Acuity Test (FrACT). Graefes Arch Clin Exp Ophthalmol 247:137–142

    Article  CAS  PubMed  Google Scholar 

  26. Lee SH, Tseng SC (1997) Amniotic membrane transplantation for persistent epithelial defects with ulceration. Am J Ophthalmol 123:303–312

    Article  CAS  PubMed  Google Scholar 

  27. Lindberg K, Brown ME, Chaves HV, Kenyon KR, Rheinwald JG (1993) In vitro propagation of human ocular surface epithelial cells for transplantation. Invest Ophthalmol Vis Sci 34:2672–2679

    CAS  PubMed  Google Scholar 

  28. Maru DS, Bruce RD, Basu S, Altice FL (2008) Clinical outcomes of hepatitis C treatment in a prison setting: feasibility and effectiveness for challenging treatment populations. Clin Infect Dis 47:952–961

    Article  PubMed  Google Scholar 

  29. Meller D, Fuchsluger T, Pauklin M, Steuhl KP (2009) Ocular surface reconstruction in graft-versus-host disease with HLA-identical living-related allogeneic cultivated limbal epithelium after hematopoietic stem cell transplantation from the same donor. Cornea 28:233–236

    Article  PubMed  Google Scholar 

  30. Meller D, Kruse F (2001) Ex-vivo Expansion kornealer Stammzellen Experimentelle Grundlagen und erste klinische Ergebnisse. Ophthalmologe 98:811–817

    Article  CAS  PubMed  Google Scholar 

  31. Meller D, Pires RT, Tseng SC (2002) Ex vivo preservation and expansion of human limbal epithelial stem cells on amniotic membrane cultures. Br J Ophthalmol 86:463–471

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Meller D, Tseng SC (1999) Conjunctival epithelial cell differentiation on amniotic membrane. Invest Ophthalmol Vis Sci 40:878–886

    CAS  PubMed  Google Scholar 

  33. Miri A, Said DG, Dua HS (2011) Donor site complications in autolimbal and living-related allolimbal transplantation. Ophthalmology 118:1265–1271

    PubMed  Google Scholar 

  34. Nakamura T, Inatomi T, Sotozono C, Ang LP, Koizumi N, Yokoi N, Kinoshita S (2006) Transplantation of autologous serum-derived cultivated corneal epithelial equivalents for the treatment of severe ocular surface disease. Ophthalmology 113:1765–1772

    Article  PubMed  Google Scholar 

  35. Nakamura T, Inatomi T, Sotozono C, Koizumi N, Kinoshita S (2004) Successful primary culture and autologous transplantation of corneal limbal epithelial cells from minimal biopsy for unilateral severe ocular surface disease. Acta Ophthalmol Scand 82:468–471

    Article  PubMed  Google Scholar 

  36. Nakamura T, Koizumi N, Tsuzuki M, Inoki K, Sano Y, Sotozono C, Kinoshita S (2003) Successful regrafting of cultivated corneal epithelium using amniotic membrane as a carrier in severe ocular surface disease. Cornea 22:70–71

    Article  PubMed  Google Scholar 

  37. Pauklin M, Fuchsluger TA, Westekemper H, Steuhl KP, Meller D (2010) Midterm results of cultivated autologous and allogeneic limbal epithelial transplantation in limbal stem cell deficiency. Dev Ophthalmol 45:57–70

    Article  PubMed  Google Scholar 

  38. Pauklin M, Steuhl KP, Meller D (2009) Characterization of the corneal surface in limbal stem cell deficiency and after transplantation of cultivated limbal epithelium. Ophthalmology 116:1048–1056

    Article  PubMed  Google Scholar 

  39. Pellegrini G, Traverso CE, Franzi AT, Zingirian M, Cancedda R, De Luca M (1997) Long-term restoration of damaged corneal surfaces with autologous cultivated corneal epithelium. Lancet 349:990–993

    Article  CAS  PubMed  Google Scholar 

  40. Prabhasawat P, Barton K, Burkett G, Tseng SC (1997) Comparison of conjunctival autografts, amniotic membrane grafts, and primary closure for pterygium excision. Ophthalmology 104:974–985

    Article  CAS  PubMed  Google Scholar 

  41. Puangsricharern V, Tseng SC (1995) Cytologic evidence of corneal diseases with limbal stem cell deficiency. Ophthalmology 102:1476–1485

    Article  CAS  PubMed  Google Scholar 

  42. Rama P, Bonini S, Lambiase A, Golisano O, Paterna P, De Luca M, Pellegrini G (2001) Autologous fibrin-cultured limbal stem cells permanently restore the corneal surface of patients with total limbal stem cell deficiency. Transplantation 72:1478–1485

    Article  CAS  PubMed  Google Scholar 

  43. Rama P, Matuska S, Paganoni G, Spinelli A, De Luca M, Pellegrini G (2010) Limbal stem-cell therapy and long-term corneal regeneration. N Engl J Med 363:147–155

    Article  CAS  PubMed  Google Scholar 

  44. Rendal-Vazquez ME, San-Luis-Verdes A, Yebra-Pimentel-Vilar MT, Lopez-Rodriguez I, Domenech-Garcia N, Andion-Nunez C, Blanco-Garcia F (2012) Culture of limbal stem cells on human amniotic membrane. Cell Tissue Bank 13:513–519

    Article  CAS  PubMed  Google Scholar 

  45. Robson MC, Krizek TJ, Koss N, Samburg JL (1973) Amniotic membranes as a temporary wound dressing. Surg Gynecol Obstet 136:904–906

    CAS  PubMed  Google Scholar 

  46. Sangwan VS, Basu S, Vemuganti GK, Sejpal K, Subramaniam SV, Bandyopadhyay S, Krishnaiah S, Gaddipati S, Tiwari S, Balasubramanian D (2011) Clinical outcomes of xeno-free autologous cultivated limbal epithelial transplantation: a 10-year study. Br J Ophthalmol 95:1525–1529

    Article  PubMed  Google Scholar 

  47. Sangwan VS, Matalia HP, Vemuganti GK, Fatima A, Ifthekar G, Singh S, Nutheti R, Rao GN (2006) Clinical outcome of autologous cultivated limbal epithelium transplantation. Indian J Ophthalmol 54:29yth

    Article  Google Scholar 

  48. Schwab IR (1999) Cultured corneal epithelia for ocular surface disease. Trans Am Ophthalmol Soc 97:891–986

    CAS  PubMed  PubMed Central  Google Scholar 

  49. Shortt AJ, Secker GA, Notara MD, Limb GA, Khaw PT, Tuft SJ, Daniels JT (2007) Transplantation of ex vivo cultured limbal epithelial stem cells: a review of techniques and clinical results. Surv Ophthalmol 52:483–502

    Article  PubMed  Google Scholar 

  50. Shortt AJ, Secker GA, Rajan MS, Meligonis G, Dart JK, Tuft SJ, Daniels JT (2008) Ex vivo expansion and transplantation of limbal epithelial stem cells. Ophthalmology 115:1989–1997

    Article  PubMed  Google Scholar 

  51. Song E, Yang W, Cui ZH, Dong Y, Sui DM, Guan XK, Ma YL (2005) Transplantation of human limbal cells cultivated on amniotic membrane for reconstruction of rat corneal epithelium after alkaline burn. Chin Med J 118:927–935

    CAS  PubMed  Google Scholar 

  52. Sorsby A, Haythorne J, Reed H (1947) Further experience with amniotic membrane grafts in caustic burns of the eye. Br J Ophthalmol 31:409–418

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Tan DT, Ficker LA, Buckley RJ (1996) Limbal transplantation. Ophthalmology 103:29–36

    Article  CAS  PubMed  Google Scholar 

  54. Thoft RA, Friend J (1983) The X, Y, Z hypothesis of corneal epithelial maintenance. Invest Ophthalmol Vis Sci 24:1442–1443

    CAS  PubMed  Google Scholar 

  55. Tsai RJ, Li LM, Chen JK (2000) Reconstruction of damaged corneas by transplantation of autologous limbal epithelial cells. N Engl J Med 343:86–93

    Article  CAS  PubMed  Google Scholar 

  56. Tseng SC (1989) Concept and application of limbal stem cells. Eye (Lond) 3(Pt 2):141–157

    Article  Google Scholar 

  57. Tseng SC, Prabhasawat P, Lee SH (1997) Amniotic membrane transplantation for conjunctival surface reconstruction. Am J Ophthalmol 124:765–774

    Article  CAS  PubMed  Google Scholar 

  58. Tsubota K, Toda I, Saito H, Shinozaki N, Shimazaki J (1995) Reconstruction of the corneal epithelium by limbal allograft transplantation for severe ocular surface disorders. Ophthalmology 102:1486–1496

    Article  CAS  PubMed  Google Scholar 

  59. Wan P, Wang X, Ma P, Gao N, Ge J, Mou Y, Wang Z (2011) Cell delivery with fixed amniotic membrane reconstructs corneal epithelium in rabbits with limbal stem cell deficiency. Invest Ophthalmol Vis Sci 52:724–730

    Article  CAS  PubMed  Google Scholar 

  60. Wang J, Li W, Min J, Ou Q, Chen J, Song E (2004) Intrasplenic transplantation of allogeneic hepatocytes modified by BCL-2 gene protects rats from acute liver failure. Transplant Proc 36:2924–2926

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Zentrum für Augenheilkunde, Klinik für Erkrankungen des vorderen Augenabschnitts, Universität Duisburg-Essen, 45147, Essen, Deutschland

    S.L. Scholz, H. Thomasen, K. Hestermann, D. Dekowski, K.-P. Steuhl &  D. Meller FEBO

Authors
  1. S.L. Scholz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H. Thomasen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. Hestermann
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D. Dekowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. K.-P. Steuhl
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. D. Meller FEBO
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to D. Meller FEBO.

Ethics declarations

Interessenkonflikt

S.L. Scholz, H. Thomasen, K. Hestermann, D. Dekowski, K.-P. Steuhl und D. Meller geben an, dass kein Interessenkonflikt besteht. Unterstützt mit Drittmitteln der Deutschen Forschungsgemeinschaft (DFG, Bonn, ME 1623/5-1).

Alle im vorliegenden Manuskript beschriebenen Untersuchungen am Menschen wurden mit Zustimmung der zuständigen Ethik-Kommission, im Einklang mit nationalem Recht sowie gemäß der Deklaration von Helsinki von 1975 (in der aktuellen, überarbeiteten Fassung) durchgeführt. Von allen beteiligten Patienten liegt eine Einverständniserklärung vor.

Additional information

Teile der Ergebnisse dieser Arbeit wurden auf dem DOG-Kongress 2013 in Berlin präsentiert.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Scholz, S., Thomasen, H., Hestermann, K. et al. Langzeitergebnisse zur autologen Transplantation von ex vivo kultiviertem Limbusepithel bei limbaler Stammzellinsuffizienz. Ophthalmologe 113, 321–329 (2016). https://doi.org/10.1007/s00347-015-0110-y

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00347-015-0110-y

Schlüsselwörter

Keywords

Search

Navigation