Zusammenfassung
Zielsetzung: Die Möglichkeiten einer zeitversetzten autologen dermoepidermalen Transplantationsstrategie als Ausgangspunkt für eine nicht invasive Rekonstruktion von ausgedehnten, akuten Weichteildefekten unterschiedlicher Genese sollen demonstriert und kritisch diskutiert werden.
Methoden: Nach Indikationsstellung wird bei Patienten eine Hautbiopsie zur Zellkultivierung entnommen. Die auf einer dreidimensionalen, biokompatiblen, aus Hyaluronsäureester bestehenden Matrix gezüchteten autologen Fibroblasten werden als „Neo-Dermis” auf die konditionierten Weichteildefekte übertragen. Nach Inkorporation der dermalen Substitute findet die Transplantation der subkonfluenten, hoch proliferativen autologen Keratinozyten auf einem Trägersystem, bestehend aus einer laserperforierten Hyaluronsäureestermembran, statt. Zehn Tage danach wird eine autologe, 0,2 mm dünne Mesh-graft-Transplantation (1 : 6) als definitve biomechanisch stabile Defektdeckung durchgeführt.
Ergebnisse: Die Übertragung von in vitro kultivierten autologen Fibroblasten resultiert in einem gut vaskularisierten dermalen Gewebsersatz. Nach Keratinozytentransfer ist eine Epithelialisierung nachweisbar. Ein definitiver, ästhetisch ansprechender Wundverschluss mit normoelastischen Gewebseigenschaften wird nach dünner Mesh-graft-Transplantation erreicht.
Schlussfolgerungen: Vorlä ufige Ergebnisse mit beschriebener Methode erscheinen viel versprechend. Klinischer Verlauf sowie histologische und immunhistochemische Resultate nach Behandlung von fünf ausgedehnten Weichteildefekten werden diskutiert. Wie in allen Bereichen des „tissue engineering” sind klinische Langzeitstudien sowie Kosten-Nutzen-Analysen erforderlich.
Abstract
Aim: The opportunities of autologous dermal and epidermal grafting as starting point for non-invasive reconstruction of extensive soft tissue defects will be demonstrated and discussed.
Methods: Skin biopsies for cell cultivation were taken from patients with extensive acute soft tissue defects of different origin. Cultured autologous fibroblasts grown on threedimensional biocompatible scaffolds made up of benzylester of hyaluronan were transplanted as “neo-dermis” on debrided and conditioned wound sites. After incorporation of the dermal equivalents grafting of subconfluent proliferative keratinocytes on hyaluronan based laserperforated membranes was performed. Ten days later a 0.2 mm thin, 1 : 6 meshed autograft to create definite biomechanical stability was overlaid.
Results: Grafting of in vitro cultured autologous fibroblasts revealed a good vascularized dermal tissue substitute. After keratinocyte-transfer formation of thin epithelium was visible. Final closure of the defects with aesthetic and normo-elastic tissue properties was achieved after thin mesh-grafting.
Conclusions: Preliminary results seem to be very promising. Clinical follow-up as well as histological and immunohistochemical outcome in the treatment of five extensive soft-tissue defects are discussed. As in all fields of tissue engineering, long-tem studies and cost-benefit analyses are required.
Schlüsselwörter
Weichteilrekonstruktion - Hyaluronsäure - Hautersatz
Key words
Soft-tissue reconstruction - hyaluronan - skin substitutes
Literatur
1
Abatangelo G, Martelli M, Vecchia R P.
Healing of hyaluronic acid-enriched wounds: Histological observations.
J Surg Res.
1983;
35
410-416
2
Chen W YJ, Abatangelo G.
Functions of hyaluronan in wound repair.
Wound Rep Reg.
1999;
7
79-89
3
Cooper M L, Hansbrough J F, Spielvogel R L, Cohen R, Bartel R, Noughton G.
In vivo optimization of a living dermal substitute employing cultured human fibroblasts on a biodegradable polyglycolic acid or polyglactin mesh.
Biomaterials.
1991;
12
243-249
4
Coulomb B, Lebreton C, Dubertret L.
Influence of human dermal fibroblasts on epidermalization.
J Invest Dermatol.
1989;
92
122-125
5
Cuono C, Langdon R, McGuire J.
Use of cultured epidermal autografts and dermal allografts as skin replacement after burn injury.
Lancet.
1986;
1
1123-1124
6
Donnersmarck von G H, Mülbauer W, Höfter E, Hartinger A.
Die Verwendung von Keratinozytenkulturen in der Schwerbrandverletztenbehandlung: bisherige Erfahrungen, Ausblicke zur weiteren Entwicklung.
Unfallchirurg.
1995;
98
229-232
7
Ejim O S, Blunn G W, Brown R A.
Production of artificial-oriented mats and strands from plasma fibronectin: a morphological study.
Biomaterials.
1993;
14
743-749
8
Hansbrough J F.
Current status of skin replacements for coverage of extensive burn wounds.
J Trauma.
1990;
30 (Suppl)
155-160
9
Hansbrough J F, Boyce S T, Cooper M L, Foreman T J.
Burn wound closure with cultured autologous keratinocytes and fibroblasts attached to a collagen- glycosaminoglycan substrate.
JAMA.
1989;
262
2125-2130
10
Hansbrough J F, Morgan J, Greenleaf G.
Advances in wound coverage using cultured cell technology.
Wounds.
1993;
5
174-194
11
Heck E, Bergstresser P, Baxter C.
Composite skin graft: frozen dermal allografts support engraftment and expression of autologous epidermis.
J Trauma.
1985;
25
106-112
12
Heimbach D, Luterman A, Burke J.
Artificial dermis for major burns. A multi-center randomized clinical trial.
Ann Surg.
1988;
208
313-320
13
Hollander D A, Stein M, Bernd A, Windolf J, Pannike A.
Autologous keratinocytes cultured on benzylester hyaluronic acid membranes in the treatment of chronic full- thickness ulcers.
J Wound Care.
1999;
8
351-355
14
Hollander D A, Soranzo C, Falk S, Windolf J.
Extensive traumatic soft tissue loss: Reconstruction in severely injured patients using cultured hyaluronan-based three-dimensional dermal and epidermal autografts.
J Trauma.
2001;
50
1125-1136
15
Kaiser H W, Stark G B, Kopp J.
Cultured autologous keratinocytes in fibrin glue suspension, exclusively and combined with STS-allograft (preliminary clinical and histological report of a new technique).
Burns.
1994;
20
23-29
16
Koyano T, Minoura N, Nagura M, Kobayashi K I.
Attachment and growth of cultured fibroblast cells on PVA/chitosan-blended hydrogels.
J Biomed Mater Res.
1998;
39
486-490
17
Krant D, Eckhardt M, Patton M L.
Combined simultaneous application of cultured epithelial autograft and Alloderm.
Wounds.
1995;
7
137-142
18
Krejci N C, Cuono C B, Langdon R C, McGuire J.
In vitro reconstruction of skin: fibroblasts facilitate keratinocyte growth and differentiation on acellular reticular dermis.
J Invest Dermatol.
1991;
97
843-848
19
Langer R, Vacanti J P.
Tissue engineering.
Science.
1993;
260
920-926
20
Moriarty K P, Crombleholme T M, Gallivan E K, O'Donnal C.
Hyaluronic acid-dependent pericellular matrices in fetal fibroblasts: implication for scar-free wound repair.
Wound Rep Reg.
1996;
4
346-352
21
Murphy G F, Orgill D P, Yannas I V.
Partial dermal regeneration is induced by biodegradable collagen-glycosaminoglycan grafts.
Lab Invest.
1990;
62
305-313
22
O'Connor N E, Mulliken J B, Banks-Schlegel S.
Grafting of burns with cultured epithelium from autologous cells.
Lancet.
1981;
1
75-78
23
Poumay Y, Pittelkow M R.
Cell density and culture factors regulate keratinocyte commitment to differenziation and expression of suprabasal K1/K10 keratins.
J Invest Dermatol.
1995;
104
271-276
24
Presti D, Scott J.
Hyaluronan-mediated protective effect against cell damage caused by enzymatically produced hydroxyl radicals is dependent on hyaluronan molecular mass.
Cell Biochem Function.
1994;
12
281-288
25
Prunieras M, Regnier M, Schlotter M.
A new method to culture human epidermal cells on allogeneic or xenogeneic dermis: preparation of recombined grafts.
Ann Chir Plast.
1979;
24
357-362
26
Rennekampff H O, Kiessig V, Hansbrough J F.
Current concepts in the development of cultured skin replacements.
J Surg Res.
1996;
62
288-295
27
Rheinwald J G, Green H.
Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells.
Cell.
1975;
6
331-344
28
Trabucchi E, Andreassi L, Malcovati M.
Ultramicroscopic observations of cultured epithelial sheets before and after grafting for major human burns.
Wounds.
1991;
3
83-88
29
West D C, Hampson I N, Arnold F, Kumar S.
Angiogenesis induced by degradation products of hyaluronic acid.
Science.
1985;
228
1324-1326
30
Zacchi V, Soranzo C, Cortivo R, Radice M. et al .
In vitro engineering of human skin-like tissue.
J Biomed Mater Res.
1998;
40
187-194
PD Dr. med. D. A. HollanderOberarzt der Unfallchirurgischen Klinik
Universitätsklinikum der RWTH Aachen
Pauwelsstraße 30
52074 Aachen
Phone: 02 41/8 08 93 50
Fax: 02 41/8 88 84 89
Email: dhollander@ukaachen.de