Biological and Ultrastructural Properties of Acelagraft, a Freeze-Dried γ-Irradiated Human Amniotic Membrane

Abstract

To compare the biological and ultrastructural properties of a commercially available decellularized and dehydrated human amniotic membrane (DDHAM) product with cryopreserved human amniotic membrane (CHAM) and to demonstrate the feasibility of DDHAM transplant in a case of chronic ocular surface disease. Methods: Histologic examination, immunohistochemical examination for extracellular matrix molecules and growth factors, transmission and scanning electron microscopy, and atomic force microscopy were performed on both DDHAM and CHAM specimens. A DDHAM transplant was performed in a patient with chronic bullous keratopathy and ocular surface instability. Results: Histologic examination and transmission electron microscopy revealed the disruption of the trilaminar structure of the basement membrane compared with CHAM, and immunohistochemical examination confirmed the loss of collagen IV and VII, laminin, and fibronectin in DDHAM. Lower levels of several growth factors were also seen in DDHAM compared with CHAM. Clinical transplant of DDHAM was, however, successful, with rapid reepithelialization. Conclusion: Significant differences in composition and ultrastructure exist between DDHAM and CHAM but do not appear to compromise cell survival in vivo. Clinical Relevance: The ease of storage and handling of DDHAM make it potentially valuable in ocular surface surgery, but its biological properties have not been well characterized, and there are also few data on its clinical application.

Comparison of the impact of preservation methods on amniotic membrane properties for tissue engineering applications

“…Delamination of the stroma was also observed in D-hAM. This disruption is commonly induced by decellularization processes [30,40,53]. In our study, except for fibronectin, labeling of the other proteins was still observed after the decellularization of hAM, although it was sometimes slightly or highly reduced.…”

“…Treatment of hAM raises issues regarding its biological and mechanical properties. Several studies have already reported some damage in the expression or the distribution of the extracellular matrix and basement membrane components of hAM after preservation [23,30,40]. In this study, the morphology of the hAM and the protein distribution were altered by the three preservation methods tested.…”

Comparison of the impact of preservation methods on amniotic membrane properties for tissue engineering applications

“…Delamination of the stroma was also observed in D-hAM. This disruption is commonly induced by decellularization processes [30,40,53]. In our study, except for fibronectin, labeling of the other proteins was still observed after the decellularization of hAM, although it was sometimes slightly or highly reduced.…”

“…Treatment of hAM raises issues regarding its biological and mechanical properties. Several studies have already reported some damage in the expression or the distribution of the extracellular matrix and basement membrane components of hAM after preservation [23,30,40]. In this study, the morphology of the hAM and the protein distribution were altered by the three preservation methods tested.…”

Hyperdry human amniotic membrane is useful material for tissue engineering: Physical, morphological properties, and safety as the new biological material

“…In the present study, the structure of the FD-AM (which has been reported to be clinically useful in several medical fields [40][41][42][43] ) was much more fragile than that of the HD-AM, because the FD-AM had rich powder for the rupture of collagen bundles in connective tissues. The photograph of HD-AM stained with hematoxylin/eosin shows morphology similar to that of fAM.…”

Augmented Dried versus Cryopreserved Amniotic Membrane as an Ocular Surface Dressing

“…However, our immunofluorescence data confirm our finding that lyoprotection of AM with trehalose or raffinose prevents loss of soluble biological factors and this is consistent with the protein array data. In contrast, an immunohistochemical analysis of AM extracellular matrix molecules has indicated reduced intensity following drying compared to cryopreservation[55]. This may be attributed to radiation damage.…”

Comparison of the impact of preservation methods on amniotic membrane properties for tissue engineering applications

“…Delamination of the stroma was also observed in D-hAM. This disruption is commonly induced by decellularization processes [30,40,53]. In our study, except for fibronectin, labeling of the other proteins was still observed after the decellularization of hAM, although it was sometimes slightly or highly reduced.…”

“…Treatment of hAM raises issues regarding its biological and mechanical properties. Several studies have already reported some damage in the expression or the distribution of the extracellular matrix and basement membrane components of hAM after preservation [23,30,40]. In this study, the morphology of the hAM and the protein distribution were altered by the three preservation methods tested.…”

Hyperdry human amniotic membrane is useful material for tissue engineering: Physical, morphological properties, and safety as the new biological material

“…In the present study, the structure of the FD-AM (which has been reported to be clinically useful in several medical fields [40][41][42][43] ) was much more fragile than that of the HD-AM, because the FD-AM had rich powder for the rupture of collagen bundles in connective tissues. The photograph of HD-AM stained with hematoxylin/eosin shows morphology similar to that of fAM.…”

Augmented Dried versus Cryopreserved Amniotic Membrane as an Ocular Surface Dressing

“…However, our immunofluorescence data confirm our finding that lyoprotection of AM with trehalose or raffinose prevents loss of soluble biological factors and this is consistent with the protein array data. In contrast, an immunohistochemical analysis of AM extracellular matrix molecules has indicated reduced intensity following drying compared to cryopreservation[55]. This may be attributed to radiation damage.…”

Comparison of the impact of preservation methods on amniotic membrane properties for tissue engineering applications

“…Delamination of the stroma was also observed in D-hAM. This disruption is commonly induced by decellularization processes [30,40,53]. In our study, except for fibronectin, labeling of the other proteins was still observed after the decellularization of hAM, although it was sometimes slightly or highly reduced.…”

“…Treatment of hAM raises issues regarding its biological and mechanical properties. Several studies have already reported some damage in the expression or the distribution of the extracellular matrix and basement membrane components of hAM after preservation [23,30,40]. In this study, the morphology of the hAM and the protein distribution were altered by the three preservation methods tested.…”

Hyperdry human amniotic membrane is useful material for tissue engineering: Physical, morphological properties, and safety as the new biological material

“…In the present study, the structure of the FD-AM (which has been reported to be clinically useful in several medical fields [40][41][42][43] ) was much more fragile than that of the HD-AM, because the FD-AM had rich powder for the rupture of collagen bundles in connective tissues. The photograph of HD-AM stained with hematoxylin/eosin shows morphology similar to that of fAM.…”

Augmented Dried versus Cryopreserved Amniotic Membrane as an Ocular Surface Dressing

“…However, our immunofluorescence data confirm our finding that lyoprotection of AM with trehalose or raffinose prevents loss of soluble biological factors and this is consistent with the protein array data. In contrast, an immunohistochemical analysis of AM extracellular matrix molecules has indicated reduced intensity following drying compared to cryopreservation[55]. This may be attributed to radiation damage.…”