Preservation, sterilization and de-epithelialization of human amniotic membrane for use in ocular surface reconstruction

Riau, Andri K.; Beuerman, Roger W.; Lim, Laurence Shen; Mehta, Jodhbir S.

doi:10.1016/j.biomaterials.2009.09.034

Cited by 184 publications

(188 citation statements)

References 153 publications

(177 reference statements)

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“…Given that AM can be prepared with various storage medium, including glycerol/DMEM, 22 dimethyl sulfoxide/ RPMI, 14,23,24 and Hank's solution, 17 we sought to evaluate the influence of the storage solution on MSC adherence on the stromal side of AM. To develop a tissue-engineered product for the clinic, the process of production shall be as simple as possible.…”

Section: Impact Of the Conditions Of Am Cryopreservation On Msc Adhermentioning

confidence: 99%

“…These treatments could degrade basal membrane if the technique is not controlled 36 and it does not ensure complete cell removal. In the same way, procedures for denuding AM differ among studies, 14,17,18,37 suggesting that biological properties of AM could be variable. The advantage of our procedure is that MSC were loaded directly on the stromal side of intact AM without any treatment technique.…”

mentioning

confidence: 99%

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In VitroCharacterization of Patches of Human Mesenchymal Stromal Cells

Roux

Bodivit

Bartis

et al. 2015

Tissue Engineering Part A

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Stem cells may represent an excellent strategy to improve the healing of skin ulcers. Today the administration mode of stem cells to skin defects remains unsatisfactory. Delivering stem cells with topical treatments represents a new strategy and answering the patients' need. Mesenchymal stromal cells (MSC) have been shown to improve wound healing of cutaneous lesions and amniotic membrane (AM) is known to represent a natural scaffold for cells. The aim of this study is to develop a tissue-engineered product combining MSC and AM for clinical use. In this work we investigated whether the stromal matrix of intact human AM could constitute a scaffold for human MSC derived from either bone marrow (BM) or adipose tissue (AT). For this purpose, clinical-grade AM, MSC, and culture medium were used. We performed experiments of short-term adherence and proliferation for 15 days after the seeding of the cells. Morphological aspects and secretion profiles of MSC onto AM were studied, respectively, by scanning electron microscopy and Luminex analysis. Results demonstrated that the stromal matrix allow the adherence in much greater amount of MSC from BM or AT compared to 2D material. Experiments of proliferation showed that both kinds of MSC could proliferate on the stromal matrix and remain viable 15 days after the seeding of the cells. The 3D analysis of MSC culture demonstrated that both types of MSC invaded the stromal matrix and grew in multiple layers while retaining their fibroblastic morphology. By studying the secretion profile of MSC onto the stromal matrix, we found that both kinds of MSC secrete important cytokines and growth factors for wound healing of cutaneous lesions, such as vascular endothelial growth factor, hepatocyte growth factor, and basic fibroblast growth factor. In conclusion, these results suggest that the stromal matrix of AM seeded with MSC represents a bioactive scaffold that should be evaluated in patients with a nonhealing cutaneous wound.

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Section: Impact Of the Conditions Of Am Cryopreservation On Msc Adhermentioning

confidence: 99%

mentioning

confidence: 99%

In VitroCharacterization of Patches of Human Mesenchymal Stromal Cells

Roux

Bodivit

Bartis

et al. 2015

Tissue Engineering Part A

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“…4,5 Since the AM must bear the load of pressure from the amniotic fluid during gestation, it has also the mechanical strength important to withstand the stress induced during the growth of tissue. 6 Besides the attractive biological properties, the AM is easy to obtain and is readily available.…”

Section: Introductionmentioning

confidence: 99%

Amniotic Membrane Scaffolds Enable the Development of Tissue-Engineered Urothelium with Molecular and Ultrastructural Properties Comparable to that of Native Urothelium

Jerman

Veranič

Kreft

2014

Tissue Engineering Part C: Methods

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The amniotic membrane (AM) is a naturally derived biomaterial that possesses biological and mechanical properties of great importance for tissue engineering. The aim of our study was to determine whether the AM enables the formation of a normal urinary bladder epithelium-urothelium-and to reveal any differences in the urothelial cell (UC) growth and differentiation when using different AM scaffolds. Cryopreserved human AM was used as a scaffold in three different ways. Normal porcine UCs were seeded on the AM epithelium (eAM), denuded AM (dAM), and stromal AM (sAM) and were cultured for 3 weeks. UC growth on AM scaffolds was monitored daily. By using electron microscopy, histochemical and immunofluorescence techniques, we here provide evidence that all three AM scaffolds enable the development of the urothelium. The fastest growth and the highest differentiation of UCs were demonstrated on the sAM scaffold, which enables the development of tissue-engineered urothelium with molecular and ultrastructural properties comparable to that of the native urothelium. Most importantly, the highly differentiated urothelia on the sAM scaffolds provide important experimental models for future drug delivery studies and developing tissue engineering strategies considering that subtle differences are identified before translation to the clinical settings.

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“…15,16 Retrieved from donor placenta to transplantation, HAM must undergo a series of preoperative preparation. 8,17,18 One important step is the de-epithelialization to uncover the basement membrane for epithelial cell attachment and proliferation. Prior studies have shown that the denuded HAM promotes a better growth and expansion of isolated LEC, while intact HAM is more suitable for limbal explant; however, this discrepancy is yet to be explained.…”

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confidence: 99%

The Effect of Amniotic Membrane De-Epithelialization Method on its Biological Properties and Ability to Promote Limbal Epithelial Cell Culture

Zhang

Yam

Riau

et al. 2013

Invest. Ophthalmol. Vis. Sci.

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Citation: Zhang T, Yam GH-F, Riau AK, et al. The effect of amniotic membrane de-epithelialization method on its biological properties and ability to promote limbal epithelial cell culture. Invest Ophthalmol Vis Sci. 2013;54:3072-3081. DOI:10.1167/ iovs.12-10805 PURPOSE. We characterized the de-epithelialized human amniotic membrane (HAM), and compared cell attachment and proliferation efficiencies.METHODS. HAM was de-epithelialized by 20% ethanol (AHAM), 1.2 U/mL Dispase (DHAM), 0.02% EDTA (EHAM), 0.25% trypsin-EDTA (THAM), and 5 M urea (UHAM), respectively, followed by gentle scraping with a #15 blade. Surface topology, extracellular matrix (ECM), and growth factor content were characterized and compared to intact HAM by electron microscopies (EM), atomic force microscopy (AFM), immunohistochemistry, and Western blotting. Primary human limbal epithelial cells (LEC) attachment and proliferation efficiencies were assayed. Statistical significance was calculated by SPSS and Fisher's least significant difference test.RESULTS. EHAM, THAM, and UHAM had intact basal lamina and smooth basement membrane surface shown under transmission and scanning EM, and AFM. Cell remnants stayed on AHAM. Disrupted basement membrane and stroma was found in DHAM. Immunostaining intensity quantification and hierarchical clustering revealed that ECM composition of EHAM and UHAM resembled intact HAM. In contrast, DHAM and THAM had drastic loss of ECM and growth factor content. LEC attachment efficiency at 24 hours after seeding was the highest in EHAM (51% as on conventional culture surface), followed by UHAM and AHAM. However, cell proliferation indices at day 10 of culture were similar among different HAM substrates, suggesting repair of ECM and basement membrane by growing epithelial cells.CONCLUSIONS. Urea denudation preserved the basement membrane integrity, ECM, and growth factor composition, and had higher cell attachment and proliferation efficiencies. With its short processing time, urea treatment offers a novel alternative for HAM de-epithelialization.

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Preservation, sterilization and de-epithelialization of human amniotic membrane for use in ocular surface reconstruction

Cited by 184 publications

References 153 publications

In VitroCharacterization of Patches of Human Mesenchymal Stromal Cells

In VitroCharacterization of Patches of Human Mesenchymal Stromal Cells

Amniotic Membrane Scaffolds Enable the Development of Tissue-Engineered Urothelium with Molecular and Ultrastructural Properties Comparable to that of Native Urothelium

The Effect of Amniotic Membrane De-Epithelialization Method on its Biological Properties and Ability to Promote Limbal Epithelial Cell Culture

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