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J. Kim, J. Kim, D. Oh, Y. Chun; Histological Changes and Clinical Effects of Noble Decellularized Bovine Amniotic Membrane Using the Freeze-Thaw-Centrifuge Method. Invest. Ophthalmol. Vis. Sci. 2010;51(13):387.
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To investigate the histological and wound healing effects of noble freeze-thaw-centrifuge-decellularized bovine amniotic membranes (BAMs) and to compare them with those with classical chemical enzyme-decellularized BAMs
Fresh BAMs with no treatment, freeze-thaw-centrifuge- decellularized BAMs, and chemical enzyme-decellularized BAMs were all examined using H&E staining, immunohistochemical staining for basic fibroblast growth factor (bFGF), and bFGF quantification with ELISA. We induced corneal alkali burns with 1N NaOH in the eyes of 30 rabbits and then performed temporary patching using fresh BAMs, freeze-thaw-centrifuge-decellularized BAMs, and chemical enzyme-decellularized BAMs (10 eyes for each treatment). We then investigated corneal epithelial healing time, corneal opacity, corneal thickness, and the infiltration of inflammatory cells and apoptotic cells in the cornea by evaluating H&E staining and TUNEL immunohistochemical staining.
Decellularization occurred successfully in both freeze-thaw-centrifuge-treated BAMs and chemical enzyme-treated BAMs. Chemical enzyme-decellularized BAMs lost bFGF and were thinner compared to fresh BAMs. Freeze-thaw-centrifuge-decellularized BAMs were similar to fresh BAMs with respect to thickness, and well preserved bFGF. There were no differences in corneal epithelial healing time among the three corneal alkali burn groups. However, four weeks after the alkali burns, the improvements in corneal opacity and corneal thickness in the chemical enzyme-decellularized BAM group were less pronounced compared to those in the other two groups. There were more inflammatory cells and apoptotic cells infiltrating the cornea in the chemical enzyme-decellularized BAM group than there were in the other two groups three days after alkali burn induction.
Noble decellularized bovine amniotic membrane with freeze-thaw-centrifuge is more likely to maintain the characteristics of the original bovine amniotic membrane. Its clinical effects on wound healing and scar inhibition are greater than those seen with chemical enzyme-decellularized BAMs.
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