Application of benzonase in preparation of decellularized lamellar porcine corneal stroma for lamellar keratoplasty.
benzonase
bioengineered cornea
decellularized cornea
keratoplasty
Journal
Journal of biomedical materials research. Part A
ISSN: 1552-4965
Titre abrégé: J Biomed Mater Res A
Pays: United States
ID NLM: 101234237
Informations de publication
Date de publication:
11 2019
11 2019
Historique:
received:
16
10
2018
revised:
09
07
2019
accepted:
12
07
2019
pubmed:
23
7
2019
medline:
4
9
2020
entrez:
23
7
2019
Statut:
ppublish
Résumé
This study was to develop anovel and efficient method using endonuclease (benzonase) to preparedecellularized lamellar porcine corneal stroma (DLPCS). The DLPCS was preparedfrom native lamellar porcine corneal stroma (NLPCS) and was treated with 1000 U/ml benzonase for 5hours. We conducted the following measurements and animal transplantation tocompare DLPCS and NLPCS. The residual DNA was decreased significantly from 367.13 ± 19.96 ng/mg to 15.41 ± 0.65 ng/mg after treatment of benzonase by the detection of fluorescentnucleic acid stain. The residual benzonase was also less than detection limit.There was no significant difference in light transmittance of DLPCS comparedwith NLPCS. The extracts of DLPCS did not inhibit cell proliferation of human cornealepithelial cells, mouse fibroblast (L-929) and African green monkey kidney cell(Vero cell). The DLPCS was transplanted into the corneas of rabbit by lamellarkeratoplasty. There was no corneal melting and graft rejection been observedwithin 12 months. The images demonstrated that the repairment of corneal nervesand keratocytes of DLPCS were in indentical shape and reflection compared withnormal cornea, and no obvious inflammatory cells were observed postoperation, byin vivo confocal microscopy. We provided novel evidence that the application ofbenzonase may improve the quality of DLPCS.
Identifiants
pubmed: 31330094
doi: 10.1002/jbm.a.36760
pmc: PMC6771539
doi:
Substances chimiques
Endodeoxyribonucleases
EC 3.1.-
Endoribonucleases
EC 3.1.-
Serratia marcescens nuclease
EC 3.1.30.2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2547-2555Subventions
Organisme : National Key R&D program of China
ID : 2018YFA0107304
Pays : International
Informations de copyright
© 2019 The Authors. Journal of Biomedical Materials Research Part A published by Wiley Periodicals, Inc.
Références
Exp Eye Res. 2014 Dec;129:151-60
pubmed: 25281830
Cornea. 1995 Jan;14(1):43-8
pubmed: 7712736
Cell Tissue Bank. 2017 Sep;18(3):403-411
pubmed: 28455604
Vaccine. 2018 May 24;36(22):3153-3160
pubmed: 28729020
Curr Eye Res. 2016 Oct;41(10):1316-1325
pubmed: 26863271
Cornea. 2017 Mar;36(3):295-299
pubmed: 27861305
Gene Ther. 2011 Feb;18(2):135-44
pubmed: 20668485
Cell Tissue Bank. 2015 Jun;16(2):249-59
pubmed: 25163609
Cornea. 2011 Jun;30(6):692-701
pubmed: 21242785
Biomed Opt Express. 2014 Apr 14;5(5):1494-511
pubmed: 24877011
PLoS One. 2015 Mar 17;10(3):e0119934
pubmed: 25781319
Lasers Surg Med. 2007 Aug;39(7):597-604
pubmed: 17868101
Cornea. 2011 Apr;30(4):371-8
pubmed: 21099407
Am J Transplant. 2015 Apr;15(4):1068-75
pubmed: 25762108
Lancet. 2012 Feb 18;379(9816):672-83
pubmed: 22019026
Am J Transl Res. 2016 Mar 15;8(3):1412-25
pubmed: 27186268
J Biomed Mater Res A. 2019 Nov;107(11):2547-2555
pubmed: 31330094
Brain Res Bull. 2010 Feb 15;81(2-3):198-210
pubmed: 19481138
Biomaterials. 2011 Apr;32(12):3233-43
pubmed: 21296410
J Int Med Res. 2012;40(3):1149-55
pubmed: 22906289