Chromatin supraorganization, DNA content, nuclear glycoprotein amount, and immunophenotypic characterization of limbal epithelial cells cultured on intact or denuded amniotic membrane versus scaffold composed of fucoidan-alginate-chitosan

• Grant number: 13/01494-7
• Support Opportunities: Regular Research Grants
• Start date: July 01, 2013
• End date: September 30, 2015
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Clinics and Surgery
• Principal Investigator: José Luiz Laus
• Grantee: José Luiz Laus
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil
• City of the host institution: Jaboticabal

Abstract

At the end 1990, the transplantation of substrates colonized by corneal limbal epithelial stem cells emerged as a promising therapeutic option for reconstructing the ocular surface and restoring the vision in patients with limbal deficiency. After 15 years of clinical practice, several aspects related to the security and the standardization of the procedures used for manipulating these substrates sill remain unclear. For example, it is not totally clear if the ideal substrate to cultivate limbal stem cells would be obtained from intact or denuded amniotic membrane. Also, it is not known whether scaffolds constructed with polysaccharides, such as fucoidan, alginate and chitosan, used for the manufacture of bioactive dressings that stimulate the epidermal epithelial cell migration and accelerate the skin wound healing process, could be used for cultivating limbal stem cells and repairing corneal lesions. Many authors have tried to evaluate the effects of intact and denuded amniotic membrane on the expansion of limbal stem cells. Additionally, in view of the increasing use of limbal stem cells, many groups attempted to create new substrates for cell culture. Inadvertently, however, all the research used repetitive or identical analysis methods for evaluating these multidisciplinary biomedical questions. Previous studies have evaluated only the differential expression of molecules correlated with the differentiation course of limbal stem cells. No study was designed for evaluating the supraorganization and the expression of nuclear macromolecules that control the biological activities of corneal limbal epithelial cells expanded ex vivo. In this way, the purpose of this research is to determine the functional status of the chromatin, the DNA and the nuclear glycoproteins of corneal limbal epithelial cell cultivated on intact amniotic membrane, denuded amniotic membrane, and scaffolds composed of fucoidan-alginate-chitosan. (AU)

Scientific publications

(7)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

HERENCIA-BUENO, KARINA E.; ALDROVANI, MARCELA; CRIVELARO, ROBERTA M.; THIESEN, ROBERTO; BARROS-SOBRINHO, ALEXANDRE A. F.; CLAROS-CHACALTANA, FLOR D. Y.; PADUA, IVAN R. M.; SANTOS, DANIELA M.; LAUS, JOSE L.. Reduction in Histone H3 Acetylation and Chromatin Remodeling in Corneas of Alloxan-Induced Diabetic Rats. CORNEA, v. 37, n. 5, p. 624-632, MAY 2018. (13/01494-7, 12/17308-5)

VALDETARO, GISELE P.; ALDROVANI, MARCELA; PADUA, IVAN R. M.; CRISTOVAM, PRISCILA C.; GOMES, JOSE A. P.; LAUS, JOSE L.. Supra-organization and optical anisotropies of the extracellular matrix in the amniotic membrane and limbal stroma before and after explant culture. BIOMEDICAL OPTICS EXPRESS, v. 7, n. 12, p. 4982-4994, DEC 1 2016. (13/25533-1, 12/17308-5, 13/01494-7)

ALDROVANI, MARCELA; BARROS SOBRINHO, ALEXANDRE A. F.; MAIROS, FERNANDA SANTOS; LAUS, JOSE LUIZ. Short-Term Effects of Y-27632, a Rho-Associated Protein Kinase Inhibitor, on Chromatin Supraorganization and DNA Amount in Epithelial Cells of the Rat Cornea and Limbus. CORNEA, v. 36, n. 7, p. 845-853, JUL 2017. (12/17308-5, 13/01494-7)

ALDROVANI, MARCELA; FILEZIO, MARCELLA ROSA; LAUS, JOSE LUIZ. A supramolecular look at microenvironmental regulation of limbal epithelial stem cells and the differentiation of their progeny. Arquivos Brasileiros de Oftalmologia, v. 80, n. 4, p. 268-272, JUL-AUG 2017. (12/17308-5, 13/01494-7)

KOBASHIGAWA, KARINA KAMACHI; ALDROVANI, MARCELA; BARROS SOBRINHO, ALEXANDRE A. F.; SANTO SILVA, PALOMA DO ESPIRITO; MARCUSSO, PAULO F.; MARINHO-NETO, FAUSTO A.; MARTINES PADUA, IVAN R.; LAUS, JOSE LUIZ. Fenótipos clínicos e anisotropias ópticas da córnea em coelhos com deficiência de células tronco limbais. Arquivos Brasileiros de Oftalmologia, v. 81, n. 5, p. 384-392, SEP-OCT 2018. (14/18007-4, 12/17308-5, 13/01494-7)

CRIVELARO, ROBERTA M.; WERTHER, KARIN. THE EYE OF CRAB-EATING FOX (CERDOCYON THOUS): ANATOMICAL CHARACTERISTICS AND NORMATIVE VALUES OF SELECTED DIAGNOSTIC TESTS, MORPHOMETRY OF CORNEAL TISSUE, AND ARRANGEMENTS OF CORNEAL STROMAL COLLAGEN FIBERS. JOURNAL OF ZOO AND WILDLIFE MEDICINE, v. 51, n. 2, p. 280-289, JUN 2020. (13/01494-7, 12/17308-5)

ALDROVANI, MARCELA; FILEZIO, MARCELLA ROSA; LAUS, JOSE LUIZ. A supramolecular look at microenvironmental regulation of limbal epithelial stem cells and the differentiation of their progeny. Arquivos Brasileiros de Oftalmologia, v. 80, n. 4, p. 5-pg., 2017-07-01. (12/17308-5, 13/01494-7)

Filed patent(s) as a result of this research project