Use of blends of bioabsorbable poly(L-lactic acid)/poly(hydroxybutyrate- co-hydroxyvalerate) as surfaces for Vero cell culture.
Vero cells, a cell line established from the kidney of the African green monkey (Cercopithecus aethiops), were cultured in F-10 Ham medium supplemented with 10% fetal calf serum at 37 degrees C on membranes of poly(L-lactic acid) (PLLA), poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and their blends in different proportions (100/0, 60/40, 50/50, 40/60, and 0/100). The present study evaluated morphology of cells grown on different polymeric substrates after 24 h of culture by scanning electron microscopy. Cell adhesion was also analyzed after 2 h of inoculation. For cell growth evaluation, the cells were maintained in culture for 48, 120, 240, and 360 h. For cytochemical study, the cells were cultured for 120 or 240 h, fixed, processed for histological analysis, and stained with Toluidine blue, pH 4.0, and Xylidine ponceau, pH 2.5. Our results showed that cell adhesion was better when 60/40 and 50/50 blends were used although cells were able to grow and proliferate on all blends tested. When using PLLA/PHBV (50/50) slightly flattened cells were observed on porous and smooth areas. PLLA/PHBV (40/60) blends presented flattened cells on smooth areas. PLLA/PHBV (0/100), which presented no pores, also supported spreading cells interconnected by thin filaments. Histological sections showed that cells grew as a confluent monolayer on different substrates. Cytochemical analysis showed basophilic cells, indicating a large amount of RNA and proteins. Hence, we detected changes in cell morphology induced by alterations in blend proportions. This suggests that the cells changed their differentiation pattern when on various PLLA/PHBV blend surfaces.
Santos
A R
AR
Departamento de Biologia Celular, Instituto de Biologia, Universidade Estadual de Campinas, Campinas, SP, Brazil. arsantosjr@yahoo.com.br
Ferreira
B M P
BM
Duek
E A R
EA
Dolder
H
H
Wada
M L F
ML
D000818:Animals; D001672:Biocompatible Materials; D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods*; D002522:Chlorocebus aethiops; D006651:Histocytochemistry; D006885:Hydroxybutyrates; D019344:Lactic Acid; D008567:Membranes, Artificial; D008855:Microscopy, Electron, Scanning; D011091:Polyesters; D011108:Polymers; D016062:Porosity; D014709:Vero Cells / Q000166:cytology* / Q000648:ultrastructure
D001672:Biocompatible Materials; D006885:Hydroxybutyrates; D008567:Membranes, Artificial; D011091:Polyesters; D011108:Polymers; D019344:Lactic Acid; C033616:poly(lactide)