A silk platform that enables electrophysiology and targeted drug delivery in brain astroglial cells.
Astroglial cell survival and ion channel activity are relevant molecular targets for the mechanistic study of neural cell interactions with biomaterials and/or electronic interfaces. Astrogliosis is the most typical reaction to in vivo brain implants and needs to be avoided by developing biomaterials that preserve astroglial cell physiological function. This cellular phenomenon is characterized by a proliferative state and altered expression of astroglial potassium (K(+)) channels. Silk is a natural polymer with potential for new biomedical applications due to its ability to support in vitro growth and differentiation of many cell types. We report on silk interactions with cultured neocortical astroglial cells. Astrocytes survival is similar when plated on silk-coated glass and on poly-D-lysine (PDL), a well known polyionic substrate used to promote astroglial cell adhesion to glass surfaces. Comparative analyses of whole-cell patch-clamp experiments reveal that silk- and PDL-coated cells display depolarized resting membrane potentials (-40 mV), very high input resistance, and low specific conductance, with values similar to those of undifferentiated glial cells. Analysis of K(+) channel conductance reveals that silk-astrocytes express large outwardly delayed rectifying K(+) current (K(DR)). The magnitude of K(DR) in PDL- and silk-coated astrocytes is similar, indicating that silk does not alter the resting K(+) current. We also demonstrate that guanosine- (GUO) embedded silk enables the direct modulation of astroglial K(+) conductance in vitro. Astrocytes plated on GUO-embedded silk are more hyperpolarized and express inward rectifying K(+) conductance (K(ir)). The K(+) inward current increases and this is paralleled by upregulation and membrane polarization of K(ir)4.1 protein signal. Collectively these results indicate that silk is a suitable biomaterial platform for the in vitro studies of astroglial ion channel responses and related physiology.
Benfenati
Valentina
V
Consiglio Nazionale delle Ricerche (CNR), Istituto per lo Studio dei Materiali Nanostrutturati (ISMN), Bologna, Italy. v.benfenati@bo.ismn.cnr.it
Toffanin
Stefano
S
Capelli
Raffaella
R
Camassa
Laura M A
LM
Ferroni
Stefano
S
Kaplan
David L
DL
Omenetto
Fiorenzo G
FG
Muccini
Michele
M
Zamboni
Roberto
R
D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000502:physiology*; D012831:Bombyx / Q000737:chemistry; D001921:Brain / Q000166:cytology*; D049109:Cell Proliferation / Q000187:drug effects; D016503:Drug Delivery Systems; D055724:Electrophysiological Phenomena / Q000187:drug effects*; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels / Q000378:metabolism; D006151:Guanosine / Q000494:pharmacology; D007211:Indoles / Q000378:metabolism; D015640:Ion Channel Gating / Q000187:drug effects; D018613:Microscopy, Confocal; D019579:Neocortex / Q000166:cytology; D011107:Polylysine / Q000494:pharmacology; D051381:Rats; D047011:Silk / Q000494:pharmacology*
D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels; D007211:Indoles; D047011:Silk; D006151:Guanosine; D011107:Polylysine; C007293:DAPI