In vitro characterization of TGF-beta1 release from genetically modified fibroblasts in Ca(2+)-alginate microcapsules.
This study was undertaken to develop an in situ source of transforming growth factor-beta1 (TGF-beta1), one of several molecules potentially useful for a tissue-engineered bioartificial cartilage. Primary human fibroblasts and murine NIH 3T3 cells were genetically modified via viral transfection to express human TGF-beta1. Two viral constructs were used, one expressing a gene encoding for the latent and the other for the constitutively active form of the growth factor. Unmodified cells served as controls. Four genetically modified cohorts and two controls were separately encapsulated in a 1.8% alginate solution using a vibrating nozzle and 0.15M calcium chloride crosslinking bath. Diameter of the spherical capsules was 410 +/- 87 microm. In vitro release rate measured over 168 hours varied with cell types and ranged from 2-17 pg/(milligram of capsules x 24 h) or 2-17 ng/(10(6) cells x 24 h). None of the formulations exhibited a large initial bolus release. Even when serum-supplemented medium was not replenished, cell viabilities remained over 55% after 1 week for all cell types. Microencapsulated genetically modified cells were capable of a constitutive synthesis and delivery of biologically significant quantity of TGF-beta1 for at least 168 hours and thus are of potential utility for artificial cartilage and other orthopedic tissue engineering applications.
Paek
Hyun J
HJ
Center for Biomedical Engineering, Brown University, Providence, RI 02912, USA.
Campaner
Anelisa B
AB
Kim
Jennifer L
JL
Aaron
Roy K
RK
Ciombor
Deborah M
DM
Morgan
Jeffrey R
JR
Lysaght
Michael J
MJ
D000464:Alginates / Q000737:chemistry*; D000818:Animals; D002214:Capsules / Q000737:chemistry*; D018929:Cell Culture Techniques; D002461:Cell Line, Transformed; D002470:Cell Survival; D002472:Cell Transformation, Viral; D005347:Fibroblasts / Q000378:metabolism*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D012190:Retroviridae / Q000235:genetics; D012867:Skin / Q000166:cytology; D013997:Time Factors; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D000464:Alginates; D002214:Capsules; D006603:Hexuronic Acids; D016212:Transforming Growth Factor beta; D020723:Glucuronic Acid