Microencapsulated cells genetically modified to overexpress human transforming growth factor-beta1: viability and functionality in allogeneic and xenogeneic implant models.
This study explores the suitability of using encapsulated genetically modified fibroblasts for orthopedic tissue engineering by examining cell survival and persistence of human transforming growth factor-beta (hTGF-beta) overexpression in xenogeneic and allogeneic implant models. Human wild-type fibroblasts, modified to produce a latent form of hTGF-beta, and murine mutant-type fibroblasts, engineered to release a constitutively active form of hTGF-beta, were encapsulated separately in Ca2+ -alginate microcapsules. Following a percentage viability assessment by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) test, microcapsules were implanted into either the subcutaneous or intraperitoneal cavities of mice. Explanted encapsulated cells were characterized for percentage viability and subjected to a release study and a viability test 1 week and 3 weeks following implantation, a time frame consistent with the requirement for orthopedic tissue engineering application of this growth factor. On average, percentage viabilities of encapsulated cells were 64%at implantation, 52% at explantation, and 56%after 1 week following either 1- or 3-week explantation. hTGF-beta release declined following in vivo implantation, more so for xenogeneic than allogeneic models, but remained in the clinically attractive range of 2 to 30 ng/(10(6) implanted cells 24 h). This technical platform for hTGF-beta is very encouraging for cartilage regeneration using orthopedic tissue engineering, and further evaluation is warranted.
Paek
Hyun J
HJ
Center for Biomedical Engineering, Brown University, Providence, Rhode Island, USA. jpaek@tissuegenesis.com
Campaner
Anelisa B
AB
Kim
Jennifer L
JL
Golden
Louis
L
Aaron
Roy K
RK
Ciombor
Deborah M
DM
Morgan
Jeffrey R
JR
Lysaght
Michael J
MJ
D000464:Alginates; D000818:Animals; D001705:Bioprosthesis; D002214:Capsules; D002470:Cell Survival; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D015870:Gene Expression; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D051381:Rats; D017207:Rats, Sprague-Dawley; D023822:Tissue Engineering; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000235:genetics; D053773:Transforming Growth Factor beta1; D014183:Transplantation, Heterologous; D014184:Transplantation, Homologous
D000464:Alginates; D002214:Capsules; D006603:Hexuronic Acids; C508960:TGFB1 protein, human; C508961:Tgfb1 protein, mouse; C508962:Tgfb1 protein, rat; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D020723:Glucuronic Acid