Sonic hedgehog gene-enhanced tissue engineering for bone regeneration

Paul Edwards, S. Ruggiero, J. Fantasia, R. Burakoff, S. M. Moorji, E. Paric, P. Razzano, D. A. Grande, J. M. Mason

Research output: Contribution to journalArticle

88 Citations (Scopus)

Abstract

Improved methods of bone regeneration are needed in the craniofacial rehabilitation of patients with significant bone deficits secondary to tumor resection, congenital deformities, and prior to prosthetic dental reconstruction. In this study, a gene-enhanced tissue-engineering approach was used to assess bone regenerative capacity of Sonic hedgehog (Shh)-transduced gingival fibroblasts, mesenchymal stem cells, and fat-derived cells delivered to rabbit cranial bone defects in an alginate/collagen matrix. Human Shh cDNA isolated from fetal lung tissue was cloned into the replication-incompetent retroviral expression vector LNCX, in which the murine leukemia virus retroviral LTR drives expression of the neomycin-resistance gene. The rat β-actin enhancer/ promoter complex was engineered to drive expression of Shh. Reverse transcriptase-polymerase chain reaction analysis demonstrated that the transduced primary rabbit cell populations expressed Shh RNA. Shh protein secretion was confirmed by enzyme-linked immunosorbent assay (ELISA). Alginate/type I collagen constructs containing 2 × 106 Shh-transduced cells were introduced into male New Zealand White rabbit calvarial defects (8 mm). A total of eight groups (N = 6) were examined. unrestored empty defects, matrix alone, matrix plus the three cell populations transduced with both control and Shh expression vectors. The bone regenerative capacity of Shh gene enhanced cells was assessed grossly, radiographically and histologically at 6 and 12 weeks postimplantation. After 6 weeks, new full thickness bone was seen emanating directly from the alginate/collagen matrix in the Shh-transduced groups. Quantitative two-dimensional digital analysis of histological sections confirmed statistically significant (P <0.05) amounts of bone regeneration in all three Shh-enhanced groups compared to controls. Necropsy failed to demonstrate any evidence of treatment-related side effects. This is the first study to demonstrate that Shh delivery to bone defects, in this case through a novel gene-enhanced tissue-engineering approach, results in significant bone regeneration. This encourages further development of the Shh gene-enhanced tissue-engineering approach for bone regeneration.

Original languageEnglish (US)
Pages (from-to)75-86
Number of pages12
JournalGene Therapy
Volume12
Issue number1
DOIs
StatePublished - Jan 2005
Externally publishedYes

Fingerprint

Bone Regeneration
Hedgehogs
Tissue Engineering
Genes
Bone and Bones
Rabbits
Collagen
Hedgehog Proteins
Murine Leukemia Viruses
Neomycin
Collagen Type I
Reverse Transcriptase Polymerase Chain Reaction
Mesenchymal Stromal Cells
Adipocytes
Population
Actins
Tooth
Fetus
Rehabilitation
Complementary DNA

Keywords

  • Fat-derived stem cells
  • Gene-enhanced bone regeneration
  • Gingival fibroblasts
  • Periosteal-derived cells
  • Retroviral expression vector
  • Sonic hedgehog

ASJC Scopus subject areas

  • Genetics

Cite this

Edwards, P., Ruggiero, S., Fantasia, J., Burakoff, R., Moorji, S. M., Paric, E., ... Mason, J. M. (2005). Sonic hedgehog gene-enhanced tissue engineering for bone regeneration. Gene Therapy, 12(1), 75-86. https://doi.org/10.1038/sj.gt.3302386

Sonic hedgehog gene-enhanced tissue engineering for bone regeneration. / Edwards, Paul; Ruggiero, S.; Fantasia, J.; Burakoff, R.; Moorji, S. M.; Paric, E.; Razzano, P.; Grande, D. A.; Mason, J. M.

In: Gene Therapy, Vol. 12, No. 1, 01.2005, p. 75-86.

Research output: Contribution to journalArticle

Edwards, P, Ruggiero, S, Fantasia, J, Burakoff, R, Moorji, SM, Paric, E, Razzano, P, Grande, DA & Mason, JM 2005, 'Sonic hedgehog gene-enhanced tissue engineering for bone regeneration', Gene Therapy, vol. 12, no. 1, pp. 75-86. https://doi.org/10.1038/sj.gt.3302386
Edwards P, Ruggiero S, Fantasia J, Burakoff R, Moorji SM, Paric E et al. Sonic hedgehog gene-enhanced tissue engineering for bone regeneration. Gene Therapy. 2005 Jan;12(1):75-86. https://doi.org/10.1038/sj.gt.3302386
Edwards, Paul ; Ruggiero, S. ; Fantasia, J. ; Burakoff, R. ; Moorji, S. M. ; Paric, E. ; Razzano, P. ; Grande, D. A. ; Mason, J. M. / Sonic hedgehog gene-enhanced tissue engineering for bone regeneration. In: Gene Therapy. 2005 ; Vol. 12, No. 1. pp. 75-86.
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