title: BMP-silk composite matrices heal critically sized femoral defects.
abstract: Clinical drawbacks of bone grafting prompt the search for alternative bone augmentation technologies such as use of growth and differentiation factors, gene therapy, and cell therapy. Osteopromotive matrices are frequently employed for the local delivery and controlled release of these augmentation agents. Some matrices also provide an osteoconductive scaffold to support new bone growth. In this study, silkworm-derived silk fibroin was evaluated as an osteoconductive matrix for healing critical sized mid-femoral segmental defects in nude rats. Four treatment groups were assessed over eight weeks: silk scaffolds (SS) with recombinant human BMP-2 (rhBMP-2) and human mesenchymal stem cells (HMSC) that had been pre-differentiated along an osteoblastic lineage ex vivo (Group I; pdHMSC/rhBMP-2/SS); SS with rhBMP-2 and undifferentiated HMSCs (Group II; udHMSC/rhBMP-2/SS); SS and rhBMP-2 alone (Group III; rhBMP-2/SS); and empty defects (Group IV). Bi-weekly radiographs revealed a progressive and similar increase in Group I-III mean defect mineralization through post-operative week (POW) 8. Radiographs, dual energy x-ray absorptiometry, and micro-computed tomography confirmed that Groups I-III exhibited similar substantial and significantly (p<0.05) greater defect mineralization at POW 8 than the unfilled Group IV defects which remained void of bone. No significant differences in Groups I-III defect healing at POW 8 were apparent using these same assays or mechanical testing. Histology at POW 8 revealed moderately good bridging of the parent diaphyseal cortices with woven and lamellar bone bridging islands of silk matrix in Groups I and III. Group II defects possessed comparatively less new bone which was most abundant adjacent to the parent bone margins. Elsewhere the silk matrix was more often enveloped by poorly differentiated loose fibrous connective tissue. Group IV defects showed minimal new bone formation. None of the treatment groups attained the mean mineralization or the mean biomechanical strength of identical defects implanted with SS and pdHMSCs alone in a previous study. However, addition of rhBMP-2 to SS prompted more bone than was previously generated using udHMSC/SS or SS alone. These data imply the clinical potential of silk scaffolds and rhBMP-2 as composite osteopromotive implants when used alone or with select stem cell populations. Additional studies in larger species are now warranted.
authors.affiliation: Tufts Cummings School of Veterinary Medicine, North Grafton, MA, USA.
authors.forename: C
authors.initials: C
authors.lastname: Kirker-Head
authors.forename: V
authors.initials: V
authors.lastname: Karageorgiou
authors.forename: S
authors.initials: S
authors.lastname: Hofmann
authors.forename: R
authors.initials: R
authors.lastname: Fajardo
authors.forename: O
authors.initials: O
authors.lastname: Betz
authors.forename: H P
authors.initials: HP
authors.lastname: Merkle
authors.forename: M
authors.initials: M
authors.lastname: Hilbe
authors.forename: B
authors.initials: B
authors.lastname: von Rechenberg
authors.forename: J
authors.initials: J
authors.lastname: McCool
authors.forename: L
authors.initials: L
authors.lastname: Abrahamsen
authors.forename: A
authors.initials: A
authors.lastname: Nazarian
authors.forename: E
authors.initials: E
authors.lastname: Cory
authors.forename: M
authors.initials: M
authors.lastname: Curtis
authors.forename: D
authors.initials: D
authors.lastname: Kaplan
authors.forename: L
authors.initials: L
authors.lastname: Meinel
mesh_terms: D015502:Absorptiometry, Photon; D000818:Animals; D012831:Bombyx; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000235:genetics / Q000378:metabolism*; D001861:Bone Regeneration / Q000502:physiology*; D016025:Bone Transplantation; D005269:Femur / Q000000981:diagnostic imaging / Q000473:pathology* / Q000601:surgery; D006801:Humans; D020007:Implants, Experimental; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology; D010012:Osteogenesis / Q000502:physiology; D051381:Rats; D011923:Rats, Nude; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D047011:Silk / Q000378:metabolism*; D013314:Stress, Mechanical; D014057:Tomography, X-Ray Computed; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism*
chemical_list: C525718:BMP2 protein, human; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D011994:Recombinant Proteins; D047011:Silk; D016212:Transforming Growth Factor beta