Large-scale bicortical skull bone regeneration using ex vivo replication-defective adenoviral-mediated bone morphogenetic protein-2 gene-transferred bone marrow stromal cells and composite biomaterials.
OBJECTIVE
Bone marrow stromal cells (BMSCs) have great potential in bone repair. We developed an animal model to test the hypothesis that ex vivo gene transfer of human bone morphogenetic protein (BMP)-2 to BMSCs via a replication-defective (E1A-deleted) adenovirus vector (AdV) with appropriate biopolymers would enhance autologous bone formation during repair of a large-scale skull defect.


METHODS
Eighteen miniature swine were treated with AdV BMP-2-transduced BMSCs in biopolymer (group 1), BMSCs in biopolymer (group 2), or biopolymer alone (group 3). After 6 months, the swine were killed, and the skull repair was examined by gross pictures, histology, 3-dimensional computed tomography, and biomechanical study.


RESULTS
Group 1 showed complete solid bone formation after 6 months, and hematoxylin and eosin staining demonstrated the presence of mature, woven, well-mineralized bone. Computed tomography showed wholesome repair of the skull defect. Statistical analysis demonstrated a significant difference in bone thickness between groups 1 and 2. Biomechanical testing showed a statistically significant difference in the stiffness of new bone formed in group 1 compared with group 2.


CONCLUSIONS
The Ad5 E1A-deleted AdV may be the optimal starting vector in ex vivo gene therapy for benign skeletal diseases. Additionally, the use of the gelatin/tricalcium phosphate ceramic/glutaraldehyde biopolymer with AdV BMP-2 gene transfer strongly enhances the bony healing of critical-size bicortical craniofacial defects. This method can be used by modifying the delivery of constructs to malunion treatment, in regional osteoporosis therapy, and spinal fusion.
Chang
Sophia Chia-Ning
SC
Department of Plastic Surgery, China Medical University Hospital, and School of Medicine, China Medical University, Taichung, Taiwan, Republic of China.
Lin
Tsung-Min
TM
Chung
Hui-Ying
HY
Chen
Philip Kuo-Ting
PK
Lin
Feng-Huei
FH
Lou
Jeuren
J
Jeng
Long-Bin
LB
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry / Q000627:therapeutic use; D001704:Biopolymers / Q000737:chemistry / Q000627:therapeutic use; D016026:Bone Marrow Transplantation / Q000379:methods*; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics*; D001861:Bone Regeneration / Q000235:genetics*; D018786:Bone Substitutes; D002130:Calcium Phosphates / Q000737:chemistry / Q000627:therapeutic use; D002478:Cells, Cultured; D003399:Craniotomy / Q000009:adverse effects / Q000379:methods; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D005976:Glutaral / Q000737:chemistry / Q000627:therapeutic use; D023421:Models, Animal; D012886:Skull / Q000166:cytology / Q000502:physiology / Q000601:surgery; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation*; D034421:Sus scrofa; D016896:Treatment Outcome; D014779:Virus Replication / Q000235:genetics; D014945:Wound Healing / Q000502:physiology
C525718:BMP2 protein, human; D001672:Biocompatible Materials; D001704:Biopolymers; D055396:Bone Morphogenetic Protein 2; D018786:Bone Substitutes; D002130:Calcium Phosphates; C018392:tricalcium phosphate; D005976:Glutaral