New source of muscle-derived stem cells with potential for alveolar bone reconstruction in cleft lip and/or palate patients.
Cleft lip and palate (CLP), one of the most frequent congenital malformations, affects the alveolar bone in the great majority of the cases, and the reconstruction of this defect still represents a challenge in the rehabilitation of these patients. One of the current most promising strategy to achieve this goal is the use of bone marrow stem cells (BMSC); however, isolation of BMSC or iliac bone, which is still the mostly used graft in the surgical repair of these patients, confers site morbidity to the donor. Therefore, in order to identify a new alternative source of stem cells with osteogenic potential without conferring morbidity to the donor, we have used orbicular oris muscle (OOM) fragments, which are regularly discarded during surgery repair (cheiloplasty) of CLP patients. We obtained cells from OOM fragments of four unrelated CLP patients (CLPMDSC) using previously described preplating technique. These cells, through flow cytometry analysis, were mainly positively marked for five mesenchymal stem cell antigens (CD29, CD90, CD105, SH3, and SH4), while negative for hematopoietic cell markers, CD14, CD34, CD45, and CD117, and for endothelial cell marker, CD31. After induction under appropriate cell culture conditions, these cells were capable to undergo chondrogenic, adipogenic, osteogenic, and skeletal muscle cell differentiation, as evidenced by immunohistochemistry. We also demonstrated that these cells together with a collagen membrane lead to bone tissue reconstruction in a critical-size cranial defects previously induced in nonimmunocompromised rats. The presence of human DNA in the new bone was confirmed by PCR with human-specific primers and immunohistochemistry with human nuclei antibodies. In conclusion, we showed that cells from OOM have phenotypic and behavior characteristics similar to other adult stem cells, both in vitro and in vivo. Our findings suggest that these cells represent a promising source of stem cells for alveolar bone grafting treatment, particularly in young CLP patients.
Bueno
Daniela Franco
DF
Institute of Biosciences, University of São Paulo, São Paulo, Brazil.
Kerkis
Irina
I
Costa
André Mendonça
AM
Martins
Marília T
MT
Kobayashi
Gerson Shigeru
GS
Zucconi
Eder
E
Fanganiello
Roberto Dalto
RD
Salles
Felipe T
FT
Almeida
Ana Beatriz
AB
do Amaral
Cássio Eduardo Raposo
CE
Alonso
Nivaldo
N
Passos-Bueno
Maria Rita
MR
D000818:Animals; D001861:Bone Regeneration; D001842:Bone and Bones / Q000473:pathology*; D019070:Cell Lineage; D002469:Cell Separation; D002971:Cleft Lip / Q000628:therapy*; D002972:Cleft Palate / Q000628:therapy*; D004247:DNA / Q000378:metabolism; D005347:Fibroblasts / Q000473:pathology; D005434:Flow Cytometry; D006801:Humans; D016130:Immunophenotyping; D007223:Infant; D009132:Muscles / Q000166:cytology*; D010012:Osteogenesis; D051381:Rats; D017208:Rats, Wistar; D019651:Plastic Surgery Procedures / Q000379:methods*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
D004247:DNA