Human levator veli palatini muscle: A novel source of mesenchymal stem cells for use in the rehabilitation of patients with congenital craniofacial malformations

Abstract

Background. Bone reconstruction in congenital craniofacial differences, which affect about 2-3% of newborns, has long been the focus of intensive research in the field of bone tissue engineering. The possibility of using mesenchymal stem cells in regenerative medicine protocols has opened a new field of investigation aimed at finding optimal sources of multipotent stem cells that can be isolated via non-invasive procedures. Here we analysed whether levator veli palatini muscle fragments, which can be readily obtained in non-invasive manner during surgical rehabilitation of cleft p­­atients during palatoplasty, represent a novel source of MSCs with osteogenic potential. Methods. We obtained levator veli palatini muscle fragments, in non-invasive procedure during surgical rehabilitation of 5 unrelated cleft palate patients (palatoplasty surgery). The levator veli palatini muscle fragments was used to obtain the mesenchymal cells using pre-plating technique in a clean rooms infrastructure and all procedures were performed at good practices of manipulation conditions. To prove that levator veli palatini muscle are mesenchymal stem cells they were induced to flow cytometry analysis and to differentiation into bone, cartilage, fat and muscle. To demonstrate the osteogenic potential of these cells in vivo a bilateral full thickness calvarial defect model was made in immunocompentent rats.Results. Flow cytometry analysis showed that the cells were positive for mesenchymal stem cell antigens (CD29, CD73, CD90), while negative for hematopoietic (CD45) or endothelial cell markers (CD31). Moreover, these cells were capable of undergoing chondrogenic, adipogenic, osteogenic and skeletal muscle cell differentiation under appropriate cell culture conditions characterizing them as mesenchymal stem cell. Defects treated with CellCeramTM scaffolds seeded with levator veli palatini muscle cells showed significantly greater bone healing compared to defects treated with acellular scaffolds. Conclusion. We have demonstrated that cells derived from levator veli palatini muscle have phenotypic characteristics similar to other mesenchymal stem cells, both in vitro and in vivo. Our findings suggest that these cells may have clinical relevance in the rehabilitation of patients with cleft palate and other craniofacial anomalies characterized by significant bone deficit.

The potential applications of artificially modified exosomes derived from mesenchymal stem cells in tumor therapy

“…In recent reports, transferring mimics and inhibitors of ncRNAs into tumor cells through MSC-exosomes has been a very promising strategy in tumor treatment. Here, we summarize the (16) BMSCs, bone marrow-derived mesenchymal stem cells; AMSCs, adipose-derived mesenchymal stem cells; UVMSCs, umbilical vein-derived mesenchymal stem cells; UCBMSCs, umbilical cord blood-derived mesenchymal stem cells; OPN, osteopontin; OCN, osteocalcin; LDL, low-density lipoprotein; fMSCs, fetal liver-derived mesenchymal stromal cells; SMMSCs, synovial membranederived mesenchymal stem cells; AFMSCs, amniotic fluid-derived mesenchymal stem cells; PMSCs, placenta-derived mesenchymal stem cells; HUCPV-MSCs, human umbilical cord perivascular-derived mesenchymal stem cells; PDLSCs, periodontal ligament stem cells; DP-MSCs, dental pulp-derived mesenchymal stem cells; Amnion MSCs, amnion-derived mesenchymal stem cells; CMSCs, chorion-derived mesenchymal stem cells; Human levator veli palatini muscle MSCs, human levator veli palatini muscle-derived mesenchymal stem cells. application of MSC-exosomes carrying noncoding RNAs or their inhibitors in tumor treatment.…”

“…MSCs have unique advantages in tumor treatment. First, MSCs can be ubiquitously found in many tissues, such as bone marrow (3); adipose (4); umbilical vein (5); umbilical cord blood (6); fetal liver (7); synovial membrane (8); amniotic fluid (9); placenta (10); Wharton's Jelly (11); human umbilical cord perivascular (12); periodontal ligament (13); dental pulp (14); amnion (15); chorion (15); and human levator veli palatini muscle (16), and are not restricted to tissues of mesodermal origin (Table 1). In addition, MSCs can be recruited to injured, inflamed, and hypoxic tissues and the tumor microenvironment, which is termed homing (17)(18)(19)(20).…”