Is There a Noninvasive Source of MSCs Isolated with GMP Methods with Better Osteogenic Potential?

Abstract

Background A new trend in the treatment for alveolar clefts in patients with cleft lip and palate involves the use of bone tissue engineering strategies to reduce or eliminate the morbidity associated with autologous bone grafting. The use of mesenchymal stem cells—autologous cells obtained from tissues such as bone marrow and fat—combined with various biomaterials has been proposed as a viable option for use in cleft patients. However, invasive procedures are necessary to obtain the mesenchymal stem cells from these two sources. To eliminate donor site morbidity, noninvasive stem cell sources such as the umbilical cord, orbicularis oris muscle, and deciduous dental pulp have been studied for use in alveolar cleft bone tissue engineering. In this study, we evaluate the osteogenic potential of these various stem cell types. Methods Ten cellular strains obtained from each different source (umbilical cord, orbicularis oris muscle, or deciduous dental pulp) were induced to osteogenic differentiation in vitro, and the bone matrix deposition of each primary culture was quantified. To evaluate whether greater osteogenic potential of the established mesenchymal stem cell strains was associated with an increase in the expression profile of neural crest genes, real-time qPCR was performed on the following genes: SRY-box 9, SRY-box 10, nerve growth factor receptor, transcription factor AP-2 alpha, and paired box 3. Results The mesenchymal stem cells obtained from deciduous dental pulp and orbicularis oris muscle demonstrated increased osteogenic potential with significantly more extracellular bone matrix deposition when compared to primary cultures obtained from the umbilical cord after twenty-one days in culture (p = 0.007 and p = 0.005, respectively). The paired box 3 gene was more highly expressed in the MSCs obtained from deciduous dental pulp and orbicularis oris muscle than in those obtained from the umbilical cord. Conclusion These results suggest that deciduous dental pulp and orbicularis oris muscle stem cells demonstrate superior osteogenic differentiation potential relative to umbilical cord-derived stem cells and that this increased potential is related to their neural crest origins. Based on these observations, and the distinct translational advantage of incorporating stem cells from noninvasive tissue sources into tissue engineering protocols, greater study of these specific cell lines in the setting of alveolar cleft repair is indicated.

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

“…The mesenchymal/multipotential nature of LVPMDSCs was con rmed by the high expression of adhesion and MSC markers in these cells, together with their multilineage differentiation potential. These phenotypic hallmarks were similar to the ones seen in primary MSCs obtained from other sources, such as fat, dental pulp, bone marrow, fallopian tube, and umbilical cord vein (12,13,(15)(16)(17)(18)(19)(20)(21)(22).…”

“…Surgeons who treat congenital cream facial differences always try to decrease the number of operations in children requiring complex or staged bone reconstruction in the process of their craniofacial rehabilitation (10). The potential to minimize the numbers of required surgeries and, by extension, surgical morbidity by using MSCs seeded onto various scaffolds to generate new bone formation is an exciting prospect (10,11,13,27).…”

“…However, these surgical procedures may be subject to complications such as donor area morbidity, postsurgical reabsorption and infections (7,8). To circumvent these problems, researchers have focused on the development of bone tissue engineering strategies and osteogenic materials that may offer alternative methods with comparatively minimal or no donor site morbidity (9,10,11,12,13). We previously reported that orbicularis oris muscle (OOM) fragments, obtained during cheiloplasty of cleft lip and palate (CL/P) patients, are a rich source of mesenchymal stem cells (MSCs) that may be useful for According to regulatory local committee and under Brazilian Laws and resolution to regulate advanced cell therapies (National Sanitary Vigilance Agency -ANVISA -RDC n214, February 8th 2018) the Sírio-Libanês Hospital Laboratory facilities have recommended clean rooms infrastructure including air particulate control (HEPA lter) and air ow.…”

Mesenchymal Stromal Cell Immunology for Efficient and Safe Treatment of Osteoarthritis

“…The accessibility, frequency, and properties of MSCs may thus differ, requiring more attention in the choice of the source of MSCs (Busser et al, 2015). Moreover, it is essential to find non-invasive cell sources to avoid donor site morbidity (Pinheiro et al, 2019). In addition to BM, MSCs have also been isolated from adipose tissue, synovial membrane, fetal tissues, and dental pulp (Leyendecker et al, 2018b).…”

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

“…After xation, the wells were stained with 0.2% alizarin red S solution (pH 4.2; Sigma-Aldrich, St. Louis, MO) for 30 minutes. For the nal wash, each well was washed with PBS (Gibco Invitrogen, Grand Island, NY) three times (15).…”