The Effect of Mesenchymal Stem Cells of Umbilical Cord on the Treatment of Cleft Palate in Children

Cleft palate or harelip is one of the most common congenital defects in humans and harelip with cleft lip in whites is estimated 1 out of every 7000 to 1000 births. In recent years, surgeons and doctors have tried to resolve this problem immediately after the birth by operation, but sometimes we encounter things that cannot be easily solved by a surgical procedure. Our goal is to use stem cells to eliminate the disease and prevent operation, because children are often afraid of surgery and pain later on. Also, the need for general anesthesia for bone remodeling involves complications, such as long-term pain and nerve disorders. By the advent of the use of alternative stem extraction techniques, a better alternative to the invasive method of cleft lip and palate therapy has been developed.


Introduction
The cleft lip is a controversial issue that has led the world's leading medical and educational centers to achieve significant growth and prosperity. The oral-pink cleft is not the exception as the most common congenital anomaly in the craniofacial region. Bone graft as an alveolar cleft is an inseparable part of the process of treating patients with a one-way or two-way cleft lip and palate. The

Materials and Methods
Separation of tissues containing MSSCS from the umbilical cord.

Human
The umbilical cord samples were collected from the Bandar Abbas Hospital in accordance with the ethical requirements of the research and with the consent of the mother after child birth [3].
The umbilical cord sample collected from the Bandar Abbas Hospital in accordance with the ethical requirements of the research and with the consent of the mother after childbirth and after transfer to sterilization room, the cultivating room was studied and examined.
To eliminate pollution, we place 70 percent of the cord in alcohol for 30 seconds. Then, to ensure that there is no blood in the specimens, we rinse it again by PBS. The cord is divided into 5 mm 2 pieces by scissors, and then the pieces are arranged inside the plate (10 cm 2 Brand) [4]. CellBio On the samples, 10 ml of the culture medium (Dulbecco's Modified Medium:DMEM) was poured and plates were incubated in a BINDER incubator at 37˚C and 5% CO 2 (Figure 1) [5].
The samples were examined each day under invert microscope ( Figure 2). On the fifth day, they were replaced by half of their culture, and every three days thereafter, the substitution was carried out. On the twentieth day, cells filled 80% of the container. After this step, the cells were counted with Trypsin-EDTA, using Trypan Blue and Neubauer Lam, and transferred to new containers [6].

MSSCs Differentiation into Jawbone Cells
In order to confirm the mesenchymal nature of the cells investigated in this study, the distinctive property of differentiating cells into osteoblast cell washing by Alizarin Red was used. Third passage cells were selected and added to a number of flasks of the osteoblast differentiation medium instead of the DMEM medium. Their environment was changed every three days. The painting was carried out after 3 weeks and then examined (Figure 3).

Results
MSSCs were successfully isolated and cultured from human umbilical cord and injected into 10 infants over an intermittent therapy. After a year, this complication improved to a large extent, and the gap between the two lips was resolved, and with the continuity of treatment, the cleft palate of these babies was improving. According to the tests and studies, the extraction and culture of the mesenchymal stem cells from the cord takes less time. The amount of cells produced by this method is treated in this way up to 90% and without any side effects ( Figure 4).

Discussion
This study, for the first time, investigates the amount of bone produced by autogenous bone tissue engineering. The results of the study indicated that although autogenous bone graft still has a golden standard of treatment for bone defects,    including the alveolar gap, now it is the usual way to treat these abnormalities by using another part of the body, such as the pelvic bone, leg or rib. It has side ef-fects, such as infection, walking disorders, sensory disturbances and defects in other organs. However, tissue engineering techniques have limited access to autogenous bone or a significant morbidity may occur in that place. This can be used as an alternative technique with the desired clinical success and bone formation.

Conflicts of Interest
The author declares no conflicts of interest regarding the publication of this paper.