Introduction: Mesenchymal cells of the teeth, including mesenchymal cells of the third molars, are a very rich source of dental pulp cells that after collection, culture and passage can be found in special laboratory conditions that these cells are located in all individuals (adults, children and even fetuses). They have the possibility of using these cells in any period without any restrictions. The hidden point that can be found in the use of these cells is the optimal function of these cells in medical science, because of all the harms and problems that can be predicted in implants and ... It is far.
Methods: Survey in people 18-25 years old Fourteen specimens were collected from the pulp and six specimens from the third molar's dental follicles in adults between the ages of 18 and 25 years. These teeth had no previous caries or restoration, and all patients were healthy and had no systemic disease. They were taken and transferred to the Molecular Cell Laboratory at 4 ° C. To reveal the pulp chamber of the teeth, they were cut from the enamel-cement joint with a carbide disk and a handpiece, and thenThe pulp was separated from the pulp chamber by a fine file of the pulp; Then, for cell culture of pulp and follicle tissue, these cells were divided into smaller pieces by surgical blade No. 10, and then they were placed in Falcon containing 4 mg / ml of collagen type solution (sigma I), 104 mg / ml. Gibco type dipase solution with a ratio of 1/1 was exposed to 37 ° C for 45 minutes and then added to the lysed tissue of the culture medium and centrifuged at 600 g for 10 minutes. Mixed cultures were transferred to a suitable zvt in an incubator at 37 ° C, 5 atmospheres and 2% CO2. This culture medium was changed every two days until 70% of the plate bottom was filled with cells when the plate bottom reached 70% filling. Were passed using trypsin _EDTA
Finally, flow cytometric analysis was used to evaluate the phenotypic profile of surface markers and the nature of stem cells from the pulp and follicular tissue of the third molar. With a concentration of 1,000,000. The cells were then divided into 6 tubes and PE 5µI was added to each tube with antibodies. The tubes were then placed in a dark environment at 4 ° C for 30 minutes, after which time the cells were washed with 1 ml of wash buffer. They were centrifuged at 1200 MPR for 5 minutes, after which each cell sample was suspended in 300 l to 500 l of washing buffer and analyzed by flow cytometry.
The second experiment in children
In this experiment, the pulp of children's deciduous teeth that were extracted for dental reasons was used. After removing the teeth and collecting them and transferring them to the laboratory environment, the teeth were first rinsed with distilled water and phosphate buffer solution. They were made of polymer and can be ironed, where the teeth were centrifuged and the surface of the Falcon was discarded.
The teeth were then removed from the falcons and rinsed thoroughly with distilled water. It was pink and very thin. It was cut into several pieces by the surgical file inside the culture medium.
Because the cell separation process was very time consuming, two separate methods were used to accelerate the cell separation process:
1) Crushed particles with trypsin and collagen {, both of which are considered body lubricants and have a powdery structure and must be dissolved to add to the composition. These two substances must be refrigerated to be always active, and if They are out of the refrigerator for more than half an hour and lose their lubricating properties. They entered the Falcon. The Falcon containing pulp and trypsin tissue was placed in the refrigerator for 8 hours. Falcon contents slipped into a small flask under the hood.
2) In another falcon, the pulp was placed directly in the refrigerator without adding trypsin and collagen. The test was performed and after emptying the contents of the falcon, it entered the incubator in small flakes. 90 ° C and 2% carbon dioxide and 32% humidity were stored for a period of time until the cells separated.
The flakes were placed under an inverted microscope once a day and the condition of the tissues was examined. When the first cell lines were separated from the tissue, the cells were transferred to a 2 ml flask by passage. These cells were called HDPCs and after expiration About 6 months after the separation of pulp tissue inside each bifurcation, the first cell lines were observed. These cells were completely elongated and had a needle-shaped appearance. After filling the bottom of the dish, cell passage was performed with trypsin and finally, depending on the number Cells from each cell passage sample were taken.For the next period, the cells were used in the fourth passage in an environment containing 27 DMSO and 47%. Mesenchymal stem cells were transferred to a nitrogen tankThey were then differentiated into fat and bone using differentiated culture media to prove that these cells were stem cells.
Results: According to the mentioned cases, it can be said that in adult cells in adults, there are also mesenchymal stem cells of dental pulp, because after a dental injury, dentin is repaired by making and depositing dentin matrix to repair the damaged area. This restorative process takes place throughout a person's life, which indicates the presence of mesenchymal cells in the pulp of adult teeth and the ability to make odontoblasts under the influence of appropriate signals.But the whole potential of adult stem cells is not as great as that of embryonic and childhood stem cells(Which is extracted from the gums, especially from the posterior gums) or the use of mesenchymal stem cells in deciduous teeth. These cells, as mentioned, are not limited to any age group and even the ability to donate these cells from one person to another is possible. Is acceptable and the condition for donating teeth:
The complete health of the person and the donor's teeth, which can be done even in adulthood when most people have lost their deciduous teeth (except occult deciduous teeth) from the mesenchymal stem cell gene of children or fetuses to repair and restore teeth. In the laboratory method, mesenchymal stem cells in adults, although they can be repaired or regenerated, but after they are separated from the patient, they must first go to the laboratory and after strengthening and differentiating them, they are injected into another person as a plum or xenophore recipientHowever, due to the many problems that occur during amplification, injection, long time and less potential and compatibility of these cells, problems such as premature tooth decay, tooth loss, immune system attack on these cells through the immune response, etc. may occur.
In this regard, it can be said that the use of secondary methods (experiments: laboratory culture of cells, mesenchymal cells of deciduous teeth) have more function and efficiency in this field.
And given the problematic factors that exist in adult mesenchymal cells, the protocol for this ideal for humans is relatively far from being applicable. In dental restorations, it can be reported that a shorter course of treatment in dental restorations using stem cells has taken place (because in the two experiments mentioned above, restorations and restorations of teeth have taken a relatively long period).
By activating stem cells, these cells can send messages to stem cells that greatly help activate repair and amplification cells (this is also possible with low-power lasers), resulting in the formation of dentin cells.( Hard tooth tissue) helps a lot.
Conclusion: So it can be said that tooth regrowth is a fact, not an ideal, and given that the tooth is made up of two different types of tissue, logically making a tooth requires communication and cooperation with epithelial cells and odontogenic mesenchyme.
Recombinant epithelial tissue and dental mesenchyme to form teeth in both the laboratory and the living environment allow the combined cells to organize and form individual layers and also to be able to differentiate odontoblasts and amyloblasts.
In order to make a complete tooth that also has enamel and dentin, epithelial and mesenchymal cells are inserted into the collagen gel solution, respectively, and then implanted inside the oral cavity, and with this technique, the presence of all dental structures such as odontoblasts, amyloilasts, pulp, Blood vessels, crown, root, periodontal ligament and alveolar bone are observed, so implantation of this dental mass (mesenchymal + epithelial cells) leads to the development of puberty and regrowth of teeth.
Therefore, it can be said that using modern engineering and methods, mesenchymal stem cells extracted from tooth pulp and deciduous teeth can be used to repair tooth tissues under tissues that have mesenchyme and connective tissue infrastructure. Any age is able to regrow its teeth using the mesenchymal stem cell gene and it is worth noting that the use of mesenchymal gene is not limited from one person to another(No age limit for people) and another point is that the use of this method if the mesenchymal gene used is healthy unlike implants has no restrictions and no harm and replacement of this Methods Instead of modern methods, dentists can use it more economically and efficiently.