KR101788447B1 - Mini-dental implant - Google Patents

Abstract

The present invention relates to the production and placement of mini-dental implants for animal models. Since the mini-dental implant according to the present invention can be applied to a mouse, which is the smallest animal model, it can be used for an implant experiment using animal models of various sizes, It is anticipated that it can be used effectively in research on the relationship between various diseases and / or various materials and implants because it can create a tooth-like environment.

Description

Mini-dental implant < RTI ID = 0.0 >

The present invention relates to the production and placement of mini-dental implants for animal models.

A dental implant, also called an artificial tooth or a third tooth, is a fixture (placed in the gums and gums) that is placed in or on the jaws for the purpose of restoring a missing tooth, Is a common surgical procedure that is widely used.

In general, the success of implant surgery is closely related to the patient 's condition, although it is widely used in various patients. If you have periodontal disease, you must be treated before implant surgery. Systemic diseases, infectious diseases, progression of other diseases, and intraoral lesions also affect the results of implant surgery. For example, there is a history of cardiovascular disease (angina pectoris, myocardial infarction, congestive heart failure, endocarditis, heart valve disease, etc.) or an endocrine system disease (diabetes, hyperthyroidism, hypothyroidism, hyperthyroidism (Cushing's syndrome) (Including osteoporosis, osteomalacia, and the like), a site where the implant is implanted, and a region where the implant is implanted. It can affect long-term maintenance (adhesion of the gum bone and implants). In addition, there is no study on the relationship between pregnancy and implant surgery in women with a high possibility of pregnancy, and it may interfere with the adhesion of the implant to the bone in the case of osteoporosis or systemic disease.

However, there are no studies on the relationship between these various diseases and the success of implant surgery or postoperative side effects. In order to do this, there are some studies that use cadavers, pigs, and dogs. However, there are limitations in performing various studies such as lack of application model, time, and economics (Journal of periodontal & Implant Science (2006) 36 (4): 913-923). Experiments using a rat larger than a mouse are occasionally performed, but the implant is placed in a large bone of a thigh portion of a mouse, There is almost no method. Therefore, the development of a small animal model that has various disease models and is easy to experiment with implant surgery is urgently required

SUMMARY OF THE INVENTION It is an object of the present invention to provide a method of manufacturing mini-dental implants of a human-like shape in an animal model.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

Hereinafter, various embodiments described herein will be described with reference to the drawings. In the following description, for purposes of complete understanding of the present invention, various specific details are set forth, such as specific forms, compositions, and processes, and the like. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well-known processes and techniques of manufacture are not described in any detail, in order not to unnecessarily obscure the present invention. Reference throughout this specification to "one embodiment" or "embodiment" means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in at least one embodiment of the invention. Accordingly, the appearances of the phrase " in one embodiment "or" an embodiment "in various places throughout this specification are not necessarily indicative of the same embodiment of the present invention. In addition, a particular feature, form, composition, or characteristic may be combined in any suitable manner in one or more embodiments.

The mini-dental implant of the present invention is used for researching the relationship between an implant and various diseases, materials, etc., placed in an animal model. The mini-dental implant is used for an animal model, There are no limitations on the type of implants used in general human operation, and the shape of the implants can be arbitrarily modified in accordance with the shape of animal teeth in order to easily put them into an animal model.

The present invention relates to an animal model in which a human PDL cell sheet (human periodontal ligament cell sheet) or a human cementoblast cell sheet is coated on a mini-dental implant according to a tooth size of a laboratory animal model Mini-tooth implants.

The present invention also relates to a mini-dental implant adapted to the tooth size of a laboratory animal model, comprising a human cementoblast cell sheet, a human umbilical vein endothelial cell sheet (HUVEC sheet) ) And a human PDL cell sheet (human periodontal ligament cell sheet) are sequentially coated.

In one embodiment of the present invention, the mini-dental implant is preferably made of a material which can be generally used for an implant, such as titanium and gold, but may be made of a material that can be used for manufacturing an implant But is not limited thereto.

In another embodiment of the present invention, the mini-dental implant is preferably coated with hydroxyapatite, but the present invention is not limited thereto, so long as it is a coating method for improving the efficiency of success of the implant.

In another embodiment of the present invention, the mini-dental implant may include a human umbilical vein endothelial cell sheet (HUVEC sheet) between the mini-dental implant and the coated cell sheet, Such as a cell, a peptide, or a compound, which helps the implantation of the implant into the implant.

In another embodiment of the present invention, the mini-dental implant is preferably a human PDL cell, which can be replaced with an ihPDL cell (immortalized human periodontal ligament cell) and a human Cem cell with an immortalized human cementoblast cell , And an ERM cell sheet (epithelial cell rest of Malassez cell sheet). However, it is generally acceptable to apply it to the mini-dental implant of the present invention, and it is a cell which can be produced in the form of a cell sheet, There is no limit to the type of cells that are present in the periodontal ligament (PDL) space.

The present invention also provides a method of preparing a mini-dental implant, comprising the steps of: (a) preparing a mini-dental implant adapted to a tooth size of a laboratory animal model; And (b) coating a human PDL cell sheet or a human cementoblast cell sheet on the prepared mini-dental implant. ≪ / RTI >

In one embodiment of the invention, the mini-dental implant may further comprise coating hydroxyapatite and / or coating a human umbilical vein endothelial cell sheet (HUVEC sheet) .

In another embodiment of the present invention, the mini-dental implant is preferably a human PDL cell, which can be replaced with an ihPDL cell (immortalized human periodontal ligament cell) and a human Cem cell with an immortalized human cementoblast cell , And an ERM cell sheet (epithelial cell rest of Malassez cell sheet). However, it is generally acceptable to apply it to the mini-dental implant of the present invention, and it is a cell which can be produced in the form of a cell sheet, There is no limit to the type of cells that are present in the periodontal ligament (PDL) space.

The present invention also provides a method of preparing a mini-dental implant, comprising the steps of: (a) preparing a mini-dental implant adapted to a tooth size of a laboratory animal model; (b) coating a human cementoblast cell sheet on the prepared mini-dental implant; (c) coating a human umbilical vein endothelial cell sheet (HUVEC sheet) on the coated mini-dental implant; And (d) coating a human PDL cell sheet with the double-coated mini-dental implant. The present invention also provides a method of making mini-dental implants for animal models.

In one embodiment of the present invention, the manufacturing method may further include coating hydroxyapatite on the surface of the mini-dental implant manufactured after step (a).

In another embodiment of the invention, the cell sheet is preferably obtained by scraping cells cultured in a culture dish with a cell scraper, more preferably by inoculating the cells in an UpCell culture dish and incubating at 37 [deg.] C And then lowering the temperature to 20 占 폚. However, the present invention is not limited thereto, as long as it is a method for producing a cell sheet that can be coated on the surface of an implant.

In another embodiment of the present invention, the step of coating the cell sheet may be coated by placing the cell sheet on the surface of the implant using micro forceps, The method of coating the sheet is not limited thereto.

In another embodiment of the present invention, the mini-dental implant is preferably a human PDL cell, which can be replaced with an ihPDL cell (immortalized human periodontal ligament cell) and a human Cem cell with an immortalized human cementoblast cell , And an ERM cell sheet (epithelial cell rest of Malassez cell sheet). However, it is generally acceptable to apply it to the mini-dental implant of the present invention, and it is a cell which can be produced in the form of a cell sheet, There is no limit to the type of cells that are present in the periodontal ligament (PDL) space.

The present invention also relates to a method of making a tooth, comprising: (a) extracting teeth of an animal model; (b) breeding the extracted site so that the alveolar bone can be filled; (c) making an implant socket with the drill bit on the generated two-tooth bone; And (d) placing the mini-dental implant of any one of claims 1 to 7. A method of placing mini-dental implants for animal models, the method comprising: However, in general, the method of implanting the implant is not limited thereto.

Since the mini-dental implant according to the present invention can be applied to a mouse, which is the smallest animal model, it can be used for an implant experiment using animal models of various sizes, Because it can create an environment similar to teeth, it is possible to study the relationship between implant treatment and disease by using animal models with diverse diseases such as diabetes, obesity, and osteoporosis, as well as various substances such as therapeutic agents and accelerators It is expected that it can be used effectively in various experiments related to implants because it can investigate the relation with implant treatment and it is expected that it will be applied to human in future with high efficiency.

1 is a view illustrating a mini-dental implant according to an embodiment of the present invention.
2 is a view illustrating a drill and a driver according to an embodiment of the present invention.
3 is a view illustrating mini-dental implants placed in a mouse according to an embodiment of the present invention.
4 is a view illustrating a mini-dental implant to which a cell sheet according to an embodiment of the present invention is applied.
FIG. 5 is a view showing a result of implanting a cell sheet and a mini-dental implant according to an embodiment of the present invention into a mouse.
FIG. 6 is a diagram showing results of placing two types of cell sheets and mini-dental implants in a mouse according to an embodiment of the present invention.
FIG. 7 is a view showing results of implanting three kinds of cell sheets and mini-dental implants into a mouse according to an embodiment of the present invention.
Figure 8 is a general overview of mini-dental implant experiments of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Example 1: Preparation and municipalization of mini-dental implants for animal models

In order to create the environment most similar to the dental implant currently applied to humans, mini-dental implants were fabricated with titanium material, which is the most commonly used implant material, and mini-dental implants were used in mice The surface of the prepared mini implants was coated with hydroxyapatite having the same biochemical structure as human bones, teeth, and so on. The prepared mini-dental implant is shown in Fig. A drill and a driver corresponding thereto were produced together (FIG. 2).

For mini-dental implant placement, the first molar of 3-week-old mice was extracted and the extracted molar was bred for 6 weeks to fill the alveolar bone. Then, a drill made on a low speed handpiece is tightened to create a socket for implanting a mini-dental implant in the two-piece bone produced, and a mini-dental implant is implanted using the manufactured driver. Respectively. After 4 weeks of incubation, teeth were observed using a micro-CT (microCT). The results are shown in Fig.

As shown in FIG. 3, it was confirmed that osseointegration was formed between the two-day-old bone of the mouse and the mini-dental implant. From the above results, it can be confirmed that the mini-dental implant of the present invention is normally placed on the tooth of the smallest sized animal among the animal models used in the experiment. It was confirmed that a mini-tooth implant can be applied.

Example 2: Application of a cell sheet to a mini-dental implant

In order to enhance the osseointegration efficiency and to provide an environment similar to a natural tooth of a human, mini-teeth prepared in the same manner as in Example 1 so that periodontal ligament (PDL) can be regenerated between a mini-tooth implant and two- A cell sheet was introduced into the implant. For cell sheet preparation, DMEM (Dulbeco's Modified Eagle's Medium) supplemented with 10% fetal bovine serum (FBS) was added to UpCell culture dish (Nunc ^® ) and inoculated with ihPDL cells (immortalized human periodontal ligament cell) In addition, MEM (Minimum Essential Medium) alpha was added to the same culture dish and inoculated with ihCem cells (immortalized human cementoblast cell). Then, the cells were cultured in a condition of 37 ° C and 5% CO ₂ so that the cells could be filled in the culture dish, and then the culture dish was transferred to a 20 ° C incubator to separate the resulting cell sheet from the culture dish surface. The separated cell sheets were cut using a No. 15 surgical blade to fit the size of the mini-dental implants so that they could be used for the experiment, and the cut cell sheets were wrapped in mini-dental implants, Mini-tooth implants were inserted together into the two-day bone (Fig. 4). After 8 weeks of raising the mice, H & E (hematoxylin and eosin) staining revealed normal periodontal ligament or cementum regeneration. The results are shown in Fig.

As shown in FIG. 5, the ihPDL cell sheet or ihCem cell sheet normally adheres between the mini-dental implant and the two-day bone, and the cells fill the gap between the spiral structures of the implant. As a result, the mini-dental implant of the present invention It was confirmed that the mouse was successfully positioned in the jawbone of the mouse.

Example  3: Application of vascular endothelial cells to mini-dental implants

In the case of mini-dental implants performed by the method of Example 2, the ihPDL cell sheet or ihCem cell sheet was normally transferred between the mini-dental implant and the two-day bone, but the arrangement of the periodontal ligament is not limited to mini-dental implant , And it was confirmed that the cementum was partially regenerated. Therefore, in order to overcome this and to create an environment more similar to a human tooth, a human umbilical vein endothelial cell sheet (HUVEC sheet) was introduced into a cell sheet. Human vascular endothelial cell sheet, ihPLD cell sheet, and ihCem cell sheet were prepared in the same manner as in Example 2, respectively. Human vascular endothelial cell sheet and ihPLD cell sheet, and human vascular endothelial cell sheet and ihCem cell The sheets were wound and placed together. The results are shown in Fig.

As shown in Fig. 6, when the ihPLD cell sheet or ihCem cell sheet is located in the two-bone direction and the human vascular endothelial cell sheet is positioned in the direction of the mini-dental implant, the cell sheet is normally immobilized between the mini-dental implant and the two- And it was confirmed that the cementum was regenerated more. However, when the sequence of the cell sheets was reversed, when the human vascular endothelial cell sheet was positioned in the two-bone direction, it was confirmed that the cell sheet was not normally transferred and large blood vessels were formed in the two-day bone. Through the above results, it was confirmed that the success of the implant treatment was determined according to the order of the cell sheets. The introduction of the cell sheet and the human vascular endothelial cells into the mini-dental implant of the present invention, I can confirm that I can do it.

Example  4: Application of various cell sheets to mini-dental implants

The ihCem cell sheet, the ihPDL cell sheet, and the human vascular endothelial cell sheet were both introduced into mini-dental implants for a more environmentally similar composition to human natural teeth. The results are shown in Fig.

As shown in Fig. 7, the ihCEM cell sheet was positioned in the mini-dental implant direction, the human vascular endothelial cell sheet was located, and finally the ihPDL cell sheet was positioned. And the mini - dental implant was implanted, it was confirmed that the environment similar to the human natural teeth was regenerated. When the implants were placed in the above order, the arrangement of the periodontal ligaments was regenerated in the same arrangement as the natural teeth, instead of the partial arrangement of the mini-dental implants. The vessels were normally placed inside the regenerated periodontal ligament, And remodeling of the two bones was observed. In addition, the mini-dental implants of the present invention, which are tightly coupled between the placed mini-dental implants and the regenerated cell tissue, demonstrate not only high binding capacity in animal models, but also an environment similar to human natural teeth I could confirm. From the above results, it can be seen that the mini-dental implant of the present invention can be applied not only to a mouse, which is the smallest animal model, but also to an environment similar to human natural teeth according to the type and / And thus it can be applied to various animal models.

As shown in FIG. 8, the mini-dental implant of the present invention forms a tooth-like tissue similar to a natural tooth of a person in which cementum is formed around the implant and cementum and surrounding bone are connected to a periodontal ligament (PDL) It is expected that various studies will be conducted in animal models to obtain similar experimental results to those in humans.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (25)

Mini-dental implants for animal models, coated with a human PDL cell sheet or a human cementoblast cell sheet, to a tooth size of a laboratory animal model, As tooth implants,
Wherein the mini-dental implant comprises a human umbilical vein endothelial cell sheet (HUVEC sheet) between the mini-dental implant and the coated cell sheet. The method according to claim 1,
Wherein said mini-dental implant is made of titanium or gold. ≪ RTI ID = 0.0 > 15. < / RTI > The method according to claim 1,
Wherein said mini-dental implant is coated with a hydroxyapatite surface. delete The method according to claim 1,
Wherein said human PDL cells are immortalized human periodontal ligament cells (ihPDL cells). The method according to claim 1,
Wherein said human Cem cell is an immortalized human cementoblast cell. The method according to claim 1,
The mini-dental implant for an animal model, wherein the mini-dental implant for an animal model further comprises an ERM cell (epithelial cell rest of Malassez cell sheet). A human cementoblast cell sheet, a human umbilical vein endothelial cell sheet (HUVEC sheet), and a human PDL cell (human umbilical vein endothelial cell sheet) were placed in a mini-dental implant according to a tooth size of a laboratory animal model Mini-dental implants for animal models, with sheets of human periodontal ligament cell sheets coated in order. 9. The method of claim 8,
Wherein said mini-dental implant is made of titanium or gold. ≪ RTI ID = 0.0 > 15. < / RTI > 9. The method of claim 8,
Wherein said mini-dental implant is coated with a hydroxyapatite surface. 9. The method of claim 8,
Wherein said human PDL cells are immortalized human periodontal ligament cells (ihPDL cells). 9. The method of claim 8,
Wherein said human Cem cell is an immortalized human cementoblast cell. 9. The method of claim 8,
The mini-dental implant for an animal model, wherein the mini-dental implant for an animal model further comprises an ERM cell (epithelial cell rest of Malassez cell sheet). (a) fabricating a mini-dental implant to the tooth size of a laboratory animal model;
(b) coating hydroxyapatite on the surface of the prepared mini-dental implant;
(c) coating a human umbilical vein endothelial cell sheet (HUVEC sheet) on the coated mini-dental implant surface; And
(d) coating a human PDL cell sheet or a human cementoblast cell sheet on the mini-dental implant coated with the human vascular endothelial cell sheet. How to make mini-dental implants. delete delete 15. The method of claim 14,
Wherein the human PDL cell is an immortalized human periodontal ligament cell (ihPDL). 15. The method of claim 14,
Wherein the human Cem cells are immortalized human cementoblast cells. 15. The method of claim 14,
Wherein the mini-dental implant for animal models further comprises an ERM cell (epithelial cell rest of Malassez cell sheet). (a) fabricating a mini-dental implant to the tooth size of a laboratory animal model;
(b) coating a human cementoblast cell sheet on the prepared mini-dental implant;
(c) coating a human umbilical vein endothelial cell sheet (HUVEC sheet) on the coated mini-dental implant; And
(d) coating a human PDL cell sheet on the double coated mini-dental implant. < Desc / Clms Page number 20 > 21. The method of claim 20,
Further comprising the step of coating hydroxyapatite on the surface of the mini-dental implant produced after step (a). 21. The method of claim 20,
Wherein the human PDL cell is an immortalized human periodontal ligament cell (ihPDL). 21. The method of claim 20,
Wherein the human Cem cells are immortalized human cementoblast cells. 21. The method of claim 20,
Wherein the mini-dental implant for animal models further comprises an ERM cell (epithelial cell rest of Malassez cell sheet). (a) extracting teeth of an animal model;
(b) breeding the extracted site so that the alveolar bone can be filled;
(c) making an implant socket with the drill bit on the generated two-tooth bone; And
(d) placing the mini-dental implant of any one of claims 1-8.

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