CN113749802A

CN113749802A - Improved porous implant loaded with exosomes derived from Schwann cells

Info

Publication number: CN113749802A
Application number: CN202111154694.0A
Authority: CN
Inventors: 张亮; 王佳乐; 何妍媛; 王欣然; 陈彧洋; 王双成; 边楠雁; 刘云舒; 刘津邑; 孙昊瑞
Original assignee: Sichuan University
Current assignee: Sichuan University
Priority date: 2021-09-29
Filing date: 2021-09-29
Publication date: 2021-12-07

Abstract

The invention discloses an improved porous implant loaded with exosomes, which is suitable for oral implantation and comprises a compact layer, a porous layer and a functional layer; the inner core is a compact layer, and the outer layer of the inner core compact layer is a communicated porous layer, so that the bone tissue can grow in the inner core compact layer, and a support is provided for the functional layer; the outer layer of the porous layer is provided with threads, and the upper section of the porous layer is provided with Morse taper connection, so that the micro-gap at the joint of the implant and the abutment can be reduced, the stability of the implant is improved, and the bone resorption is effectively reduced; the pores of the porous layer are filled with a functional layer, the functional layer is internally provided with a gelatin compound loaded with exosomes from Schwann cells, and the exosome-gelatin compound can uniformly release the exosomes from Schwann cells to the peripheral bone tissues and has the functions of repairing damaged nerves, regulating tissue inflammation and promoting bone regeneration. The invention has the functions of repairing damaged nerves, regulating tissue inflammation and promoting bone regeneration, effectively relieves the problem of bone absorption after implantation, and can improve the stability of the initial stage of implantation to a certain extent.

Description

Improved porous implant loaded with exosomes derived from Schwann cells

Technical Field

The invention relates to the technical field of medical instruments, in particular to an improved porous implant loaded with exosomes derived from Schwann cells.

Background

The oral implant treatment is a tooth loss repairing means realized by implant materials. The current commonly used planting material in clinic is titanium and titanium alloy material, and has the advantages of light weight, relatively low elastic modulus, strong shock absorption capability, higher hardness, excellent corrosion resistance and the like. However, the titanium and titanium alloy materials still have the following problems in application: 1. the titanium and titanium alloy materials are biological inert materials, are difficult to be connected with surrounding tissues to form a whole, generally need 3-6 months to realize osseointegration, and have long combination time. 2. The elastic modulus of titanium is far higher than that of surrounding bone tissues, so that a stress shielding effect is easily generated around the implant, bone absorption and bone atrophy around the implant are caused, and finally the implant is loosened and falls off.

The porous scaffold material can reduce the overall elastic modulus of the material on one hand, and on the other hand, the formed hole structure can guide bone tissues to grow into a firmer embedding structure. Is beneficial to cell growth, extracellular matrix deposition, nutrient and oxygen entry and metabolite discharge, and provides conditions for bone tissue growth. The Morse taper connection structure can generate larger friction and help the retention, and reduce the stress. The porous titanium implant is combined with the Morse taper connection, so that the stability of the implant can be obviously improved.

Exosomes can transfer their rich proteins and RNA to damaged nerve cells to enhance axonal regeneration of damaged nerves to effect repair of peripheral nerve injury. The existing research finds that Schwann cells can play a role in regulating inflammation around the implant, effectively reduce the degeneration and absorption of soft and hard tissues of the implant, and the exosomes derived from Schwann cells play a great role in regulating inflammation. In addition, the exosome from Schwann cells can also accelerate the repair capacity of damaged cartilage tissues, so the exosome also has the promotion effect on the regeneration of damaged and atrophic bone tissues around the implant.

Therefore, if the Schwann cell exosome is combined with the porous material and is combined with the Morse taper connecting structure to be used for modifying the titanium alloy implant, the osseointegration can be effectively promoted, the inflammatory reaction around the implant is improved, and the stability of the implant is obviously improved.

Disclosure of Invention

The invention aims to overcome the functional defects of the existing implant and provide an improved porous implant loaded with exosomes from Schwann cells, which has the advantages of simple structure and stable planting, and is beneficial to improving the problems of peringitis, bone resorption and the like in the application of the implant.

In order to achieve the purpose, the technical scheme of the invention is as follows:

an improved porous implant loaded with schwann cell-derived exosomes includes a dense layer, a porous layer, and a functional layer. The porous layer is positioned on the surface of the implant and between the threads, the compact layer is coated by the porous layer, and the functional layer is filled between pores of the porous layer.

Preferably, the dense layer is made of a titanium alloy material.

Preferably, the porous layer has a rough surface, and the pore structure is favorable for filling the functional layer.

Preferably, the functional layer carries a bio-filler material that can be released uniformly.

The invention also provides a construction method of the biological filling material in the technical scheme, the biological filling material comprises a gelatin solution and an exosome solution, and the construction method comprises the following steps:

1. exosome extraction

Exosome EXOs is derived from Schwann cell strain RSC96 CRL-2765, and the extraction comprises the following steps:

1) culturing Schwann cells SCs for 24h, taking cell supernatant, centrifuging for 10min at 300 Xg, 10min at 2000 Xg, 30min at 10000 Xg, removing precipitate, and collecting supernatant;

2) centrifuging at 100000 × g for 70min by ultracentrifugation, precipitating, resuspending with PBS, centrifuging at 100000 × g for 70min, and removing supernatant to obtain EXOs.

2. Filling of biological material

1) Carrying out acid etching on the implant to obtain an acid-etched implant;

2) soaking the implant subjected to acid etching in the step 1) in a gelatin solution for 10-15 min, and cleaning to obtain a gelatin-loaded implant;

the method comprises the steps of soaking the acid-etched implant in a gelatin solution for 10min, preferably washing for 2 times, preferably washing for 1min each time, and preferably rinsing.

3) Soaking the implant loaded with gelatin obtained in the step 2) in an exosome solution for 10-15 min, and cleaning to obtain an implant loaded with exosomes;

the method comprises the steps of soaking the gelatin-loaded implant in an exosome solution for 10min, preferably washing for 2 times, preferably washing for 1min each time, and preferably rinsing.

4) After the implant loaded with the exosomes is obtained, the method preferably repeats the step 2) and the step 3) for 4-5 times to obtain the improved implant.

Preferably, the implant and the abutment are connected by a Morse taper structure.

Through the technical scheme, the invention has the beneficial effects that:

the inner core is hard and not easy to break, the porous structure on the surface increases the roughness of the implant and promotes the integration of the implanted bone, and the exosome-gelatin compound between the threads on the surface of the implant can uniformly release exosomes from Schwann cells to the surrounding bone tissues, so that the exosome-gelatin compound has the functions of repairing damaged nerves, adjusting tissue inflammation and promoting bone regeneration, and the Morse taper connection improves the stability of the implant and effectively reduces bone resorption by reducing the micro-gap at the joint of the implant and the base station.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

Fig. 1 is a schematic view of the novel implant-abutment;

figure 2 is a view of the novel implant-abutment connection;

FIG. 3 is a schematic view of the novel implant profile;

fig. 4 is a cross-sectional side view of the novel implant;

fig. 5 is a cross-sectional straight view of the novel implant.

Reference numerals

1-a dense layer; 2-a porous layer; 3-a functional layer; 4-implant; 5-screw thread.

Detailed Description

In order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further explained below by combining the specific drawings.

Referring to fig. 1, an improved porous implant loaded with exosomes derived from schwann cells comprises a dense layer 1, a porous layer 2, and a functional layer 3. The porous layer is positioned on the surface of the implant 4 and between the threads 5, the compact layer 1 is coated by the porous layer 2, and the functional layer 3 is filled between the pores of the porous layer 2.

Preferably, the dense layer is made of a titanium alloy material.

1. exosome extraction

2. Filling of biological material

1) Carrying out acid etching on the implant to obtain an acid-etched implant;

2) soaking the implant subjected to acid etching in the step 1) in a gelatin solution for 10-15 min, and cleaning to obtain the gelatin-loaded implant.

The method comprises the following steps of soaking the acid-etched implant in a gelatin solution for 10min, preferably washing for 2 times, preferably washing for 1min each time, and preferably rinsing in a washing mode;

3) soaking the implant loaded with gelatin obtained in the step 2) in an exosome solution for 10-15 min, and cleaning to obtain the implant loaded with exosomes.

The method comprises the steps of soaking the implant loaded with gelatin in an exosome solution for 10min, preferably washing for 2 times, preferably washing for 1min each time, and preferably rinsing in a washing mode;

Preferably, the implant 4 is connected with the abutment 5 in a morse taper structure.

The foregoing shows and describes the general principles and broad features of the present invention and advantages thereof. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.

Claims

1. An improved porous implant loaded with schwann cell-derived exosomes, comprising: the high-strength composite material comprises a compact layer (1), a functional layer (3), an implant (4) and a thread (5), and is characterized in that a porous layer (2) is arranged on the surface of the compact layer (1).

2. The improved porous implant loaded with exosomes derived from schwann cells as claimed in claim 1, characterized in that the pores of said porous layer (2) are filled with a functional layer (3).

3. The modified porous implant loaded with exosomes derived from schwann cells according to claim 1, wherein said steps comprise:

(1) extracting exosomes EXOs derived from Schwann cell SCs; (2) and (4) filling the biological material.

4. The modified porous implant loaded with exosomes derived from schwann cells as claimed in claim 2, wherein step (1) comprises: performing grouped culture on SCs for 24h, taking cell supernatant, centrifuging for 10min at 300 Xg, centrifuging for 10min at 2000 Xg, centrifuging for 30min at 10000 Xg, and removing precipitate to collect supernatant; centrifuging at 100000 × g for 70min by ultracentrifugation, precipitating, resuspending with PBS, centrifuging at 100000 × g for 70min, and removing supernatant to obtain EXOs.

5. The modified porous implant loaded with exosomes derived from schwann cells as claimed in claim 2, wherein step (2) comprises: carrying out acid etching on the implant to obtain an acid-etched implant; soaking the acid-etched implant in a gelatin solution, wherein the soaking time of the acid-etched implant in the gelatin solution is preferably 10min, the cleaning frequency is preferably 2 times, the cleaning time is preferably 1min each time, and the cleaning mode is preferably rinsing; soaking the obtained gelatin-loaded implant in an exosome solution, soaking the gelatin-loaded implant in the exosome solution for 10min, preferably washing for 2 times, preferably washing for 1min each time, and preferably rinsing in a washing mode; after the implant loaded with the exosomes is obtained, gelatin soaking and exosome soaking are preferably repeated for 4-5 times to obtain the improved implant.

6. The improved porous implant loaded with exosomes derived from schwann cells as claimed in claim 1, wherein the implant (4) is connected with the abutment (5) in a Morse taper configuration.