阅读说明：本技术 一种超顺磁性纳米颗粒骨修复膜及其制备方法 (Superparamagnetic nanoparticle bone repair membrane and preparation method thereof ) 是由 孙剑飞 李政 顾宁 于 2020-05-25 设计创作，主要内容包括：本发明公开了一种超顺磁性纳米颗粒骨修复膜及其制备方法,所述超顺磁性纳米颗粒修饰的骨修复膜,是从牛腱中提取的以酶解法得到的胶原蛋白为基质,组装上超顺磁性纳米颗粒(γ-Fe<Sub>2</Sub>O<Sub>3</Sub>)。在骨创伤外科手术后,由于骨愈合过程中由于结缔组织增生速度较骨组织快,增生的结缔组织过早的占据骨缺损区域从而阻碍了骨组织的长入,导致骨不连或愈合缓慢。因此在骨缺损处放置超顺磁性纳米颗粒修饰的骨修复膜,该膜可作为一种机械屏障,控制周围组织长入骨缺损区,为骨细胞的生长提供一个引导空间；其次,骨创伤手术手术术后,在外加特定磁场的作用下超顺磁性纳米颗粒具有的电磁效应和其他间接效应,能促进骨组织损伤的修复,达到加速康复的作用。(The invention discloses a superparamagnetic nanoparticle bone repair membrane and a preparation method thereof, wherein the superparamagnetic nanoparticle modified bone repair membrane is prepared by taking collagen extracted from bovine tendon and obtained by an enzymolysis method as a matrix and assembling superparamagnetic nanoparticles (gamma-Fe) 2 O 3 ). After the surgical operation of bone trauma, since the proliferation speed of the connective tissue is faster than that of the bone tissue in the bone healing process, the proliferated connective tissue occupies the bone defect area too early to block the growth of the bone tissue, thus leading to bone nonunion or slow healing. Therefore, a bone repair membrane modified by superparamagnetic nano particles is placed at the bone defect part, and the membrane can be used as a mechanical barrier to control the peripheral tissues to grow into the bone defect areaProviding a guide space for the growth of bone cells; and secondly, after the bone trauma surgery, the superparamagnetic nanoparticles have electromagnetic effect and other indirect effects under the action of an external specific magnetic field, so that the repair of the bone tissue injury can be promoted, and the effect of accelerating recovery is achieved.)

1. A superparamagnetic nanoparticle bone repair membrane is characterized in that: the bone repair membrane is a natural biological inactivated acellular dermal matrix collagen membrane with gamma-Fe on the surface₂O₃A coating of superparamagnetic nanoparticles.

2. The superparamagnetic nanoparticulate bone repair film according to claim 1, wherein: the bone repair membrane is prepared by extracting collagen from tendon of cattle with enzymolysis as matrix, and assembling superparamagnetic nanoparticle (gamma-Fe) on the surface₂O₃）。

3. A preparation method of a superparamagnetic nanoparticle bone repair membrane is characterized by comprising the following steps: the method comprises the following steps:

step 1: removing sarcolemma and fascia from adult bovine achilles tendon, washing with normal saline for 8-12 times, soaking in 75% alcohol solution for 15-25min, freezing in a low temperature refrigerator of-60 deg.C, cutting into 0.6-1.2mm slices, and soaking in acetic acid solution for 1-3 hr;

step 2: repeatedly washing the slices with PBS for 20-30 times and soaking for 1-2 h, washing with distilled water for 20-30 times, further digesting with trypsin, ultrasonic treating for 44-52 h, washing with PBS for 20-30 times, and washing with distilled water for 30-40 times;

and step 3: placing the cell-removed slice into formaldehyde solution for 2-3 h, taking out, washing with PBS for 30-35 times, and washing with distilled water for 20-30 times;

and 4, step 4: after the above steps are completed, the collagen membrane is placed on the gamma-Fe₂O₃And (3) putting the nano-particle colloidal solution into a freeze dryer for freeze drying at the temperature of 30-40 ℃ for 15-20 min, and taking out the nano-particle colloidal solution for cobalt-60 (Co) irradiation disinfection.

4. The superparamagnetic nanoparticulate bone repair film according to claim 1, wherein: the using method of the bone repair membrane comprises the following steps: the superparamagnetic iron oxide magnetic nano particle is assembled on a bone repair membrane, and can generate an electromagnetic effect and other indirect effects under the action of an external magnetic field to promote the repair of bone tissue damage.

Technical Field

The invention belongs to the field of medical materials, and particularly relates to a superparamagnetic nanoparticle bone repair membrane and a preparation method thereof.

Background

Periosteum is a dense connective tissue membrane covering the surface of bone (except joint surfaces), and fresh periosteum is pink and rich in blood vessels, nerves and osteoblasts. The periosteum is divided into an inner layer and an outer layer, and the fibrous layer contains fibroblasts, collagen and elastic fibers, and a network of nerves and capillaries, so that the mechanical stability of the periosteum is provided. The cambium stores a large number of different types of cells, providing a source of cells for bone formation and repair. The role of natural periosteum in maintaining normal skeletal blood circulation and metabolism has been widely recognized by anatomical, physiological, pathological and clinical orthopedics medicine. Clinically, the protection and reconstruction of periosteum directly affect the success or failure of bone trauma or bone disease treatment. Aiming at defective bone grafting, the application of a bone repair membrane guided regeneration technology can effectively prevent connective tissues from occupying a bone defect area too early and promote bone healing. The artificial periosteum can effectively prevent connective tissues from growing into a junction area between the bone grafting material and the host bone defect by wrapping the bone defect part with the artificial periosteum, and creates good conditions for guiding the bone tissues with slower growth speed to migrate into the bone defect area.

The magnetic field is a non-invasive physical factor and can directly act on organisms to generate corresponding biological effects, and when the magnetic field of the organisms is changed, the activity of cell molecules and the activity of the organisms are influenced. A tissue of a living body contains a large number of free electrons, and these free electrons are stationary in a natural state. However, when the organism tissue is in an electromagnetic environment, the free electrons in the organism tissue are excited to move continuously, so that the organism in the changing electromagnetic field can generate a very strong response under very small stimulation. At present, the research on treating bone diseases by utilizing the electromagnetic effect is carried out in China, and the feasibility of the electromagnetic effect for exerting the treatment effect is proved. The magnetic nanoparticles are a chemical compound formed by combining nanoparticles and magnetic elements such as iron, nickel and the like, and have the advantages of special magnetic effect, magnetic guidance, good biocompatibility, low toxicity, long blood circulation time, specific advantages due to small size and the like. In one aspect, the gamma-Fe used in the present invention₂O₃The iron oxide particles have passed the safety certification of the ISO international quality control system and have low cytotoxicity to organisms; on the other hand, the remanence Mr and the coercive force Hc of the superparamagnetic nano-particles basically tend to be zero,resulting in only very weak magnetic dipole interaction between particles and a stable magnetic fluid can be formed. At present, the iron oxide nanoparticles are widely applied biomedical nano materials, have good biocompatibility, in-vivo stability and special electromagnetic properties, can generate a plurality of unique effects under an external electromagnetic field, and are increasingly researched and paid attention to magnetic drug carriers, tumor thermotherapy and brain deep magnetic stimulation.

Disclosure of Invention

In order to solve the problems, the invention discloses a superparamagnetic nanoparticle bone repair membrane and a preparation method thereof.

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

a superparamagnetic nanoparticle bone repair film is a natural biological inactivated acellular dermal matrix collagen film with gamma-Fe on the surface₂O₃A coating of superparamagnetic nanoparticles.

Further, the bone repair membrane is collagen extracted from bovine tendon and obtained by enzymolysis as matrix, and superparamagnetic nano-particles (gamma-Fe) are assembled on the surface of the collagen₂O₃）。

A preparation method of a superparamagnetic nanoparticle bone repair membrane comprises the following steps:

step 1: removing sarcolemma and fascia from adult bovine achilles tendon, washing with normal saline for 8-12 times, soaking in 75% alcohol solution for 15-25min, freezing in a low temperature refrigerator of-60 deg.C, cutting into 0.6-1.2mm slices, and soaking in acetic acid solution for 1-3 hr.

Step 2: washing the slices with PBS for 20-30 times and soaking for 1-2 h, washing with distilled water for 20-30 times, further digesting with trypsin, ultrasonic treating for 44-52 h, washing with PBS for 20-30 times, and washing with distilled water for 30-40 times.

And step 3: and (3) placing the cell-removed slice into a formaldehyde solution for 2-3 h, taking out, washing with PBS for 30-35 times, and washing with distilled water for 20-30 times.

And 4, step 4: after the above steps are completed, the collagen membrane is placed on the gamma-Fe₂O₃And (3) putting the nano-particle colloidal solution into a freeze dryer for freeze drying at the temperature of 30-40 ℃ for 15-20 min, and taking out the nano-particle colloidal solution for cobalt-60 (Co) irradiation disinfection.

The invention has the beneficial effects that:

the superparamagnetic nanoparticle acellular dermal matrix collagen membrane is implanted into a bone injury part, and performs a specific action on bone tissues by utilizing the electromagnetic effect and other indirect effects of the magnetic nanoparticles under an externally applied electromagnetic field, such as a magnetocaloric effect, a magnetoelastic effect and the like. Because the magnetic material can gather the electromagnetic field, the action intensity of the electromagnetic field is locally enhanced in the bone tissue, the electromagnetic action energy at the interface of the magnetic nanoparticle assembly structure and the tissue cells is greatly increased, and the point can ensure that a stronger electromagnetic effect can be generated at the interface of the nano material even if only the electromagnetic field with lower intensity exists in the external environment. Meanwhile, the electromagnetic field generated by the polarization or magnetization of the nanoparticle interface is quickly attenuated along with the increase of the distance, so that the electromagnetic effect is a local electromagnetic effect, the in-vivo electromagnetic effect is only acted on the local part of the bone injury tissue, and the simultaneous influence on multiple parts of the whole body is avoided.

Detailed Description

The present invention will be further illustrated below with reference to specific embodiments, which are to be understood as merely illustrative and not limitative of the scope of the present invention.

The bone repair membrane is a natural inactivated acellular dermal matrix collagen membrane, can be collagen extracted from bovine tendon and obtained by enzymolysis, and is assembled with superparamagnetic nanoparticles (gamma-Fe) on the surface₂O₃）。

The application method of the superparamagnetic nanoparticle bone repair membrane comprises the following steps: the superparamagnetic iron oxide magnetic nano particle is assembled on a bone repair membrane, and can generate an electromagnetic effect and other indirect effects under the action of an external magnetic field to promote the repair of bone tissue damage.

The superparamagnetic nanoparticle bone repair membrane provided by the invention has the preparation method that