WO2014185600A1 - Bone cement composition and preparation method therefor - Google Patents

Abstract

The present invention relates to a bone cement composition and a preparation method therefor, and more specifically, to a bone cement composition including natural antibiotics, and a preparation method therefor.

Description

Bone Cement Composition and Method for Making the Same

The present invention relates to a bone cement composition and a method for producing the same, and more particularly, to a bone cement composition comprising a natural antibiotic and a method for producing the same.

Conventional bone filling compositions, that is, bone cement compositions, are widely used in the world as adhesives for fixing a bone defect or a metal artificial joint such as an artificial hip joint with surrounding bone. The bone cement composition is not only used in the method of inserting and inserting an artificial body into the body due to breakage of the spine and bone, but also a total knee arthroplasty and reoperation due to various joints, spine and bone damage. There is a bone cement composition is used to fix the position of the articulated body in the body for the role of filling bone defects during this operation.

Such bone cement composition is usually divided into a powder part and a liquid part, and is kneaded just before application during surgery or the like to initiate the polymerization reaction of the monomer, and leave the kneaded material to a certain high viscosity state. It has been used as being applied to the application target site by the handling operation at the time it became.

In addition, in general, bone cement composition is also used as a method of treating various spinal diseases such as vertebral compression fractures caused by osteoporosis, by filling and maintaining the empty space of the spine.

Looking at the related art related to bone cement, "bone cement composition and its manufacturing method" (Korean Patent Publication No. 10-2013-002 8405, Patent Document 1), β-3 calcium phosphate (β-tricalcium phosphate; ß- TCP), a bone cement composition comprising ammonium monophosphate (NH ₄ H ₂ PO ₄ ), diammonium phosphate ((NH ₄ ) ₂ HPO ₄ ) and water, the starting powder being β-3 calcium phosphate powder Preparing (step 1); Preparing a purification solution by mixing and stirring the first ammonium phosphate, the second ammonium phosphate and water (step 2); The step of introducing the cured liquid prepared in step 2 to the ß-3 calcium phosphate powder prepared in step 1 and stirring (step 3); is disclosed a configuration comprising a. This technique has extended the curing time so that it can be used in a three-dimensional molding process and the like. However, this bone cement has a problem that it is difficult to identify the shape or location when taking a picture using radiation after the procedure.

As a technique to solve this problem, "bone cement system for bone reinforcement" (Korean Patent Publication No. 10-2013-0028056, Patent Document 2) is a solid component of about 38 to about 42% by weight of the solid component Contrast agent; A polymerization initiator in the range of about 0.3 to about 0.5 weight percent of the solid component; Bone replacement material comprising hydroxyapatite in the range of about 14 to about 16 weight percent of the solid component, and poly (methylacrylate-co-methylmethacrylate), in the range of about 43 to about 46 weight percent of the solid component, Comprising at least one of poly (meth) acrylate, polymethyl (meth) acrylate, poly (methylmethacrylate) and / or poly (methylmethacrylate-co-styrene) polymers, blends, mixtures or copolymers Examples are disclosed which consist of solid polymers and comprise zirconium dioxide, barium sulfate and / or titanium dioxide and mixtures and blends thereof to form the contrast agent.

An example of using a contrast agent in the range of 30 to 60% by weight in the powder component is disclosed as an example of the use of the contrast agent. In particular, in the case of barium sulfate also has the role of improving the mechanical strength of the bone cement has the advantage of providing excellent strength after the procedure.

On the other hand, a technique for the purpose of treating inflammation at the site of bone sivent injection is described in "Bone Cement Composition with Fiber-Reinforced and / or Increased Fluidity" (Korean Patent Publication No. 10-1226811, Patent Document 3). It has been published.

The said technique is a (meth) acrylate type polymer large diameter supplier whose average particle orientation is 10-60 micrometers, (meth) acrylate type polymer small diameter particle | grains whose average particle diameter is 0.1-2.0 micrometers, and a (meth) acrylate type monomer And a polymerization initiator, and the content ratio of the (meth) acrylate-based polymer small-diameter particles is 5 to 30 with respect to the total amount of the (meth) acrylate-based polymer small-diameter particles and the (meth) acrylate-based polymer large-diameter particles. It is characterized by mass%, having a calcium-salt containing component, a glycolide repeating unit, discontinuous fibers to provide fluidity of bone cement, and the addition of antibiotics therein is disclosed.

Patent document 3 as described above is characterized in that it uses a wide range of antibiotics. However, depending on the type of antibiotics there is a problem that can not match the components for providing the fluidity of the bone sheet to reduce the fluidity, excessively lower the polymerization time or lower the physical properties of the bone cement.

 The present invention is to solve the problems occurring in the prior art as described above, the present invention includes a natural ingredient that can reduce the side effects due to the use of conventional antibiotics, while having an antimicrobial function and minimize the side effects on the human body, It is possible to provide a bone cement composition having good flowability without using a polymerization inhibitor and having a shorter polymerization time (setting time) than conventionally.

The present invention includes a liquid phase including methyl methacrylate (MMA), dimethyl-p-toluidine (DMPT) and caffeic acid phenethyl ester (CAPE); And a powder comprising polymethyl methacrylate (PMMA), zirconia, and benzoyl peroxide.

According to a preferred embodiment of the present invention, the liquid phase may include 150 to 200 parts by weight of methyl methacrylate and 1 to 10 parts by weight of dimethylphytoluidine based on 1 part by weight of caffeic acid ester, and the powder is caffeic acid It may include 200 to 300 parts by weight of polymethyl methacrylate, 100 to 150 parts by weight of zirconia and 5 to 20 parts by weight of benzoyl peroxide based on 1 part by weight of ester.

According to another preferred embodiment of the present invention, the liquid phase may include 170 to 180 parts by weight of methyl methacrylate and 5 to 6 parts by weight of dimethylphytoluidine per 1 part by weight of caffeic acid ester, and the powder It may include 250 to 280 parts by weight of polymethyl methacrylate, 110 to 130 parts by weight of zirconia and 10 to 15 parts by weight of benzoyl peroxide based on 1 part by weight of phosphate ester.

According to another preferred embodiment of the present invention, the average particle diameter of the polymethyl methacrylate may be 10 ~ 100 ㎛.

In addition, the present invention is a liquid phase including methyl methacrylate (methyl methacrylate; MMA), dimethyl p-toluidine (DMPT) and caffeic acid phenethyl ester; And preparing a bone cement raw material of polymethyl methacrylate (PMMA), a powder comprising zirconia and benzoyl peroxide, and mixing the liquid and powder bone cement raw materials. It provides a bone cement manufacturing method comprising the step of.

According to a preferred embodiment of the present invention, the liquid phase may include 150 to 200 parts by weight of methyl methacrylate and 1 to 10 parts by weight of dimethylphytoluidine based on 1 part by weight of caffeic acid ester, and the powder is caffeic acid It may include 200 to 300 parts by weight of polymethyl methacrylate, 100 to 150 parts by weight of zirconia and 5 to 20 parts by weight of benzoyl peroxide based on 1 part by weight of ester.

According to another preferred embodiment of the present invention, the reaction time may be 5 to 15 minutes.

According to another preferred embodiment of the present invention, the mixing may be performed at 20 ~ 25 ℃.

The present invention includes a natural antibiotic, caffeic acid phenethyl ester, has antibacterial properties, has less side effects on the human body, has good flowability without using a separate polymerization inhibitor, and shorter polymerization time (setting time) than before. Eggplant may provide a bone sivent composition and a method of making bone cement.

In addition, the bone cement composition of the present invention is a separate antibiotic by containing caffeic acid ester which is an active ingredient of natural propolis with antibacterial and anti-inflammatory effect confirmed stability problems such as reverse mutation, micronucleus test, etc. that may occur when administered in the human body It is possible to prevent the occurrence of fungus, which is a factor of secondary infection that occurs during surgery, and furthermore can provide a bone cement composition that can not use the polymerization inhibitor because caffeic acid ester functions as a polymerization inhibitor. .

Figure 1 shows a part implanted in the femoral portion of the rabbit to examine the properties appearing in vivo implantation of the bone cement composition of the present invention.

Figure 2 shows the results of histological examination of the femoral twelve weeks after implanting the bone cement composition of the Examples and Comparative Examples of the present invention in the rabbit thigh.

3 shows the injection position for testing intradermal safety for bone cement compositions of Examples and Comparative Examples of the present invention.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail. However, the best mode for carrying out the invention described below is provided to those skilled in the art to fully understand the present invention, and may be modified in many different forms, and the scope of the present invention is as follows. It is not limited to the described embodiment.

In the present invention, it was found that the antimicrobial effect was suppressed without the addition of a separate polymerization inhibitor such as hydroquinone as a result of mixing the caffeic acid ester as a natural antibiotic in the liquid phase and mixed with the powder.

The caffeic acid ester is added as an antibacterial component and a polymerization inhibitor and is one of active ingredients of propolis, which is a kind of flavonoid-based component known as vitamin P having an antimicrobial effect, and mainly acts as an antibacterial anti-inflammatory agent in propolis. It is known. The molecular formula of the caffeic acid ester is C ₁₇ H ₁₆ 0 _{4 And the} chemical structure is shown in the formula (1) below.

Formula 1

The caffeic acid ester has an antibacterial, antifungal and antiviral effect in the human body. In addition, the caffeic acid esters inhibit inflammation by blocking the secretion of enzymes that cause inflammation. In particular, because it is water-soluble, its range of use is diverse, and even if ingested excessively characterized by nontoxic.

When the human body becomes acidic and the waste accumulates in the body, the waste accumulates around the cells, and the cells begin to melt by this acid residue. Cytotoxicity such as PG is produced, causing inflammatory reactions such as fever and soreness. In the process, enzymes such as lipoxygenase and cyclouxinase are required to make cytotoxic components, ie, leucotriene (LT) and prostaglandin (PG), which cause inflammation.

Caffeic acid ester, an active component of propolis, exhibits anti-inflammatory activity because it simultaneously inhibits two enzymes, lipoxygenase and cyclooxygenase, which produce leucotriene (LT) and prostaglandin (PG). It is more potent than regular painkillers.

The bone cement composition of the present invention contains a liquid comprising metal methacrylate, dimethyl phytoluidine and caffeic acid phenethyl ester and a powder containing polymethyl methacrylate, zirconia, and benzoyl peroxide, The patient can be used by mixing the liquid and powder during the procedure.

In the bone sieve composition of the present invention, when the powder and the liquid are mixed, the polymerization initiator and the liquid acrylate monomer in the powder react to perform the polymer polymerization reaction of the acrylate monomer, and the polymerization accelerator is polymerized at room temperature. You can accelerate the reaction so that this happens better.

In addition, the bone cement composition of the present invention comprises a caffeic acid ester in the liquid phase, the liquid phase comprises 150 to 200 parts by weight of methyl methacrylate and 1 to 10 parts by weight of dimethyl phytoluidine relative to 1 part by weight of caffeic acid ester It is preferable.

If the ratio of caffeic acid ester is added higher than the above range, the function of the caffeic acid ester as a polymerization inhibitor is strengthened, so that the polymerization time is increased so that it may be difficult to use as a bone cement composition, and caffeic acid ester Although the ratio of is lower than the above range, even if the caffeic acid ester is added, there may be a problem in that it does not exhibit antimicrobial efficacy.

Furthermore, more preferably, a liquid phase containing 170 to 180 parts by weight of methyl methacrylate and 5 to 6 parts by weight of dimethylphytoluidine may be used based on 1 part by weight of caffeic acid ester.

In addition, the powder in the bone cement composition of the present invention is not particularly limited as long as it is a mixing ratio to increase the strength of the bone filling portion, preferably 200 to 300 parts by weight of polymethyl methacrylate relative to 1 part by weight of caffeic acid ester, It may comprise 100 to 150 parts by weight of zirconia and 5 to 20 parts by weight of benzoyl peroxide, more preferably 250 to 280 parts by weight of polymethyl methacrylate, 110 to 130 parts by weight of zirconia and 1 part by weight of caffeic acid ester and It may include 10 to 15 parts by weight of benzoyl peroxide.

[Acrylate polymer]

The acrylate polymer included in the bone cement composition of the present invention is polymethyl methacrylate (PMMA; Polymethyl methacrylate), poly methylacrylate (poly methylacrylate), polystyrene, and copolymers thereof Preferably, the acrylate polymer is preferably in the form of beads of 10 to 100 μm because the acrylate polymer is pre-dissolved in the liquid phase.

[(Meth) acrylate monomer]

The acrylate monomer contained in the liquid phase of the bone cement composition of the present invention constitutes a component for forming a base material of bone cement, and the bone cement composition is cured by polymerizing the acrylate monomer, which is the polymerizable monomer, and as a result, a cured product. Is obtained.

As an acrylate-type monomer of this invention, methyl (meth) acrylate, ethyl (meth) acrylate, isobutyl (meth) acrylate, hexyl (meth) acrylate, octyl (meth) acrylate, dodecyl (meth) At least one selected from the group consisting of a) acrylate, N-isopropyl (meth) acrylamide, hydroxyethyl methacrylate and acrylonitrile is preferred.

In the present invention, the acrylate monomer is preferably included in 150 to 200 parts by weight based on 1 part by weight of the caffeic acid ester.

[Polymerization Initiator]

As the polymerization initiator contained in the bone cement composition of the present invention, benzoyl peroxide, tert-butyl peroxide, lauroyl peroxide, azobisisobutyronitrile and the like can be used. Moreover, in these, since the polymerization reaction of the acrylate-type monomer contained in the said liquid phase starts quickly, and it is easy to continue the reaction, it is preferable to use benzoyl peroxide.

It is preferable that the powder of the bone cement composition of this invention contains 5-20 weight part of benzoyl peroxides which are the said polymerization initiators with respect to 1 weight part of caffeic acid esters.

If the polymerization initiator is included in an amount of less than 5 parts by weight with respect to 1 part by weight of the caffeic acid ester, a problem may occur in that the polymerization reaction of the acrylate monomer may not proceed, and the polymerization initiator may be included in 1 part by weight of the caffeic acid ester. If it contains an amount exceeding 20 parts by weight, a problem may occur that the polymerization initiator remains in the polymer formed by the polymerization of the acrylate monomer.

In addition, benzoyl peroxide added as the polymerization initiator is added to the powder, and when the liquid and the powder is kneaded immediately before the operation is mixed with the liquid metal methacrylate monomer to allow the polymerization to be initiated.

[Filler and molding agent]

Fillers may include inorganic materials such as titanium dioxide, calcium phosphate (hydroxyapatite, tricalcium phosphate), barium sulfate, silicon oxide (silica), aluminum oxide (alumina), zirconium oxide (zirconia), and the like. Alternatively, a combination of two or more kinds can be used. Among them, zirconium oxide (zirconia) having an X-ray contrast effect is preferable because it improves radiopacity as compared with barium sulfate used as a conventional molding agent. As described above, the radiopacity is increased so that the position and shape of the X-ray or the like after imaging in the body can be understood.

Zirconium oxide (zirconia) of the present invention may be included in 100 to 150 parts by weight based on 1 part by weight of the caffeic acid ester.

[Polymerization accelerator]

The bone cement composition of the present invention preferably contains a polymerization accelerator together with a polymerization initiator in order to advance the polymerization reaction of the metal methacrylate monomer more quickly.

The polymerization accelerator may include dimethyl-p-toluidine, 2,4,6-tris (dimethylaminomethyl) phenol, and the like. In these, since the polymerization reaction of an acrylate-type monomer advances rapidly, it is preferable to use dimethyl pituluidine.

The dimethyl pituluidine is preferably included 1 to 10 parts by weight based on 1 part by weight of caffeic acid ester.

If the polymerization accelerator is included in an amount less than 1 part by weight based on 1 part by weight of caffeic acid ester, it may be difficult to cause the polymerization reaction of the acrylate monomer to proceed. When it contains an amount exceeding 10 parts by weight, a problem may occur that causes a side effect of the polymerization accelerator remaining in the cured product formed by polymerization of the acrylate monomer.

In addition, the present invention is a liquid phase containing methyl methacrylate (methyl methacrylate), dimethyl p-toluidine (dimethyl-p-toluidine) and caffeic acid phenethyl ester; And a powder comprising polymethyl methacrylate, zirconia, and benzoyl peroxide; preparing bone cement raw material; and reacting after mixing the liquid and powder bone cement raw material. It provides a bone cement manufacturing method comprising a.

The liquid phase is not particularly limited as long as it contains methyl methacrylate, dimethylphytoluidine and caffeic acid ester, but preferably 150 to 200 parts by weight of methyl methacrylate and 1 based on 1 part by weight of caffeic acid ester. It may comprise from 10 parts by weight of dimethyl phytoluidine, more preferably from 170 to 180 parts by weight of methyl methacrylate and 5 to 6 parts by weight of dimethyl phytoluidine relative to 1 part by weight of caffeic acid ester.

Further, the powder is not particularly limited as long as it contains polymethyl methacrylate, zirconia and benzoyl peroxide, but preferably 200 to 300 parts by weight of polymethyl methacrylate, based on 1 part by weight of caffeic acid ester, It may comprise 100 to 150 parts by weight of zirconia and 5 to 20 parts by weight of benzoyl peroxide, more preferably 250 to 280 parts by weight of polymethyl methacrylate, 110 to 130 parts by weight of zirconia and 1 part by weight of caffeic acid ester and It may include 10 to 15 parts by weight of benzoyl peroxide.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the present invention is not limited by the following examples.

Example 1 Preparation and Mixing of Antibacterial Bone Cement Composition

First, 240 g of PMMA constituting the powder part, 107.46 g of zirconia, and 10.75 g of benzoyl peroxide were weighed, and stirred for 3 hours in a tube mixer so that each component could be sufficiently mixed to prepare a powder. Thereafter, the powder was packed in 20g portions. The packaged powder was sterilized by gamma-ray.

Liquid phase was prepared by mixing MMA 155.47g, DMPT 4.81g (3% by weight relative to the total weight of the liquid), 0.91g (20mM) of caffeic acid ester (CAPE).

Weigh the raw materials of the MMA, DMPT, caffeic acid ester in each%, and after the antimicrobial filtering (0.2㎛) so as not to contain impurities, packed 8.5g each, the packaged liquid portion is ethylene oxide (Ethylene oxide, EO) Sterilized by gas.

20 g of the powder was improved and prepared, and 9 g of the liquid phase was improved and prepared, followed by mixing at 23 ± 2 ° C. to measure the polymerization reaction time. The polymerization reaction time was measured to 12 minutes 10 seconds to facilitate the polymerization, it was confirmed that the appropriate time.

As a result of the compression test of the bone cement prepared as described above, the compressive strength was measured to be 123 MPa, and when the ISO 5833 compressive strength standard is 70 MPa, the bone cement manufactured according to the present invention meets the strength standard. I could confirm that.

Comparative Example 1

Bone cement was prepared in the same manner as in Example 1, except that hydroquinone was added to the liquid portion as a polymerization inhibitor.

As a result of measuring the polymerization reaction time by mixing the liquid phase and the powder of the bone cement composition of Comparative Example 1, the polymerization reaction time was measured to be 20 minutes or more, and the polymerization reaction was also poor. Furthermore, as a result of the compression test, the compressive strength was measured at 60 MPa, and it can be confirmed that the compressive strength is significantly lower than that of the bone cement composition of Example 1.

Experimental Example 1 Aseptic Experiment

Aseptic experiment was performed on the powder and liquid phase prepared in Example 1 at the Korea Institute of Chemical Fusion Testing.

The suitability and validation of the medium were verified according to the method suggested by the Korean Pharmacopoeia Sterility Test Method. Specifically, the sterile test was performed by immersing the prepared powder and liquid completely in sterile distilled water, washing with ultrasonic waves for 5 minutes, filtering through a filter, and culturing the filter paper in soybean case digestion medium and liquid thioglycolic acid medium. .

When soy casein digestion medium and liquid thioglycolic acid medium were tested with a test strain as shown in Table 1 below, bacteria were cultured within 3 days and fungi were evaluated within 5 days, but the results were evaluated as shown in Table 2. As described above, when the filter paper for the liquid and the powder prepared in Example 1 were immersed and cultured, the microorganisms did not proliferate for 14 days, so that the sterility of the bone cement composition of the present invention could be confirmed.

Table 1

badge	Test strain	Inoculation amount ()	Culture condition	Judgment
Liquid Thioglycolic Acid Medium	S. aureus	≤ 100	Aerobic culture (32.5 ± 2.5) ℃, 3 days	Growth
	P. aeruginosa	≤ 100	Aerobic culture (32.5 ± 2.5) ℃, 3 days	Growth
	C. sporogenes	≤ 100	Aerobic culture (32.5 ± 2.5) ℃, 3 days	Growth
Soybean Casein Digestive Medium	B. sub / Jlis	≤ 100	Aerobic culture (22.5 ± 2.5) ℃, 3 days	Growth
	C. albicans	≤ 100	Aerobic culture (22.5 ± 2.5) ℃, 5 days	Growth
	A. niger	≤ 100	Aerobic culture (22.5 ± 2.5) ℃, 5 days	Growth

TABLE 2

badge		Test start date (2013-01-04)	Intermediate inspection date (2013-01-11)	Last inspection date (2013-01-18)	Judgment
Liquid Thioglycolic Acid Medium	Specimen	-	No growth	No growth	fitness
	Voice control	-	No growth	No growth
Soybean Casein Digestive Medium	Specimen	-	No growth	No growth	fitness
	Voice control	-	No growth	No growth

Experimental Example 2 Animal Transplantation Experiment-Histological Evaluation

The bone sieve prepared in Example 1 of the present invention was bone-grafted to rabbits, and the local influence of the biological tissues that appeared thereafter was evaluated.

This experiment was conducted in accordance with the "Common Standards Standard for Biological Safety of Medical Devices Announced No. 2011-58 of the Korea Food and Drug Administration" and the Korea Chemical Convergence Act based on the Animal Protection Law No. 10995 (2011-08-04, all amended). The test was conducted in compliance with the regulations on the use and management of laboratory animals (Laboratory Animal Ethics Committee, 2010-12-10).

The powder and liquid phase of the bone cement composition prepared in Example 1 and Comparative Example 1 were mixed and cured, then processed into specimens and implanted into the femur of rabbits.

The test animals were four healthy New Zealand White rabbits weighing 3 kg or more, and after depilation on the day before transplantation, each test animal was spaced about 1 cm in the right femur as shown in FIG. 1. The specimen prepared in Comparative Example 1 was implanted into three sites, and the specimen prepared in Example 1 was implanted into three sites in the same manner to the left femur.

After drilling, the specimen was implanted into the femur using a dental drill (AEU-7000MG Implant / Surgery system), and the sutures were used to suture the muscles and skin.

Experimental Example 2-1 Measurement of Weight Change after Transplantation

Body weights were measured before transplantation, at 4 weeks, 8 weeks after the transplant, and at the end of the test. As shown in Table 3, all four animals showed normal weight gain.

TABLE 3

Animal number	Week (s) after implantation
	0	12
M1	3342.0	3734.8
M2	3595.9	3925.1
M3	3457.1	4013.4
M4	3599.2	4367.8
Mean	3498.6	4009.9
Standard deviation	123.6	265.1

Experimental Example 2-2 Evaluation of Transplantation Site

Experimental Example 2-2-1: Visual Evaluation

After the test, grafts were visually evaluated. At 12 weeks after transplantation, each graft was observed using a magnifying glass at autopsy. No specific gross findings due to the test substance were observed. Ten test samples and 10 control samples were identified at autopsy and were not denatured.

Experimental Example 2-2-2: Histological Evaluation

Twelve weeks after transplantation, the femur was removed for histological evaluation. Implantation sites were taken with adjacent tissues and fixed in 10% neutral buffer formalin, followed by deliming (Rapidcal ™, BBC Biochemical) for 3 days. Tissue samples were prepared by Hema toxyl in & Eosin staining for histopathological examination.

Figure 2 shows the histopathology test results of the transplantation site of the experimental animal 1 12 weeks after transplantation. The site where the specimens of Example 1 and Comparative Example 1 were implanted was slightly different depending on the individual, but it was judged that the dense bone contacted with the control sample and the test sample was generally regenerated.

Experimental Example 3 Investigation of Intradermal Reactivity

Intradermal reactivity of the bone cement composition prepared in Example 1 was carried out in accordance with "Common Reference Standard for Biological Safety of Medical Device of Food and Drug Administration Notice No. 2011-58".

The powder and liquid phase prepared in Example 1 were mixed and cured, washed with water, and left to stand at room temperature for 24 hours, and eluted with sterile physiological saline and cottonseed oil at 70 ° C. for a long time at a rate of 20 mL per 4 g of sample.

Two New Zealand White rabbits were injected intravenously with 0.2 mL of sterile physiological saline and eluate oil eluate, and then evaluated for local reactivity immediately after injection and at 24, 48 and 72 hours post injection. Sterile physiological saline and brine oil were used as a control and injected into the site shown in FIG. Local irritation was observed immediately after the injection and 24 hours, 48 hours and 72 hours after the injection. As a result, no erythema, swelling and skin were observed at the injection site, and the cement composition prepared in Example 1 did not cause an intradermal reaction. It was confirmed that.

Experimental Example 4. Acute Toxicity Test

In order to investigate the acute toxicity of the bone cement composition prepared in Example 1 after the intravenous and intraperitoneal administration of the eluate prepared in Experiment 3 to 50 mL / Kg BW to the ICR mice as shown in Table 4 below After 72 hours, general symptoms and body weight change were evaluated. As a result, general symptoms and deaths related to eluate administration were not observed.

Table 4

Group	Dosing solution	Sex	Animal number	Clinical signs				Mortality (dead / total)
				4h	12h	48h	72h
G1	Negalive Conlrol (Sterile saline)	Male	1101	NAD	NAD	NAD	NAD	0% (0/5)
			1102	NAD	NAD	NAD	NAD
			1103	NAD	NAD	NAD	NAD
			1104	NAD	NAD	NAD	NAD
			1105	NAD	NAD	NAD	NAD
G2	Treatment (Sterile saline extract)	Male	1201	NAD	NAD	NAD	NAD	0% (0/5)
			1202	NAD	NAD	NAD	NAD
			1203	NAD	NAD	NAD	NAD
			1204	NAD	NAD	NAD	NAD
			1205	NAD	NAD	NAD	NAD
G3	Negalive Conlrol (Cotton seed oil)	Male	1301	NAD	NAD	NAD	NAD	0% (0/5)
			1302	NAD	NAD	NAD	NAD
			1303	NAD	NAD	NAD	NAD
			1304	NAD	NAD	NAD	NAD
			1305	NAD	NAD	NAD	NAD
G4	Treatment (Cotton seed oil extract)	Male	1401	NAD	NAD	NAD	NAD	0% (0/5)
			1402	NAD	NAD	NAD	NAD
			1403	NAD	NAD	NAD	NAD
			1404	NAD	NAD	NAD	NAD
			1405	NAD	NAD	NAD	NAD

In Table 4, NAD is an abbreviation of No Abnormalily Detecled.

Experimental Example 5. Microbial Return Mutation Test

For microbial return mutation test for the bone cement composition prepared in Example 1, sterile physiological saline as a polar solvent, DMSO as a nonpolar solvent was used, and the following strain was used as a test strain.

Base-substituted: Salmonella typhimurium TA100, TA1535 and

Escherichia coli WP2 uvr A

Frame-shift type: Salmonella typhimurium TA98, TA1537

After mixing and curing the liquid and powder prepared in Example 1 and washed with water and left for 24 hours at room temperature, sterile physiological saline and DMSO were added at a rate of 20 mL per 4g respectively and eluted at 70 ℃ for 24 hours. The eluted stock solution was diluted in 50%, 25%, 12.5%, and 6.25%, and used as the test solution in a total of 5 levels. No contamination by microorganisms was observed in the test solution and S9 mix.

In addition, compared with the negative control group in all strains treated with the test solution, there was no increase in the number of reverse mutation colonies, and growth inhibition could not be observed. In the positive control group, the reverse mutation of the negative control group was observed to be significantly increased than the number of colonies for each strain, so that the test was appropriately performed. The eluate of the test substance was determined not to cause the reverse mutation in all strains.

Claims (7)

1. Liquid phase including methyl methacrylate (MMA), dimethyl-p-toluidine (DMPT) and caffeic acid phenethyl ester; And

   Powders including polymethyl methacrylate (PMMA), zirconia and benzoyl peroxide;

   Bone cement composition containing.
2. According to claim 1, The liquid phase contains 150 to 200 parts by weight of methyl methacrylate and 1 to 10 parts by weight of dimethyl phytoluidine per 1 part by weight of caffeic acid ester,

   The powder is a bone cement composition comprising 200 to 300 parts by weight of polymethyl methacrylate, 100 to 150 parts by weight of zirconia and 5 to 20 parts by weight of benzoyl peroxide based on 1 part by weight of caffeic acid ester.
3. According to claim 1, wherein the average particle diameter of the polymethyl methacrylate is 10 ~ 100 ㎛ bone cement composition.
4. Liquid phase including methyl methacrylate (MMA), dimethyl-p-toluidine (DMPT) and caffeic acid phenethyl ester; Preparing a bone cement raw material; and a powder comprising polymethyl methacrylate (PMMA), zirconia, and benzoyl peroxide; and

   Reacting after mixing the bone cement raw material of the liquid and powder

   Bone cement manufacturing method comprising a.
5. The method according to claim 4, wherein the liquid phase comprises 150 to 200 parts by weight of methyl methacrylate and 1 to 10 parts by weight of dimethylphytoluidine per 1 part by weight of caffeic acid ester,

   The powder is a bone cement production method comprising 200 to 300 parts by weight of polymethyl methacrylate, 100 to 150 parts by weight of zirconia and 5 to 20 parts by weight of benzoyl peroxide based on 1 part by weight of caffeic acid ester.
6. The method of claim 4, wherein the reaction time is 5 to 15 minutes.
7. According to claim 4, The mixing is bone cement manufacturing method is performed at 20 ~ 25 ℃.

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