JPS58148828A - Agent and kit for root canal treatment - Google Patents

Abstract

PURPOSE:To provide a root canal treatment agent containing a protease as a main component, and capable of effectively removing the odontoblastic layer remaining on the inner wall of the root catal after pulpecotomy, biochemically, without using a mechanical means. CONSTITUTION:To objective root canal treatment agent is obtained by using a protease (e.g. an enzyme such as trypsin, collagenase, etc.) as a main component, and dissolving the protease in a buffer solution (e.g. phosphate buffer solution, tris-buffer solution, borate buffer solution, etc.) optionally containing a protease-activating substance (e.g. calcium ion, magnesium ion, etc.). The activities of the enzymes in the treatment agent are >=100 Units/ml, preferably 300-10,000 Units/ml for collagenase and >=500 Units/ml, preferably 10<3>-8X10<5> Units/ml for trypsin.

Description

[Detailed description of the invention]

The present invention relates to a 1m root canal treatment agent in dental treatment, - a treatment kit or a composite chrysanthemum). (I
11 Odontoblasts remaining attached to the inner wall of the medullary canal
A treatment method that is often used is to remove as much of the layer as possible by mechanical means such as a microengine and then perform subsequent treatment. When this method is used, the judgment of the extent of odontoblast removal must rely on the intuition of the practitioner.The removal of the odontoblasts is often incomplete, and as a result, lines in the remaining odontoblasts Lyson enzyme
Oal Enzyme) etc.
This can lead to problems such as abscess formation in the pottery root. In view of the above-mentioned problems in dental treatment in the prior art, the main purpose of the present invention is to biochemically remove the odontoblast layer that remains adhered to the inner wall of #1 root canal even after pulp extraction without the need for mechanical means. V? - A new root canal treatment agent that can perform extremely effective removal treatment - t1! The purpose of the present invention is to provide kits for root canal treatment. The above ni! of the present invention! is a narrow tube treatment agent containing various proteases, especially collagenase as a main component, and supplementary components such as a buffer solution and #activating substance, or various combinations of the present invention described below for providing a simple treatment at the time of treatment. Chrysanthemum) @Ic is achieved more effectively. However, the present inventors have discovered for the first time that postcanal Ii fixed dentinal cells can be very effectively digested and removed by the action of protease, particularly collagenase, and have thus arrived at the present invention. The processing agent of the present invention, its composition, usage/dose, effects, etc., will be explained in more detail below. The enzymes that can be used in the present invention, such as WIX for odontoblast digestion and withdrawal treatment, are unmodified. Collagen substrates (specifically acting collagenase and other proteases such as trypsin, chymotrypsin, pepsin, hahain, etc. (hereinafter referred to as ``general proteases'')
It is broadly divided into According to the findings of the present inventors, dentin bud mj1 is fixed to immature dentin via so-called collagen fibers,
Therefore, by allowing collagenase to act on this, it can be very effectively and specifically detached from immature dentin. On the other hand, in the case of groteases in general,
Not only collagen fiber but also thin M! i! It is thought to act on walls, etc., and its mechanism of action is different from collagenase in this respect, but it is a highly active bulky liquid.

[When used, almost complete digestion and removal of odontoblasts is possible with these alone. On the other hand, in order to make more effective use of these differences in the action mechanisms of the two, there is a method in which the root canal after pulp extraction is predigested using general grotease as much as possible, and then completely removed using collagenase. May be suggested. King Nawa too,
When using this method (σ), the pre-digestion treatment accelerates the penetration of collagenase into collagen fibers, and as a result, rapid and reliable treatment can be achieved. Although commercially available enzymes are usually sufficient as the enzymes that can be used in the present invention, their properties, buffers, etc. are summarized as follows. 1 Collagenase Clositridium himtolyticum
, M. tuber-culosis, Bacter
oides rnelaninogenica. Streptomyces madurae, Tri
chophytonachoenleiniL Amp
Typical enzymes include collagenases produced by various microorganisms such as S. ergilla oryzae, and collagenases present in various animal tissues such as mouse fibroblasts. It is clear that a wide variety of other collagenases that can act on the enzyme can be used as appropriate. These collagenases usually contain their activators (
Calcium ions or magnesium ions are required as activators, and the optimum pH thereof varies depending on the production source, but is generally within the range of pH 6.5 to 9. Therefore, in the treatment agent of the present invention, collagenase lyophilized powder or tablets are usually combined with calcium chloride (CaC).
1g) *Used for practical use by dissolving in a buffer containing an activator such as calcium carbonate (Ca Cox ). 2. Proteases in general, trypsin, chymotrypsin. It mainly contains so-called endopeptidases such as cathepsin, ficin, papain, and b-line, and in addition to these so-called endopeptidases such as aminopeptidase and carboxypeptidase, as long as the action conditions such as pH < 1 are approximately the same and the action is not inhibited. Can be used in combination with exopeptidase. All of these enzymes can be used as commercially available products. On the other hand, when used in the pre-digestion treatment described above, the optimal pH value can be almost the same as that of a commercially available collagenase agent.
Trypsin is highly preferred. 3. Buffers Phosphate, boron salt, Tris-HCl buffer, etc. - Various well-known mild *gv as long as they are therapeutically acceptable.
It can be selected and used as appropriate. Furthermore, since the processing agent of the present invention can be used for a relatively short period of time, its toxicity and other effects on the brain are practically not a problem as long as it is washed immediately after the reaction is completed. Particularly suitable examples include N-phantom-N& phosphate buffers and the like. Collagenase usually uses Ca''+ or M as its activator.
g! As mentioned above, these are the ones that require the first prize, and these are CaC1a. CaCO5, CI (CHICOO)! , MgC1
1 in the form of various inorganic or organic salts such as m, MgC01, etc.
It is sufficient to add a trace amount of about 01 to 101S. Even if a phosphate-based buffer solution is used, for example, if 1g of CaC is used, about 5 x 10" will stably exist, so it can be used satisfactorily. Dosage and administration, each MM #Activity unit of element The activity unit of each srs element used in the present invention is manufactured by Nguma Co., Ltd. (P, 0.BQX 1450 m, ST. LOUI S, MO, 63178U, S, M,)
In accordance with the regulations, it is defined as follows@. a) One unit of collagenase has the activity of releasing an amount of amino acids equivalent to 10 μmole of leucine by ninhydrin coloring when acting on undenatured collagen for 5 hours at pH 7, 4, and 37°C in the presence of Cal/Lam ions. and (defined as 2).b) Hydrolyze 1.OA mole of α-N-pensoyl-L-arginine ethyl ester (BAEE) per minute at pH 7, 6, and 25°C. C) One unit of other non-specific norotease can be defined as the activity of pH 7,
When treated with casein at 4.37°C for 5 hours, ninhydrin coloration showed 1. Amino acid corresponding to leunon in O1imol@? Defined as the activity released. 2. Dosage/Usage Is the non-inventive treatment agent 5 koji? r! After removing the pulp, the root canal is cleaned with light, then one amount of the treatment agent is injected and filled into the C. used in It is desired that the time required for dental treatment L, -F to react with the equipment is as short as possible, and in reality, it is usually within one hour, more preferably within a few minutes to 30 minutes. It should be. From this point of view, the activities of various enzymes in the treatment agent should be set as follows: a) -7 lagenase: at least 1 OOUnit
s/d, more preferably 300 to 10.000 Units
s/d. b) Trypsin: at least 500 Units/
aJ. More preferably 10" to 8 x 10' Unit
s/117°C) Other non-specific proteases: at least 500 Units/aZ, more preferably 1.500-50.000 Units/11j. It should be noted that the above-mentioned dosage regulations can be said to be appropriate even in the case of the aforementioned two-step treatment method, but they can also be used with lower activity. Father, the volume of one root canal is usually about 1 to 5 μl, and the number of root canals is about 1 to 4 per W11, so 1
The amount of treatment agent O) required for each treatment will depend on the number of teeth to be treated, but will generally be about 20 to 100 μl. Although it is practical, it can be stored stably for about 1 month at 0 to 4°C, so it may be possible to prepare a larger amount in advance and store it in a cooled state. Φ (), it is particularly advantageous for the treatment agent of the present invention to be provided in the form of a kit. Some typical examples of the kits of the present invention are summarized below. Instead of an ampoule, the above amount may be dispensed in a larger capacity vial when used. In addition, other grotease kits are also provided in the same manner as above. 3 ml [2 , "Protease Shell Kit" was used as the first kit for pretreatment, and "IHI I Collagenase Kit" was used as the second kit for main treatment.Hereinafter, the present invention will be explained in more detail with reference to experimental examples. ] The #1 crown of an adult cow, from which unnecessary tissue such as dead meat adhering to the surrounding area has been completely removed, is cut transversely with a grinder to extract the produced tooth pulp, and then cut longitudinally with a grinder to remove the pulp. Thoroughly wash the sample with saline and prepare it as a raw sample.
T)'pe v-S; 136 Units /N
t) at pH 7,4,0,067M-phosphate buffer (0
A treated sample tooth was prepared by immersing the tooth in a test tube at 37° C. for 60 minutes in a treatment solution of collagenase activity 6 J300 Unit Hatake/- obtained by dissolving the tooth in a solution containing 1 m of CaCl. Next, after thoroughly washing this treated sample tIi with physiological saline, p
It was diluted with H7,4,0,067M phosphate buffer and immersed in a 2% glutaraldehyde solution to prepare a sample for an electronic microscope. Attached figures 1 and 2 are magnified at 1,000x and 10,000x, respectively.
This is a scanning electron microscopy photograph of an untreated sample (sealing wax) at a magnification of 1,000 times; on the other hand, Figures 3 and 4 are greatly enlarged *i, o
oo and 10.000x magnification of the above collagenase treatment and electronic microscopic photograph oo As can be seen by comparing these, in the collagenase-treated sample*, not only odontoblasts but also collagen Confirm that Φ fibers are also clearly removed under extreme pressure. Experimental Example 2 Tryp/N (Sigma grade, Type
Tryptopin activity 4 obtained by dissolving eM, 8.500 Units+/"f) in pH 7, 4,0067M phosphate buffer (containing 0,005% CaC1z)
．． 25 X I O' Units/Ill fermented confectionery liquid was used.
Samples for electron microscopy were obtained by processing I. Figures 5 and 6 are electron micrographs of the treated sample teeth (1,000 yen each).
0 and 10,000 times). As is clear from the figure, in the sample treated with trypone, the odontoblast #i was almost completely removed, but it was observed that some collagen φ fibers remained undigested. Experimental Example 3 Four types were used in Experimental Example 2, except that the pHl1 of the buffer solution was adjusted to the optimum pHl value for each protease, and the sample Akira was mixed with chymotrypone, pepsin, and papain solutions (each protease activity: 140.000). Units/') K and processed to obtain a sample for electron microscopy. The results were determined to be approximately equivalent to the trypne treatment. Experimental Example 4 A raw sample was treated with tryptonium solution as described above and 1ii1.
℃ for 10 minutes, followed by optical washing with physiological saline, and embedding at 37℃ for 10 minutes in the same collagenase treatment solution as above. As a result of electron microscopy, dentinal buds were removed.
It was confirmed that both the collagen and fibers were removed by the Vc specimen. Experimental Example 5 Under the same conditions as Experimental Example 1 above, the raw sample was immersed in Y Test W, and the optical density at 280 nm1 of the treatment solution was
,D) and protein-containing t (Folin-Lowr)
'Modified method: Salmon temporal change of bovine serum albumin standard) Y #j was added. The results are shown in Figures 7 and 8. As is clear from the figure, the processing VC! under the conditions in question! + indicates that the predetermined purpose is achieved after 10 to 15 minutes. Experimental Example 6 1 Under the same conditions as in Experimental Example 1, a human sample * (male, 26 o'clock; body part, lower left No. 8; symptom C-2) was treated to obtain a sample for electron microscopy. As a result of electron microscope observation, it was confirmed that both odontoblasts and collagen fibers were completely removed. The above also applies when using a sample that has been cut and extracted with a microengine as a normal treatment without being longitudinally cut, and the treatment solution is injected into the root canal (llil medullary cavity) and treated. The same results were obtained. 2 Human leprosy samples were treated according to Experimental Examples 2 to 4 above,
As a result of electron microscopic observation of the treatment *8M, it was found that a result of approximately 4th grade was obtained compared to the case of 10-. Experimental Example 7 Raw samples *V were immersed in collagenase treatment solutions with various activities, and the optical density at 280 nmK (0, D ) of the treated waves was measured.
Change over time ()0. D) is hooked to form the mold shown in Figure 9.
Reaction-! I got it. From the figure, the minimum activity of the treatment solution that is substantially effective for removing odontoblasts is approximately 100 Uni.
It is recognized as sad/-jiii degree. In addition, the treatment solution used is Nogma Collagenase (136
Units/)) each 1 to 10"f'k, PH7,4,
0,05M-) Lis Hydrochloric Acid Buffer & (0,005 Whisper CaC
1*Contains)lIljKll! It was prepared by On the other hand, experiments similar to those described above were conducted for trypsin and other proteases 1ij, and the doses (activities) that were substantially useful in treatment were determined as described above.

[Brief explanation of drawings]

Attached FIGS. 1 to 6 are electron micrographs of experimental examples of the present invention, and FIGS. 7 to 9 are explanatory diagrams of the experiments. Percentage Applicant Atto Co., Ltd. Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 [0,0] Figure 9 Procedural Amendment (Method) July 1982 28th, Mr. Kazuo Wakasugi, Commissioner of the Office of Special Correction 1. Indication of the case: 1982 Patent No. 29816 2. Name of the invention: Root canal treatment agent and kit for the same treatment 3. Relationship with the amendment case: Patent applicant's address: 103 4-5-7 Kobune-cho, Nihonbashi, Chuo-ku, Tokyo Date of amendment order: June 11, 1980 (Date of dispatch: June 29, 1988) 5. Subject of amendment 6. Statement of contents of amendment No. 20 14. Brief explanation of the drawings J on page and page 21 is corrected as follows: [4. Brief explanation of the drawings Figures 1 to 9 are explanatory diagrams of experiments in experimental examples of the present invention. , Figure 1 is an electron micrograph of the root canal of an untreated sample at a magnification of 1,000x, Figure 2 is an electron micrograph of the root canal section at a magnification of 10,000x, and Figure 30 is an electron micrograph at a magnification of 1.
．． Figure 4 is an electron micrograph of the root canal treated with collagenase at a magnification of 10,000x. Figure 5 is an electron micrograph of the root canal treated with trypsin at a magnification of 1.000x. Figure 6 is an electron microscope photograph (2) magnified 10,000 times, and Figure 7 is a graph showing changes in optical density (0, D) over time. Figure 8 is a graph showing changes in protein content over time;
FIG. 1 is a graph showing a dose-response curve of an enzyme reaction. ”

Claims (1)

[Claims] A root canal treatment agent containing Tll protease as a main component. (2) The processing agent for ie* according to claim (1), wherein the protease is an enzyme selected from the group consisting of trypsin, chymotrypsin, pepsin, and papain. (3) %# that the protease is collagenase! F. The processing agent according to claim (1). (4) Claim No. 3 in which collagenase is dissolved in mild rtlK containing its activator if necessary.
) The processing agent according to item 1. (where the collagenase activity is at least 100 [JB
itB/m.
The processing agent according to item 41. (6) Its collagenase activity is 300~i o, o
o. The processing agent according to claim 1), which is Units/117. (7) Claim No. (4), characterized in that the activating substance is calcium or magnesium ion.
The processing agent according to any one of items (6) to (6). (8) A root canal treatment kit consisting of a combination of a collagenase agent and a buffer solution for maintaining it at its optimum pH. (9) Add a predetermined amount of the collagenase agent to the
When dissolved in III[, its collagenase activity is low (
Claim number 100 Units/width (
The kit according to paragraph (transition). Trout Its collagenase activity is 300-10,000 Un
The kit according to claim 9, which is ~. 11TJ 4I#! where the buffer is one of a phosphate buffer, a Tris buffer, and a borate buffer. The kit according to any one of claims (8) to (F). The kit according to any one of claims 8 to 00, further comprising a collagenase activator, if necessary. σ) The kit according to claim 03, wherein an α3 predistribution activator is added in advance to the collagenase agent and/or the buffer i[. [alpha]hiro [sigma]) Pre-luki kit according to claim [alpha][1], wherein the activator is Cal/lam salt or magnesium salt. 0 The first part of the preliminary treatment which has a protease agent other than collagenase as a main ingredient) and the second part consisting of the kit according to any one of claims 14 to 14 A composite kit for root canal treatment comprising a combination of the following: The composite kit according to claim 1, wherein the U*V protease is any one of triglyceride, chymotrypone, pepsin, and ]()<in. The composite kit according to claim 3 or 3, wherein the first kit comprises the protease agent and a buffer solution containing the protease agent at its optimum pH. U When the protease agent is dissolved in a predetermined amount of pre-balanced buffer, its protease activity is 500 to 8
O' Units/-. (2) The composite kit according to claim 11, wherein the protease agent is a trinosine agent.