WO2003047645A1

WO2003047645A1 - Sponge-like porous apatite-collagen composite, sponge-like superporous apatite-collagen composite and process for producing the same

Info

Publication number: WO2003047645A1
Application number: PCT/JP2002/012782
Authority: WO
Inventors: Masayuki Okazaki; Yasuhiro Yoshida
Original assignee: Japan Science And Technology Agency
Priority date: 2001-12-07
Filing date: 2002-12-05
Publication date: 2003-06-12
Also published as: JP2003169845A

Abstract

In the field of regeneration therapy engineering, there have been vigorously made research and development of apatite-collagen composites. However, existing composites cannot always satisfy various requirements such as holding a physiologically active substance or stem cells such as osteoblasts for a definite period of time, accelerating the infiltration of cells, and being replaced by a new tissue through spontaneous metabolism. Under these circumstances, it is intended to provide composites fulfilling these requirements. Namely, composites of apatite carbonate, which is highly close to bone in chemical composition and crystalline structure, and collagen having an excellent compatibility with biological tissues are formed. These composites have a porous or superporous sponge-like structure, which makes it possible to surely hold a physiologically active substance or stem cells for a definite period of time. Thus, a composite having excellent cell infiltration properties and metabolic properties can be obtained.

Description

Description: Sponge-like porous apatite / collagen composite, sponge-like superporous apatite / collagen composite, and methods for producing them

Technical field

The present invention relates to a novel superporous biomaterial for the purpose of developing a scaffold for replacing hard tissue, that is, a superporous composite of collagen and carbonate apatite. In particular, the present invention relates to a superporous composite having excellent cell infiltration properties, comprising a metabolic carbonate abatite having a composition and a crystal structure similar to that of a living bone or a tooth, and collagen, and a method for producing the same.

Background art

In recent years, research on collagen and apatite complexes has been actively conducted.

Japanese Patent Application Laid-Open Publication No. 2000-266628 discloses orthopedic, neurosurgery and plastic surgery comprising various bioabsorbable organic materials including collagen and calcium phosphate compounds including carbonate-containing apatite. A composite type bone filling material used for repairing a bone defect in a subject or the like is described. Also, Japanese Patent Application Laid-Open No. 8-187254 in accordance with the invention of the present inventors discloses that a collagen layer and an apatite / collagen layer are appropriately formed so as to be able to cope with both soft and hard tissues. A laminated composite film formed by laminating in combination is described. Further, Japanese Patent Application Laid-Open No. 8-336559 discloses a container for an artificial organ for newly generating an autologous organ tissue for treating a lesion of the organ tissue, and at least the container. It describes an artificial organ container whose inner wall is made of a material mainly composed of apatite containing an organic substance such as collagen. U.S. Pat. No. 5,071,4,336 describes that a sponge having hydroxyapatite and gel-like collagen is used as a bone replacement material for plastic surgery. Also, U.S. Patent Nos. 5,776,193 and 6,187,047 state that porosity is imparted in the design of bone replacement materials. Have been. Further, U.S. Pat. No. 6,201,039 describes various medical artificial bones comprising a hydroxyapatite starting from a hydroxypatite precursor, which is treated with collagen, and which is similar to a living bone tissue. It states that the organization will be obtained. In these documents, it is true that apatite is used as a material constituting the composite, and although carbonate apatite is exemplarily described therein, carbonate apatite is described therein. The chemical composition is very similar to the biological bone tissue, and the crystallographic relationship is also very similar (this point will be described later with reference to FIGS. 3 and 4). It is a material that has excellent affinity for it and is a particularly preferred material, or it is a particularly preferred material as a medical scaffold material, because carbonate apatite has a different composition from apatite among the apatites. There is no statement that this is positively suggested. In addition, he suggests and mentions the design concept of the complex of the present invention, to increase the infiltration of cells, and to set the complex to a sponge-like high porosity as a means for that purpose. There is no description. On the other hand, today, the rapid progress of regenerative medical engineering has led to the introduction of osteoblasts and stem cells together with bioactive materials into hard tissue substitutes, thereby enabling rapid formation of living cells including bone formation. The attempt to encourage Therefore, it is required that the material to be used has properties as a high-level biofunctional material. That is, bioactive substances and stem cells must be maintained for a certain period of time, sustained release should be exhibited, and they should have excellent cell invasiveness and be metabolized and replaced by new tissues. For various demands, those with characteristics that can meet these requirements are required. On the other hand, the conventional composite is as described above, and it cannot be said that it is a material that can sufficiently respond to the various requests as described above. The present invention seeks to develop and provide a scaffold material for hard tissue replacement that meets the above-mentioned demands, and as a result of intensive research, among various known apatites, carbonate apatite is particularly close to living bone tissue. It has been found that the material has a composition and crystallinity and is excellent in affinity for living tissue, and in addition, this apatite is complexed with collagen, so-called bioregenerating medical apatite. In order to obtain a collagen composite, if the material design of the composite can be designed to be porous and high porosity sponge-like, it is excellent in terms of, for example, enhancing the infiltration of new tissues. We conducted intensive research from the viewpoint that it is preferable as a regenerative medical engineering material. As a result, the sponge-like high porosity, porous composite was successfully obtained by treating both the carbonate carbonate and collagen materials by a specific process. It has been confirmed that the composite material obtained as a result of the above can be sufficiently used as a regenerative medical engineering material, and that the intended purpose can be achieved by this. is there.

The present invention has been made based on the above series of findings and the like. Disclosure of the invention

That is, the present invention solves the above-mentioned problems, and provides the following sponge-like porous apatite-collagen composite, sponge-like superporous apatite.collagen composite, and methods for producing them.

(1) A sponge-like porous body for replacing living hard tissue, which is a composite for replacing living hard tissue, comprising a carbonate carbonate and collagen, wherein the tissue structure of the complex is a porous sponge. Carbonated abatite-collagen complex.

(2) The sponge-like porous carbon dioxide apatite collagen for substituting a living hard tissue according to the above (1), wherein the complex has a porosity of 5 to 90 vol%. Complex.

(3) The composite according to the above (2), wherein the composite has a porosity of 50 to 90 vol%. Complex.

(4) The living tissue replacement according to the above (1), wherein the complex contains 95 wt% or less of carbonate apatite and has a porosity of 5 to 90 vol%. Sponge-like porous carbon apatite / collagen complex.

(5) The sponge-like porous carbonic acid according to the above (1), wherein the carbonic acid apatite contains a trace component similar to that contained in the biological apatite. Apatite / collagen composite body.

(6) The sponge-like porous carbon dioxide apatite-collagen complex for substituting a living hard tissue according to the above (5), wherein the trace component is Mg, Fe, Zn, F, or CI.

(7) Collagen is dissolved in acid to prepare an acidic collagen solution, gelled by adding alkali, and then mixed with apatite carbonate particles to obtain a water-containing composite gel composed of apatite and collagen. The water-containing composite gel thus obtained is centrifuged to remove water appropriately, frozen, and freeze-vacuum-dried to obtain a sponge-like porous carbonated apatite-collagen complex, which is a substitute for living hard tissue. For producing spongy porous carbonated apatite / collagen composites.

(8) Collagen is dissolved in acid to prepare an acidic collagen solution, gelled by adding alkali, and then mixed with apatite carbonate particles to form a hydrous composite gel composed of apatite and collagen. The resulting hydrous composite gel is centrifuged to remove water as appropriate, frozen, freeze-dried and vacuum-dried to obtain sponge-like porous apatite collagen, and then insolubilized. A method for producing a sponge-like porous carbonated apatite / collagen complex for replacing living hard tissue.

(9) The method for producing a sponge-like porous carbonate apatite-collagen complex for replacing living hard tissue according to the above (8), wherein the insolubilization treatment is performed by irradiation with UV light. (10) The method for producing a sponge-like porous carbonate apatite-collagen complex for substituting a living hard tissue according to the above (8), wherein the insolubilization treatment is a chemical treatment. That is, a first solution of the present invention is a living hard tissue replacement complex comprising carbonate apatite and collagen, characterized by exhibiting a porous sponge shape. This is a sponge-like porous carbonated apatite / collagen composite. A second solution of the present invention is a sponge-like porous material for replacing a living tissue, wherein the composite according to the first solution has a porosity of 5 to 90 vol%. It is a carbonate apatite / collagen complex. A third solution of the present invention is a sponge-like ultraporous material for substituting a living hard tissue, wherein the composite according to the second solution has a porosity of 50 to 90 vol%. This is a complex of carbonic acid apatite and collagen. A fourth solution of the present invention is characterized in that the complex according to the first solution contains 95% by weight or less of carbonate apatite and has a porosity of 5 to 90% by volume. It is a sponge-like porous carbon apatite / collagen complex for replacing living hard tissue. A fifth solution of the present invention is the substitute for living hard tissue, characterized in that the carbonate apatite in the first solution contains a trace component similar to that contained in the living body apatite. Sponge-like porous carbonate It is a patite-collagen complex. A sixth solution of the present invention is a sponge-like porous material for substituting a living hard tissue, wherein the trace component in the fifth solution is Mg, Fe, Zn, F, or CI. It is a carbonate apatite / collagen complex. The seventh solution of the present invention is to prepare an acidic collagen solution by dissolving collagen in an acid, gelling by adding an alkaline solution, and then mixing the carbonated apatite particles, A water-containing composite gel consisting of collagen and collagen is obtained. The obtained water-containing composite gel is centrifuged to remove water as needed, and then frozen and freeze-vacuum dried to form a sponge-like porous material. A method for producing a sponge-like porous carbonated apatite-collagen complex for replacing a living hard tissue, characterized by obtaining a carbonated apatite-collagen composite.

The eighth solution of the present invention is to prepare an acidic collagen solution by dissolving collagen in an acid, gelling by adding an alkaline solution, then mixing the carbonated apatite particles, and mixing with the carbonated apatite. A water-containing composite gel comprising collagen was obtained, and the obtained water-containing composite gel was centrifuged to appropriately remove water, and then frozen and freeze-vacuum-dried to form a sponge-like porous carbonate. This is a method for producing a sponge-like porous carbonate-collagen complex for replacing living hard tissue, which comprises obtaining an apatite-collagen complex and then insolubilizing the complex. A ninth solution of the present invention is a sponge-like porous carbon dioxide for replacing living hard tissue, wherein the eighth insolubilization treatment is performed by irradiation with UV light. This is a method for producing a patite-collagen complex. A tenth solution of the present invention is a method for producing a sponge-like porous carbonated apatite-collagen complex for substituting a living hard tissue, wherein the eighth insolubilization treatment is a chemical treatment. The usual apertype is C ai. (Ρ θ4) 6 (OH) is represented by 2, carbonate Apatai you want to use in the present invention, the chemical composition of the general formula; C a ₁₀ one X (PO 4) 6- γ ( CO s) γ ( OH) 2-z, where 0 ≤ X ≤ 3 to 5, 0 <Y 2 to 4, 0 ≤ Z ≤ 1 to 2, and its crystallinity is very similar to the crystal structure of living bone ing.

The synthesis method has been reported in various papers [eg, "Biomaterials" 1990, Volll, Octorber, pp. 568-572, M. Okazaki and four others, "Insolubilized properties of UV-irradiated C03-apatite — collagen composites J". And is known. The reason why carbonate apatite is particularly used in the present invention has already been described. Synthetic carbonate apatite has a chemical composition and crystal structure close to that of a living body, and thus has a high affinity for living tissue. And familiarity.

That is, a comparison of the synthetic carbonate apatite with the bones of the human body by X-ray diffraction patterns is as shown in FIGS. 3 and 4. The crystal structures of both are almost similar and cannot be distinguished. In addition, the requirement for setting the complex to be sponge-like porous or sponge-like super porous is that, by this, the bioactive substance to be introduced and osteoblasts can be retained, and the cells can be infiltrated. And ensure landing It has an action. The sponge-like porous composite is defined by the porosity and is selected according to the porosity in use.However, the range in which the porosity is set or selected depends on the purpose of use. The decision will be taken into account. The selection will be clarified through further experiments. In any case, the complex of the present invention is unique in that it is set in a sponge-like state as compared with the conventional complex, and thus the complex is recognized. On the other hand, the composite based on the previous design, as described in the present invention, may be described as being porous even if the description is actually porous, or a description suggesting this or a guide to that effect. There is no intentional statement up to the point where a sponge-like one is aimed. Rather, they tend to shy away in terms of strength, and therefore, the direction of development has not been taken into account, even at the expense of strength, to aggressive porous sponge bodies. On the other hand, the present invention has a sponge shape and a porosity of 5 to 90 vol%. As the porosity is reduced, that is, in other words, as the density of the composite (the packing density of the apatite particles) is increased, the sponge does not appear and its properties are lost. The lower limit of 5 vol% is that if it is less than 5 vol%, it will not be possible to meet the above-mentioned requirements such as cell invasiveness. The reason for setting the upper limit to 90 vol% is beyond this—in this case, the strength of a biomaterial support is the limit. This is what we stipulated. INDUSTRIAL APPLICABILITY The present invention can freely set the porosity in a wide range, thereby meeting a wide range of usage modes and demands. In any case, the void portion plays an important role in regenerative medical engineering as a void for holding a bioactive substance and a living cell, or as a void for implantation and infiltration of cells. In particular, the composite set at 50 to 90 vol% has a porosity that accounts for more than half of the volume of the composite, and can be said to be a superporous composite. Which range of porosity is to be set or selected depends on its use in regenerative medical engineering, that is, a support for bone regeneration (for osteoblast culture), a support for cell culture, and an artificial organ. It is set appropriately according to various uses such as a tissue culture support or a bone filling material, and referring to clinical examples. If the purpose is to emphasize not only cell invasiveness but also strength, it can be used in combination with other support mechanisms. Next, collagen will be described. Collagen is commercially available, and the collagen used in the present invention is usually commercially available and sufficient, and there is no particular limitation on its quality and the like. The significance of collagen in the present invention is that, in producing a sponge-like porous composite, it plays a role as a binder for carbonic acid apatite crystal particles and also adjusts porosity. Also plays an important role together with other factors. That is, the sponge-like porous composite is produced by the above-described process, but depends on various conditions in a series of processes such as the concentration of the collagen solution, the amount of mixed carbon apatite, and the centrifugation operation for separating water. The porosity, etc., of the resulting composite varies and is affected. In the process of producing a spongy porous composite, collagen Is dissolved in an acid and then neutralized to neutrality or alkalinity in order to dissolve and gelate. That is, collagen is dissolved by an acid and gelled when an alkali is added to the acidic solution and the pH is adjusted to 7 or more. The mixing operation of carbonated apatite is performed after neutralization, because if mixed at the stage when the solution is acidic, the apatite crystals may be partially eluted or altered, or the crystal structure of the particles may be altered. This may have some effect on the situation, and this should be avoided. In the region having a pH of 7 or more, the collagen in the acidic collagen solution gels. Mix apatite immediately after neutralization. In other words, if the gelation progresses due to the neutralization, the viscosity increases due to the gelation and it becomes difficult to mix, so it is preferable to carry out immediately after the neutralization.However, the gelation is adjusted to an appropriate viscosity, and As an embodiment, the mixing amount of the apa-titer particles is appropriately controlled and agitated, thereby controlling the dispersion state and the like of the two in the composite. It is not something that must be done. After the mixing operation is completed, centrifugation operation and freeze-vacuum drying treatment are performed. The significance is that the hydrogel composite obtained by mixing has not yet formed a porous state in this state, and this operation process As a result, water is removed, and the gel body becomes a sponge-like porous body.

After that, the insolubilization treatment is that, as it is, the collagen gel body is soluble, is easily metabolized by living tissue, cannot maintain its form, and causes inconvenience. is there. In (4) of the disclosure of the invention, the complex may be a carbonate apatite. The point that the content was 95% by weight or less and the complex was specified only based on the upper limit of carbonate apatite is not limited to the lower limit, that is, when carbonate apatite S is substantially close to 0 wt%. (Collagen 100 wt%) is also possible. Regarding the upper limit set at 95 wt%, when the carbonate apatite exceeds 95 wt%, the composite becomes dense and it becomes difficult to obtain the desired porosity. In addition, it is specified because the amount of collagen decreases and it becomes difficult to maintain the form of the complex by retaining carbonate carbonate crystals.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is an electron micrograph showing the structure of the sponge-like superporous carbonate apatite / collagen complex of the present invention.

Fig. 2 is an electron micrograph showing the structure of a conventional apatite-collagen complex.

FIG. 3 is an X-ray diffraction diagram of the synthetic carbonate apatite.

FIG. 4 is an X-ray diffraction pattern of a human bone.

Fig. 5 is an electron micrograph showing the bone tissue of the sample not irradiated with UV.

FIG. 6 is an electron micrograph showing the bone tissue of the sample irradiated with UV.

BEST MODE FOR CARRYING OUT THE INVENTION

The sponge-like porous carbonated abatite-collagen composite of the present invention has a sponge shape and a high porosity. As described above, when used as a cell culture support, it has good retention of the introduced bioactive substance and good cell infiltration and metabolism, as described above. Not only can it be used for the same purpose as the normal apatite-collagen complex, but it can also be used in the fields, applications, or cell invasiveness that were previously considered difficult by taking advantage of the characteristics shown above. It is considered one of the embodiments to carry out the invention specifically for the support for cell culture, and its use is more versatile in that it can be used in a wider range of applications than the complex of the previous embodiment. It is unique in that it is compatible with specialized uses that emphasize the abundance, implantation and invasiveness of cells, and can be used for any purpose in any case. It can be said that the present invention includes various embodiments. Example

The present invention will be described based on examples described below. This embodiment discloses a specific example for describing the present invention, and the present invention is not limited to the embodiment.

[Procedure for preparing sponge-like porous carbonate apatite / collagen complex] [Production of carbonate apatite];

1. Prepare a 3 molar CH 3 COO NH4 buffer aqueous solution, and further add 0.5 mL of a 1 00 mmol aqueous Ca (CHsCOO) 21120 aqueous solution, including 60 millimol (NH4) 2 CO 3 60 Millimol NH4H2PO4 aqueous solution Prepare 0.5 liter.

While stirring these Ca-side and P-side solutions in a buffer solution, monitor and control them so that the reaction temperature is 60 ± 1 ° C and the pH is 7.4 ± 0.2 ° C. The whole amount was added while gradually supplying at a constant speed with a microphone mouth tube pump. The control of the pH value is based on the ammonia water and the pH control. Adjusted with a trawler. As a result of the reaction, the supplied Ca / P molar ratio; 100/60; 1.67, and the obtained crystal particles were analyzed. The chemical composition of the starting mixture and the obtained carbonate apatite was analyzed. between the, fixed relationship is established, the starting mixture is reacted almost whole amount in stoichiometric relationships, so that the above-mentioned general formula of _{interest; C a 10 - X (P} 0 4) 6 i γ (CO ₃ ) γ (OH) 2—z, where 0≤X≤3 ~ 5, 0≤Y2 ~ 4, 0≤Z≤ :! It was found that carbonate apatite in the composition range represented by No. 2 was generated and recovered. The obtained carbonate apatite crystal grains are separated, washed with water, and dried, and used as a raw material for the carbonate apatite-collagen complex targeted in the present invention. Note that the example disclosed here is merely an embodiment, and the present invention is not limited to this. In other words, the composition of carbonate apatite, as shown in the general formula, has values of x, y, and z each within a predetermined range. It is possible.

During the reaction and when adjusting the pH value of the end point, the pH value is set to 7 or more, that is, to the near side. The reaction temperature may be in the range of about 40 ° C. to 80 ° C., but keeping the temperature as low as possible at 60 ° C. is particularly preferable for obtaining uniform crystals.

[Preparation of collagen solution and production of carbonate apatite / collagen hydrous composite gel];

Freeze vacuum drying § terrorist collagen from cowhide [Corporation two Tsubi, Japan] was dissolved in 1 0 _ ³ N hydrochloric acid solution, or commercially available collagen solution CC ELL GEN Co. Koken, Japan] Was used to prepare a 0.5 wt% collagen solution. To this collagen solution, 0.05 N NaOH solution was added dropwise, and when the whole amount was neutralized, the above carbonate abatite was immediately mixed. Further, NaOH solution was added dropwise to gel the collagen, and the collagen was gelled. As a result, a sponge-like porous carbonate and collagen precursor, which is a precursor of the collagen complex, was obtained.

This gelation reaction is caused by the acidity of the acidic collagen solution, regardless of the presence or absence of apatite. In other words, the phenomenon is based on the change in the molecular structure of the collagen itself. The concentration of the collagen solution prepared for the production of the complex is preferably 0.1 to 2.0% by weight.

Due to the difference in the concentration of the collagen solution, the state of the hydrous composite gel obtained after gelation differs in viscosity and water content. That is, the moisture content can be freely controlled by the difference in the concentration. By controlling the water content freely, that is, by increasing the water content, a composite having a high porosity can be obtained, and conversely, from the water-containing composite gel having a low water content, the porosity can be obtained. Thus, a material having a low porosity can be obtained, thereby controlling the porosity and the porosity in a target state. As described above, it is extremely important to adjust the concentration of the collagen solution. However, in the above examples, 0.1 to 2.0 wt% is used. Therefore, the lower limit was set because the desired composite could not exhibit its function and the shape and strength of the target composite were too weak, which hindered use. 2. If it exceeds O wt%, the viscosity at the time of gelation becomes strong, and the amount suitable for forming a bulky porous body for carbonated apatite particles is uniformly dispersed and mixed. In this case, the mixing operation and the subsequent dehydration operation for making the material porous are also difficult.

[Preparation of composite sponge];

In this step, water is separated and removed from the water-containing composite gel obtained by the above-mentioned process by a specific removing means. Through which a porous material is formed. That is, an intended sponge-like porous composite having a high porosity is realized and obtained.

At this time, the gelation reaction was performed in a mold having a predetermined shape, whereby a water-containing composite gel having a certain shape was obtained, and then processed into a predetermined shape by separating and removing water. Obtaining a sponge-like porous composite is also one of the embodiments.

Needless to say, it can be processed into an appropriate shape by arbitrarily cutting, bonding, or the like later, and this excellent workability is also one of the features of the present invention. The operation of separating and removing water in the composite sponge preparation process is as follows. First, the water-containing composite gel obtained in the previous step is centrifuged to preliminarily remove water that has liberated in a substantial amount, and then removes it. The unretained water is removed by freezing the gel as it is at 180 ° C. or with liquid nitrogen for 2 hours, followed by freeze vacuum drying, and sublimation and separation of the frozen water. By performing this moisture separation operation, the mixture becomes porous, and when pressed with a finger, an elastic sponge-like porous composite having a high porosity can be obtained.

(Insolubilization treatment)

Next, the sponge-like porous composite obtained by the above-mentioned process dissolves and swells in the living tissue as it is, gradually loses its form, and can no longer maintain the desired form. And may cause inconvenience. In particular, in the case of a sponge-like porous composite having a high porosity as in the present invention, the sponge-like porous composite has a large surface area as compared to a solid one, and is in a state where solubility is easily promoted. Therefore, it is desirable to perform insolubilization. Examples of the insolubilization treatment include ultraviolet irradiation and chemical treatment. The result does not change much.

(Example of insolubilization treatment) Place the apatite-collagen complex obtained by the above-mentioned process in a sterile box at a distance of 10 cm under a UV lamp (10 W, ultraviolet light with a wavelength of 253.7 nm). And irradiated with UV light for 2 hours on both sides.

As a result of comparing this sample with a sample without UV irradiation based on animal experiments, the sample without UV irradiation (Fig. 5) swells and peels easily, but the sample with UV irradiation (Fig. 6) shows a constant value. The shape of the bone was maintained, and good bonding with the bone was shown.

As the insolubilization means other than the UV irradiation, a treatment using a chemically treated gamma ray using Daltar aldehyde 0.1 to 5 wt% is also used.

Insolubilization by these means, in the case of UV irradiation, by one CN = N-bridge between adjacent amino groups molecules of the protein are formed, and when by glutaric aldehyde arsenide de shows an N-CH ₂ - It is believed that this is due to the formation of N-bridge bonds. That is, as described above, the present invention obtains composite gels having different viscosities and different water contents depending on the difference in collagen concentration and the like, thereby producing a sponge having an appropriate porosity and pore diameter according to the purpose. It is in. In the present invention, since the water content of the gel can be freely changed, the porosity and the pore diameter can be freely adjusted by controlling the water content, thereby obtaining a sponge-like composite having an extremely high porosity. Can be done. Although the pore diameter has not been described in detail, it is generally said that the invasiveness of cells depends on the pore diameter, and the pore diameter into which cells can easily enter is 200 to 3 • 0 microns. In other words, this value serves as a reference index, but the optimal range will be awaiting further research. Another feature of the present invention is that, since the present invention is in the form of a sponge, it can be easily formed. The biological apatite has trace elements of trace elements such as Mg, Fe, Zn, F, CI, etc., and it is possible to approach this in the present invention. The addition of trace elements during production is included as an embodiment of the present invention. When the high porosity and sponge-like porous carbonate-collagen complex of the present invention obtained as described above is observed by an electron micrograph, this complex shows that apatite particles are present in a collagen-sparse fibrous tissue. It shows a structure in which pups are scattered in tufts in some places, and it is observed that cavities and voids occupy most of the total volume of the complex, and that it has a very sparse sponge shape (Fig. ).

On the other hand, in the conventional apatite-collagen complex (Fig. 2), the apatite particles are more densely packed than in the sample in Fig. 1, and the difference in the structure and structure between the two is evident. Industrial applicability

As described above, according to the present invention, carbonate apatite having a very similar chemical composition and crystal structure to the bone of a living body is used, and this is composited with collagen, so that it has an affinity for biological tissue. In addition to being able to obtain a complex with good quality, the structure of the complex is made of sponge-like porous or ultra-porous, so that physiologically active substances and stem cells can be effectively and reliably retained for a certain period of time, and cell infiltration It provides a metabolic complex with excellent properties, and its significance is extremely large.

Claims

The scope of the claims

1. A living body hard tissue replacement complex comprising a carbonate apatite and collagen, wherein the tissue structure of the complex has a porous sponge shape. Carbonated abatite / collagen complex.

2. The sponge-like porous carbon apatite-collagen composite for living hard tissue replacement according to claim 1, wherein the composite has a porosity of 5 to 90 vol%. body.

3. The sponge-like ultra-porous carbonate apatite collagen for replacing living hard tissue according to claim 2, wherein the composite has a porosity of 50 to 90 vol%. Complex.

4. The substitute for living hard tissue according to claim 1, wherein the composite contains 95 wt% or less of carbonate apatite and has a porosity of 5 to 90 vol%. Sponge-like porous carbon apatite / collagen complex.

5. The sponge-like porous carbon apatite for substituting a living hard tissue according to claim 1, wherein the carbonate apatite contains a trace component similar to that contained in the biological apatite. · Collagen complex.

6. The sponge-like porous apatite-collagen complex for substituting a living hard tissue according to claim 5, wherein the trace component is Mg, Fe, Zn, F, CI.

7. Collagen is dissolved in acid to prepare an acidic collagen solution, gelled by adding alcohol, and then mixed with apatite carbonate particles to obtain a hydrous composite gel composed of apatite and collagen. The obtained water-containing composite gel is centrifuged to remove water appropriately, frozen, and freeze-vacuum-dried to obtain a sponge-like porous carbonated apatite-collagen complex, which is a substitute for living hard tissue. For producing a sponge-like porous carbonated apatite-collagen composite for use.

8. Collagen is dissolved in acid to prepare an acidic collagen solution, gelled by adding alcohol, and then mixed with carbonated apatite particles to obtain a hydrous composite gel composed of carbonated apatite and collagen. The obtained water-containing composite gel is centrifuged to remove water appropriately, frozen, freeze-vacuum-dried to obtain sponge-like porous carbonated abatite collagen, and then subjected to insolubilization treatment. A method for producing a sponge-like porous carbonated apatite-collagen complex for hard tissue replacement.

9. The method for producing a sponge-like porous carbonated carbonate-collagen complex for replacing living hard tissue according to claim 8, wherein the insolubilization treatment is performed by UV light irradiation.

10. The method according to claim 8, wherein the insolubilization treatment is performed by a chemical treatment.