TW201821057A - Resorbable crosslinked form stable membrane - Google Patents

Abstract

The invention relates to: - A resorbable crosslinked form stable membrane which comprises a composite layer of collagen material and inorganic ceramic particles containing 1.5 to 3.5 weight parts of inorganic ceramic for 1 weight part of collagen material, sandwiched between two layers of elastic pretensed collagen material, the collagen material comprising 50-100 % (w/w) collagen and 0-50 % (w/w) elastin, - the above crosslinked resorbable crosslinked form stable membrane for use as an implant to support bone formation, bone regeneration, bone repair and/or bone replacement at a non-containing dental bony defect site in a human or animal, - a process for preparing the above crosslinked resorbable crosslinked form stable membrane comprising the steps of: (a) Preparing a composite layer of inorganic ceramic particles and collagen material, optionally crosslinking that composite layer, (b) Assembling and glueing the composite layer of collagen material and inorganic ceramic particles between two layers of collagen material submitted to tensioning leading to a stretching of the collagen material in the linear region of the stress-strain curve, thereby giving a composite layer of collagen material and inorganic ceramic particles sandwiched between two layers of elastic pretensed collagen material, and (c) Crosslinking that composite layer of collagen material and inorganic ceramic particles sandwiched between two layers of elastic pretensed collagen material, followed by a hydrophilic making treatment.

Description

Resorbable crosslinked morphologically stable membrane

The present invention relates to a novel resorbable cross-linked morphologically stable film for oral cavity, relating to a method for preparing a film and supporting bone formation and bone regeneration as a defect site in a human or animal bone-free defect The use of implants for bone repair and/or bone replacement.

To regenerate bone-free defects by bone formation (such as horizontal or vertical expansion in the maxilla or mandible), mechanical stabilization of the defect is required (Bendkowski 2005 "Space to Grow" The Dentist: 3 Merli, Migani et al. 2007 Int. J. Oral Maxillofac. Implants 22(3): 373-82; Burger 2010 J. Oral Maxillofac. Surg. 68(7): 1656-61; Louis 2010 Oral Maxillofac. Surg. Clin North Am. 22(3): 353-68). In fact, oral tissue suffers from complex mechanical forces during chewing, swallowing, tongue movement, speech, tooth movement, and orthodontic treatment. Especially during wound healing after surgery, internal and external forces can occur, and stress, shear and bending moments are generated in the regeneration device and the newly formed structure. A morphologically stable membrane that resists their forces and achieves a useful component for mechanical stabilization. Titanium mesh, titanium plates or titanium-reinforced PTFE morphologically stable films are known for this purpose, which must be removed during secondary surgery after bone regeneration. An example of a commercially available titanium-reinforced morphologically stable film is the Cytoplast® film sold by Osteogenics. However, it has been reported that there is a high probability of cracking or other complications when using an expanded reinforced titanium membrane (Strietzel 2001 Mund Kiefer Gesichtschir. 5(1): 28-32; Merli, Migani et al. 2007 supra; Rocchietta , Fontana et al. 2008 J. Clin. Periodontol. 35 (8th Supplement): 203-15). Prior to the introduction of resorbable collagen membranes in 1996, unreinforced PTFE membranes were commonly used, but disappeared rapidly after introduction of the collagen membranes. Resorbable morphologically stable membranes have received attention in order to avoid the necessity of removing morphologically stable membranes or meshes during secondary surgery. Several resorbable morphologically stable films or webs made substantially of PLA (polylactic acid) or PLGA (glycolic acid copolymer) have been described. The examples to be noted are (1) "Sonic Weld RX®" and "Resorb-X®" from KLS Martin; (2) "Guidor®" from Sunstar Americas; (3) "Inion GTR SystemTM" from Curasan And (4) "RapidSorb®" from Depuy Synthes. The disadvantages of these membranes are that during the in vivo hydrolysis degradation of such membranes, their release causes tissue irritation and histological identification of lactic acid and/or glycolic acid that interfere with wound healing (Coonts, Whitman et al. 1998 Biomed. Mater). Res. 42(2): 303-11; Heinze 2004 Business briefing: Lobal Surgery: 4; Pilling, Mai et al. 2007 Br J. Oral Maxillofac. Surg. 45(6): 447-50). To overcome PLGA/PLA associated with wound healing problems, use autologous bone blocks from patients and partially or fully purified bone blocks (such as Bio-Oss® Block (Geistlich Pharma AG) or Puros® Allograft Block (RTI Surgical) Inc.)) is generally accepted. The inadequacy of autologous bone masses is that they are collected from the second site, resulting in greater pain (Esposito, Grusovin et al. 2009 Eur. J Oral Implantol. 2(3): 167-84). In order to be able to use the collected autologous bone fragments during surgery (usually combined with xenograft bones), the so-called bone shield technique using autologous cortical bone from the mandible was developed (Khoury, Hadi et al. 2007 "Bone Augmentation" In Oral Implantology", London, Quintessence). The disadvantage of this procedure is that it is highly technically sensitive and related to the incidence of the second site and is more painful. In addition, the bone shield is only applied laterally, so no mechanical protection is given from the coronal aspect of the defect. The term "bone shield" is used to advertise PLA/PLGA and partially demineralized cortical bone shields (Semi-Soft and Soft Lamina Osteobiol® from Tecnoss). The disadvantage of this demineralized bone shield is that the curved bone shield must be fixed at all times, and the curved bone shield is relatively thick compared to, for example, the titanium-reinforced PTFE membrane, and it has only the coronal aspect of the bone defect. The circle of the curved edge appears. For dentists, the 6 mm to 8 mm wide pings in the coronal aspect of the ridge will be very good (Wang and Al-Shammari 2002 Int. J. Periodontics Restorative Dent. 22(4): 335- 43). The resorbable morphologically stable collagen membrane disclosed in US-8353967-B2 attempts to combine smooth healing and morphological stability, the resorbable morphologically stable collagen membrane from 5% of the collagen-containing suspension in the mold to 25% ethanol/water was prepared by freeze-drying and heating at 100 ° C to 140 ° C. This film is manufactured by Osseous Technologies of the United States and sold under the trade name "Zimmer CurV Preshaped Collagen Membrane" by Zimmer. The commercially available film has a weak form stability and a thickness of about 1.5 mm, which rises to about 2.3 mm after incubation in brine; this can result in a higher risk of cracking. In summary, current solutions therefore do not adequately address the needs of dentists or patients. There is a higher risk of requiring secondary surgery and/or wound healing. A solution that is not associated with a higher risk of unhealed wound healing is not a non-morphologically stable membrane that requires a second surgery and has other disadvantages. US 2013/0197662 discloses a process for the manufacture of biological materials comprising: a) by applying a controlled amount of a gel comprising collagen to a bonding surface of a non-porous collagen-based material and allowing the porous collagen-based material The surface is in contact with a gel applied to the bonding surface to partially hydrate a portion of the porous material at the interface between the materials such that the porous collagen-based material joins the non-porous collagen-based material; b) dries the gel to Bonding the materials together; and c) crosslinking the collagen in the bonding layer. The obtained biomaterial obtained can be combined with the mineralized [0042], [0048] porous collagen-based material and the mechanically strong non-porous collagen-based material, thereby providing a load-bearing tissue for regeneration (especially The structure of the meniscus, articular cartilage, tendons and ligaments, the manufactured biomaterial has both porosity and mechanical strength, that is, it can withstand compressive forces and tensile forces. Nothing is disclosed for resisting the bending moment of the combined biological material, or for the composite of the mineralized porous collagen-based material. US 2014/0193477 teaches that in the manufacture of collagen mats from procollagen proteins, stretching the collagen prior to its crosslinking increases its mechanical strength, especially the ultimate tensile strength (UTS), hardness and modulus of elasticity (Yang Modulus) (Refer to [0109], [0110] for details). Langdon, Shari E et al. Biomaterials 1998, 20(2), 137-153 CODEN and Chachra, Debbie et al, Biomaterials 1996, 17(19), 1865-1875 CODEN reveals that stretching the pericardium source film before film cross-linking improves Its tensile strength and hardness.

It is an object of the present invention to provide a cross-linkable morphologically stable film for oral resorption which is suitable for resisting stress, shear and bending moments to support bone formation, bone regeneration, bone at a bone-free defect site Repair and/or bone replacement, especially in the maxilla or mandible, is amplified horizontally or vertically, which does not have the above disadvantages. This object is achieved by the invention as defined in the appended claims.

The present invention provides a resorbable cross-linked morphologically stable film for oral cavity comprising 1.5 to 3.5 parts by weight of an inorganic ceramic collagen material and a composite layer of inorganic ceramic particles per 1 part by weight of the collagen material. The composite layer of the collagen material and the inorganic ceramic particles is sandwiched between two layers of elastic pre-tensioned collagen material, the collagen material comprising 50% to 100% (w/w) collagen and 0% to 50% ( w/w) elastin. The term "collagen material" as used herein means a collagen-based material comprising 50% to 100% (w/w) collagen and 0% to 50% (w/w) elastin. In this context, elastin content is determined by the determination of the known method involving hydrolysis and RP-HPLC by means of a linear/iso-chainin assay (see, for example, Journal of Chromatography, Guida E. et al. 1990).Development And Validation Of a High Performance Gas chromatography Method For The Determination Of Desmosines In Tissue Or Rodriguqe P 2008 in The Open Respiratory Medicine JournalQuantification Of Mouse Lung Elastin During Prenatal Development ). To determine the elastin/iso-chainin content of dry elastin, the elastin of the sponge was subjected to an elastin separation procedure as described by Starcher and Galione in 1976 (Analytical Biochemistry)Purification And Comparison Of Elastin From Different Animal Species ). The collagen material is suitable for tissue derived from a natural source containing collagen and elastin in this ratio. Examples of such tissues include vertebrates, in particular the peritoneal or pericardium of mammals (eg, pigs, cows, horses, sheep, goats, rabbits), placental membranes, small intestinal submucosa (SIS), dermis, dura mater , ligaments, tendons, diaphragm (thoracic diaphragm), omentum, muscle or organ fascia. Such tissues are preferably pigs, cows or horses. The tissue of interest is the pig, cow or horse peritoneum. Usually collagen is mainly type I collagen, type III collagen or a mixture thereof. Collagen may also include a proportion of any type II, IV, VI or VIII collagen or any combination of these or any collagen type. Preferably, the collagen material contains 70% to 90% (w/w) collagen and 30% to 10% (w/w) elastin. An example of a suitable starting material for the preparation of such a collagen material is a collagen film prepared by a process similar to that described in the "Examples" of EP-B1-1676592, which is a porcine, bovine or equine periplasm or pericardium. Or the membrane Geistlich Bio-Gide® (available from Geistlich Pharma AG, Switzerland) prepared by the porcine peritoneum by this process. Preferably, the collagen material is derived from the porcine, bovine or equine pericardium or pericardium, small intestinal mucosa (SIS) or muscle fascia. The collagen material is generally and preferably a fibrous collagen material having a natural fibrous structure or as a sheared collagen fiber. However, if the collagen material has sufficient mechanical stabilization and maximum tensile strength in terms of elastic modulus, it may also be in a composite layer of collagen material and inorganic ceramic particles, or in elastic pre-tensioned collagen material. Non-fibrous collagen materials (such as fibrils reconstituted from molecular collagen or cross-linked collagen fragments with sufficient biocompatibility and resorbability) are used in the layers (see below). The term "resorbable" as used herein means that the crosslinked morphologically stable membrane can be resorbed in vivo, particularly via the activities of collagenase and elastase. The controlled in vivo resorbability of the crosslinked morphologically stable membrane is essential for the healing without excessive inflammation or cracking. Detailed description of Clostridium histolyticum from the following detailed description (Clostridium Histolicum The enzymatic degradation test of collagenase (Example 4, Example 3) gave an excellent prediction of resorbability in vivo. All tested prototypes of the resorbable cross-linked morphologically stable membranes of the invention tested exhibited at least 10% collagen degradation after 4 hours (as assessed by DC Protein assay using type I collagen as a standard), The rate of collagen degradation (below the rate of the Geistlich Bio-Gide® membrane) depends on the crosslinking conditions used. The term "crosslinking" means that the resorbable morphologically stable membrane has been subjected to at least one crosslinking step (usually chemical crosslinking (using, for example, EDC and NHS) or by dehydration heat treatment (DHT) crosslinking, usually by The step of chemically crosslinking (using, for example, EDC and NHS) or by means of dehydration heat treatment (DHT) of the collagen composite material sandwiched between two layers of elastic pre-tensioned collagen material and the assembled composite layer of inorganic ceramic particles. Optionally, the composite layer of collagen material and inorganic ceramic particles has been crosslinked by chemical crosslinking or by dehydration heat treatment (DHT) prior to its assembly into the film of the invention. The term "cross-linking morphologically stable membrane for reabsorbable oral cavity" means that the resorbable crosslinked membrane can be mechanically stabilized by providing a defect (ie, resisting stress, shear and bending moments occurring in the oral cavity). Support bone formation, bone regeneration, bone repair and/or bone replacement at the defect site of the human or animal without bone. The morphological stability of the film of the present invention was evaluated by a 3-point uniaxial bending test as described in detail below (in Example 4.2): the test is similar to the method described in EN ISO 178 and ASTM D6272-10, the film of the present invention. Immerse in PBS at a pH of 7.4 and 37 °C. This test demonstrates that the film of the present invention provides substantially greater stabilization than the comparative PLA film Resorb-X® (KLS Martin). In general, in the 3-point uniaxial bending test, the resorbable crosslinked morphologically stable film resists at least 0.20 N, preferably at least 0.30 N, at 8 mm strain. The term "elastic pre-tensioned collagen material layer" means that the layer of collagen material has been tensioned prior to cross-linking of the layers such that the initial size of the layer of collagen material elongates or pulls from the toe region of the stress-strain curve. Extending into a linear (also known as elastic) zone (see, Blayne A. Roder et al, 2002, Journal of Biomechanical Engineering, 124, 214-222, especially Figure 3, page 216, or Figure 5 of the present application). In this linear region, the modulus of elasticity is the highest and thus the highest hardness is achieved. The sheet of collagen material can be radially tensioned, for example, by a spring. The force to be applied such that the collagen material is elongated or stretched into the linear region of the stress-strain curve depends on the collagen material. When the collagen material is derived from a peritoneal membrane of pig, cow or horse, the linearity of the stress-strain curve of the collagen material can be radially generated on the collagen material sheet by a spring tensioned between 1 N and 3 N. The tension of the zone is such that 40% to 100% of the initial size of the layer of collagen material is stretched or stretched. The term "elastic pre-tensioned collagen material" thus means a collagen material that has been stretched so as to be in the linear (elastic) region of the stress-strain curve. Elastic modulus of elastic pre-tensioned collagen material (also known as Young's modulus) (ie, the slope of the linear region of the stress-strain curve expressed in MPa) is generally from 1 MPa to 1000 MPa, preferably from 2 MPa to 150 MPa, especially from 5 MPa to 80 MPa. It seems that there is a need to have two layers of "elastic pre-tensioned collagen material" sandwiched between a collagen material and a composite layer of inorganic ceramic particles to protect the composite layer from fracture when the film is subjected to tensile, compressive, shear and bending moments. . Preferably, one of the layers of elastic pre-tensioned collagen material comprises pores from 5 μm to 500 μm. When the film is placed in place, the perforated layer of elastic pre-tensioned collagen material will be oriented toward the bone defect, which allows the bone forming cells to easily invade into the inorganic ceramic collagen composite. Inorganic ceramics are biocompatible materials that promote bone regeneration, such as hydroxyapatite or natural bone minerals. A well-known natural bone mineral that promotes bone growth in tooth, periodontal and maxillofacial bone defects is Geistlich Bio-Oss® available from Geistlich Pharma AG. The skeletal-based mineral material based on hydroxyapatite is made of natural bone by the process described in U.S. Patent No. 5,167,961, which maintains the trabecular structure and nanocrystalline structure of the natural bone. Preferably, the inorganic ceramic is a natural bone mineral based on hydroxyapatite, such as Geistlich Bio-Oss®. The inorganic ceramic particles generally have a size of from 50 μm to 600 μm, preferably from 150 μm to 500 μm, especially from 250 μm to 400 μm. The composite of the collagen material and the inorganic ceramic particles contains 1.5 parts by weight to 3.5 parts by weight, preferably 2.0 parts by weight to 3.0 parts by weight, of the inorganic ceramic per part by weight of the collagen material. In fact, it has been unexpectedly found that, as less than 1.5 parts by weight of the inorganic ceramic per 1 part by weight of the collagen material, or more than 3.5 parts by weight of the inorganic ceramic per 1 part by weight of the collagen material, as defined above It was evaluated by the 3-point uniaxial bending test described in detail below (in Example 4.2): the film was not "morphologically stable". When the composite of the collagen material and the inorganic ceramic particles contains 2.0 parts by weight to 3.0 parts by weight of the inorganic ceramic of 1 part by weight of the collagen material, the morphological stability is particularly high. The resorbable cross-linked morphologically stable film of the present invention is hydrophilic and is typically completely wetted by PBS in 5 minutes to 10 minutes. The cell adhesion characteristics of the resorbable crosslinked morphologically stable membrane of the present invention are similar to those of Geistlich Bio-Gide®, which is a good healing property due to its low cracking rate or low excessive inflammatory rate. Well known. This indicates good healing properties without adverse conditions such as cracking or excessive inflammation. This good healing property has been observed when implanting the crosslinked morphologically stable membrane of the present invention to protect bone defects caused in rabbit skulls. The thickness of the resorbable crosslinked morphologically stable film of the present invention is usually from 0.5 mm to 2.5 mm, preferably from 1.0 mm to 2.0 mm, especially from 1.2 mm to 1.8 mm. Typical shapes and typical dimensions of the resorbable crosslinked morphologically stable film of the present invention are presented in FIG. The present invention also relates to the above resorbable crosslinked morphologically stable membrane for use in implants supporting bone formation, bone regeneration, bone repair and/or bone replacement at a defect site in a human or animal bone-free defect. Things. The invention also relates to a method of preparing a resorbable cross-linked morphologically stable film as defined above, the resorbable cross-linked morphologically stable film comprising collagen sandwiched between two layers of elastic pre-tensioned collagen material a composite layer of material and inorganic ceramic particles, the method comprising the steps of: (a) preparing a composite layer of collagen material and inorganic ceramic particles, optionally crosslinking the composite layer, and (b) subjecting the two layers of collagen to be tensioned A composite layer of the collagen material and the inorganic ceramic particles is assembled and glued between the protein materials, the tensioning causes the collagen material to be stretched in a linear region of the stress-strain curve; thereby obtaining a sandwich of the two layers of elasticity Tighten the composite layer of collagen material and inorganic ceramic particles between the collagen materials, and (c) cross-link the collagen material and the inorganic ceramic particles sandwiched between the two layers of elastic pre-tensioned collagen material Union, followed by the process of manufacturing hydrophilicity. Step (a) can be carried out by: - producing hydroxyapatite bone mineral particles as inorganic ceramic particles from cortical or cancellous bone by a process similar to that described in US-A-5,417,975, or alternatively Geistlich Bio-Oss Small Granules (available from Geistlich Pharma AG) is ground into smaller particles and subjected to sieving of the particles in the desired range (eg, 150 μm to 500 μm or 250 μm to 400 μm) Thereby, the sieved hydroxyapatite bone mineral particles are obtained. - Preparation of a fibrous collagen material by: o subjecting a collagen-rich tissue from a pig, cow or horse peritoneum or pericardium to a process similar to that described in the example of EP-B1-1676592; or Starting from the Geistlich Bio-Gide membrane (available from Geistlich Pharma AG) obtained from the pig peritoneum, or the intermediate product obtained before the industrial production of the Geistlich Bio-Gide membrane is sterilized (this article refers to Starting with the sterilizing Geistlich Bio-Gide film, o (for example, using scissors), the collagen fiber tissue thus obtained is cut into pieces, and the pieces of the cut collagen fiber tissue are mixed with dry ice using a chopper. Thus, the sheared collagen fibers are obtained, o the collagen fiber tissue is cut into pieces by a screen shearing machine, thereby obtaining sieved fragments of the collagen fiber fragments. - Preparation of a composite layer of fibrous collagen material and hydroxyapatite bone mineral particles by: o mixing and shaking 0 wt% to 40 wt% of sheared collagen fibers and 60 wt in phosphate buffered saline PBS The fraction of the sieved collagen fiber fragments obtained above 100% by weight is shaken, o the above obtained from 1.5 parts by weight to 3.5 parts by weight (especially 2.0 to 3.0 parts by weight) of the sieved hydroxyapatite The bone mineral particles are added to 1 part by weight of the fibrous collagen obtained in the above paragraph, and centrifuged at 2000 x g to 6000 x g, preferably 3000 x g to 5000 x g, and the obtained centrifugal block is poured into a rectangular model using a spatula. Form the board. The composite layer of the obtained fibrous collagen material and hydroxyapatite bone mineral particles was dried in a vacuum oven. It is not necessary to crosslink the dry composite layer of collagen material and inorganic ceramic particles after the end of (a), but the crosslinking has the benefit of facilitating the disposal of the composite layer during step (b). Crosslinking can be carried out using chemicals or by dehydration heat treatment (DHT). Crosslinking using the chemical can be carried out using any pharmaceutically acceptable crosslinking agent capable of providing the desired mechanical strength to the crosslinked morphologically stable film. Suitable such crosslinking agents include: glutaraldehyde, glyoxal, formaldehyde, acetaldehyde, 1,4-butane diglycidyl ether (BDDGE), N-sulfobutanediamine-6-(4 '-Azido-2'-nitrophenylamino)hexanoate, hexane diisocyanate (HMDC), cyanamide, diphenylphosphonium azide, genipin , EDC (1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide) and a mixture of EDC and NHS (N-hydroxysuccinimide). Crosslinking using chemicals is preferably carried out using a mixture of NHS and EDC. In this case, the dried composite layer of the above-obtained fibrous collagen material and hydroxyapatite bone mineral particles may be at a pH of 5.5 containing 10 mM to 400 mM EDC and 13 mM to 520 mM NHS 0.1 M MES ( 2-(N-morpholinyl)-ethanesulfonic acid) and 40% ethanol solution were crosslinked at room temperature for 1 hour to 3 hours. Subsequent to 0.1 M Na at pH 9.5₂ HPO₄ The prototype was incubated twice in the buffer for 1 hour to 3 hours to terminate the reaction. The polar residue can be removed by incubating the prototype in 1 M sodium chloride solution for 1 hour and incubating twice in 2 M sodium chloride solution for 1 hour. The chemically crosslinked prototype can be washed in distilled water for a total of 8 times from 30 minutes to 60 minutes. It can then be immersed in ethanol for 15 minutes for a total of 5 times, followed by 5 minutes of diethyl ether for 5 minutes and then dried overnight at 10 mbar and 40 ° C, or by lyophilization (below below -5 ° C) Drying is carried out by freezing and drying by a conventional lyophilization treatment. Alternatively, crosslinking is carried out by dehydration heat treatment (DHT) at 0.1 mbar to 10 mbar and at 80 ° C to 160 ° C for 1 to 4 days. In this case, a subsequent drying method is not required. Step (b) can be carried out by: - preparing a collagen fiber gel by: o using a high pressure homogenizer of 1500 bar to 2000 bar at a pH of 3.5 and a concentration of 3% H₃ PO₄ The sieved fragments of the above collagen fragments are mixed in an aqueous solution, and the mixture is repeatedly mixed several times. o The obtained slurry is neutralized to pH 7.0 by adding a sodium hydroxide solution, and the collagen is concentrated by lyophilization and by a chopper. Homogenize the latter, o by heating to 60 ° C until no other visible particles, from the obtained slurry to prepare a pH of 7.4 phosphate buffered saline PBS collagen fiber gel containing 2% to 10% solution, and - use (For example) a device similar to the device of Figure 2, subjecting the two pre-wetting layers of collagen material to tension, causing the collagen material to stretch in the linear region of the stress-strain curve, thereby obtaining two layers of wetting Elastic pre-tensioning of the collagen material, inserting a composite layer of the collagen material and the inorganic ceramic particles obtained in (a) infiltrated with the above-mentioned collagen fiber glue into the two layers of the wet elastic pre-tensioned collagen material Using two devices, such as a device similar to the device of Figure 3, to press the two layers of the wet elastic pre-tensioned collagen material against the collagen material and the inorganic ceramic particles infiltrated with the collagen fiber cement. Composite layer and drying of collagen material and inorganic ceramic particles sandwiched between two layers of wet elastic pre-tensioned collagen material under reduced pressure (for example, 20 mbar to 1 mbar) at a temperature of 35 ° C to 45 ° C The composite layer. In the procedure described above, one of the pre-wetting layers of collagen material may have been needle-punched to include pores from 5 μm to 500 μm. In step (c), the collagen material and the inorganic ceramic sandwiched between the two layers of elastic pre-tensioned collagen material may be cross-linked using a chemical (for example, using EDC and NHS) or by dehydrating heat treatment of DHT. a composite layer of particles. Chemical crosslinking can be carried out using any pharmaceutically acceptable crosslinking agent capable of providing the necessary mechanical strength to the crosslinked morphologically stable film. Suitable such crosslinking agents include: glutaraldehyde, glyoxal, formaldehyde, acetaldehyde, 1,4-butane diglycidyl ether (BDDGE), N-sulfobutanediamine-6-(4 '-Azido-2'-nitrophenylamino)hexanoate, hexane diisocyanate (HMDC), cyanamide, diphenylphosphonium azide, genipin, EDC ( 1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide) and a mixture of EDC and NHS (N-hydroxysuccinimide). Crosslinking using chemicals is preferably carried out using a mixture of EDC and NHS. In this case, the dried composite layer of the above-obtained fibrous collagen material and hydroxyapatite bone mineral particles may be at a pH of 5.5 containing 10 mM to 400 mM EDC and 13 mM to 520 mM NHS 0.1 M MES ( 2-(N-morpholinyl)-ethanesulfonic acid) and 40% ethanol solution were crosslinked at room temperature for 1 hour to 3 hours. Subsequent to 0.1 M Na at pH 9.5₂ HPO₄ The prototype was incubated twice in the buffer for 1 hour to 3 hours to terminate the reaction. The polar residue can be removed by incubating the prototype in 1 M sodium chloride solution for 1 hour and incubating twice in 2 M sodium chloride solution for 1 hour. The chemically crosslinked prototype can be washed in distilled water for a total of 8 times from 30 minutes to 60 minutes. It can then be immersed in ethanol for 15 min for a total of 5 times, followed by diethyl ether treatment for 5 minutes three times, followed by drying at 10 mbar and 40 ° C for 30 minutes; or by lyophilization (below -10) It is frozen at ° C and dried by a conventional lyophilization treatment without dehydration and drying by means of solvent treatment. Alternatively, crosslinking is carried out by dehydration heat treatment (DHT) at 0.1 mbar to 10 mbar and at 80 ° C to 160 ° C for 1 to 4 days. In this case, a subsequent drying method is not required. The process for producing hydrophilicity of step c) generally comprises hydrophilically immersing the crosslinked composite layer of the collagen material and the inorganic ceramic particles sandwiched between the two layers of elastic pre-tensioned collagen material to a physiologically acceptable salt. In solution, such as a sodium chloride solution, preferably from 100 g/l to 300 g/l, especially from 150 g/l to 250 g/l sodium chloride solution. Preferably, the process of producing hydrophilicity comprises hydrophilically immersing the crosslinked composite layer of the collagen material and the inorganic ceramic particles sandwiched between the two layers of elastic pre-tensioned collagen material into the sodium chloride solution. The resorbable crosslinked morphologically stable film of the present invention can be sterilized by X-ray, β-ray or gamma radiation. The following examples illustrate the invention without limiting its scope.Instance 1 Preparation of raw materialsPreparation size is 250 μ m to 400 μ m It Hydroxyapatite fine particles ( A ) Hydroxyapatite bone mineral fine particles are produced from cortical or cancellous bone using an additional screening step between 250 μιη and 400 μιη as described in Examples 1 to 4 of US-A-5,417,975. Or produce hydroxyapatite bone mineral fine particles by: grinding Geistlich Bio-Oss® Small Granules by careful impact using an impact gun and an additional screening step between 250 μm and 400 μm Geistlich Pharma AG, CH-6110, Switzerland). The hydroxyapatite bone mineral fine particles (A) having the size of 250 μm to 400 μm prepared above were stored in a glass bottle until use.Preparation of collagen fibers ( B ) As described in the "Examples" of EP-B1-1676592, the peritoneum from young pigs is completely free of meat and fat by mechanically washing under running water and treated with 2% NaOH solution for 12 hours. It was then washed under running water and the membrane was acidified with 0.5% HCl. After acidification through the entire thickness of the material (for about 15 minutes), the material was washed with water until a pH of 3.5 was obtained. Subsequently, shrink with 7% saline solution, using 1% NaHCO₃ The solution is neutralized and the material is washed under running water. Subsequently, it was dehydrated with acetone and degreased with n-hexane and dried using ethyl ether. The collagen film thus obtained was cut into 2 cm × 2 cm pieces by hand using scissors. Alternatively, a 2 cm x 2 cm Geistlich Bio-Gide® patch block (available from Geistlich Pharma AG) was manually cut using scissors. The 1 g 2 cm × 2 cm collagen membrane piece obtained above was mixed with 200 ml of dry ice and mixed in a chopper (Retsch® Grindomix) at 5000 rpm until no obstruction occurred. The speed was then increased to 6000 rpm, 7000 rpm, 9000 rpm, and 10,000 for 20 seconds to 30 seconds with 50 ml of dry ice added each time. The dry ice was vaporized, and the thus obtained collagen fibers (B) were stored in a Minigrip plastic package until further use.Preparation of shearer collagen fiber fragments ( C ) The 2 cm × 2 cm collagen fiber pieces obtained above were sheared at 1500 rpm in a shearing machine with a 0.8 mm sieve to obtain sieved fragments of the shearer collagen fiber fragments (C ).Preparation of collagen fiber glue ( D ) The sieved fragments (C) of the shearer collagen fiber fragments are mixed in water to obtain a 3% solution by adding phosphoric acid H₃ PO₄ The pH was set to 3.5, and the suspension was homogenized at a high pressure of 1500 to 2000 bar, which was repeated 3 to 5 times. The resulting slurry was neutralized to about pH 7 by the addition of sodium hydroxide solution NaOH and gelled overnight at 4 °C. After freezing at -40 ° C for 4 hours, the collagen was concentrated by lyophilization at -10 ° C and 0.310 mbar and homogenized by a chopper. Collagen fiber gel (D) was prepared from the obtained slurry of phosphate buffered saline containing 2% to 10% solution of pH 7.4 by heating to 60 ° C until no other visible particles were present.Instance 2 Preparation of cross-linked hydroxyapatite/collagen plate (E) 4 g of collagen fiber (B) prepared in Example 1 and 6 g of shear collagen fiber fragment (C) were buffered with 140 g of phosphate. The brine was mixed and shaken in a mixer mixer. In another example, the collagen fibers are completely replaced by shearer collagen fiber fragments. 20 g of hydroxyapatite fine particles (A) prepared in Example 1 were added and mixed artificially. This 34.14 g mixture was centrifuged at 7000 g (7000 times gravitational acceleration) for 2 minutes. Pour the pellet into a flat rectangular shape of 8 cm × 12 cm between two polyamine meshes (with a pore size of 21 μm and a total open structure of 17%) and concentrate the material by removing excess water with a laboratory spoon. . The obtained plate was compressed at a pressure of 1 kPa to 1.7 kPa, and dried in a vacuum oven at 30 ° C / 50 mbar for 2 hours, followed by drying at 30 ° C / 10 mbar for 8 hours. Remove the polyamine mesh.Cross-linked hydroxyapatite collagen plate as appropriate To facilitate disposal of the hydroxyapatite collagen plate, the latter is chemically crosslinked or crosslinked by dehydration heat treatment (DHT). Chemical cross-linking of collagen with EDC/NHS improves the overall stability of the hydroxyapatite collagen plate. Subsequently, the plate was dried at room temperature at pH 5.5 with 0.1 M MES (2-(N-morpholinyl)-ethanesulfonic acid) and 40% ethanol containing 10 mM to 400 mM EDC and 13 mM to 520 mM NHS. Cross-linking for 2 hours. By 0.1 mol/l Na at pH 9.5₂ HPO₄ The prototype was incubated twice in buffer for one hour to stop the reaction. The polar residue can be removed by incubating the prototype in 1 mol/l sodium chloride solution for 1 hour and incubating twice in 2 mol/l sodium chloride solution for 1 hour. The chemically crosslinked prototype can be washed in distilled water for a total of 8 times from 30 minutes to 60 minutes, followed by dehydration by immersing it in ethanol for 15 minutes for a total of 5 times. This is carried out by treating the diethyl ether three times for 5 minutes and then drying at 10 mbar and 40 ° C for 30 minutes, or by lyophilization (freezing at less than -10 ° C and drying by conventional lyophilization). Drying treatment. Alternatively, crosslinking is carried out by dehydration heat treatment (DHT) at 0.1 mbar to 10 mbar and at 80 ° C to 120 ° C for 1 to 4 days. In this case, a subsequent drying method is not required.Instance 3 A resorbable cross-linked morphologically stable film (M) is prepared by assembling and gluing two layers of elastic pre-tensioned collagen layers on two opposite faces of a hydroxyapatite/collagen plate (E) by reference 2 and Figure 3 will better understand the following description. The assembly of a flat or U-shaped prototype requires the use of a fixed or bendable frame that can tension the layer of collagen material.Form flat or U shape prototype ( F ) Figure 2 is a schematic illustration of an apparatus suitable for tensioning a layer of collagen material prior to assembly into a flat or U-shaped morphologically stable film of the present invention. The device consists of a frame (a) which can be made of any suitable material, such as steel or aluminum. The main purpose of the frame is to anchor the spring (b) that tensions the two wet collagen layers (c). The hydroxyapatite/collagen plate (E) is positioned between the two collagen layers (c). If a U-shaped resorbable crosslinked morphologically stable film is required, a negative model (e) for bending the collagen plate (E) and a frame with a hinge (f) are used, thus obtaining a U-shaped linear prototype. By stretching or stretching 40% to 100% of the initial length, by tensioning each spring 2 N to 3 N, the layer of collagen material of the unsterilized Geistlich Bio-Gide Collagen layer is pre-tensioned so as to be in collagen The linear region of the stress curve of the protein material. In this linear region, the modulus of elasticity is highest and thus the highest hardness is achieved. Due to the viscoelastic nature of the collagenous tissue, the wetted and taut material is held in tension for approximately 30 minutes. Due to the relaxation of the pre-tensioned collagen membrane, the spring is again tensioned 1 N to 3 N in the linear region of the stress curve of the collagen material. Two round pieces of collagen with a diameter of 10 cm cut from the unsterilized Geistlich Bio-Gide® collagen membrane, one of which contains a needle of 50 needles with a shaft diameter of 0.88 mm per square centimeter Tube perforation. The two round pieces of collagen are wetted and tensioned in a radial manner by 12 springs each tensioned to 1 N to 3 N, resulting in a 40% elongation of the collagen pieces from the initial size. 100%. After completing this step, both sides of the hydroxyapatite/collagen plate (E) are wetted with collagen fiber glue (C), and then the hydroxyapatite/collagen plate is placed in two elastic pre- Tighten between the collagen layers. A central rod (e) and a hinge (f) are required to produce a U-shaped prototype (see below). The elastic pretensioned film was placed on a hot plate and preheated to 40 °C. The crosslinked Bio-Oss plate (E) obtained in Example 2 was briefly immersed in the preheated fiber gel (D) and placed between two elastic pre-tensioned collagen films. Polyamide mesh and sponge (made of polyurethane, 5 cm thick, density approximately 20 mg/cm³ Up to 25 mg/cm³ , with interconnected micropores) placed on both sides, compressing 50% to 95%, resulting in compressive stresses up to 120 kPa. Referring to Figure 3, it shows the assembly of a flat morphologically stable membrane: (1) is a steel plate, (2) is a compressed polyurethane sponge, (3) is a polyamine mesh, and (4) is an elastic pre-tensioned collagen. The protein layer, and (5) is a crosslinked hydroxyapatite collagen plate. Subsequently, the construct was dried in a vacuum oven at 40 ° C, with air pressure dropping smoothly to 10 mbar over a total of 32 hours.form U shape prototype By bending the construct on a suitable negative model and replacing one of the sponges with a thinner polyurethane foam or a fiber-free paper towel, those skilled in the art will readily adapt the apparatus of Figures 2 and 3 and the above. The method is to form a U-shaped straight or curved prototype.Cross-linking flat or U shape prototype ( G ) Use a pair of scissors or a small circular saw to cut a flat or U-shaped prototype (F) to the desired size. Subsequently, the prototype is crosslinked by chemical crosslinking or by dehydration heat treatment (DHT). Chemical cross-linking was carried out in 0.1 mol/L MES buffer having a pH of 40 Vol-%, EDC and NHS concentrations of 10 mM to 400 mM and 13 mM to 520 mM, respectively, at a pH of 5.5. The prototype concentration in the cross-linking solution was 10%. For uniform crosslinking, the plates were initially treated under vacuum (<40 mbar) and the crosslinking reaction was carried out at 4 °C for 2 hours, and all buffers were pre-cooled to this temperature. By 0.1 mol/l Na at pH 9.5₂ HPO₄ The prototype was incubated twice in buffer for one hour to stop the reaction. The polar residue was removed by incubating the prototype in 1 mol/l NaCl solution for 1 hour and incubated twice in 2 mol/l NaCl solution for 1 hour. The prototype was washed in distilled water for a total of 8 times from 30 minutes to 60 minutes. Subsequent to 5 times of ethanol treatment for 15 minutes and diethyl ether treatment for 5 minutes and subsequent drying at 10 mbar and 40 ° C overnight or until the product is completely dry, or by conventional freeze-drying (below -10) Dehydration and drying treatment are carried out by freezing at ° C and drying by conventional lyophilization treatment without the solvent treatment. Alternatively, crosslinking is carried out by dehydration heat treatment (DHT) at from 0.1 mbar to 10 mbar at from 80 ° C to 160 ° C for 1 to 4 days. In this case, a subsequent drying method is not required. The prototype obtained by the above method was wetted in saline or PBS for one hour or two hours. To allow wetting within 10 min, the prototype was pre-wetted in distilled water for approximately 1 hour to 2 hours. At this time, it is also possible to perforate one side with the above-mentioned syringe. Sodium chloride was coated by incubating the prototype three times in a 200 g/l NaCl solution for 40 min. Sodium chloride (H) was precipitated as described below.Dry cross-linking flat or U shape prototype ( H ) The crosslinked prototype was dehydrated by immersing in ethanol for 15 minutes for a total of 5 times. Subsequently, it is dried by solvent (three times of diethyl ether treatment for 5 minutes and subsequent drying at 10 mbar and 40 ° C), or by conventional lyophilization (freezing at less than -10 ° C and by conventional means The lyophilized treatment is dried to dry the crosslinked prototype. The crosslinked morphology of the different prototypes in the wet state stabilizes the film from 1.0 mm to 2.0 mm, most of which is from 1.2 mm to 1.8 mm. The dried prototype can be sterilized by x-ray radiation from 27 kGy to 33 kGy as appropriate.Instance 4 Characteristics of resorbable crosslinked morphologically stable film The following characteristics of the resorbable crosslinked morphologically stable film obtained in Example 3 were determined: (1) wettability in PBS, (2) mechanical strength, (3) Use fromClostridium Enzymatic degradation of collagenase, and (4) cell adhesion, (5) measurement of the elongation of the elastic pre-tensioned collagen material layer, (6) measurement of the thickness of the collagen hydroxyapatite plate and the final prototype value.( 1 ) PBS middle Wetability For different prototypes of resorbable cross-linked morphologically stable membranes, the time to complete wetting in PBS (phosphate buffered saline) was observed as between 5 minutes and 10 minutes as assessed by the naked eye, depending on the time. Treated with sodium chloride before dehydration with ethanol and drying.( 2 ) Mechanical strength The morphological stability of the film of the present invention was evaluated by a 3-point uniaxial bending test similar to the method described in EN ISO 178 and ASTM D6272-10, and the film of the present invention was immersed at pH7 . 4 and the temperature was 37 ° C in PBS. This test is considered very useful because it is designed to mechanically stabilize each of the morphologically stable membranes without bone defect sites to be affected by the bending moment. Thus, 3 or 4 point bends can be used as tests to characterize the materials used and additionally to compare, for example, different products having different thicknesses. For material characterization, the flexural modulus is the most suitable parameter. However, to compare products with different thicknesses, the maximum force after indentation of 8 mm to 10 mm is more relevant and therefore used to characterize the product. In the 3-point uniaxial bending test used, the specimen was cut to a size of 50 mm × 13 mm and incubated in PBS at 37 ° C as observed by the naked eye until complete wetting. Mechanical testing was performed at 5 mm per minute in a 3-point bending device with a span width of 26 mm and a radius of 5 mm for each support structure. The bending module was calculated to be within 1% and 5% bending strain. After reducing the intermediate indentation between 8 mm and 10 mm, the maximum force is read. A film of the present invention having a thickness of 1.5 mm crosslinked by EDC/NHS, a film of the present invention having a thickness of 1.6 mm crosslinked by DHT, and a PLA film Resorb-X having a thickness of 0.137 mm from KLS Martin ® for testing. Figure 4 shows the change in force as a function of the stress of the films, showing the invention by EDC/NHS cross-linking (8 mm strain about 0.65 N) or by DHT cross-linking (8 mm strain about 0.40 N) The mechanical stabilization of the film is substantially better than the mechanical stabilization of the PLA film Resorb-X® (8 mm strain about 0.10 N). The membrane of the invention will thus better stabilize bone-free defect sites.( 3 ) Use from Clostridium Enzymatic degradation test In humans, collagen is degraded by human tissue matrix metalloproteinases (MMPs), cathepsins, and presumably by partial serine proteases. Since collagenase is the most important enzyme for the direct degradation of collagen, the most studied are MMP (especially MMP-1, MMP-8, MMP-13 and MMP-18) (Lauer-Fields et al. 2002)Matrix Metalloproteinases And Collagen Catabolism In Biopolymers - Peptide Science Section and Song et al. 2006Matrix Metalloproteinase Subject dependent And Independent Collagen Deposition In Frontiers in Bioscience). The collagenase capacity of the degraded collagen tissue and membrane depends on the substrate flexibility and collagen type, MMP active site and MMP external site. The collagenase is aligned at the triple helix collagen, unwinding the triple helix collagen and subsequently decomposing it (Song et al. 2006, supra). To overcome the difference in the degradation of different types of collagen, it is often used from a higher catalytic rate.Clostridium Collagenase to assess collagen degradation by collagen (Kadler et al. 2007)Collagen At a Glance In J Cell Sci). In general, natural collagen products degrade faster than chemically crosslinked collagen products. In this test, a collagen product (a sample of a resorbable cross-linked morphologically stable membrane of 1 mg/ml collagen) and 50 units/ml were cultured in a calcium-containing buffer at 37 °C.Clostridium (One unit is defined as: in the ninhydrin coloration, at 37 ° C, pH 7.4, in the presence of calcium ions, the release from the burdock tendon equivalent to 1.0 mol of leucine in 5 hours Peptide); degradation of the collagen matrix was measured by the naked eye and with "DC Protein Assay" from Bio-Rad Laboratories (Hercules, USA, order number 500-0116) using type I collagen as a reference material. The collagen concentration was determined using a microplate spectrometer (Infinite M200, available from Tecan). All prototypes of the resorbable cross-linked morphologically stable membranes of the present invention exhibit at least 10% collagen degradation after 4 hours (as assessed by DC Protein assay using type I collagen as standard), collagen degradation rate ( The rate below the Geistlich Bio-Gide® film depends on the crosslinking conditions used.( 4 ) Cell adhesion 8mm membrane punching with 100,000 human gingival fibroblasts previously labeled with fluorescent, lipophilic dyes was first inoculated, incubated in PBS for 24 hours at 37 ° C, and removed by washing the membrane in PBS. Non-adherent cells, lysing adherent cells and quantifying the cells by measuring fluorescence at 485 nm to assess cell adhesion of different membranes. Fluorescence was normalized to a standard curve established by punching the membrane of the inoculated cells that were not washed prior to lysis. The results obtained for the morphologically stable resorbable film are presented in Figure 5, which is a horizontally oriented, percentage-dependent adhesion of different types of dental membranes, the resorbable cross-linked morphologically stable membrane of the present invention. , and a histogram of % of cells on the Cystoplast® PTFE membrane (Keystone Dental). Figure 5 shows that the resorbable cross-linked morphologically stable film adhered to the present invention is about 10.5%, which is closer to the Geistlich Bio-Gide® film (about 13%) than the Cystoplast® PTFE film (about 4%). Geistlich Bio-Gide® membranes are well known for their good healing properties with low cracking rates (Zitzmann, Naef et al. 1997; Tal, Kozlovsky et al. 2008) or no excessive inflammation (Jung, 2012). The measurement of the adhesion of the human gingival fibroblasts to the resorbable cross-linked morphologically stable membrane of the present invention is a predictor of soft tissue healing without adverse conditions such as excessive inflammation or cracking.( 5 ) Measuring the elongation of the elastic pre-tensioned collagen material layer To determine the amount of tension in the collagen layer, a dry collagen layer was attached to the tensioning ring (Fig. 2, part a) using a spring that has not been tensioned (Fig. 2, part b). At least 4 points spaced apart from each other by a few centimeters are marked in the middle of the film using a pencil or a pen. Use a ruler to measure the distance between each point. The measured distance is defined as the initial length between each point. Immerse the collagen layer in water and tighten to the desired force. The collagen layer was incubated in water for 30 minutes. Due to the viscoelastic nature of most collagen layers, the tension is reduced. Therefore, the collagen layer needs to be tightened again. After incubation for 30 minutes to 40 minutes, the distance between each point was measured with a ruler. The percent strain is determined by subtracting the initial length from the length after tensioning, dividing by the initial length, and multiplying by 100. For unsterilized Geistlich Bio-Gide, a typical result in the linear region of the stress-strain curve is between 40% and 100% strain (elongation, elongation). The strain value measured by this method is not directly equivalent to the strain value obtained in the uniaxial elongation test.( 6 ) Measuring the thickness of the collagen hydroxyapatite plate and the final prototype The thickness of the final prototype or collagen/hydroxyapatite sheet "E" can be measured as described above or by using a sliding caliper.( 7 ) Analysis of mechanical properties of different collagen layers ( Figure 5 ) To compare collagen layers from different sources and evaluate their mechanical properties, wet the sample using standard uniaxial tension. The general settings for this analytical method are described in ASTM D882-09 "Standard Test Method for Tensile Properties of Thin Plastic Sheeting". Due to the high cost of the collagen membrane used, several parameters of the test can be adapted. The sample was cut into, for example, a 2 cm × 1 cm rectangular sheet, pre-wetted in isotonic phosphate buffered saline and mounted to a tension tester with a distance of 1 cm between each sample holder. The sample was tensioned at a constant speed of 33% of the initial length per minute. The pre-force recorded at 100% initial length is typically set to 50 kPa. The distance between the two sample holders is used to calculate the elongation of the sample. Therefore, the stress-strain curve of Fig. 5 is obtained. The present invention has been described and illustrated in detail in the drawings and the description herein. Other variations of the disclosed embodiments can be understood and effected by the practice of the invention as claimed in the appended claims. In the scope of application for patents, the word "comprising" does not exclude other elements; the definite article "a/an" does not exclude the plural.

1, 1'‧‧‧ flat

2, 2'‧‧‧U-shaped straight line

3, 3'‧‧‧ U-shaped arc

1‧‧‧ steel plate

2‧‧‧Compressed polyurethane sponge

3‧‧‧ Polyamide network

4‧‧‧Elastic pre-tensioned collagen layer

5‧‧‧Crosslinked hydroxyapatite collagen plate

BRIEF DESCRIPTION OF THE DRAWINGS The present invention will now be described in further detail with reference to the preferred embodiments of the preferred embodiments of the invention, in which: Figure 1 shows a typical shape of a resorbable crosslinked morphologically stable film in accordance with the present invention. And typical size. The membranes may be flat corresponding to the alveolar cavities of one to three teeth (incisors, canines, premolars or molars) positioned at the left or right side of the front or back of the denture (1) ), (1'), U-shaped linear (2), (2') or U-shaped curved (3), (3'). The size of the anterior product is similar to the size of the posterior product and the radius of curvature is the radius of the alveolar ridge. Typical dimensions are: a = 5 nm to 20 nm, b = 8 mm to 20 mm, c = 6 mm to 10 mm, d = 25 mm to 40 mm, e = 15 mm, f = 20 mm to 40 mm. 2 is a schematic illustration of an apparatus for tensioning a polymer layer prior to assembly of the polymer layer into the flat or U-shaped morphologically stable film of the present invention. Figure 3 shows the assembly of a flat morphologically stable membrane: (1) is a steel plate, (2) is a compressed polyurethane sponge, (3) is a polyamine mesh, and (4) is an elastic pre-tensioned collagen layer. And (5) is a crosslinked hydroxyapatite collagen plate. Figure 4 shows the force versus strain change in a 3-point bending analysis test of the resorbable morphologically stable film of the present invention crosslinked by EDC/NHS or DHT compared to PLA film Resorb-X® (KLS Martin). Figure 5 presents a stress-strain curve of a few commercially available wet and sterile collagen materials that can be used in the elastic pre-tensioned collagen material layer of the resorbable cross-linked morphologically stable film according to the present invention. , that is, Geistlich Bio-Gide® collagen membrane derived from pig peritoneum (Geistlich Pharma AG), Jason® collagen membrane derived from porcine pericardium (aap Biomaterials/Botiss), and Dynamatrix® collagen derived from porcine SIS Membrane (Cook Biotech Inc.), and the stress-strain curve of collagen material derived from muscle fascia. In each of these stress curves, there is a toe region characterized by a large strain based on a minimum stress value, a linear or elastic region characterized by a linear increase in strain per unit stress, and characterized by fracture of the polymer fiber. The failure zone. In the stress-strain curve presented in this figure, the elastic modulus (or Young's modulus, or the slope of the linear region of the stress-strain curve) of the Geistlich Bio-Gide® film is about 8 MPa, Jason film The modulus of elasticity is about 64 MPa, the modulus of elasticity of the Dynamatrix® film is about 54 MPa, and the modulus of elasticity of the collagen material derived from the muscle fascia is about 56 MPa. Figure 6 is a bar graph of % of human gingival fibroblasts that have adhered to the membrane after incubation for 24 hours at 37 ° C in PBS, which are Geistlich Bio-Gide® collagen membranes, delivered by DHT (FRM) A prototype of the resorbable morphologically stable membrane of the present invention and a histogram of a Cystoplast® PTFE membrane (Keystone Dental).

Claims (15)

A cross-linkable morphologically stable film for respirable oral cavity, comprising a composite layer of collagen material and inorganic ceramic particles containing 1.5 parts by weight to 3.5 parts by weight of inorganic ceramic per 1 part by weight of collagen material, the collagen material And a composite layer of inorganic ceramic particles sandwiched between two layers of elastic pre-tensioned collagen material (ie, a collagen material that has been stretched into an elastic region in a stress-strain curve), the collagen material comprising 50% Up to 100% (w/w) collagen and 0% to 50% (w/w) elastin. The resorbable morphologically stable film according to claim 1, wherein the composite layer of the collagen material and the inorganic ceramic particles contains 2.0 parts by weight to 3.0 parts by weight of the inorganic ceramic per 1 part by weight of the collagen material. A cross-resorbable morphologically stable membrane according to claim 1 or 2, wherein the collagen material comprises 70% to 90% (w/w) collagen and 10% to 30% (w/w) elastin . The resorbable cross-linked morphologically stable film according to any one of claims 1 to 3, wherein the collagen material is derived from a tissue of a natural source selected from the group consisting of a mammalian peritoneum or pericardium, a placental membrane, Small intestinal submucosa (SIS), dermis, dura mater, ligament, tendon, diaphragm (eg chest diaphragm), omentum and fascia of muscle or organ. The resorbable cross-linked morphologically stable film according to any one of claims 1 to 3, wherein the collagen material is derived from a pig, cow or horse peritoneal or pericardium, small intestinal mucosa (SIS) or muscle fascia. The resorbable crosslinked morphologically stable film according to any one of claims 1 to 4, wherein the elastic pre-tensioned collagen material has an elastic modulus of from 2 MPa to 150 MPa. The resorbable cross-linked morphologically stable film of any one of claims 1 to 4, wherein one of the layers of the elastic pre-tensioned collagen material layer comprises pores of from 5 μm to 500 μm. The resorbable crosslinked morphologically stable film according to any one of claims 1 to 7, wherein the inorganic mineral particles have a size of from 150 μm to 500 μm. The resorbable crosslinked morphologically stable film according to any one of claims 1 to 8, wherein the inorganic ceramic is hydroxyapatite. The resorbable crosslinked morphologically stable film according to any one of claims 1 to 9, wherein the inorganic ceramic is a hydroxyapatite bone mineral. The resorbable crosslinked morphologically stable film according to any one of claims 1 to 10, which is chemically crosslinked. The resorbable crosslinked morphologically stable film according to any one of claims 1 to 10, which is subjected to dehydration heat treatment for DHT crosslinking. A method for producing a crosslinked resorbable crosslinked morphologically stable film according to any one of claims 1 to 12, which comprises the steps of: (a) preparing a composite layer of inorganic ceramic particles and collagen material, as the case may be The composite layer is crosslinked, (b) assembling and gluing a composite layer of the collagen material and the inorganic ceramic particles between the two layers of collagen material subjected to tension, the tensioning stretching the collagen material in stress-strain a linear region of the curve; thereby obtaining a composite layer of collagen material and inorganic ceramic particles sandwiched between two layers of elastic pre-tensioned collagen material, and (c) sandwiching the two layers of elastic pre-tensioned collagen material The composite layer of the collagen material and the inorganic ceramic particles is crosslinked, followed by a process for producing hydrophilicity. The method of claim 13, wherein the process of producing hydrophilicity comprises immersing the crosslinked composite layer of the collagen material and the inorganic ceramic particles sandwiched between the two layers of elastic pre-tensioned collagen materials in a sodium chloride solution. . A resorbable cross-linked morphologically stable film according to any one of claims 1 to 12, which is useful for supporting bone formation, bone regeneration, bone repair and/or at a defect-free site in a human or animal. Or bone replacement implants.