DE102005041414A1 - Glass sponge collagen obtained by gradually corroding glass sponge basal spicule in alkaline solution; dialyzing the obtained extract and subsequently lyophilizing, useful for the production of e.g. biological material and bullets - Google Patents

Abstract

Glass sponge collagen, obtained by gradually corroding glass sponge basal spicule in alkaline solution; dialyzing the obtained extract and subsequently lyophilizing. Independent claims are included for: (1) the preparation of the glass sponge collagen comprising crushing and washing the glass sponge of basal spicule in alkaline solution at 35-37[deg]C and gradually corroding; dialyzing to isolate the glass sponge collage and subsequently lyophilizing; and (2) a procedure for in-vitro-production of nano-structured composite and materials, which consist of sea-glass sponge collagen and silicon dioxide, comprising: lyophilizing the solution in alkaline buffer solution; mineralizing through a mixture of the solved collagen with silane solution as silicate precursor; growing and agglomerating the silicate nano particle in room temperature; and air drying the obtained product in room temperature for a period of at least 4 days.

Description

The The invention relates to glass sponge collagen, process for its preparation and its use. The collagens obtained from marine glass sponges can be used for Production of a variety of biomaterials based on Collagen and silica are used.

invertebrates Animals of the deep sea, including Glass sponges, have a big one Number of exceptional Skeletal structures on. Various representatives of glass sponges of The genus Hexactinellida (Porifera) are characterized by their basal spicules known.

Hyalonema glass sponges are also known as "glass sponge" because of the twisted basal spicules, which anchor the sponge in the soft ground sediment and can grow up to a meter long.This type of stem of spicules can be affected by water flow bend and curve and thus easily adapt to the change in the flow direction or strength. 1 shows a marine glass sponge Hyalonema sieboldi, a typical member of the family Hyalonematidae. Presumably, the flexibility of these spicules results from the presence of organic networks.

The Origin and the possible Role of the organic matrix in the midst of these skeletons containing silicon are still unknown. The investigation of the biochemical composition and the microstructure of the spicules of marine glass sponges as Representative of natural Structured biomaterials is of fundamental scientific nature and practical meaning. The real understanding of the role of the organic matrix the hexactinellida, which structurally affects the silica mineralization works, is possibly the key for the development of new biomimetic silicates Biomaterials for biomedical and technical applications.

Around to understand the role of the organic matrix in basal spicules However, before the isolation of the same is necessary.

So far based on the usual Technique of desilication of sponge spicules on use of hydrofluoric acid (See also Uriz et al., Siliceous Spicules and Skeleton Frameworks in Sponges: Origin, Diversity, Ultrastructural Patterns, and Biological Functions, Microscopy Research and Technique 62: 279-299 (2003)). Recently became This method is considered too aggressive, the drastic changes the structure of possibly stored proteins can cause (See, e.g., Croce et al., Structural characterization of siliceous spicules from marine sponges, Biophysical Journal 86: 526-534 (2004)).

A further biogenic dissolution method is based on treatment with 0.5 M NaOH and subsequent heating 85 ° C (cf. also Conley, D.J. An interlaboratory comparision for the measurement of biogenic silica in sediments, Marine Chemistry 63: 39-48 (1998)). It is stored because of the associated denaturation Proteins also not acceptable.

collagen becomes common from skin, tendons or other parts of animals (bovine, swine, Poultry) obtained in processes that extractions in acid and / or by enzymes. Fundamental spoken, include collagen preparation methods the purification of crude collagen by extraction with dilute organic acids, the precipitation with salts, gelation and / or lyophilization, filtration, etc. After removal of connective tissue, fat and impurities will be the tissue of a moderate treatment with proteolytic enzymes, such as. Pepsin, after which the collagen is neutral pH is deposited. Then it is solved again and remaining Impurities are deposited in an acidic environment (e.g., U.S. 6,548,077).

Mammalian collagens are potentially risky in that they can transmit pathogens (prions, viruses), while marine collagens are less risky in this regard. Collagens are isolated from skin (see Japanese patents no. JP8283665 . JP2000050811 . JP2003301144 . JP2003092997 . JP2004269478 . JP2005053847 , WO2004035625), Schuppens (see Japanese patents no. JP5155900 . JP5093000 . JP11266803 . JP2003023970 . JP2003327599 ) and fins (see Takeshi Nagai & Nobutaka Suzuki, Isolation of collagen from fish waste material - skin, bone and fins, Food Chemistry 68 (3): 277-281 (2000)) of various fish species, as well as of various marine invertebrates. (e.g. US 5,714,582 . US 6,916,910 ; US 20042007318 A1 ) including jellyfish (cf. JP10084916 . JP2003321497 . JP2004099513 . US 6,894,029 ), Molluscs (eg JP61000100 ; Nagai et al., Isolation and characterization of collagen from the outer skin waste material of cuttlefish (Sepia lycidas), Food Chemistry 72 (4): 425-429 (2001); Takeshi Nagai & Nobutaka Suzuki, Preparation and partial characterization of collagen from paper nautilus (Argonauta argo, Linnaeus) outer skin, Food Chemistry 76: 149-153 (2002)), corals (see, eg, Tillet-Barret et al., Characterization of heterotrimeric collagen molecules in a sea pen (Cnidaria, Octocorallia), European Journal of Biochemistry 203: 179-184 (1992)), and sea urchins ( JP2003082152 ).

So far published regulations for the isolation of sponge collagen Laboratory character, involve many steps and are therefore bad for one Scale magnification suitable (see, for example, Jungua et al., Biochemical and morphological studies on the collage of horny sponges. Ircina filaments compared to spongines, Connective Tissue Research 2: 193-203 (1974); Swatchek et al., Marine sponge collagen: isolation, characterization and effects on the skin parameters surface-pH, moisture and sebum, European Journal of Pharmaceutics and Biopharmaceutics 53: 107-117 (2002)).

Travis et al., 1967, used 0.3 M EDTA at pH 7.8 and 4 ° C for demineralization the spicules of a sea sponge (Genus Scypha: Calcarea: Porifera) and hydrogen fluoride as a gas for demineralization of silicic spicules of Euplectella (Hexactinellida: Porifera) with the aim of isolating collagenous proteins (Travis et al., Comparative Studies of Organic Matrices of Invertebrates Mineralized Tissues, Journal of Ultrastructure Research 18: 519-550 (1967)). They were analyzed by transmission electron microscopy collagenous fibrillar Structures detected, but due to lack of molecular biological and biochemical methods not as a defined type of collagen could be identified.

Ethanol-preserved material of the horn sponge Chondrosia reniformis was washed three times under running water, cut into small pieces and homogenized while a Tris-HCl buffer mixture (100 mM, pH 9.5, 10 nM EDTA, 8 M urea, 10 mM 2-mercaptoethanol) was added. After stirring at room temperature for 24 hours, the viscous extract was centrifuged. The pellet was discarded and collagen was precipitated from the supernatant by adjusting the pH to 4.0 with acetic acid and finally recovered by centrifugation ( EP 1259120 A ).

The known methods for collagen extraction from marine sponges are unsatisfactory and, above all, is for the isolation of collagen made of glass sponges (Hexactinellida) no method known yet. So there is no doubt Need for improved methods for isolating collagen high Quality, that for the Development of novel biomaterials based on collagen-silica is usable.

From Of particular interest are biomaterials of silicon dioxide structures, by living sponges be generated, as well as the possibilities the utilization of the corresponding formation mechanisms for the synthesis of these remarkable biomaterials.

silicon is often used for the production of poly-siloxane-containing materials such as semiconductors, glasses, ceramics, Plastics, optical fibers, etc. The production of these materials usually requires high temperatures or the use of corrosive substances.

in the Contrast runs The biological production of amorphous silica completely under Physiological conditions starting with a variety of well-structured Shells, thorns, fibers and spheres in many protista, diatomae, sponges, Mollusks and also higher ones Plants can be produced (See E.G. Weaver & Morse, 2003; Perry C.C & Keeling-Tucker T., Biosilification: the role of the organic matrix in structure control, Journal of Biological and Inorganic Chemistry 5: 537-550 (2000)).

Becoming present undertaken a series of biotechnologically oriented experiments the molecular mechanism of poly-siloxane synthesis under physiological Decipher conditions, what the prospect of new, environmentally sound strategies for synthesis and structuring of these important materials.

Around exploiting the advantage of marine organisms, the relatively large quantities of biogenic silica, have molecular biologists started using such genes and proteins (except for collagen) ultimately isolating the biosynthesis and optionally nanofabrication of silicon dioxide.

Morse et al., 2003 ( US 6,670,438 ) developed special techniques for the polymerization of silicon dioxide and silicon networks including a silicon alkoxide substrate and a catalyst, which leads at about neutral pH to the assembly, hydrolysis and subsequent condensation of these substrates, that is structurally effective. A catalyst in the context of this invention is generally a protein or polypeptide. Preferred proteins or polypeptides are silicatein filaments (but not collagen or collagen-like proteins), silicatein subunits, cysteine homopolymers, etc.

Silicatein filaments and their subunits chemically form the axial core structures of silicon spicules in marine sponges of the genus Demospongia: Porifera. In vitro they control spatially the formation of polymeric silica and silicon networks from the corresponding alkoxide substrates under physiological Conditions. Usually would be here acid or base catalysis necessary. The silicatein filaments are out with acids and hypochlorite purified silica spicules of sponge Tethya aurantia obtained by dissolving the sponge in buffered hydrofluoric acid.

Similar results were reported by Müller et al. 2004 ( EP 1 320 624 B1 ) with recombinant silicatein.

Just few publications regarding collagen-silica composites are known. Thus, Coradin et al., 2002 (Coradin et al. A novel route to collagen-silica biohybrids, in: Organic / Inorganic Hybrid Materials, C. Sanchez, R. M. Laine, S. Yang and C.J. Brinker, eds, Materials Research Society Symposium Proceedings 726: 79-83 (2002)) and Eglin et al., 2004 (Eglin et al., Collagen-silica hybrid Sodium silicate and sodium chloride effects on type I collagen fibrillogenesis, Bio-Medical Materials and Engineering 21: 1-8 (2004)) collagen-silica composites supported on collagen gel matrices were based. They were made from rat tail tendon collagen and sodium silicate produced by co-gelation. The resulting materials showed a very good biocompatibility, but their mechanical properties were the limiting factor for one wide use.

task The present invention therefore provides methods of isolation collagen from marine glass sponges and methods for the biomimetic production of composites and materials on the Base of collagen and silica.

According to the invention Problem solved by a glass sponge collagen, which can be obtained by slow etching of Glass sponge basal spicules in alkaline solution, dialysis of the thus obtained Extract and subsequent Lyophilization.

• 1. Zerkleinern und Waschen von Glasschwamm-Basal-Spiculen
• 2. langsames Ätzen in alkalischer Lösung, in geschlossenen Behältnissen bei 35 bis 37°C
• 3. Dialyse zur Isolation des Glasschwammkollagens aus dem alkalischen Extrakt
• 4. Trocknen (Lyophilisierung)

According to the invention, the glass sponge collagen is produced from glass sponge basal spicules by the following process steps:

• 1. Mincing and washing glass sponge basal spicules
• 2. slow etching in alkaline solution, in closed containers at 35 to 37 ° C
• 3. Dialysis to isolate the glass sponge collagen from the alkaline extract
• 4. Drying (lyophilization)

Prefers are glass sponges of the genus Hexactinellida, in particular of the genera Hyalonema sieboldi, H. affine, Monorhaphis chuni, which is particularly due to the size of her Basal spicules and their glass fibers of a few millimeters in diameter up to 3 m in length suitable.

When alkaline solution becomes a 2.5M aqueous NaOH solution (pH 12) or an alkaline adjusted to pH 8 1% sodium dodecyl sulfate or used an alkaline pH adjusted to pH 8 1% rhamnolipid biosurfactant solution. The alkaline NaOH serves in each case to dissolve the amorphous silicate network. According to the invention is slow, over a Period of at least 10 days etched. The extraction of collagen from the existing silicate networks is determined by the sodium dodecylsulfate and rhamnolipid biosurfactant solution additionally promoted.

in the Dialysis dialysis remains remaining inorganic residues of the extracted organic portions (collagen) separately.

The glass sponge collagen according to the invention has the following features:
The fibril diameter is about 20 nm. The collagen has a white color and is resistant to acid and pepsin treatment. In slightly basic media homogeneous highly concentrated collagen suspensions can be prepared. The collagen is easily cleaved with trypsin. Unlike calfskin collagen leaves the glass sponge collagen silicate easily.

to Invention belongs also the use of the glass sponge collagen according to the invention for the production of biomaterials such as spheres, fibers, rods, powders, granules, scaffolds, Tile.

• 1. Lösung des Lyophilisats in alkalischer Puffer-Lösung
• 2. Mineralisation durch Mischung des gelösten Kollagens mit Silan-Lösung als Silikat-Precursor
• 3. Wachstum und Agglomeration bei Raumtemperatur
• 4. Lufttrocknung bei Raumtemperatur über einen Zeitraum von mindestens 4 Tagen

The in-vitro production of nanostructured composites and materials consisting of marine glass sponge collagen and silicon dioxide is carried out with the following process steps:

• 1. Solution of the lyophilisate in alkaline buffer solution
• 2. Mineralization by mixing the dissolved collagen with silane solution as silicate precursor
• 3. Growth and agglomeration at room temperature
• 4. Air dry at room temperature for at least 4 days

Of the First step is the solution of the purified and dried marine glass sponge collagen in Tris-HCl buffer solution (100 mM; pH 8.5). Unlike other animal collagens that only in acidic solutions soluble are and insoluble in basic solutions, Marine Glass Sponge Collagen has the remarkable property in acidic solutions insoluble and in basic solutions soluble to be.

The next step is to mix the dissolved collagen as an organic template for the silica mineralization with the silica-containing substrate. The substrate is preferably tetramethoxysilane (TMOS, (Si (OCH ₃ ) ₄ ) because of its stability at neutral pH values At higher pH values, TMOS hydrolyzes in silica (Si (OH) ₄ ) Monomeric silicic acid molecules condense under the influence of collagen Microfibrils to nanoparticles and form amorphous silica.

hydrolysis

condensation

Formation of silica nanoparticles

The next Steps are the growth and agglomeration of amorphous silica nanoparticles into layers on the nanometer scale, which deposit along and around the collagen fibrils. there it is a non-catalytic self-organization. Ultrastructural analysis this self-assembled, collagen-silica composite shows that amorphous silica deposited on the surface of collagen fibrils and has formed "nano-pearl necklaces" that very strong to the nanoparticulate Structures of natural Remember spicules.

Of the last step is a slow air drying (no fast freeze-drying) the samples obtained as described above, which after some A compact material with increased strength and elasticity (comparable with natural equivalents) forms. To form needles and scaffolds is the drying carried out in a suitably shaped container. to Production of balls, the collagen suspension in the form of a Drop into the TMOS solution Pipette.

Based enclosed Representations and embodiments the invention will be closer explained. Showing:

1 Images of a Hyalonema sieboldi glass sponge (a) whose flexible basal spicules can be bound to a full circle (b).

2 SEM images of a natural H. sieboldi basal spicule (a) and after treatment with the alkaline solution (2.5 M NaOH) (b).

3 SEM image (a) and Confocal Laser Scanning Microscopy (LSM) image (b) of H. sieboldi basal spicule treated with alkaline solution.

4 REM (a), TEM (b) and AFM (c) Images of the microfibrils obtained after the slow alkaline demineralization treatment of the basal spicule of H. sieboldi.

• * Datenbank: MSDB, Imperial College London (http://www.matrixscience.com)
• ** Protein-Scores größer als 61 sind signifikant (p<0.05)

5 Mascot search results for the fibrillar protein of H. sieboldi spicules, which was isolated by the alkaline treatment.

• * Database: MSDB, Imperial College London (http://www.matrixscience.com)
• ** Protein scores greater than 61 are significant (p <0.05)

6a , b SEM image of a Monorhaphis chuni basal spicule treated with alkaline solution (2.5 M NaOH). It shows that the multifibrillar organic matrix serves as a template for silicate mineralization

7 TEM image of the microfibrils obtained after the slow alkaline demineralization treatment of the basal spicule of Monorhaphis chuni.

8th SEM images: The image (a) shows nanoparticles of amorphous silicate, which were deposited in vitro on the previously isolated from spicules collagen from a silica solution. The observed structures are very similar to those of natural glass sponge spicules (b).

9 Biomimetic rod-shaped collagen-silicate composites (top) that resemble the natural spicules of Monorhaphis chuni (below).

10 Biomimetic spherical collagen-silicate composite material

11 Biomimetic-made collagen-silicate scaffolds

Embodiment 1

manufacturing of collagen fibrils from Hyalonema sieboldi basal spicules

1b shows a highly flexible Hyalonema sieboldi basal spicule, tied to a full circle. Dried basal sponge sponges from Hyalonema sieboldi (length 50 cm, diameter 1.5 mm) are washed three times with distilled water, cut into 2 to 5 cm pieces and placed in a 10 ml plastic flask containing 5 ml a 2.5 M aqueous NaOH solution. To avoid evaporation, the vessel was sealed and thermostated for a period of 14 days with gentle shaking at 35 ... 37 ° C.

The effectiveness of this slow alkaline etching was confirmed by scanning electron microscopy (SEM) ( 2a , Federation 3a ) and Confocal Laser Scanning Microscopy (LSM) ( 3b ) outside at different points of the spicules and within the cross section of the same.

2a shows an SEM image of a natural H. sieboldi basal spicule. 2 B shows an SEM image of the same basal spicules after treatment with the alkali solution (2.5 M NaOH).

3a and 3b show pictures of a H. sieboldi basal spicule, which was treated with alkaline solution. The SEM image ( 3a ) shows that the multifibrillar organic matrix serves as a template for silicate mineralization. 3b shows a photograph with a laser scanning microscope, wherein the fluorescent fibrillar network is visible in the alkaline treated H. sieboldi basal spicule.

The SEM images after the slow etching process show very clearly that a fibrillar organic matrix is the biomineralization in terms of structure or morphology, ie a kind of template represents.

The microfibrils with diameters of about 20 ... 30 nm are present in bundles with a total diameter of between 1 μm and 5 μm, which can be easily determined by means of SEM ( 4a ), TEM (Transmission Electron Microscopy) ( 4b ) and AFM (Atomic Force Microscopy) ( 4c ) can be visualized.

The contained in the plastic containers Solution, which are now the extracts of Hyalonema spicules, so also fibrillar protein was taken with a pipette, in dialysis membranes Roth / Germany with an exclusion limit (MWCO) of 14 kD and 48 hours at 4 ° C dialyzed against deionized water.

The Organic material obtained by the dialysis was in a vacuum dried (Christ lyophilizer / Germany), whereby a powdery, whitish Lyophylisate was obtained.

The Lyophilisate was fractionated by HPLC. An acetonitrile gradient was used for this purpose (0-60%) in 0.1% Trifluoroacetic. Only one main peak was observed.

The approximate Molecular weights of the proteins in the lyophilisate were determined by gel electrophoresis (Sodium dodecyl sulfate, SDS, Laemmli, Great Britain 1970) to 10% and 12% gel plates determined. This was done using a set of molecular weight markers (Silver stain SDS molecular standard mixtures, Sigma, USA). The lyophilizate was dissolved in buffer (1M Tris-HCl, pH 6.8, 2.5% SDS, 10% glycerol, 0.0125% bromophenol blue), incubated for 5 min at 95 ° C and subsequently added to the 10% or 12% SDS-polyacrylamide gels. After Electrophoresis (75 V over 1.5 hours), the gels were stained. To visualize the proteins, GelCode Silver was used for the 10% gel SNAP Stain Kit II (Pierce, USA) for the 12% Coomassie Brilliant Blue R250 used.

Around Certainty about the nature of the proteins isolated from the spicules of glass sponges Coomassie blue stained electrophoresis gels were added to the Determination of the amino acid sequence by Mass spectrometric sequencing techniques (MALDI and Finnigan LTQ) (described in: Shevchenko et al., Mass Spectrometric Sequencing of Proteins from Silver-stained Polyacrilamide Gels. Analytical Chemistry 68: 850-858 (1996)).

Four electrophoretic bands were excised ( 5 ) and the protein material contained in the gel was removed by means of trypsin. Then the trypsin protein mixture was treated with Finnigan LTQ and analyzed by MALDI peptide fingerprinting. A subsequent comparison of the MSDB Protein Database (MSDB, Imperial College London (http://www.matrixscience.com) led to the identification of collagen alpha 1 and 2 in three out of four bands (high molecular weight) ( 5 ). There are similarities with canine collagen alpha 1, rat collagen, mouse collagen and bovine collagen alpha 2.

Embodiment 2

manufacturing of collagen fibrils from Hyalonema sieboldi basal spicules

dried basal sponge glass sponges from Hyalonema sieboldi (length 50 cm, Diameter 1.5 mm) washed three times with distilled water, in 2 ... 5 cm long pieces cut and placed in a 10 ml plastic jar containing 5 ml of a 1% sodium dodecyl sulfate solution (SDS, adjusted to pH 8.0 with 0.1M NaOH).

All subsequent steps were carried out analogously to Embodiment 1.

The Investigation of the collagen thus obtained shows that this Way collagen of the same quality as in embodiment 1, but in larger quantities win.

Embodiment 3

manufacturing of collagen fibrils from Hyalonema sieboldi basal spicules

dried basal sponge glass sponges from Hyalonema sieboldi (length 50 cm, Diameter 1.5 mm) are washed three times with distilled water, in 2 ... 5 cm long pieces cut and placed in a 10 ml plastic jar containing 5 ml of a 1% rhamnolipid biosurfactant solution (adjusted to pH 8.0 with 0.1M NaOH). All subsequent steps were carried out analogously to embodiment 1.

The Investigation of the collagen thus obtained shows that this Way collagen of the same quality as in embodiment 2, but in larger quantities win.

Embodiment 4

manufacturing collagen fibrils from basal sponge glass sponges from Hyalonema affine

dried basal glass sponge sponges of Hyalonema affine (length 50 cm, Diameter 1.5 mm) washed three times with distilled water, in 2 ... 5 cm long pieces cut and placed in a 10 ml plastic jar containing 5 ml of a 2.5M aqueous NaOH solution or a 1% sodium dodecyl sulfate solution (SDS, adjusted to pH 8.0 with 0.1M NaOH) or a 1% rhamnolipid biosurfactant solution (adjusted to pH 8.0 with 0.1M NaOH).

All subsequent steps were carried out analogously to Embodiment 1.

The thus obtained collagen was analyzed. The analysis results showed that in this way collagen of the same quality as in the isolation from Hyalonema sieboldi lets win. Equally different also in this embodiment Methods in quantity of the isolated collagen depending on used Extraction solution.

Embodiment 5

Production of collagen fibrils Basal glass sponge spicules from Monorhaphis chuni

dried basal glass sponge spicules of Monorhaphis chuni (length 50 cm, Diameter 5 mm) are washed three times with distilled water, in 2 ... 5 cm long pieces cut and put in a 10ml plastic jar containing 5 ml of a 2.5 M aqueous NaOH solution or 1% sodium dodecyl sulfate solution (adjusted to pH 8.0 with 0.1M NaOH) or rhamnolipid biosurfactant solution (adjusted to pH 8.0 with 0.1M NaOH).

Around To avoid evaporation, the tube was closed and placed over a Thermostated for 14 days with gentle shaking at 35 ... 37 ° C.

The effectiveness of this slow alkaline etching was confirmed by scanning electron microscopy (SEM) ( 6a , b) checked on the outside at different points of the spicules and within the cross-section thereof.

The SEM images after the slow etching process show very clearly that a fibrillar, egg-like organic matrix ( 6b ) determines the biomineralization in terms of structure or morphology, ie represents a kind of template.

The microfibrils with diameters of approx. 20 ... 30 nm can be analyzed by means of TEM ( 7 ) are made visible.

extracts the spicules of Monorhaphis chuni containing fibrillar protein became dialyzed against deionized water. The membranes (Roth / Germany) had an exclusion limit (MWCO) of 14 kDa. Dialysis was on 48 hours at 4 ° C. The recovered material was dried in vacuo (Christ Lyophilizer / Germany). (The further procedure is the same as described in Example 1)

So far we know this work, the first, the results are clear the presence of collagen in the spicules of Monorhaphis chuni clues.

To embodiment 5 leaves collagen of the same quality as in the isolation of Hyalonema sieboldi and Hyalonema affine win. Due to the approximately 4 times larger diameter could be significantly increased Achieve yields in the context of collagen extraction.

Equally different also in this embodiment Methods in quantity of the isolated collagen depending on used Solvent.

Embodiment 6

manufacturing of collagen-silicate composites

Collagen from the marine glass sponges Hyalonema sieboldi, Hyalonema affine and Monorhaphis chuni was isolated as described above (Examples 1 ... 5). A collagen aliquot (0.1 ... 100 mg dry matter per ml Tris-HCl pH 8.5) is placed in a plastic covered container and placed in 5 ... 10 ml Tris-HCl buffer (100 mM, pH 8, 5), at room temperature and with vigorous shaking by Vortex Genius 2 for 15 minutes. A corresponding aliquot (0.1... 10 ml) of the collagen suspension obtained in this way is placed in a plastic vessel which, by its shape, determines the geometry of the composite material obtained later, containing a 0.1 to 10 M tetramethoxysilane solution (TMOS). contains (ABCR GmbH, Germany). After 40 ... 80 sec of intensive shaking at room temperature, again on the Vortex Genie 2, the contents of the vessel become hard. The vessel is opened to allow the volatile reaction products to escape. This starts the slow air drying process. After 4 to 5 days of air drying, the original diameter of the material has shrunk from about 10 to 4 ... 6 mm ( 8th above).

In this way, collagen-silicate composites can not only be in the form of rods ( 9 above), but also in the form of spheres ( 10 ), Scaffolds ( 11 ) getting produced.

Claims (7)

Glass sponge collagen, obtainable by slow etching of Glass Sponge basal Spiculen in alkaline solution, Dialysis of the extract thus obtained and subsequent lyophilization. Glass sponge collagen according to claim 1, characterized that the basal spicules the glass sponges the class of Hexactinellida be used. Process for producing glass sponge collagen made of glass sponge basal spicules characterized in that comminuted and washed glass sponge basal spicules in alkaline solution, in closed containers at 35 to 37 ° C etched slowly and that the extract obtained for the isolation of glass sponge collagen dialyzed and lyophilized. Method according to claim 3, characterized that glass sponges of the class of Hexactinellida. Use of glass sponge collagen for manufacturing of biomaterials such as spheres, fibers, rods, powders, granules, scaffolds, Tile. Process for the in vitro production of nanostructured Composites and materials made from marine glass sponge collagen and silicon dioxide, with the following process steps: - Solution of the Lyophilisate in alkaline buffer solution - Mineralization by mixing of the solved Collagen with silane solution as a silicate precursor - Growth and agglomeration of the silicate nanoparticles at room temperature - air drying at room temperature a period of at least 4 days Method according to Claim 6, characterized that a Tris-HCl buffer is used as buffer.