The present invention relates to a biological tissue repair material which is used for the purpose of local treatment and tissue repair in intracorporeal and extracorporeal injuries; damaged skin, muscle and bone tissues such as cuts, burns and fractures.

Background of the Invention

Today, studies are usually done on uses of materials derived from natural silk on the basis of silks derived from Bombyx mori -popularly known as silkworm- and materials obtained from these silks. Besides the fact that superior physical, biological, chemical and pharmacological properties of these natural silks are remarkable; spider silks have different structures which are very rich in nano structure contents and protein contents because the purposes of spiders’ to spin a web are very different. Therefore, studies should be developed by focusing on these natural materials which can differ by structure-property and are quite applicable for biotechnological gains.

While spider silks are used for obtaining some tissue repair materials, information about spider species and silk forms (baseline, cocoon and hiding net silks) are not included. However, spider species and silk forms may have very different structural contents. In addition, currently-used conventional silk types are costly in an economic sense, it does not seem possible for spiders to live for a long time and to return to their natural life. For this reason, it is needed to create suitable laboratory conditions and equipment in order to obtain economical and high- quality spider silks and tissue repair materials, without harming spiders and their natural habitats, and to determine productive spider species which are suitable for farm life environments and periodic strippings.

The United States patent document no. US2015056256, an application in the state of the art, discloses treatment of spider silk thread for use as a thread of a composition in industrial applications such as medicine, textile or bio-artificial cell tissue. The spider silk thread used is obtained from genetically modified organisms.

The United States patent document no. US2013190786, another application in the state of the art, discloses a neural implant consisting of spider silk, especially natural spider silk fibres. In particular, the present invention makes available spider silk for use for the production of a pharmaceutical composition. A thread like spider silk is arranged essentially in an axial direction within a sheathing formed of spider silk threads. The sheathing can be knitted, woven, laced or braided of spider silk threads.

Summary of the Invention

An objective of the present invention is to realize a biocompatible tissue repair material which is used for the purpose of local treatment and tissue repair in intracorporeal and extracorporeal injuries; in cases with tissue losses such as skin, muscle and bone.

Another objective of the present invention is to realize an original tissue repair material through use of cocoon which has superior biophysical and biochemical properties obtained from Argiope bruennichi spider.

Another objective of the present invention is to realize a tissue repair material whereby tissue repair time is accelerated through use of cocoon sections coated with nano composites in biogel form. Another objective of the present invention is to realize a custom-design tissue repair material which has a tissue scaffold suitable for morphology of tissues such as skin, muscle, and bone by means of pattern assembly.

Detailed Description of the Invention

Stripping-winding system, pattern assembly and capillary tube used for“A Tissue Repair Material” realized to fulfil the objectives of the present invention are shown in the figure attached, in which:

Figure 1 is a view of a stripping-winding system whereby the silk stripping required for the inventive tissue repair material is carried out. Figure 2 is a view of a pattern assembly enabling to obtain the inventive tissue repair material in a form suitable for morphology of skin, muscle and bone tissues.

Figure 3 is a view of examples of winding orders to a pattern assembly which indicates obtaining the inventive tissue repair material in a form suitable for a desired tissue morphology.

Figure 4 is a view of a capillary tube system which is used for coating the inventive tissue repair material.

The inventive tissue repair material which is used for the purpose of local treatment and tissue repair in intracorporeal and extracorporeal injuries; damaged skin, muscle and bone tissues such as cuts, bums and fractures comprises at least one starting material composed by the silks in the form of cocoon and baseline created in laboratory environment {in vitro ) by gathering female members of adult Argiope bruennichi spider which is commonly found at coastal area of the Eastern Black Sea Region of Turkey in August, has large population, does not have reproductive problems, can be kept alive under laboratory conditions for a long time, can also live in different climates and altitudes, is compatible with permanent settlement and generates networks with very superior physico-bio- chemical characteristics. These natural silk materials obtained from Argiope bruennichi spider have high mechanical properties, superior biocompatibility, strong bioactivity against various bacteria and fungi, and the ability to absorb harmful UV rays. Due to their permanent settlement features, optimum three Argiope bruennichi spiders can be located in the same living cabinet. A silk structure rich in alanine and glycine can be obtained as a result of feeding spiders with a protein-rich grasshopper 36 hours before stripping. As a result of feeding with grasshopper, the protein content within the silk structure and the sizes and numbers of alanine-rich crystallites leading to silk stability can be increased. In addition, numbers and sizes of glycine-rich nanoplates providing flexibility property due to feeding with grasshopper are brought under control and made suitable for standardization. Spiders, which have been kept alive in living cabinets for a long time (minimum 4-5 months), are then released back to their natural habitats after this period of time. Spiders, which have been released back to natural habitats are observed in the same natural place again throughout their survival time. In other words, a slight change occurs in their main living localizations according to different times.

Stripping the silk, which is the starting material of the inventive tissue repair material obtained from Argiope bruennichi spider, is performed by using a stripping-winding system (1). The stripping-winding system (1) comprises a regulator (2) which enables to receive the required voltage; an electric motor (3) which provides rotary motion by means of the power it receives from the regulator (2); a hollow glass tube (4) which is placed to the end of the electric motor (3) and rotates together with the electric motor (3); a spider fixing plate (5) which is the place where the live adult female Argiope bruennichi spider whereby the silks, to be wound to the glass tube (4) performing the rotary motion, will be stripped is fixed; a control unit (6) with a computer connection and software control which enables to adjust parameters such as winding number, length, speed and frequency of the stripped silks; buttons (7) which are intended for starting, pausing and stopping the stripping; and a display (8) which indicate number of winding, number of cycle termination and number of current cycle (Figure 1). To prevent the spiders from cutting the silk before the stripping, their hind legs are put together and bandaged and the bandage is fixed to the spider fixing plate (5). The stripping process is started by fixing one or two spiders to the spider fixing plate (5). The glass tube (4) fixed to the end of the electric motor (3) is rotated by the power received from the regulator (2) and thereby the silks stripped from the spider are wound here. The glass tubes (4) preferably have 15 cm length and 2-5 cm variable diameter. As a result of the winding, the silk filaments (surgical sutures) used for surgical purpose are obtained on coils in a specific diameter and length. Mechanical properties of the original filaments prepared have elongation, i.e. flexibility, value of 255,2±0,3 N/mm ² (stress) and 11,3±0,2% (strain).

In the invention, a pattern assembly is used for preparing tissue scaffolds suitable for morphology of skin, muscle, and bone tissues which generate the tissue repair material and are obtained by means of silks. Wound grafts used for surgical purpose are obtained by means of the pattern assembly from the silks wound to the glass tubes (4). The pattern assembly consists of two parts, namely an upper part which is circular and can rotate and a platform part which is created so as to maintain the balance from objects of different shapes and sizes and carries the upper part. 12 tabs numbered from 0 to 11 by equal intervals at the edges of the circular structure, are located on the upper part of the pattern assembly (Figure 2). The silks passed around the tabs of the pattern assembly by orders suitable for obtaining the tissue morphology and a graft compatible with the morphology of the tissue aimed to be repaired is obtained. 0-6-5-10-11-4-3-0-9-6-3-5-1-0-11-7- 10-2-1-11-10-9-3-4-8-7-3-2-8-9-1-0-11-3-4-10-9-5-6-8-5-4-9-1 0-3-2-11-1-8-3-6- 1-7-0-5-7-6-11 or 0-5-6-11-10-7-6-5-1-11-0-6-7-1-0-4-5-10-9-2-8-3-4-7-2-3-9-8- 4-3-8-9-1-2-8-7-3-9-1-10-2-1-8-3-7-6-10-4-11-5 can be exemplified as the order of winding (Figure 3). The said pattern assembly is created by transactions of forming by heat treatment, cutting and bonding, by using pyrex material from glasses which are resistant to corrosion with borosilicate (containing B2O3), have low expansion coefficient, resistant to high temperatures (500°C). When UV transmittance and reflectance of the graft material prepared in 20x20 mm size are measured, it is determined that amount of UV absorption is about 70% at 355-400 nm wavelength.

Tissue materials such as surgical suture (filament) and graft are obtained by winding to the inventive glass tubes (4) and using the pattern assembly. Top of the tissue materials are covered by levan nano composites that are bio polymer obtained from aloe vera and plants with moisturizing and tissue repairing feature prepared entirely by natural silk micro powders in order to strengthen their bioactivity. Homogeneous nano composite gels are obtained by combining aloe vera and levan biogels with silk micro powders -which are obtained by cleaning and grinding of cocoon- mix in certain ratios and mixing with centrifuge. The graft and the surgical sutures prepared by baseline and cocoon threads are not directly coated with aloe vera and levan biogels, they are subjected to a coating process with nanocomposites in gel form for more original and effective tissue repairs. Thereby, the healing process is further accelerated with the effect of nano structures being uniformly dispersed in moist gel structure without being limited to the pattern zones wherein only silk lines are located. Upon the silks coated to the glass tubes (4) are taken into suitable morphological structures in the pattern assembly, the grafts are coated by dripping the nano composite gels onto this assembly and impregnated by a centrifuge process. Whereas surgical sutures are coated by being passed through the nano composite which is filled such that no air gap will remain inside the capillary tube, after they are inserted into the steel needle (Figure 4). The samples used in the coating accelerate the tissue repairs and enable to maintain the natural moisture environment of the tissue by utilizing antimicrobial, antifungal and wound-healing bioactivities at the silk tissue interface by providing more effective recovery. Following the coating, the tissue repair material reaches the efficiency to provide integration with the wound by means of its mechanical property, is beneficial in wound healing process due to its electrical conductivity, becomes a natural protein source with antibacterial property which is effective in burn treatment and skin tissue repair by creating a barrier against harmful rays of the sun by means of its ability to absorb UV rays, exhibits biocompatibility with natural support materials that can be used for tissue repair such as hydroxy apatite in bone fracture or damage, and creates sub-bridge and ground for crystalline structure development.

The inventive tissue repair materials such as grafts and filaments prepared are classified according to their morphologies, patterns, baseline numbers, baseline diameters, filament lengths, protein contents, nano structure distributions rich in alanine and glycine for surgical applications and then packaged and sterilized with gamma radiation of 10 kGy before use. As a result of this sterilization performed by low dose, the structure is not damaged in molecular and nano scale.

Within these basic concepts; it is possible to develop various embodiments of the inventive tissue repair material; the invention cannot be limited to examples disclosed herein and it is essentially according to claims.