TR2021018537A2 - TETRACYLIN HYDROCHLORIDE-COLLAGEN-CHITOSAN FILLED NANO BIOMATERIAL - Google Patents
TETRACYLIN HYDROCHLORIDE-COLLAGEN-CHITOSAN FILLED NANO BIOMATERIALInfo
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- TR2021018537A2 TR2021018537A2 TR2021/018537A TR2021018537A TR2021018537A2 TR 2021018537 A2 TR2021018537 A2 TR 2021018537A2 TR 2021/018537 A TR2021/018537 A TR 2021/018537A TR 2021018537 A TR2021018537 A TR 2021018537A TR 2021018537 A2 TR2021018537 A2 TR 2021018537A2
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- infection
- biomaterial
- collagen
- wounds
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- 229920001661 Chitosan Polymers 0.000 title claims abstract description 16
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- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
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Abstract
Mevcut buluş, İçinde bulunduğumuz yüzyılda endüstriyelleşme ile kullanılan iş 5 makinelerinin artması, silah sanayindeki gelişmeler, halen sürmekte olan savaşlar, trafik kazaları komplike yaralanma sıklığını artırmakta ve bu kazalarda acil cerrahi müdahale zorunluluk hale gelmektedir. Bununla birlikte birçok hastalıkta da cerrahi müdahalelere ihtiyaç duyulmaktadır. Kazalar ve hastalıklara yapılan cerrahi müdahaleler sonucu oluşan yaralar için acil önlem alarak çevrenin olumsuz 10 etkilerinden korumak ve hızlı bir şekilde iyileşmesini sağlamak hayati önem taşımaktadır. Bu yaralarda karşılaşılan en önemli sorunlardan ikisi kanama ve enfeksiyon riskidir. Ameliyat sonrası oluşabilecek enfeksiyon riski de cerrahların karşılaştığı önemli bir sorundur. Hatta oluşan enfeksiyonun kanama ile bütün vücuda yayılması da bir başka büyük problem olmakta ve telafisi mümkün olmayan sonuçlar 15 doğurabilmektedir. Dolayısı ile kaza ve ameliyatlar sonucu oluşan açık yaraların büyük ölçüde arttığı günümüzde mümkün olduğunca hızlı bir şekilde kanamanın durdurulması ve enfeksiyon riskinin ortadan kaldırılması için yeni yöntemler ve materyaller geliştirilmesi zorunlu hale gelmiştir. Buluşumuzda, Kitosan ve PVA doku iskelesine Tetrasiklin Hidroklorür antibiyotik ve 20 Kollajen kanama durdurucu ajan yükleyerek yaraların iyileşme sürecini enfeksiyon riski olmadan hızlandıran bir biyomalzeme elde etmek ile ilgilidir.The present invention, the increase in the work machines used with industrialization in the current century, developments in the arms industry, ongoing wars, traffic accidents increase the frequency of complicated injuries and emergency surgical intervention becomes a necessity in these accidents. However, many diseases require surgical interventions. It is of vital importance to protect the wounds from the negative effects of the environment by taking urgent measures for the wounds caused by the surgical interventions to the accidents and diseases and to ensure a rapid recovery. Two of the most important problems encountered in these wounds are the risk of bleeding and infection. The risk of infection that may occur after surgery is also an important problem faced by surgeons. In fact, the spread of the infection to the whole body with bleeding is another big problem and may cause irreparable results. Therefore, it has become imperative to develop new methods and materials in order to stop bleeding as quickly as possible and to eliminate the risk of infection, as open wounds resulting from accidents and surgeries have increased to a large extent. In our invention, it is about obtaining a biomaterial that accelerates the healing process of wounds without the risk of infection by loading Tetracycline Hydrochloride antibiotic and 20 Collagen hemostatic agents on the Chitosan and PVA tissue scaffold.
Description
TARIFNAME TETRASIKLIN HIDROKLORÜR-KOLLAJEN-KITOSAN DOLGULU NANO BIYOMALZEME Teknik Alan Kazalar ve çesitli hastaliklara karsi yapilan cerrahi müdahaleler sonucu olusan yaralar için acil önlem alarak çevrenin olumsuz etkilerinden korumak ve hizli bir sekilde iyilesmesini saglamak hayati önem tasimaktadir. Bu yaralarda karsilasilan en önemli sorunlardan ikisi kanama ve enfeksiyon riskidir. Ameliyat sonrasi olusabilecek enfeksiyon riski de cerrahlarin karsilastigi baska bir önemli sorundur. Hatta olusan enfeksiyonun kanama ile bütün vücuda yayilmasi da bir baska büyük problem olmakta ve telafisi mümkün olmayan sonuçlar dogurabilmektedir. Bulus, kanama durdurucu özelligi olan kitosan ve kolajeni Tetrasiklin Hidroklorür antibiyotik kullanilarak kanama ve enfeksiyon olmadan yarayi iyilestiren bir biyomalzeme ile ilgilidir. Önceki Teknik Medikal uygulamalarda biyomalzemeler önemli yer kaplamaktadir. Basit küçük yara bantlarindan gelismis biyomalzemelere kadar birçok çalisma yapilmistir. Ancak bu çalismalarda kitosan kolajen ve antibiyotik bir arada kullanilarak kanamayi durdurarak enfeksiyon riski olmadan yarayi iyilestiren nano doku iskelesi çok az bulunmaktadir. DESCRIPTION TETRACYCLIN HYDROCHLORIDE-COLLAGEN-KITOSAN FILLED NANO BIOMATERIALS Technical Area Wounds caused by accidents and surgical interventions against various diseases To protect it from the negative effects of the environment by taking urgent measures for It is vital to ensure his recovery. The most important of these wounds Two of the problems are the risk of bleeding and infection. May occur after surgery The risk of infection is another important problem faced by surgeons. Even if you are The spread of the infection to the whole body with bleeding is another big problem. and may have irreversible consequences. Invention, astringent Bleeding using chitosan and collagen Tetracycline Hydrochloride antibiotics, which are characteristic of and a biomaterial that heals the wound without infection. Prior Art Biomaterials occupy an important place in medical applications. simple minor wound Many studies have been carried out from bands to advanced biomaterials. However, this In studies, by using a combination of chitosan collagen and antibiotics, stopping bleeding There is very little nano-scaffolding that heals the wound without the risk of infection.
Ameliyat sonrasi olusan ameliyat yaralari içinde etkili bir yöntem bulusta kullanilmistir. An effective method for post-operative surgical wounds was used in the invention.
Ayrica biyobozunur oldugu için vücuttan rahatlikla atilabilir. Hangi oranda olmasi gerektigi ve etkinligi in-vitro test ile kontrol edilmistir. basvuruda kitosan PVA ile electrospinning yöntemiyle çekilmis ancak kolajen ve antibiyotik kullanilmamistir. basvuruda kitosan kullanilmis ancak electrospinning yöntemi kullanilmamistir. Ayrica enfeksiyona karsi herhangi bir islem yapilmamistir. In addition, since it is biodegradable, it can be easily excreted from the body. In what proportion The need and effectiveness were checked by in-vitro test. At application, chitosan was extracted with PVA by electrospinning method, but collagen and No antibiotics were used. Chitosan was used in the application, but the electrospinning method was not used. Moreover No action was taken against the infection.
Bulusun Amaçlari Bulusumuzda amacimiz yaralarin kanama olmadan enfeksiyon riski teskil etmeden iyilesme sürecinin hizlanmasini saglamaktir. Yara olan bölgenin cerrahi islem sonucu alinmasiyla olusan boslugun doku iskelesi gibi doldurularak kanamayi durduran iltihaplanmadan iyilesmesini saylayan biyomalzemenin saglik malzemesi olarak kullanilmasidir. Objectives of the Invention Our aim in our invention is to heal wounds without bleeding and without risk of infection. to accelerate the healing process. Result of surgical procedure of the injured area It stops the bleeding by filling the space created by the removal of the tissue like a scaffold. As the health material of the biomaterial that counts the healing from inflammation is to be used.
Resimlerin Açiklamasi Sekil 1: Biyomalzemenin electrospinning yöntemiyle elde edilmis numunesi. Description of Pictures Figure 1: Electrospinning sample of the biomaterial.
Sekil 2: Biyomalzemenin taramali elektron mikroskobu (SEM) görüntüsü Sekil 3: Biyomalzemenin in-vitro test sonucu kontrol ve biyomalzemeli agar görüntüsü. a) Staphylococcus Aureus bakterisi 3. Gün Ekim Sonucu Olusan Kontrol ve Denek Görüntüsü. b) Pseudomonas Aeruginosa bakterisi 3. Gün Ekim Sonucu Olusan Kontrol ve Denek Görüntüsü. Figure 2: Scanning electron microscope (SEM) image of the biomaterial Figure 3: In-vitro test result of biomaterial control and biomaterial agar image. a) Staphylococcus Aureus bacteria 3rd Day Sowing Control and Subject Image. b) Pseudomonas Aeruginosa bacterium 3rd Day Sowing Control and Subject Image.
Bulusun Detayli Açiklamasi Kitosanin biyolojik olarak yenilenebilir olmasi; biyobozunur, biyouyumlu, nonantijenik, nontoksik ve biyofonksiyonel özellikleri bu polimerin ve bu polimer kullanilarak elde edilen komplekslerin yara örtü materyali ve ilaç dagitim sistemleri gibi biyomedikal uygulamalarda kullanilmasina olanak saglamistir. Bunlara ek olarak kitosanin hemostatik özelliklere sahip oldugu, makrofaj olusumunu tetikledigi ve yara iyilesmesini hizlandirdigi bilinmektedir. Tüm bu özelliklerin yani sira kitosanin antibakteriyel özelligi ile hemostatik özelligi de ilgi çeken konulardandir. Detailed Description of the Invention The fact that chitosan is biorenewable; biodegradable, biocompatible, nonantigenic, The nontoxic and biofunctional properties of this polymer are obtained by using this polymer. biomedical materials such as wound dressing material and drug delivery systems allows it to be used in applications. In addition, chitosan It has hemostatic properties, triggers macrophage formation and It is known to accelerate healing. Besides all these features, chitosan Its antibacterial and hemostatic properties are also interesting topics.
Kollajen, insanlar da dahil tüm memelilerin bag dokusu içinde bulunan bir proteindir. Collagen is a protein found in the connective tissue of all mammals, including humans.
Kollajen protein bizi bir arada tutan zamk olarak adlandirilir ve vücudumuzda %25'ten daha fazlasini olusturur. Kollajen, vücutta bulunan ve dogada canlilarin hareket sisteminin yapi taslarini, özellikle kemik, kikirdak, lif ve eklemleri olusturan proteindir. Collagen protein is called the glue that holds us together and is more than 25% in our body. creates more. Collagen, found in the body and the movement of living things in nature. It is the protein that forms the building blocks of the system, especially bone, cartilage, fiber and joints.
Bu protein birbiri üzerine sarilmis üç alfa zincirinden meydana gelir. 19 tane degisik tipi tanimlanmis olup, tip I, tip ll seklinde isimlendirilir. Bu çesitlilik moleküler yapidan kaynaklanmaktadir. Bu çalismamizda tip 1 kollajen kanama durdurucu ajan kullandik. This protein consists of three alpha chains coiled on top of each other. 19 different types defined and named as type I, type II. This diversity is due to the molecular structure originates. In this study, we used type 1 collagen hemostatic agent.
Bu kollajenin ana molekülü tropokollajendir. Tropokollajenler de hücrenin içinde üretilen prokollajenlerden olusur. Kollajen vücutta ve dogada canlilarin yapisinda bol miktarda bulundugundan biyouyumlulugu çok iyi olan bir malzemedir. Kollajen büyük ölçüde hemostatik ajan olarak sigir asil tendonundan üretilmekte olup kollajen iyi bir kanama durdurucu özellige sahiptir. Ayrica doku iskelesi olarak kullandigimiz kanama durdurucu özelliginin oldugu bilinmektedir. Kollajenden yapilan biyomateryaller birçok farkli avantaj sunmaktadir: biyoyumludurlar ve dokulara karsi toksik degildirler ve iyi bilinen yapisal, fiziksel, kimyasal, biyolojik ve immunolojik özelliklere sahiptirler. The main molecule of this collagen is tropocollagen. Tropocollagens are also inside the cell. It consists of procollagens produced. Collagen is abundant in the body and in the structure of living things in nature. It is a material with very good biocompatibility since it is present in large quantities. collagen big It is produced mainly from the sigir Achilles tendon as a hemostatic agent, and collagen is a good It has anti-bleeding properties. We also use bleeding as tissue scaffold. It is known to have a stopping feature. Biomaterials made of collagen offers different advantages: they are biocompatible and non-toxic to tissues and have good They have known structural, physical, chemical, biological and immunological properties.
Geçmis 20 yilda doku degisimi için hayvanlardan saflastirilmis kollajenlerden farkli uygulamalarda yararlanilmaktadir. Kollajen jeller, fibroblastlar ve bovin kollajenden yapilmaktadir. Kollajen süngerler, içerisinde fibroblastlarin kültürlendigi ve göç ettigi kollajen matrikslerin dondurularak kurutulmasiyla üretilmektedir. Üzerine fibroblastlarin kültürlendigi sentetik lifler naylon ya da poliglaktik asit ile kollajenin birlestirilmesiyle yapilmaktadir. Kollajen membranlar yalniz ya da epidermal katmanla birlikte kullanilmaktadir. Ayrica in vitro olarak insa edilmis deriye benzeyen kollajen bazli ürünler de mevcuttur. Different from animal-purified collagen for tissue replacement in the past 20 years used in applications. Collagen gels from fibroblasts and bovine collagen is being done. Collagen sponges, in which fibroblasts are cultured and migrated It is produced by freeze-drying collagen matrices. Fibroblasts on The synthetic fibers it is cultured from are combined with nylon or polyglactic acid and collagen. is being done. Collagen membranes alone or with the epidermal layer is used. In addition, in vitro-constructed skin-like collagen-based products are also available.
Tetrasiklin HCl bakteriostatik etkili bir ajandir; bu etkisini mikroorganizmalarin protein sentezini inhibe ederek gösterir. Tetrasiklin, penisilin ve streptomisinden çok daha genis bir spektruma sahiptir. Birçok gram (+ ) ve gram (-) mikroorganizmaya karsi etkilidir. Tetrasiklin gastrointestinal sistemden kolaylikla absorbe edilir ve degisik oranlarda kan proteinlerine baglanir; kisa sürede elde edilen kan konsantrasyonlari 6- 8 saat süreyle etkili düzeyde kalir. Tetrasiklin karaciger ve safra içinde konsantre olur ve biyolojik yönden aktif bir sekilde idrar ve feçesle itrah edilir; kan-beyin bariyerini kolaylikla geçen tetrasiklin beyin-omurilik sivisi içinde istenen düzeyde bulunur. Çok genis bir spekturumalari olsa da en az selektif antibiyotiklerdendir. Vücutta, karaciger tarafindan kan dolasimindan alinir, konsantre edilip safra yoluyla bagirsaga gönderilir. Tetracycline HCl is a bacteriostatic agent; This effect of microorganisms on protein by inhibiting its synthesis. Tetracycline is much more than penicillin and streptomycin. has a wide spectrum. Against many gram (+) and gram (-) microorganisms it is effective. Tetracycline is easily absorbed from the gastrointestinal tract and varies widely. binds to blood proteins in proportions; blood concentrations achieved in a short time 6- It remains effective for 8 hours. Tetracycline concentrates in liver and bile and is excreted in the urine and faeces in a biologically active manner; the blood-brain barrier Tetracycline, which passes easily, is present in the cerebrospinal fluid at the desired level. A lot Although they have a wide spectrum, they are among the least selective antibiotics. body, liver It is taken from the blood circulation by the bloodstream, concentrated and sent to the intestine through bile.
Buradan tekrar emilip kana geçer ve daha sonra böbreklertarafindan vücuttan atilirlar. From here, they are reabsorbed and passed into the blood, and then they are excreted from the body by the kidneys.
Polivinil alkol (PVA), biyouyumlu ve biyobozunur bir yari kristalin polimerdir ve çok iyi lifli materyal olusturabilme yetenegi gösterir. Suda çözünebilir bir hidroksi polimer olan PVA çok iyi kimyasal dayanima, fiziksel ve mekanik özelliklere ve esneklige sahiptir. Polyvinyl alcohol (PVA) is a biocompatible and biodegradable semi-crystalline polymer and very good It shows the ability to form fibrous material. a water-soluble hydroxy polymer PVA has very good chemical resistance, physical and mechanical properties and flexibility.
Sulu ortamda mükemmel elektro çekilebilirlik gösterir. It shows excellent electro-absorbability in aqueous media.
Polivinil alkol (PVA) destek polimeri ile PVA Kitosan doku yüzeyi elde ederek bu biyomalzemenin içerisine Kollajen Hemostatik Matrix ve degisik oranlarda Tetrasiklin hidroklorür antibiyotik katarak elektro çekim yöntemi ile biyomalzemeler elde edildi. By obtaining PVA Chitosan tissue surface with polyvinyl alcohol (PVA) support polymer, this Collagen Hemostatic Matrix and Tetracycline at different rates into the biomaterial Biomaterials were obtained by electrospinning method by adding hydrochloride antibiotics.
Elde edilen biyomalzemelerin yapisini incelemek için Taramali Elektron Mikroskobu (SEM) analizleri yapilarak sonuçlari görüldü. Biyomalzemenin in vitro ortamda etkinliginin incelenmesi için bakteri ekimi yapilarak olusan koloniler sayildi. Elektro çekim yöntemiyle elde biyomalzemenin SEM görüntüleri incelendiginde PVA/kitosan çözeltisinden iyi bir nano yüzey elde edildigi, liflerin genel özelliklerinin bozulmadigi görülmüstür. Scanning Electron Microscopy to examine the structure of the obtained biomaterials (SEM) analyzes were performed and the results were seen. Biomaterial in vitro In order to examine its effectiveness, the colonies formed by bacterial cultivation were counted. Electro When the SEM images of the biomaterial obtained by the extraction method are examined, PVA/chitosan A good nano surface is obtained from the solution, the general properties of the fibers are not deteriorated. has been seen.
Bulus hazirlanirken Tetrasiklinhidroklorür oraninin degismesi ile nanolif olusumu açisindan büyük degisiklikler gösterdigi saptanmistir. Nanofiber formation with the change of tetracycline hydrochloride ratio while preparing the invention It was found that there were significant changes in terms of
Biyomalzemenin üretimi için iki farkli çözelti hazirlanip bunlar daha sonra birbirlerine karistirilmistir. Birincisi agirlikça 10 g. çözelti için çözücü olarak % 2'lik asetik asit kullanilmistir. %2'lik asetik asit çözeltisinin içerisinde PVA ve Kitosan 24 saat süre ile oda sicakliginda manyetik karistiricida çözülmüstür. Ikinci olarak ise agilikça serum fizyolojik içerisinde kollajen hemostatik matrix ve tetrasiklinhidroklorür antibiyotik oda sicakliginda 2 saat süre ile manyetik karistiricida çözülmüstür. Daha sonra iki çözelti birbirine karistirilmistir. Hazirlanan tüm nanolif çekim çözeltilerinde çözelti içerisindeki toplam katki orani (PVA, Kitosan, Kollajen ve Tetrasiklin hidroklorür) her zaman %10 alinmistir. Ayrica ikinci çözeltiyi hazirlarken serum fizyolojik sabit 3.5 gr. alinmistir ve çözeltideki kati madde miktari sabit tutulmustur. Serum fizyolojik çözeltisini hazirlarken çözebildigi madde miktari ve içerisindeki NaCI dikkate alinmis ve birkaç denemeden sonra 3.5 9. serum fizyolojik çözelti için kullanilmistir. Son olarak biyomalzemenin orani asagidaki sekilde elde edilmistir. For the production of the biomaterial, two different solutions are prepared and these are then interconnected. mixed. The first is 10 g in weight. 2% acetic acid as solvent for solution used. PVA and Chitosan in 2% acetic acid solution for 24 hours dissolved in magnetic stirrer at room temperature. Secondly, agilic serum collagen hemostatic matrix and tetracycline hydrochloride antibiotic chamber in physiological It was dissolved in the magnetic stirrer for 2 hours at the temperature. Then two solutions are mixed together. In all prepared nanofiber spinning solutions, the total additive rate (PVA, Chitosan, Collagen and Tetracycline hydrochloride) is always 10% has been taken. In addition, while preparing the second solution, normal saline solution is 3.5 gr. taken and The amount of solids in the solution was kept constant. While preparing the physiological saline solution The amount of substance it can dissolve and the NaCl in it were taken into account and after a few trials after 3.5 9. saline was used for the solution. Finally, the biomaterial The ratio is obtained in the following figure.
Kitosan içerisine katilan Kollajen ve Biyomalzeme içerisindeki katki oranlari T etrasiklin Hidroklorür oranlari Bivomalzeme PVA (%) Tetrasiklin Hi drokl orür Kollajen Tetrasiklinhidroklorur 60 40 10 25 Biyomalzeme olarak kullanilabilmesi için etkinligi laboratuvar ortaminda in vitro test yapilmistir. Hazirlanan çözeltilerle gram pozitif ve negatif bakteriler üzerinde etkinligi incelenmistir. Bu bölümde ilk olarak serum fizyolojik sivisi ile hazirlanan 0,5 MCFarland bakteri solüsyonlari ile koyun kanli agarlara yapilan ekim sonucunda gram pozitif (S. Collagen added to chitosan and Additive ratios in the biomaterial Tetracycline Hydrochloride rates bivomaterial PVA (%) Tetracycline Hydrochloride Collagen Tetracycline Hydrochloride 60 40 10 25 To be used as a biomaterial, its efficacy has been tested in vitro in a laboratory setting. has been made. Efficacy on gram positive and negative bacteria with prepared solutions has been examined. In this section, 0.5 MCFarland, which is first prepared with physiological saline, As a result of sowing on sheep blood agar with bacterial solutions, gram positive (S.
Aureus) ve gram negatif bakterileri (P. Aeruginosa) üzerinde etkili oldugu görülmüstür. Aureus) and gram negative bacteria (P. aeruginosa).
Biyomalzemenin gram pozitif (S. Aureus) bakterilerine karsi gram negatif (P. Biomaterial against gram positive (S. aureus) bacteria gram negative (P.
Aeruginosa) bakterilerinden daha fazla etkili oldugu görülmüstür. Aeruginosa) bacteria were found to be more effective.
Yapilan biyomalzemenin 3.gün bakteri ekimlerinde üremeye karsi 90% basari sagladigi görülmüstür. Dolayisiyla biyouyumlu, biyobozunur, kanamayi durduran dogal yönü fazla olan bir biyomalzeme medikal sanayide sikça kullanilabilecektir. 90% success against the growth of the biomaterial in the 3rd day bacterial sowing it has been seen to provide. Therefore, it is biocompatible, biodegradable, and stops bleeding. A biomaterial with a high natural aspect can be used frequently in the medical industry.
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