Kitosan Esaslı İlaç Taşıyıcı Sistem Üretimi ve In vitro PerformansınınBelirlenmesi

Sunulan çalışmanın amacı kemik dolgu malzemesi ve ilaç taşıma sistemi olarak kullanılabilecek kitosan mikro kürelerin üretimi,karakterizasyonu ve performansının in vitro olarak belirlenmesidir. Bu amaçla; kitosan esaslı mikro küreler emülsiyon çarprazbağlama yöntemi ile üretilmiş olup, mikro kürelerin boyut, şekil ve ilaç yükleme verimliliklerine etki eden karıştırıcı hızı, çaprazbağlama ajanı gibi faktörler incelenmiştir. İlaç çalışmalarında kullanılmak üzere bakteriyel enfeksiyonların tedavisinde kullanılanantibiyotik ciprofloxacin tercih edilmiştir. Ayrıca, hazırlanan mikro kürelerin biyoaktivitelerini test etmek için yapay vücut sıvısı(SBF) hazırlanmış ve 3 hafta boyunca mikro küreler SBF içerisinde bekletilerek hidroksiapatit çökelmesi (HA) takip edilmiştir. SBFiçerisine konulan mikro küreler haftalık olarak toplanmış, Taramalı Elektron Mikroskopu (SEM) ve Fourier Dönüşümlü KızılötesiSpektroskopisi (FTIR) ile analiz edilmiştir. Işık Mikroskopu ve SEM analizlerine göre küre boyutları karıştırma hızı arttıkça azalmışve 102,27 ± 34,58 µm den 16,38 ± 3,26 µm ye kadar düşmüştür. Düşük miktarda kullanılan çapraz bağlayacı ajan glutaraldehit (GA)(NH2:CHO, 10:1 mol:mol) ise küre şekillerinde bozukluğa neden olurken, yüksek miktarda GA kullanımı (NH2:CHO, 1:10 mol:mol)kürelerin şeklinde iyileşmeye, küre boyutunda ve ilaç yükleme verimliliğininde azalmaya, ilaç salım hızında ise yavaşlamaya nedenolmuştur. SBF ile inkübe edilen mikro kürelerin SEM görüntülerine göre HA yapıları 1. haftadan sonra yüzeylerde birikmeyebaşlamış, bu yapılar 3. haftada yüzeylerde daha belirgin hale gelmiştir. FTIR analizleri de HA çökelmesini işaret eden $P{O^{-3}}_4$ gruplarının 3. haftada daha belirgin olduğunu göstermiştir. Bu sonuçlara göre antibiyotik ciprofloxacin yüklü kitosan mikro küreleremülsiyon çapraz bağlama yöntemi ile başarılı bir şekilde üretilmiş olup, karıştırma hızı ve çapraz bağlayıcı ajanı GA miktarlarınınmikro küre boyut, şekil ve salım profillerine etkisi ortaya konmuştur. Biyoktivite testlerinde kitosan mikro kürelerin, osteoentegrasyonu arttırma potansiyelinin olduğu gösterilmiştir. Dolayısıyla kitosan mikro küreler, kemik doku hasarlarında lokalolarak biyomoleküllerin salımını yapabilen dolgu malzemesi olarak veya implant malzemelerin yüzeylerinin modifikasyonundakullanılabilir.

Synthesis of Chitosan Based Drug Delivery System and Evaluation Its Performance In Vitro

The aim of the present study is to synthesize, characterize chitosan microspheres that can be used as bone-filling material and drugcarrier system, and to evaluate its performance in vitro. For this purpose; chitosan-based microspheres were synthesized by theemulsion cross-linking method and factors such as stirring rate and cross-linker that may affect the size, shape, and drug loadingefficiency of microspheres were examined. Antibiotic ciprofloxacin that is used in the treatment of bacterial infection was preferredfor drug studies. Furthermore, simulated body fluid (SBF) was prepared to determine the bioactivity of the microspheres, andhydroxyapatite (HA) precipitation on microspheres was followed for 3 weeks in SBF. Microspheres were collected from SBF weekly and characterized using light microscopy, Scanning Electron Microscopy (SEM), and Fourier Transform Infrared Spectroscopy(FTIR). According to the light microscope and SEM analysis, the sphere dimensions decreased as the stirring rate increased anddecreased from 102,27 ± 34,58 µm to 16,38 ± 3,26 µm. Low amount of cross-linking agent ( glutaraldehyde (GA)), (NH2: CHO, 10: 1mol: mol) caused distortion in the shape of microspheres, while the high amount of GA (NH2: CHO, 1:10 mol: mol) caused smoothmicrospheres, a reduction in the size and drug loading efficiency, and a slowdown in the drug release rate. According to the SEMimages of the microspheres incubated with SBF, HA structures began to precipitate on the surfaces after the first week, and thesestructures became more pronounced in the third week. $P{O^{-3}}_4$ groups attributed to HA precipitation in FTIR spectrum were moreobvious at week 3. In conclusion; antibiotic ciprofloxacin loaded chitosan microspheres were synthesized successfully by theemulsion crosslinking method, and the effect of stirring rate, crosslinking agent GA amounts on microsphere size, shape, and releaseprofiles were demonstrated. Enhanced HA precipitation was shown on chitosan microspheres in bioactivity tests. Hence, chitosanmicrospheres may be used as bone-filling material that can release biomolecules to damaged sites locally, or as coating to modify thesurfaces of implant materials.

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