Investigation of the Loofah Reinforced Chitosan-Silk Hydrogel Composite Scaffolds for Cartilage Tissue Regeneration

Abstract

Current therapies for the treatment of cartilage defects are limited due to the low self-healing capacity of cartilage. In recent years, tissue engineering has been proposed as a promising approach for cartilage regeneration. In this study, silk fibroin/chitosan hydrogels reinforced with luffa cylindrica were prepared for cartilage tissue defects. Biocompatible, biodegradable silk fibroin and chitosan polymers were cross-linked with genipin which is a natural and nontoxic cross-linking agent. Scanning electron microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FTIR) were used for the characterization of the morphology and chemical structures, respectively. Dynamic mechanical analysis was used to determine the viscoelastic properties, while compression test was applied to examine the mechanical properties of the scaffolds. The cytotoxicity, viability and proliferation of rabbit mesenchymal stem cells on the scaffolds were investigated by LDH, WST, and collagen assay. All of the produced hydrogel composite scaffolds had an interconnected microporous structure and loofah nanofibers were well-integrated within the structure. The hydrogel composite scaffold cross-linked with 0.3% wt. genipin (L-CSG3) demonstrated comparable water content (94.4±0.2%), tan δ (0,18 at 1 Hz) and compressive modulus (5,5 MPa) values to that of articular cartilage. Besides, based on the in-vitro test results, this hydrogel composite scaffold showed enhanced viability on rabbit mesenchymal stem cells. Consequently, this hydrogel composite scaffold presented a great promise for cartilage tissue regeneration.

Keywords

• Loofah
• chitosan
• silk
• hydrogel scaffold
• cartilage regeneration

Lif Kabağı Takviye Edilmiş Kitosan-İpek Hidrojel Kompozit Doku İskelelerinin Kıkırdak Doku Hasarı Tedavisinde Kullanımının Araştırılması

Abstract

Kıkırdak doku hasarlarının onarılmasındaki mevcut tedaviler, kıkırdağın kendi kendini iyileştirme kapasitesinin düşük olması nedeni ile sınırlıdır. Son yıllarda doku mühendisliği, kıkırdak rejenerasyonu için umut verici bir yaklaşım olarak önerilmektedir. Bu çalışmada, kıkırdak doku hasarları için lif kabağı ile güçlendirilmiş ipek fibroin/kitosan hidrojeller hazırlanmıştır. Biyouyumlu, biyolojik olarak parçalanabilir ipek fibroin ve kitosan polimerleri, doğal ve toksik olmayan bir çapraz bağlama maddesi olan genipin ile çapraz bağlanmıştır. Taramalı elektron mikroskobu (SEM) ve Fourier Dönüşümü Kızılötesi Spektroskopisi (FTIR) sırasıyla morfoloji ve kimyasal yapı karakterizasyonu için kullanılmıştır. Viskoelastik özelliklerin belirlenmesi için dinamik mekanik analiz cihazı (DMA) kullanılırken, iskelelerin mekanik özelliklerini incelemek için basma testi kullanılmıştır. Doku iskelelerinin sitotoksisitesi, hücre canlılığı ve çoğalması tavşan mezenkimal kök hücreleri kullanılarak LDH, WST ve kollajen testi ile araştırılmıştır. Üretilen hidrojel kompozit doku iskelelerinin tamamının birbirine bağlı mikro gözenekli bir yapıya sahip olduğu ve lif kabaklarının yapıya iyi entegre olduğu görülmektedir. Ağırlıkça %0,3 genipin ile çapraz bağlanan hidrojel kompozit doku iskelesi (L-CSG3), eklem kıkırdağıyla karşılaştırılabilir su içeriği (94,4±% 0,2), tan δ (1 Hz'de 0,18) ve basma modülü (5,5 MPa) değerleri göstermiştir. Ayrıca, in-vitro test sonuçlarına göre, bu hidrojel kompozit doku iskelesi, tavşan mezenkimal kök hücrelerinde gelişmiş canlılık göstermiştir. Sonuç olarak, bu hidrojel kompozit doku iskelesi, kıkırdak dokusu rejenerasyonu için umut vaat ettiği söylenebilir.

Keywords

• lif kabağı
• kitosan
• ipek
• hidrojel doku iskelesi
• kıkırdak rejenerasyonu

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