Abstract
Aim.To perform a comparison of three-dimensional nanostructure of porous biocompatible scaffolds made of fibroinBombix moriand recombinant spidroin rS1/9.Materials and methods.Three-dimensional porous scaffolds were produced by salt leaching technique. The comparison of biological characteristics of the scaffolds shows that adhesion and proliferation of mouse fibroblastsin vitroon these two types of scaffolds do not differ significantly. Comparative experimentsin vivoshow that regeneration of bone tissue of rats is faster with implantation of recombinant spidroin scaffolds. Three-dimensional nanostructure of scaffolds and interconnectivity of nanopores were studied with scanning probe nanotomography (SPNT) to explain higher regenerative activity of spidroin-based scaffolds.Results.Significant differences were detected in the integral density and volume of pores: the integral density of nanopores detected on 2D AFM images is 46 μm–2 and calculated volume porosity is 24% in rS1/9-based scaffolds; in fibroin-based three-dimensional structures density of nanopores and calculated volume porosity were 2.4 μm–2 and 0.5%, respectively. Three-dimensional reconstruction system of nanopores and clusters of interconnected nanopores in rS1/9-based scaffolds showed that volume fraction of pores interconnected in percolation clusters is 35.3% of the total pore volume or 8.4% of the total scaffold volume.Conclusion.Scanning probe nanotomography method allows obtaining unique information about topology of micro – and nanopore systems of artificial biostructures. High regenerative activity of rS1/9-based scaffolds can be explained by higher nanoporosity of the scaffolds.
Highlights
Three-dimensional nanostructure of scaffolds and interconnectivity of nanopores were studied with scanning probe nanotomography (SPNT) to explain higher regenerative activity of spidroin-based scaffolds
Three-dimensional reconstruction system of nanopores and clusters of interconnected nanopores in rS1/9-based scaffolds showed that volume fraction of pores interconnected in percolation clusters is 35.3% of the total pore volume or 8.4% of the total scaffold volume
3. Анализ наноструктуры матриксов из рекомбинантного спидроина: а – атомно-силовом микроскопе (АСМ)-изображение топографии поверхности блока матрикса на основе рекомбинантного спидроина rS1/9 после среза ультрамикротомом, размер скана 1,8 × 1,8 мкм, диапазон вариации высоты 20 нм; б – трехмерная реконструкция кластеров взаимосвязанных нанопор в стенке макропоры матрикса на основе рекомбинантного спидроина rS1/9 (7,54 × 3,28 × 1,0 мкм)
Summary
В сравнительных экспериментах in vivo показано, что регенерация костной ткани у крыс происходит быстрее при имплантации матриксов из рекомбинантного спидроина. При сравнительном анализе наноструктуры искусственных матриксов из регенерированного шелка и рекомбинантного спидроина, полученных методом выщелачивания, показано, что скэффолды из спидроина имеют более высокую интегральную плотность нанопор, степень пористости и объемную долю взаимосвязанных пор. Three-dimensional nanostructure of scaffolds and interconnectivity of nanopores were studied with scanning probe nanotomography (SPNT) to explain higher regenerative activity of spidroin-based scaffolds. Трехмерные пористые биосовместимые искусственные матриксы (скэффолды) являются одними из самых перспективных имплантатов-носителей в регенеративной медицине, помогая организму восстанавливать его собственные ткани в месте повреждения [6]. В нашей статье с помощью метода сканирующей зондовой нанотомографии (СЗНТ) мы сравнили нано- и микроструктуру пористых матриксов из рекомбинантного спидроина и фиброина шелка
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