Preparation and properties of artificial auricles based on PVA/BNC composites
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摘要: 耳廓再造的关键是实现植入材料与天然组织间的生物力学适配，然而迄今尚未找到一种理想的耳廓替代物。本研究将细菌纳米纤维素(Bacterial nanocellulose, BNC)匀浆加至不同浓度的聚乙烯醇(Polyvinyl alcohol, PVA)水溶液中，通过冻融法使两者物理交联形成兼具高弹柔韧性和高力学强度的PVA/BNC复合材料，并对其理化性能和细胞相容性进行表征。结果表明：该材料具有高吸水性、低溶胀比，较高的韧性、弹性和缝合强度等特点，最大压缩模量可达到6.98 ± 0.49 MPa，与天然耳廓组织的生物力学相适配。最大缝合强度可达7.06 ± 0.33 N，完全满足临床缝合需求。BNC的加入促进了细胞在材料表面的粘附、生长和增殖，使PVA/BNC复合材料具有更高的细胞密度和活力，是一种很有潜力的人工耳廓材料。Abstract: The key to auricle reconstruction is to achieve a biomechanical fit between the implanted material and the natural tissue. So far, an ideal auricle substitute has not been found. In this study, bacterial nanocellulose (BNC) homogenate was added into polyvinyl alcohol (PVA) aqueous solutions of different concentrations, and the freeze-thaw method was used to form PVA/BNC composite materials with both high elastic flexibility and high mechanical strength via physically cross-linking. The physical and chemical properties and cytocompatibility of the composites were characterized. The results show that the material has the characteristics of high water absorption, low swelling ratio, as well as high toughness, elasticity and suture strength. The maximum compressive modulus reaches 6.98 ± 0.49 MPa, which matches the biomechanics of natural auricle tissue. The maximum suture strength reaches 7.06 ± 0.33 N, which fully meets the needs of clinical suture. Addition of BNC promotes the adhesion, growth and proliferation of cells on the surface of the material, giving the PVA/BNC composite material higher cell density and vitality. All the results show that the PVA/BNC composite is a promising biomaterial for artificial auricles.
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图 1 PVA、PVA/BNC材料的宏观、微观形貌
Figure 1. Macroscopic and microscopic morphologies of PVA and PVA/BNC materials (a) Macroscopic morphology of PVA and PVA/BNC materials with different concentrations; (b) Diagram of an auricular sample; (c) Scanning electron microscope microscopic topography of materials
图 2 材料的基本理化性质
(a)不同浓度PVA、PVA/BNC材料的红外光谱图；(b)含水率；(c)吸水率；(d)饱和溶胀率；(e)X射线衍射图谱
Figure 2. Basic physical and chemical properties of materials
(a) Fourier transform infrared spectra of PVA and PVA/BNC materials with different concentrations;(b) Moisture content; (c) Water absorption rate;(d) Saturated swelling rate; (e) X-ray diffraction pattern
图 3 PVA和PVA/BNC材料的压缩力学性能与缝合强度
(a)不同浓度PVA、PVA/BNC材料的压缩强度；(b)压缩模量；(c)压缩应力-应变曲线；(d)16%PVA和PVA/BNC循环压缩应力-应变曲线；(e)循环压缩能量耗散与能量损耗系数；(f)缝合强度
Figure 3. Compressive mechanical properties and suture strength of PVA and PVA/BNC materials
(a) Compressive strength of PVA and PVA/BNC materials with different concentrations; (b) Compressive modulus; (c) Compressive stress-strain curves; (d) 16% PVA and PVA/BNC cyclic compressive stress-strain curves; (e) Cyclic compression energy dissipation and energy dissipation coefficient;(f) Suture retention
图 4 PVA、PVA/BNC材料的拉伸力学性能
(a)各PVA、PVA/BNC材料的拉伸强度；(b)拉伸模量；(c)断裂伸长率；(d)拉伸应力-应变曲线；(e)16%PVA和PVA/BNC循环拉伸应力-应变曲线；(f)循环拉伸韧性与能量损耗系数
Figure 4. Tensile mechanical properties of PVA and PVA/BNC materials
(a) Tensile strength of PVA and PVA/BNC materials; (b) Tensile modulus; (c) Elongation at break; (d) Tensile stress-strain curve; (e) 16% PVA and PVA/BNC cyclic tensile stress-strain curves; (f) Cyclic tensile toughness and energy dissipation coefficient
图 5 小鼠成纤维细胞的相容性
(a) CCK-8实验结果；(b)浸提液培养的细胞存活率；(c)浸提液培养L929细胞1、2、3天后细胞荧光图；(d)电镜显示细胞在材料上生长1、3、5天后的形态图
Figure 5. Cytocompatibility of mouse fibroblasts
(a) CCK-8 experimental results; (b) Cell viability of extract culture; (c) Cell fluorescence pattern of L929 cells after culture in extract for 1, 2 and 3 days; (d) Electron microscopy showing morphology of cells after 1, 3, and 5 days of growth on the material
表 1 孔隙率与比表面积测试结果
Table 1. Porosity and specific surface area test results
Sample Skeletal density/(g·cm−3) Apparent density/(g·cm−3) Porosity/% Specific surface area/(m2·g−1) BNC
16%PVA
16%PVA/BNC1.056 ± 0.017
1.219 ± 0.006
1.207 ± 0.0050.009
0.909
0.45599.17
25.47
62.2758.98
24.96
26.48Notes: BNC—Bacterial nanocellulose; PVA—Polyvinyl alcohol 
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表 2 材料的结晶度
Table 2. Crystallinity of materials
Sample Crystallinity/% BNC 44.73 ± 5.07 8%PVA 34.08 ± 4.36 12%PVA 34.66 ± 9.87 16%PVA 35.80 ± 8.60 20%PVA 36.15 ± 6.28 8%PVA/BNC 37.38 ± 7.52 12%PVA/BNC 38.39 ± 7.88 16%PVA/BNC 45.68 ± 3.94 20%PVA/BNC 40.07 ± 1.47
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