Non-animal derived recombinant collagen-based biomaterials as a promising strategy towards adipose tissue engineering.

Lana Van Damme, Phillip Blondeel, Sandra Van Vlierberghe
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Abstract

Adipose tissue engineering (ATE) has been gaining increasing interest over the past decades, offering promise for new and innovative breast reconstructive strategies. Animal-derived gelatin-methacryloyl (Gel-MA) has already been applied in a plethora of TE strategies. However, due to clinical concerns, related to the potential occurrence of immunoglobulin E-mediated immune responses and pathogen transmission, a shift towards defined, reproducible recombinant proteins has occurred. In the present study, a recombinant protein based on human collagen type I, enriched with arginine-glycine-aspartic acid was functionalized with photo-crosslinkable methacryloyl moieties (RCPhC1-MA), processed into 3D scaffolds and compared with frequently applied Gel-MA from animal origin using an indirect printing method applying poly-lactic acid as sacrificial mould. For both materials, similar gel fractions (>65%) and biodegradation times were obtained. In addition, a significantly lower mass swelling ratio (17.6 ± 1.5 versus 24.3 ± 1.4) and mechanical strength (Young's modulus: 1.1 ± 0.2 kPa versus 1.9 ± 0.3 kPa) were observed for RCPhC1-MA compared to Gel-MA scaffolds.In vitroseeding assays showed similar cell viabilities (>80%) and a higher initial cell attachment for the RCPhC1-MA scaffolds. Moreover, the seeded adipose-derived stem cells could be differentiated into the adipogenic lineage for both Gel-MA and RCPhC1-MA scaffolds, showing a trend towards superior differentiation for the RCPhC1-MA scaffolds based on the triglyceride and Bodipy assay. RCPhC1-MA scaffolds could result in a transition towards the exploitation of non-animal-derived biomaterials for ATE, omitting any regulatory concerns related to the use of animal derived products.

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以重组胶原蛋白为基础的非动物源生物材料是实现脂肪组织工程学的一种前景广阔的战略。
过去几十年来,脂肪组织工程(ATE)越来越受到人们的关注,它为新的和创新的乳房重建策略带来了希望。动物源性明胶-甲基丙烯酰已被应用于大量的脂肪组织工程策略中。然而,由于临床上担心可能会出现免疫球蛋白(IgE)介导的免疫反应和病原体传播,人们开始转向使用确定的、可重复的重组蛋白。在本研究中,一种以人类 I 型胶原蛋白为基础、富含精氨酸-甘氨酸-天冬氨酸(RGD)的重组蛋白被光交联甲基丙烯酰基(RCPhC1-MA)功能化,加工成三维支架,并使用聚乳酸(PLA)作为牺牲模具的间接打印方法,与经常应用的动物源明胶-甲基丙烯酰基(Gel-MA)进行比较。两种材料都获得了相似的凝胶分数 (> 65 %)和生物降解时间。此外,与 Gel-MA 相比,RCPhC1-MA 的质量膨胀率(17.6 ± 1.5 对 24.3 ± 1.4)和机械强度(杨氏模量:1.1 ± 0.2 kPa 对 1.9 ± 0.3 kPa)明显较低。体外播种试验显示,RCPhC1-MA 支架具有相似的细胞存活率(> 80%)和更高的初始细胞附着率。此外,在Gel-MA和RCPhC1-MA支架上播种的脂肪来源干细胞都能分化成成脂系,根据甘油三酯和Bodipy测定,RCPhC1-MA支架的分化趋势更优。RCPhC1-MA 支架可使 ATE 向利用非动物来源生物材料过渡,从而避免了与使用动物来源产品有关的任何监管问题。
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