Department of Stem Cell Research, TJC Life Research and Development Center, TJC Life, Seoul, Republic of Korea.
Department of Health Management, Public Health Center, Chuncheon, Republic of Korea.
Biochem Biophys Res Commun. 2020 Jan 29;522(1):213-219. doi: 10.1016/j.bbrc.2019.11.049. Epub 2019 Nov 20.
In cell-based tissue engineering, fibrin hydrogel can be utilized to produce scaffolds to treat cartilage. However, the optimal fibrin formulation for cartilage regeneration has not yet been studied. This study aimed to find the optimal fibrin formulation and determine whether fibrin optimized with human adipose-derived stem cells (hASCs) increased the in-vivo chondrogenic potential. To find the optimal formulation, fibrin constructs were divided into twelve groups with different ratios of fibrinogen (10, 20, 30, and 50 mg/mL) to thrombin (10, 50, and 100 IU/mL), following which the physical and biological properties of cell-free and cell-embedded fibrin were investigated. The results from cell-free hydrogels showed that increases in the concentrations of fibrinogen and thrombin corresponded to increases in stiffness and initial weight. Moreover, hydrogel degradation was inhibited in high-concentration formulations. In cell-embedded fibrin constructs, the variation of gel formulation did not affect cell viability. However, cell behavior depended on the gel formulation. hASCs within high-concentration fibrinogen formulation maintained a round morphology similar to natural chondrocytes. Variations in thrombin concentration had a lesser effect on cell morphology. In terms of in-vivo cartilage formation, the formulation with 30 mg/mL fibrinogen and 100 IU/mL thrombin showed the highest cartilage formation, as evidenced through collagen type II alpha 1 chain (COL2) and safranin-O, 4 weeks after implantation. The results may lead to optimally designed 3D bio-scaffolds in which we can control both cell survival and chondrogenic potential for cartilage tissue engineering. Scaffolds made with the optimal fibrin formulation can be applied to develop cell therapies with mesenchymal stem cells to treat osteoarthritis.
在基于细胞的组织工程中，纤维蛋白水凝胶可用于制造支架来治疗软骨。然而，用于软骨再生的最佳纤维蛋白配方尚未得到研究。本研究旨在找到最佳纤维蛋白配方，并确定是否用人类脂肪来源干细胞（hASCs）优化的纤维蛋白增加了体内软骨形成潜力。为了找到最佳配方，将纤维蛋白构建体分为 12 组，每组的纤维蛋白原（10、20、30 和 50mg/mL）与凝血酶（10、50 和 100IU/mL）的比例不同，随后研究了无细胞和细胞嵌入纤维蛋白的物理和生物特性。无细胞水凝胶的结果表明，纤维蛋白原和凝血酶浓度的增加对应于硬度和初始重量的增加。此外，高浓度配方抑制了水凝胶的降解。在细胞嵌入纤维蛋白构建体中，凝胶配方的变化不会影响细胞活力。然而，细胞行为取决于凝胶配方。高浓度纤维蛋白原配方中的 hASCs 保持类似于天然软骨细胞的圆形形态。凝血酶浓度的变化对细胞形态的影响较小。就体内软骨形成而言，纤维蛋白原浓度为 30mg/mL 和凝血酶浓度为 100IU/mL 的配方显示出最高的软骨形成，这可通过植入后 4 周的 II 型胶原α 1 链（COL2）和番红-O 来证明。这些结果可能导致最佳设计的 3D 生物支架，在该支架中我们可以控制细胞存活和软骨形成潜力，用于软骨组织工程。使用最佳纤维蛋白配方制造的支架可应用于开发间充质干细胞的细胞疗法，以治疗骨关节炎。
