Kung Fu-Chen
Department of Health Healing and Health Marketing, Kainan University, Taoyuan 338, Taiwan. Tel.: +886-3-341-2500 #7971; Fax: +886-3-341-4428; E-mail:
Biomed Mater Eng. 2018;29(2):241-251. doi: 10.3233/BME-171726.
Imbalance crosslink density and polymer concentration gradient is formed within the traditional alginate hydrogel using calcium chloride as a crosslinking agent in external gelation for instantaneously process. In this studying, type I collagen (Col I) blended calcium salt form of poly(γ-glutamic acid) (γCaPGA) was mixing with RGD-modified alginate with convenient gelation process and suitable for practical use. The hydrophilicity of the resulting hydrogels was evaluated through swelling tests, water retention capacity tests, and water vapor permeation tests. Mineralization was qualitatively evaluated by alizarin red dyeing at day 14, verifying the deposition of calcium. The in vitro osteogenic differentiation is monitored by determining the early and late osteocalcin (OCN) and osteopontin (OPN) markers with MG63 cells. Obtained results demonstrated that no extremely changes in mechanical properties. After 14 days of culture, hydrogels significantly stimulated OCN/OPN gene expressions and MG63 cell proliferation. Unusually, γCaPGA with RGD-modified alginate appeared better calcium deposition in 14 days than the other. However, addition of Col I can counterpoise RGD effect in blood coagulation and platelet adhesion made the hydrogel more flexibility and selectively in use. This studying provided that non-covalently crosslinked hydrogel by γCaPGA with alginate can be upgrading by RGD and Col I in water uptake capability, obviously effective for MG63 cells and are remarkably biocompatible and exhibited no cytotoxicity. Moreover, results also displayed the injectable process without complicated procedure, have high cost/performance ratio and have great potential for bone regeneration.
在传统的藻酸盐水凝胶中,使用氯化钙作为交联剂进行外部凝胶化的瞬间过程,会形成交联密度和聚合物浓度梯度的不平衡。在本研究中,将I型胶原蛋白(Col I)与聚(γ-谷氨酸)的钙盐形式(γCaPGA)混合,并与RGD修饰的藻酸盐混合,其凝胶化过程简便且适用于实际应用。通过溶胀试验、保水能力试验和水蒸气渗透试验对所得水凝胶的亲水性进行了评估。在第14天通过茜素红染色对矿化进行定性评估,以验证钙的沉积。通过用MG63细胞测定早期和晚期骨钙素(OCN)和骨桥蛋白(OPN)标志物来监测体外成骨分化。获得的结果表明力学性能没有极端变化。培养14天后,水凝胶显著刺激了OCN/OPN基因表达和MG63细胞增殖。不同寻常的是,含RGD修饰藻酸盐的γCaPGA在14天时的钙沉积比其他的更好。然而,添加Col I可以抵消RGD在凝血和血小板粘附中的作用,使水凝胶在使用中更具柔韧性和选择性。本研究表明,γCaPGA与藻酸盐形成的非共价交联水凝胶可以通过RGD和Col I在吸水能力方面得到提升,对MG63细胞明显有效,并且具有显著的生物相容性且无细胞毒性。此外,结果还显示该注射过程无需复杂程序,具有高性价比,在骨再生方面具有巨大潜力。