可注射型华通胶光交联水凝胶用于组织工程软骨再生的实验研究

Injectable Wharton's Jelly Photo-Crosslinking Hydrogel for Tissue-Engineered Cartilage Regeneration

目的探索可注射型华通胶光交联水凝胶用于构建组织工程软骨的可行性。方法制备可注射型华通胶光交联水凝胶载体材料,检测其流变特性及溶胀率;随后取兔耳软骨细胞与该水凝胶均匀混合后体外培养,验证其细胞相容性;最后将软骨细胞-华通胶光交联水凝胶复合物植入裸鼠皮下培养2周和6周,行大体和组织学观察、生物力学及生物化学定量检测。结果华通胶光交联水凝胶具有适宜的流变特性及良好的细胞相容性,最高溶胀率约为1.2。体内培养2~6周,逐渐形成富有弹性的瓷白色成熟软骨样组织,组织学(HE,Safranin-O及Ⅱ型胶原免疫组化染色)显示可逐渐形成典型的软骨特征,包括丰富的陷窝和均匀的软骨特异性基质沉积。生物力学和生物化学检测显示,随着体内培养时间由2周延长至6周,新生软骨组织逐渐成熟至与正常软骨组织相似。结论可注射型华通胶光交联水凝胶适合用于构建组织工程软骨。

Objective To explore the feasibility of injectable Wharton's jelly photo-crosslinking hydrogel to construct tissue-engineered cartilage.Methods Photo-crosslinking hydrogel was fabricated with decellularized Wharton's jelly and followed by rheological properties and swelling ratio tests.Auricular chondrocytes derived from rabbit were evenly incorporated into the hydrogel for cytocompatibility examination.Thereafter,the cell-hydrogel constructs were subcutaneously implanted into nude mice.Gross observation,histological observation and quantitative evaluation of neocartilage test were carried out after in vivo culture for 2 or 6 weeks.Results The Wharton's jelly photo-crosslinking hydrogel possessed satisfactory rheological properties and suitable cytocompatibility,and the maximum swelling ratio was about 1.2.Notably,samples gradually presented white-ivory appearance after extended in vivo culture time from 2 weeks to 6 weeks.Histological evaluation,including HE,Safranin-O,and type II collagen immunohistochemical staining gradually displayed typical cartilaginous features with abundant lacunae and homogeneous cartilage specific matrix deposition.Biomechanical and biochemistry analyses indicated that neocartilage was comparable to native cartilage after extended in vivo culture time from 2 weeks to 6 weeks.Conclusion Wharton's jelly photo-crosslinking hydrogel is a promising natural-derived material for tissue engineering cartilage regeneration.