Jia Zhilin, Ma Hailin, Liu Jiaqi, Yan Xinyu, Liu Tianqing, Cheng Yuen Yee, Li Xiangqin, Wu Shuo, Zhang Jingying, Song Kedong
State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024, China.
Department of Hematology, The First Affiliated Hospital of Dalian Medical University, Dalian 116011, China.
Materials (Basel). 2023 Feb 26;16(5):1937. doi: 10.3390/ma16051937.
Bone tissue engineering is a novel and efficient repair method for bone tissue defects, and the key step of the bone tissue engineering repair strategy is to prepare non-toxic, metabolizable, biocompatible, bone-induced tissue engineering scaffolds of suitable mechanical strength. Human acellular amniotic membrane (HAAM) is mainly composed of collagen and mucopolysaccharide; it has a natural three-dimensional structure and no immunogenicity. In this study, a polylactic acid (PLA)/Hydroxyapatite (nHAp)/Human acellular amniotic membrane (HAAM) composite scaffold was prepared and the porosity, water absorption and elastic modulus of the composite scaffold were characterized. After that, the cell-scaffold composite was constructed using newborn Sprague Dawley (SD) rat osteoblasts to characterize the biological properties of the composite. In conclusion, the scaffolds have a composite structure of large and small holes with a large pore diameter of 200 μm and a small pore diameter of 30 μm. After adding HAAM, the contact angle of the composite decreases to 38.7°, and the water absorption reaches 249.7%. The addition of nHAp can improve the scaffold's mechanical strength. The degradation rate of the PLA+nHAp+HAAM group was the highest, reaching 39.48% after 12 weeks. Fluorescence staining showed that the cells were evenly distributed and had good activity on the composite scaffold; the PLA+nHAp+HAAM scaffold has the highest cell viability. The adhesion rate to HAAM was the highest, and the addition of nHAp and HAAM could promote the rapid adhesion of cells to scaffolds. The addition of HAAM and nHAp can significantly promote the secretion of ALP. Therefore, the PLA/nHAp/HAAM composite scaffold can support the adhesion, proliferation and differentiation of osteoblasts in vitro which provide sufficient space for cell proliferation, and is suitable for the formation and development of solid bone tissue.
骨组织工程是一种针对骨组织缺损的新型高效修复方法，而骨组织工程修复策略的关键步骤是制备具有合适机械强度、无毒、可代谢、生物相容性良好且具有骨诱导性的组织工程支架。人脱细胞羊膜（HAAM）主要由胶原蛋白和粘多糖组成；它具有天然的三维结构且无免疫原性。在本研究中，制备了聚乳酸（PLA）/羟基磷灰石（nHAp）/人脱细胞羊膜（HAAM）复合支架，并对该复合支架的孔隙率、吸水率和弹性模量进行了表征。之后，使用新生Sprague Dawley（SD）大鼠成骨细胞构建细胞 - 支架复合物，以表征该复合物的生物学特性。综上所述，该支架具有大小孔复合结构，大孔径为200μm，小孔径为30μm。添加HAAM后，复合材料的接触角降至38.7°，吸水率达到249.7%。添加nHAp可提高支架的机械强度。PLA + nHAp + HAAM组的降解率最高，12周后达到39.48%。荧光染色显示细胞在复合支架上分布均匀且活性良好；PLA + nHAp + HAAM支架的细胞活力最高。对HAAM的粘附率最高，添加nHAp和HAAM可促进细胞快速粘附到支架上。添加HAAM和nHAp可显著促进碱性磷酸酶（ALP）的分泌。因此，PLA/nHAp/HAAM复合支架可在体外支持成骨细胞的粘附、增殖和分化，为细胞增殖提供足够空间，适用于实体骨组织的形成与发育。
