构建具备良好热稳定性、自组装性质及生物相容性的可食性细胞外基质(extracellular matrix,ECM)类似物支架对于制造结构化细胞培养肉制品至关重要。将羧甲基壳聚糖(carboxymethyl chitosan,CMCS)引入牛骨胶原蛋白(bovine bone collagen,...构建具备良好热稳定性、自组装性质及生物相容性的可食性细胞外基质(extracellular matrix,ECM)类似物支架对于制造结构化细胞培养肉制品至关重要。将羧甲基壳聚糖(carboxymethyl chitosan,CMCS)引入牛骨胶原蛋白(bovine bone collagen,BBC)体系中,通过光谱分析(紫外、红外、荧光光谱)发现BBC与CMCS的相互作用随着引入CMCS添加量的增加而增强,但并未影响BBC的三螺旋结构。差示扫描量热法/热重分析结果表明,CMCS的引入增强了BBC体系的热稳定性。浊度试验及扫描电子显微镜/透射电子显微镜观察结果证实了CMCS引入后胶原蛋白纤维形成度呈上升趋势,聚集行为更明显且自组装速率产生变化,呈现出更疏松扭曲的三维结构以及更大的纤维直径及更广泛的直径分布。但CMCS的引入并未明显影响BBC的D-周期性结构(胶原纤维自组装过程中形成的特征性明暗交替的周期性横纹结构)形成及其长度,且CMCS引入前后体系的细胞相容性也未呈现显著性差异。随着引入CMCS添加量增加,CMCS和BBC之间的静电作用力可能较共价相互作用和氢键更占优势。这些结果表明,CMCS的引入不影响BBC三螺旋结构完整性和生物相容性,并改善了BBC的热稳定性及体外自组装性质。这为开发新型优良可食性胶原蛋白基ECM仿生支架在细胞培养肉领域的应用以及畜禽骨副产物高值化精深加工利用提供了参考信息。展开更多
Abstract Objectives To elucidate the molecular changes of bone collagen during the development of postmenopausal osteoporosis and to investigate the molecular effects of estrogen replacement. Methods An ad...Abstract Objectives To elucidate the molecular changes of bone collagen during the development of postmenopausal osteoporosis and to investigate the molecular effects of estrogen replacement. Methods An adult ovariotomy rat model was used. Type Ⅰ collagen and matrix metalloproteinase 9 (MMP 9) expressions in bone tissues of rats treated by sham surgery (SH), bilateral ovariotomy (OVX) and OVX with estradiol (OVX E2) were analysed at mRNA level by using dot blot technique. The distribution of mRNA of these two genes in bone tissues was studied by in situ hybridization. Results The expression levels of both type Ⅰ collagen and MMP 9 in bone tissues of OVX rats were higher than those of SH group, while treated with estradiol, the expression of both genes declined to some degree. In situ hybridization showed that type Ⅰ collagen mRNA located in osteoblasts, whereas MMP 9 was mainly expressed in osteoclasts, some lining cells on bone surface, and some mononuclear cells in bone marrow. Conclusions The reduction of high bone turnover in osteoporotic bone tissues induced by estrogen replacement may result from alterations in gene expression related to bone formation and bone resorption. These alterations are consistent with the changes observed previously by histomorphometry and biochemical markers of bone metabolism on OVX animals and postmenopausal osteoporosis.展开更多
文摘构建具备良好热稳定性、自组装性质及生物相容性的可食性细胞外基质(extracellular matrix,ECM)类似物支架对于制造结构化细胞培养肉制品至关重要。将羧甲基壳聚糖(carboxymethyl chitosan,CMCS)引入牛骨胶原蛋白(bovine bone collagen,BBC)体系中,通过光谱分析(紫外、红外、荧光光谱)发现BBC与CMCS的相互作用随着引入CMCS添加量的增加而增强,但并未影响BBC的三螺旋结构。差示扫描量热法/热重分析结果表明,CMCS的引入增强了BBC体系的热稳定性。浊度试验及扫描电子显微镜/透射电子显微镜观察结果证实了CMCS引入后胶原蛋白纤维形成度呈上升趋势,聚集行为更明显且自组装速率产生变化,呈现出更疏松扭曲的三维结构以及更大的纤维直径及更广泛的直径分布。但CMCS的引入并未明显影响BBC的D-周期性结构(胶原纤维自组装过程中形成的特征性明暗交替的周期性横纹结构)形成及其长度,且CMCS引入前后体系的细胞相容性也未呈现显著性差异。随着引入CMCS添加量增加,CMCS和BBC之间的静电作用力可能较共价相互作用和氢键更占优势。这些结果表明,CMCS的引入不影响BBC三螺旋结构完整性和生物相容性,并改善了BBC的热稳定性及体外自组装性质。这为开发新型优良可食性胶原蛋白基ECM仿生支架在细胞培养肉领域的应用以及畜禽骨副产物高值化精深加工利用提供了参考信息。
文摘Abstract Objectives To elucidate the molecular changes of bone collagen during the development of postmenopausal osteoporosis and to investigate the molecular effects of estrogen replacement. Methods An adult ovariotomy rat model was used. Type Ⅰ collagen and matrix metalloproteinase 9 (MMP 9) expressions in bone tissues of rats treated by sham surgery (SH), bilateral ovariotomy (OVX) and OVX with estradiol (OVX E2) were analysed at mRNA level by using dot blot technique. The distribution of mRNA of these two genes in bone tissues was studied by in situ hybridization. Results The expression levels of both type Ⅰ collagen and MMP 9 in bone tissues of OVX rats were higher than those of SH group, while treated with estradiol, the expression of both genes declined to some degree. In situ hybridization showed that type Ⅰ collagen mRNA located in osteoblasts, whereas MMP 9 was mainly expressed in osteoclasts, some lining cells on bone surface, and some mononuclear cells in bone marrow. Conclusions The reduction of high bone turnover in osteoporotic bone tissues induced by estrogen replacement may result from alterations in gene expression related to bone formation and bone resorption. These alterations are consistent with the changes observed previously by histomorphometry and biochemical markers of bone metabolism on OVX animals and postmenopausal osteoporosis.
