异种脱蛋白组织工程骨支架材料的制备及理化特性研究

Establishment of scaffold materials of deproteinized heterogeneous bone and their physicochemical properties

目的制备及研究可供临床使用的异种脱蛋白组织工程骨支架材料。方法对猪肋骨进行一系列理化处理,制成脱蛋白骨,并对其进行组织学、扫描电镜、红外光谱、X射线衍射分析,X射线能量散射分析、微量凯氏定氮及力学性能测定。结果脱蛋白骨间质胶原纤维染色阳性,黏蛋白染色阴性,而新鲜骨组织黏蛋白及胶原染色均阳性。脱蛋白骨保持原骨组织的天然网状孔隙系统,孔径为(472.5 l±7.02)μm,孔隙率为(78.15%±6.45%),脱蛋白骨红外光谱显示胶原存在,无脂肪。新鲜骨与脱蛋白骨皆为曲线型羟基磷灰石图谱,Ca/P比分别为(1.71±0.95)、(1.68±0.76),二者差异无显著性(P>0.05),蛋白质含量分别为(26.6%±2.23%)、(19.1%±2.14%),二者差异有显著性(P<0.05)。脱蛋白骨除弹性模量比新鲜骨升高外,压缩破坏载荷及破坏极限载荷与新鲜骨差异无显著性(P>0.05)。结论制备的脱蛋白组织工程骨支架材料的理化性质及力学强度符合理想支架材料要求。其免疫原性有待于进一步实验观察。

Objective To establish scaffold materials of deproteinized heterogeneous tissue engineering bone. Methods Deproteinized bones were made through a series of physicochemical treatment from pig ribs and were analyzed with histology, scanning electron microscopy, infrared spectrum, X-ray diffraction analysis, X-ray energy scattering analysis, microdose Kjeldahl determination and mechanical analysis. Results Stain of interstitial collagen fibers was positive and mucin stain was negative in deproteinized bone, but stain of interstitial collagen fibers and mucin were all positive in fresh bone. Deproteinized bone maintained natural network pore system, its pore size was （472.51±7.02）μm, its porosity was （78.15±6.45）%. Infrared spectrum showed collogen in deproteinized bone. Fresh and deproteinized bone were all of curve style atlas of hydroxyapatite, their ratio of calcium to phosphors was （ 1.71±0.95 ） and （ 1.68±0.76） （ P 〉 0.05 ）, and their protein content was （26.6±2.23） % and （ 19.1±2.14） % （ P 〈 0.05 ）. The compression destruction load and destruction maximum load of deproteinized bone were of no significant difference with those of fresh bone （P 〉 0.05 ）, except for high elastic modulus. Conclusion Physicochemical property and mechanical strength of scaffold materials of established deproteinized tissue engineering bone meet request of ideal scaffold materials. Its immunogenicity should be further observed.