CAJ | 학술논문

目的评估聚D,L-乳酸(PDLLA)三维多孔支架的体外细胞相容性,探讨其作为骨组织工程支架类材料的可行性,为进一步进行体内试验提供依据.方法将分离、获取的第3代骨髓间充质干细胞(BMSCs)按随机数字表法分成5组(0,3,6,9,12周组),分别用含体积分数20％降解液(0,3,6,9,12周时的PDLLA降解液)的细胞培养液和成骨诱导分化培养液进行细胞培养(以不含有降解液的细胞培养液或者成骨诱导分化培养液培养组为对照组,即0周组),从BMSCs的毒性、活力、成骨分化能力等方面定量评估PDLLA支架材料的细胞相容性;将分离、获取的第3代BMSCs种植于PDLLA支架材料上,定期进行扫描电子显微镜观察,从不同时相点细胞在PDLLA材料表面的生长情况定性评估PDLLA支架材料的细胞相容性. 结果各组与对照组比较均未发现对细胞具有明显的毒性作用;各组均未发现对细胞的活力具有明显的负面影响(t3=-0.441,P=0.671;t6=1.596,P=0.154;t9=-0.492,P=0.636;t12=-1.135,P=0.283);各组细胞ALP染色和钙结节染色结果均表现为阳性,ALP活性定量检测和Ⅰ型胶原蛋白表达的定量检测与对照组之间差异均无统计学意义.扫描电镜结果显示,BMSCs在PDLLA三维多孔支架材料上呈现较好的贴壁生长.结论 PDLLA支架材料对BMSCs具有较好的体外相容性,可以作为一种骨组织工程支架类材料进行后续的体内试验.
목적 평고취D,L-유산(PDLLA)삼유다공지가적체외세포상용성,탐토기작위골조직공정지가류재료적가행성,위진일보진행체내시험제공의거.방법 장분리、획취적제3대골수간충질간세포(BMSCs)안수궤수자표법분성5조(0,3,6,9,12주조),분별용함체적분수20％강해액(0,3,6,9,12주시적PDLLA강해액)적세포배양액화성골유도분화배양액진행세포배양(이불함유강해액적세포배양액혹자성골유도분화배양액배양조위대조조,즉0주조),종BMSCs적독성、활력、성골분화능력등방면정량평고PDLLA지가재료적세포상용성;장분리、획취적제3대BMSCs충식우PDLLA지가재료상,정기진행소묘전자현미경관찰,종불동시상점세포재PDLLA재료표면적생장정황정성평고PDLLA지가재료적세포상용성. 결과 각조여대조조비교균미발현대세포구유명현적독성작용;각조균미발현대세포적활력구유명현적부면영향(t3=-0.441,P=0.671;t6=1.596,P=0.154;t9=-0.492,P=0.636;t12=-1.135,P=0.283);각조세포ALP염색화개결절염색결과균표현위양성,ALP활성정량검측화Ⅰ형효원단백표체적정량검측여대조조지간차이균무통계학의의.소묘전경결과현시,BMSCs재PDLLA삼유다공지가재료상정현교호적첩벽생장.결론 PDLLA지가재료대BMSCs구유교호적체외상용성,가이작위일충골조직공정지가류재료진행후속적체내시험.
Objective To investigate the in vitro cytocompatibility of three-dimensional porous scaffolds of poly-D,L-lactic acid (PDLLA) and discuss the feasibility of PDLLA as a scaffold for bone tissue engineering.Methods BMSCs of the third passage were seeded on osteogenetic differentiation medium or culture medium containing 20％ volume fraction degraded liquid (PDLLA degradation liquid of 0,3,6,9,and 12 weeks) according to the random number table.Osteogenetic differentiation medium or culture medium without PDLLA was used as controls.Cell viability,cytotoxicity,and osteogenic differentiation were detected for study on cytocompatibility of PDLLA.Scanning electron microscopy was used to observe the growth of BMSCs on the surface of PDLLA scaffolds.Results PDLLA scaffolds presented no significant cytotoxic on the growth of BMSCs.PDLLA scaffolds had no negative effect on cell viability compared with the controls (t3 =-0.441,P =0.671; t6 =1.596,P =0.154; t9 =-0.492,P =0.636; t12 =-1.135,P=0.283).ALP staining and calcium nodule staining were positive and there were no significant differences in ALP and collagen Ⅰ protein quantitative detection compared with the controls.BMSCs grew well on the inner surface of the PDLLA three-dimensional porous scaffolds.Conclusion Three-dimensional porous scaffolds of PDLLA present good cytocompatibility in vitro and can be used as bone tissue engineering scaffolds for subsequent in vivo research.