CAJ | 학술논문

目的：目前可注射组织工程骨或者载体支架多为凝胶状，其表面积、强度、硬度等方面无法满足复杂多变的骨、软骨缺损修复的要求。文中主要评价新型可注射微球型多孔壳聚糖／羟基磷灰石陶瓷支架的生物学性能。方法制备质量分数分别为30％、50％、70％的可注射多孔壳聚糖／羟基磷灰石复合微球，分别利用液氮的冷冻效应（液氮组）和非液氮途径（非液氮组）处理后烧结获得羟基磷灰石陶瓷球，拟做为新型组织工程骨的载体支架材料。通过扫描电镜观察支架的微细结构，测定孔隙率；通过生物力学实验测定力学性能。将人脐静脉内皮细胞（ human umbilical vein endothelial cell ， HUVECs）种植于多孔壳聚糖／羟基磷灰石陶瓷支架，通过电镜扫观察HUVECs在多孔壳聚糖／羟基磷灰石陶瓷支架内部的生长情况验证多孔壳聚糖／羟基磷灰石陶瓷支架的生物融合性。结果液氮组和非液氮组不同质量分数的陶瓷支架肉眼无差异。扫描电镜下的外观图及内部图可见：2组微球陶瓷支架均呈球形、大小均匀、形态规则。液氮处理过的微球陶瓷支架表面见大量不规则孔隙；而非液氮处理微球陶瓷支架表面比较光滑，无明显的孔隙；2组微球陶瓷支架随着壳聚糖／羟基磷灰石的质量百分比的增大，内部的孔隙随之减少，使支架内部结构变的致密。液氮组50％质量分数支架平均直径、平均压缩强度均高于30％质量分数支架[（0．48±0．11） mm vs （0．40±0．08） mm，（1．75±0．14） MPa vs （1．21±0．12） MPa，P＜0．05）]，孔隙率低于30％质量分数支架[（79±2）％ vs （87±1）％，P＜0．05]，平均孔径差异无统计学意义（P＞0．05）。电镜扫描见HUVECs在多孔三磷酸钙支架内部呈多突触样附着，生长良好。结论液氮组50％质量分数多孔壳聚糖／羟基磷灰石陶瓷支架具有较强的力学强度和较好的生物融合性，能够做为新型组织工程骨的载体支架。目的：目前可註射組織工程骨或者載體支架多為凝膠狀，其錶麵積、彊度、硬度等方麵無法滿足複雜多變的骨、軟骨缺損脩複的要求。文中主要評價新型可註射微毬型多孔殼聚糖／羥基燐灰石陶瓷支架的生物學性能。方法製備質量分數分彆為30％、50％、70％的可註射多孔殼聚糖／羥基燐灰石複閤微毬，分彆利用液氮的冷凍效應（液氮組）和非液氮途徑（非液氮組）處理後燒結穫得羥基燐灰石陶瓷毬，擬做為新型組織工程骨的載體支架材料。通過掃描電鏡觀察支架的微細結構，測定孔隙率；通過生物力學實驗測定力學性能。將人臍靜脈內皮細胞（ human umbilical vein endothelial cell ， HUVECs）種植于多孔殼聚糖／羥基燐灰石陶瓷支架，通過電鏡掃觀察HUVECs在多孔殼聚糖／羥基燐灰石陶瓷支架內部的生長情況驗證多孔殼聚糖／羥基燐灰石陶瓷支架的生物融閤性。結果液氮組和非液氮組不同質量分數的陶瓷支架肉眼無差異。掃描電鏡下的外觀圖及內部圖可見：2組微毬陶瓷支架均呈毬形、大小均勻、形態規則。液氮處理過的微毬陶瓷支架錶麵見大量不規則孔隙；而非液氮處理微毬陶瓷支架錶麵比較光滑，無明顯的孔隙；2組微毬陶瓷支架隨著殼聚糖／羥基燐灰石的質量百分比的增大，內部的孔隙隨之減少，使支架內部結構變的緻密。液氮組50％質量分數支架平均直徑、平均壓縮彊度均高于30％質量分數支架[（0．48±0．11） mm vs （0．40±0．08） mm，（1．75±0．14） MPa vs （1．21±0．12） MPa，P＜0．05）]，孔隙率低于30％質量分數支架[（79±2）％ vs （87±1）％，P＜0．05]，平均孔徑差異無統計學意義（P＞0．05）。電鏡掃描見HUVECs在多孔三燐痠鈣支架內部呈多突觸樣附著，生長良好。結論液氮組50％質量分數多孔殼聚糖／羥基燐灰石陶瓷支架具有較彊的力學彊度和較好的生物融閤性，能夠做為新型組織工程骨的載體支架。
목적：목전가주사조직공정골혹자재체지가다위응효상，기표면적、강도、경도등방면무법만족복잡다변적골、연골결손수복적요구。문중주요평개신형가주사미구형다공각취당／간기린회석도자지가적생물학성능。방법제비질량분수분별위30％、50％、70％적가주사다공각취당／간기린회석복합미구，분별이용액담적냉동효응（액담조）화비액담도경（비액담조）처리후소결획득간기린회석도자구，의주위신형조직공정골적재체지가재료。통과소묘전경관찰지가적미세결구，측정공극솔；통과생물역학실험측정역학성능。장인제정맥내피세포（ human umbilical vein endothelial cell ， HUVECs）충식우다공각취당／간기린회석도자지가，통과전경소관찰HUVECs재다공각취당／간기린회석도자지가내부적생장정황험증다공각취당／간기린회석도자지가적생물융합성。결과액담조화비액담조불동질량분수적도자지가육안무차이。소묘전경하적외관도급내부도가견：2조미구도자지가균정구형、대소균균、형태규칙。액담처리과적미구도자지가표면견대량불규칙공극；이비액담처리미구도자지가표면비교광활，무명현적공극；2조미구도자지가수착각취당／간기린회석적질량백분비적증대，내부적공극수지감소，사지가내부결구변적치밀。액담조50％질량분수지가평균직경、평균압축강도균고우30％질량분수지가[（0．48±0．11） mm vs （0．40±0．08） mm，（1．75±0．14） MPa vs （1．21±0．12） MPa，P＜0．05）]，공극솔저우30％질량분수지가[（79±2）％ vs （87±1）％，P＜0．05]，평균공경차이무통계학의의（P＞0．05）。전경소묘견HUVECs재다공삼린산개지가내부정다돌촉양부착，생장량호。결론액담조50％질량분수다공각취당／간기린회석도자지가구유교강적역학강도화교호적생물융합성，능구주위신형조직공정골적재체지가。
[Abstract ] Objective At present, the majority of injectable tissue engineering bones or carrier stents are gel , whose surface area , intensity, and hardness cannot satisfy the requirements of the repair of complex and varied bone and cartilage defects .This paper evaluated the new injectable microspherical porous chitosan/biological properties of the hydroxyapatite ceramic scaffold . Methods Injectable porous chitosan /hydroxyapatite composite microspheres with mass fractions of 30%, 50%, and 70% were prepared respectively . The hydroxyapatite ceramic ball was obtained by sintering with liquid nitrogen freezing ( liquid nitrogen group ) or without liquid nitrogen pro-cessing ( non-liquid nitrogen group ) as a new carrier of bone tissue engineering scaffold material .The microstructure of the scaffold was observed and the porosity measured under the scanning electron microscope .The mechanical properties were determined through biome-chanical experiments.Human umbilical vein endothelial cells (HUVECs) were grown in the porous chitosan/hydroxyapatite ceramic scaf-fold followed by observation of the growth of the cells and validation of the biological fusion of the scaffold . Results No difference was observed with the naked eye in the ceramic scaffold of different mass fractions in the liquid nitrogen and non -liquid nitrogen groups . Scanning electron microscopy exhibited spherical shape , uniform size, and regular morphology of the ceramic scaffolds in both groups .A large number of irregular pores were seen in the surface of the microspherical ceramic scaffolds treated with liquid nitrogen but not in the surface of those not treated .With increased mass percentage of chitosan/hydroxyapatite , the internal pores were reduced and the interior structure compacted.In the liquid nitrogen group, the scaffold of 50%mass fraction had a significantly larger diameter ([0.48 ±0.11] mm), higher compression intensity ([1.75 ±0.14] MPa), and lower porosity ([79 ±2]%) than that of 30%mass fraction ([0.40 ± 0.08] mm, [1.21 ±0.12] MPa, and [87 ±1]%) (all P<0.05).Electron microscope scanning revealed well -grown HUVECs with multiple synapses in the porous tricalcium phosphate scaffold. C onclusion The porous chitosan /hydroxyapatite ceramic scaffold of 50%mass fraction treated with liquid nitrogen , with its strong mechanical intensity and high biological fusibility , can be used as a new carrier of bone tissue engineering scaffolds .