本文選題：骨替代材料 + 可注射性　； 參考：《中國修復重建外科雜志》2017年06期

【摘要】：目的制備復合透明質酸鈉的硫酸鈣可注射材料,觀察其促進骨再生作用,為骨缺損修復提供一種可注射材料。方法將硫酸鈣分別與透明質酸鈉溶液、交聯(lián)透明質酸鈉溶液、PBS溶液,按照2∶1(W/V)比例混合,制備3種復合材料(記作CA+HA、CA+HAC以及CA)。通過將3種復合材料浸泡于PBS溶液中觀察其形變并進行X線衍射分析,觀察材料穩(wěn)定性。參照ISO10993-5制備3種復合材料浸提液,用于培養(yǎng)小鼠前成骨細胞(MC3T3-E1),并采用細胞增殖-毒性檢測試劑盒(cell counting kit-8,CCK-8)方法檢測材料的生物相容性和不同濃度浸提液促進細胞增殖的能力,以單純培養(yǎng)基培養(yǎng)細胞作為對照組。采用成骨分化培養(yǎng)基制作促進細胞增殖的最佳濃度浸提液,用于培養(yǎng)MC3T3-E1細胞,ELISA法檢測成骨分化相關蛋白ALP、Ⅰ型膠原(collagen typeⅠ,COL-Ⅰ)及骨鈣蛋白(osteocalcin,OCN)濃度。取新西蘭大白兔制作股骨髁骨缺損模型,分別植入CA+HA、CA+HAC以及CA材料,于6、12周取標本行Micro-CT掃描并計算骨組織占組織體積百分比(bone volume/tissue volume,BV/TV),然后標本切片行HE染色,觀察缺損區(qū)新骨形成情況。結果復合材料穩(wěn)定性檢測示,CA+HA及CA+HAC具有良好可注射性,在PBS溶液中不易潰散,優(yōu)于CA。生物相容性實驗顯示,培養(yǎng)6、12、24 h,CA組吸光度(A)值均低于對照組(P0.05);CA+HA組、CA+HAC組A值與對照組比較,差異無統(tǒng)計學意義(P0.05)。細胞增殖實驗顯示,25%、50%濃度浸提液培養(yǎng)5 d時CA組、CA+HA組和CA+HAC組A值均顯著高于對照組(P0.05);75%、100%濃度浸提液培養(yǎng)后,僅CA+HA組A值高于對照組(P0.05)。選擇50%濃度浸提液進行成骨分化實驗。ELISA檢測示,培養(yǎng)14、21 d時CA+HA組和CA+HAC組ALP、COL-Ⅰ、OCN濃度均顯著高于對照組、CA組(P0.05)。Micro-CT檢查示,6、12周時CA+HA組、CA+HAC組間BV/TV差異無統(tǒng)計學意義(P0.05),但均顯著高于CA組(P0.05);HE染色示,6周時CA組缺損處無成形骨組織,CA+HA組和CA+HAC組可見少量骨組織;12周時,CA組可見到少量細條索狀骨組織形成,CA+HA組和CA+HAC組均見較多條索狀骨組織,明顯優(yōu)于CA組,但兩組間無顯著差異。結論硫酸鈣與透明質酸鈉溶液及其交聯(lián)產品按照2∶1(W/V)比例混合制備的復合材料具有穩(wěn)定性和可注射性,體外能促進小鼠MC3T3-E1細胞增殖和分化,植入新西蘭大白兔體內后具有良好成骨能力,有望作為一種可注射材料用于骨缺損微創(chuàng)治療。
[Abstract]:Objective to prepare calcium sulfate injectable composite sodium hyaluronate and observe its effect on bone regeneration and provide an injectable material for bone defect repair. Methods calcium sulfate was mixed with sodium hyaluronate solution and cross-linked sodium hyaluronate solution to prepare three kinds of composite materials (CA HACA HAC and CAA) according to the ratio of 2: 1 W / V. The stability of the composites was observed by immersion in PBS solution and X-ray diffraction analysis. Three kinds of composite extractions were prepared with reference to ISO10993-5 to culture mouse preosteoblasts MC3T3-E1. The biocompatibility of the materials and the ability of different concentrations of extractions to promote cell proliferation were tested by cell counting kit-8 CCK-8 (cell proliferation-toxicity test kit). The cells were cultured in culture medium as control group. The optimal concentration extract of osteogenic differentiation medium was used to detect the concentration of osteoblast-associated proteins (ALP, collagen 鈪，

本文編號：1808183