中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
35期
6221-6227
,共7页
陈跃平%高辉%陈亮%董盼锋%尹庆水
陳躍平%高輝%陳亮%董盼鋒%尹慶水
진약평%고휘%진량%동반봉%윤경수
骨关节植入物%骨关节损伤基础实验%乙醇%股骨头坏死%脂肪细胞%大白兔%脂质代谢紊乱%脂滴%肥大%骨内压%省级基金
骨關節植入物%骨關節損傷基礎實驗%乙醇%股骨頭壞死%脂肪細胞%大白兔%脂質代謝紊亂%脂滴%肥大%骨內壓%省級基金
골관절식입물%골관절손상기출실험%을순%고골두배사%지방세포%대백토%지질대사문란%지적%비대%골내압%성급기금
bone and joint implants%basic experiment of bone injury%alcohol%femoral head necorsis%fat cells%rabbit%dyslipidosis%lipid droplet%enlargement%intraosseous pressure%provincial grants-supported paper
背景：乙醇已成为股骨头缺血性坏死的致病因素，其所致的骨髓内脂质代谢异常可能是股骨头缺血性坏死起病的重要原因，但机制尚未完全明确。<br> 目的：从分子水平观察在乙醇作用下脂肪细胞结构功能的变化，以期分析酒精性股骨头坏死的发病机制。<br> 方法：采用原代脂肪细胞体外培养技术，取大白兔股骨头髓内脂肪组织，分离获取脂肪细胞，以油红O染色行细胞表型鉴定。取传代稳定的髓内脂肪细胞，将盖玻片切割成10 mm×10 mm，种植前置入24孔培养板孔内，分为乙醇组和对照组，每组24孔，每孔为1个样本。对照组不加乙醇，乙醇组加入0.15 mol/L乙醇，分别于4，6，8，10 d 更换培养液，换液时不再加入乙醇，均培养至10 d。培养终止后，取出盖玻片行油红O染色，光镜下观察脂肪细胞形态并计数。<br> 结果与结论：随着时间延长，乙醇组脂肪细胞数量明显多于对照组(P<0.001)，2组小脂滴均逐渐增多、增大，但乙醇组更明显。培养4，6，8，10 d的髓内脂肪细胞数量乙醇组分别为(200.90±24.60)，(1102.30±76.73)，(1160.30±28.37)，(1199.70±44.74)个/cm2；对照组分别为(99.80±10.82)，(0.40±94.71)，(1000.20±41.85)，(1059.80±26.79)个/cm2，脂肪细胞数量随乙醇作用的时间延长而增多。提示乙醇能够促进髓内脂肪细胞增殖肥大，这可能是长期酗酒后股骨头骨髓内脂肪组织增多，骨内压增加，血流灌注减少，导致缺血，从而发生股骨头坏死的主要原因。
揹景：乙醇已成為股骨頭缺血性壞死的緻病因素，其所緻的骨髓內脂質代謝異常可能是股骨頭缺血性壞死起病的重要原因，但機製尚未完全明確。<br> 目的：從分子水平觀察在乙醇作用下脂肪細胞結構功能的變化，以期分析酒精性股骨頭壞死的髮病機製。<br> 方法：採用原代脂肪細胞體外培養技術，取大白兔股骨頭髓內脂肪組織，分離穫取脂肪細胞，以油紅O染色行細胞錶型鑒定。取傳代穩定的髓內脂肪細胞，將蓋玻片切割成10 mm×10 mm，種植前置入24孔培養闆孔內，分為乙醇組和對照組，每組24孔，每孔為1箇樣本。對照組不加乙醇，乙醇組加入0.15 mol/L乙醇，分彆于4，6，8，10 d 更換培養液，換液時不再加入乙醇，均培養至10 d。培養終止後，取齣蓋玻片行油紅O染色，光鏡下觀察脂肪細胞形態併計數。<br> 結果與結論：隨著時間延長，乙醇組脂肪細胞數量明顯多于對照組(P<0.001)，2組小脂滴均逐漸增多、增大，但乙醇組更明顯。培養4，6，8，10 d的髓內脂肪細胞數量乙醇組分彆為(200.90±24.60)，(1102.30±76.73)，(1160.30±28.37)，(1199.70±44.74)箇/cm2；對照組分彆為(99.80±10.82)，(0.40±94.71)，(1000.20±41.85)，(1059.80±26.79)箇/cm2，脂肪細胞數量隨乙醇作用的時間延長而增多。提示乙醇能夠促進髓內脂肪細胞增殖肥大，這可能是長期酗酒後股骨頭骨髓內脂肪組織增多，骨內壓增加，血流灌註減少，導緻缺血，從而髮生股骨頭壞死的主要原因。
배경：을순이성위고골두결혈성배사적치병인소，기소치적골수내지질대사이상가능시고골두결혈성배사기병적중요원인，단궤제상미완전명학。<br> 목적：종분자수평관찰재을순작용하지방세포결구공능적변화，이기분석주정성고골두배사적발병궤제。<br> 방법：채용원대지방세포체외배양기술，취대백토고골두수내지방조직，분리획취지방세포，이유홍O염색행세포표형감정。취전대은정적수내지방세포，장개파편절할성10 mm×10 mm，충식전치입24공배양판공내，분위을순조화대조조，매조24공，매공위1개양본。대조조불가을순，을순조가입0.15 mol/L을순，분별우4，6，8，10 d 경환배양액，환액시불재가입을순，균배양지10 d。배양종지후，취출개파편행유홍O염색，광경하관찰지방세포형태병계수。<br> 결과여결론：수착시간연장，을순조지방세포수량명현다우대조조(P<0.001)，2조소지적균축점증다、증대，단을순조경명현。배양4，6，8，10 d적수내지방세포수량을순조분별위(200.90±24.60)，(1102.30±76.73)，(1160.30±28.37)，(1199.70±44.74)개/cm2；대조조분별위(99.80±10.82)，(0.40±94.71)，(1000.20±41.85)，(1059.80±26.79)개/cm2，지방세포수량수을순작용적시간연장이증다。제시을순능구촉진수내지방세포증식비대，저가능시장기후주후고골두골수내지방조직증다，골내압증가，혈류관주감소，도치결혈，종이발생고골두배사적주요원인。
BACKGROUND:Alcohol has become pathogenic factors of avascular necrosis, and the alcohol induced <br> abnormal lipid metabolism in bone marrow may be the important reason for the onset of avascular necrosis, but the mechanism is not clear yet. <br> OBJECTIVE:To observe the changes of structure and function of fat cel s under the action of alcohol, in order to analyze the pathogenesis of alcoholic femoral head necrosis. <br> METHODS:Primary adipocytes in vitro culture technique was used to obtain rabbit femoral head intramedul ary adipose tissue, and then the fat cel s were separated, and the phenotype was identified with oil red O staining. The passaged stable intramedul ary fat cel s were col ected. Coverslip was cut into 1 cm × 1 cm in size, and placed in the 24-wel culture plate before planting. The cel s were randomly divided into alcohol group and control group, 24 holes (each hole for a sample) in each group. The control group was without alcohol, while the alcohol group was added with 0.15 mol/L alcohol. At 4, 6, 8 and 10 days, the culture medium was replaced. Medium was changed and no longer adding alcohol, and then cultured for 10 days. When the culture terminated, the coverslip was removed for oil red O staining. Final y, the morphology and the number of the fat cel s were observed under light <br> microscope. <br> RESUTLS AND CONCLUSION:With time prolonging, the number of fat cel s in the alcohol group was significantly more than that in the control group (P<0.001). The lipid droplets in the two groups were gradual y increased and enlarged, but more significant in the alcohol group. The number of intramedul ary fat cel s in the alcohol group after cultured for 4, 6, 8 and 10 days was respectively (200.90±24.60), (1 102.30±76.73), (1 160.30±28.37) and (1 199.70±44.74)/cm2;the <br> number of intramedul ary fat cel s in the control group was respectively (99.80±10.82), (0.40±94.71), (1 000.20± 41.85) and (1 059.80±26.79)/cm2, the number of fat cel s increased with the time of alcohol influence. Alcohol can promote the intramedul ary fat cel s to increase and enlarge, and this may be the main reason for femoral head necrosis, as long-term alcoholism can lead to bone marrow fat tissue increasing, intraosseous pressure increasing and perfusion reducing, thus resulting ischemia.
