CAJ | 학술논문

目的观察不同浓度壳聚糖季铵盐（HTCC）对人牙周膜成纤维细胞（HPDLC）增殖和细胞周期的影响，并与壳聚糖（CS）相比较，为HTCC的合理应用提供依据。方法原代培养人牙周膜成纤维细胞，免疫组织化学鉴定细胞来源。四甲基偶氮唑盐（MTT）法测定质量浓度分别为2?000、1?000、100、50、10、3、1.5、1、0.5和0.2?mg·L-1的HTCC和CS对HPDLC?培养1、3、5?d细胞增殖活性的影响；流式细胞术测定质量浓度为3?g·L-1和3?mg·L-1的HTCC和CS及1?g·L-1的HTCC对HPDLC细胞周期的影响。应用SSPS?13.0软件包对数据进行统计学分析。结果经免疫组织化学显示：培养的细胞波形丝蛋白染色阳性，角蛋白染色阴性，证明细胞来源于外胚层。MTT实验表明，与对照组相比，HTCC在质量浓度为2?000、1?000、100和50?mg·L-1时，光密度（OD）值降低；质量浓度为10、3、1.5、1、0.5?和0.2?mg·L-1时，OD值增高。而CS在所有实验质量浓度范围内均能促进细胞的增殖，在相对高质量浓度培养5?d?后对HPDLC的促增殖作用与对照组的差异具有统计学意义（P<0.05）。流式细胞术实验结果表明，3和1?g·L-1的HTCC可诱导细胞凋亡。质量浓度为3?g·L-1和3?mg·L-1的CS使S%降低，G2%上升，其差异具有统计学意义（P<0.01）。结论质量浓度为50~2?000?mg·L-1的HTCC抑制HPDLC的增殖，3和1?g·L-1的HTCC诱导细胞凋亡，显示出明显的细胞毒性；质量浓度为0.2~10?mg·L-1的HTCC明显促进了HPDLC增殖。HTCC和CS可通过改变细胞周期影响细胞增殖。
목적관찰불동농도각취당계안염（HTCC）대인아주막성섬유세포（HPDLC）증식화세포주기적영향，병여각취당（CS）상비교，위HTCC적합리응용제공의거。방법원대배양인아주막성섬유세포，면역조직화학감정세포래원。사갑기우담서염（MTT）법측정질량농도분별위2?000、1?000、100、50、10、3、1.5、1、0.5화0.2?mg·L-1적HTCC화CS대HPDLC?배양1、3、5?d세포증식활성적영향；류식세포술측정질량농도위3?g·L-1화3?mg·L-1적HTCC화CS급1?g·L-1적HTCC대HPDLC세포주기적영향。응용SSPS?13.0연건포대수거진행통계학분석。결과경면역조직화학현시：배양적세포파형사단백염색양성，각단백염색음성，증명세포래원우외배층。MTT실험표명，여대조조상비，HTCC재질량농도위2?000、1?000、100화50?mg·L-1시，광밀도（OD）치강저；질량농도위10、3、1.5、1、0.5?화0.2?mg·L-1시，OD치증고。이CS재소유실험질량농도범위내균능촉진세포적증식，재상대고질량농도배양5?d?후대HPDLC적촉증식작용여대조조적차이구유통계학의의（P<0.05）。류식세포술실험결과표명，3화1?g·L-1적HTCC가유도세포조망。질량농도위3?g·L-1화3?mg·L-1적CS사S%강저，G2%상승，기차이구유통계학의의（P<0.01）。결론질량농도위50~2?000?mg·L-1적HTCC억제HPDLC적증식，3화1?g·L-1적HTCC유도세포조망，현시출명현적세포독성；질량농도위0.2~10?mg·L-1적HTCC명현촉진료HPDLC증식。HTCC화CS가통과개변세포주기영향세포증식。
Objective ??To?evaluate?the?effects?of?quaternized?chitosan[N-(2-hydroxy-3-trimethylammonium)?propylchitosan?chloride]?(HTCC)?on?the?proliferation?and?cell?cycle?activities?of?human?periodontal?ligament?cells(HPDLCs)?with?different?concentrations?compared?with?chitosan(CS).?Investigating?the?mechanisms?underlying?the?use?of?HTCC?is?necessary.?Methods???Primary?cells?were?isolated?from?human?periodontal?ligament.?HPDLCs?were?cultured?and?characterized?by?immunocytochemistry.?The?effect?of?varying?concentrations(2?000,?1?000,?100,?50,?10,?3,?1.5,?1,?0.5,?and?0.2?mg·L-1)?of?HTCC?and?CS?on?HPDLCs?were?evaluated?by?methyl?thiazolyl?tetrazolium(MTT)?assay.?Flow?cytometry?(FCM)?was?used?to?determine?the?cell?cycle?states?of?HPDLCs?with?HTCC(3?g·L-1,?1?g·L-1,?and?3?mg·L-1)?and?CS(3?g·L-1?and?3?mg·L-1).?The?SPSS?13.0?software?package?was?used?for?statistical?analysis.?Results???Immunocytochemistry?results?show?that?the?cells?stained?positively?to?antibodies?against?vimentin?and?stained?negatively?to?antibodies?against?cytokeratin,?indicating?that?the?cells?were?external?embryo?mesenchymal?cells.?The?MTT?assay?showed?decreased?optical?density(OD)?values?of?HTCC?at?concentrations?of?2?000,?1?000,?100,?and?50?mg·L-1?and?increased?OD?values?at?concentrations?of?10,?3,?1.5,?1,?0.5,?and?0.2?mg·L-1.?CS?accelerated?the?proliferation?of?HPDLC?at?all?tested?concentrations.?Compared?with?the?control,?a?significant?difference?was?observed?at?5?d(P<0.01).?FCM?results?revealed?that?HTCC?at?3?g·L-1?and?1?g·L-1?induced?apoptosis,?whereas?CS?at?3?g·L-1?and?3?mg·L-1?induced?a?decrease?in?S%?and?increase?in?G2%.?A?significant?difference(P<0.01)?was?observed?compared?with?the?control.?Conclusion???HTCC?and?CS?affected?the?proliferation?of?HPDLCs?by?changing?the?cell?cycle.?HTCC?at?concentrations?of?50?mg·L-1?to?2?000?mg·L-1?inhibited?the?proliferation?of?HPDLCs?and?exhibited?strong?cytotoxicity.?HTCC?at?concentrations?of?0.2?mg·L-1?to?10?mg·L-1?stimulated?the?proliferation?of?HPDLCs.?Different?cytobiological?effects?of?HTCC?and?CS?on?cell?cycle?were?observed.