六种保存液对牙周膜成纤维细胞活性的影响

目的评价6种不同保存液对牙周膜成纤维细胞活性的影响。方法培养因正畸拔除的前磨牙分离出的牙周膜成纤维细胞,采用CCK-8法检测细胞在自来水、Hank平衡盐溶液、豆浆、生理盐水、牛奶和DMEM高糖培养基6种保存液中1h、2h、4h、8h和24h五个时间点时的细胞生物学活性。结果在五个时间点,DMEM组细胞残存率大于生理盐水组,两组差异有统计学意义(P<0.01),豆浆、HBSS和DMEM三组之间细胞残存率无统计学差异(P>0.05),在1h、4h和24h 3个时间点,牛奶组细胞残存率大于DMEM组和豆浆组,其两组差异均有统计学意义(P<0.05)。结论牛奶、豆浆、HBSS和DMEM高糖培养基4种溶液是有效的牙周膜成纤维细胞临时保存液,而生理盐水和自来水保存效果差,不适合作为保存液。     

不同离体时间和保存液对犬离体牙牙周膜细胞活力影响的实验研究

目的:探讨完全脱位牙不同离体时间和保存液对牙周膜细胞活力的影响。方法:麻醉拔除犬牙35个,首先将20个牙随机分为5组,分别为室温干燥放置0、30、60、120、240 min组,另15个牙室温干燥放置30 min后,随机分为3组,分别放入牛奶、HBSS液,100 g/L蜂胶液中浸泡2 h。各组处理完成后,采用全牙消化法获得牙周膜细胞,并通过4 g/L台盼蓝染色法检测各组牙周膜活细胞数和存活率。结果:室温干燥放置30、60、120、240 min后,牙周膜细胞存活率依次为33.6%、23.6%、18.5%、0.8%,而0 min的牙周膜细胞存活率可达95.5%。拔后30 min,经牛奶、HBSS液和100 g/L蜂胶液中保存2 h后,牙周膜细胞均有活力,其细胞存活率大小依次为100 g/L蜂胶液、HBSS液和牛奶,其中100 g/L蜂胶液与HBSS液相比无统计学差异(P>0.05),但与牛奶组相比,均有统计学差异(P<0.05)。结论:随着离体时间延长,完全脱位牙根面牙周膜细胞活力明显下降。100 g/L蜂胶液和HBSS液保存犬牙牙周膜细胞活力优于牛奶液。     

碱性成纤维细胞生长因子对人牙周膜细胞多配体蛋白聚糖-4基因表达的影响

目的 通过研究碱性成纤维细胞生长因子(bFGF)对人牙周膜细胞(PDLC)多配体蛋白聚糖-4 mRNA水平表达的影响,探讨bFGF在人PDLC迁移中的作用.方法 收集正畸拔除的12～18岁青少年健康前磨牙68颗,体外原代培养人PDLC,用外源性bFGF刺激细胞,培养24、48、72 h后,采用实时荧光定量聚合酶链反应检测细胞内多配体蛋白聚糖-4 mRNA的表达.结果 培养24h时,实验组多配体蛋白聚糖-4 mRNA表达量显著高于对照组(P＜0.01),其中1.0 ng·mL-1组升高最为显著;培养48h时,1.0 ng·mL-1组多配体蛋白聚糖-4 mRNA表达量高于对照组(P＜0.05);培养72h时,1.0 ng·mL-1组多配体蛋白聚糖-4mRNA表达量低于对照组(P＜0.05).结论 初期bFGF促进人PDLC内多配体蛋白聚糖-4mRNA合成,但随着时间的延长,bFGF抑制多配体蛋白聚糖-4mRNA合成,这种改变是促进PDLC迁移过程中一个重要的调节因素.     

低氧对人牙周膜细胞骨向分化能力的影响

目的 研究低氧对人牙周膜细胞（PDLC）的碱性磷酸酶（ALP）活性及成骨相关基因表达的影响.方法 采用组织块法体外分离培养人PDLC;取第3 ～ 5 代细胞分别在常氧和低氧条件下培养后检测其ALP 活性;并通过荧光定量聚合酶链反应（PCR）观察低氧处理后人PDLC 中成骨相关基因的表达变化.结果 低氧12、24、36、48 h 组ALP 活性均比常氧组高;其中低氧48 h 组与常氧组相比明显升高,差异有统计学意义（P ＜ 0.05）;低氧48 h 组ALP mRNA 表达比常氧组高,差异具有统计学意义（P ＜ 0.05）;4 个时间点低氧组骨钙蛋白（OCN）mRNA 表达均比常氧组高,且差异有统计学意义（12、36 h：P ＜ 0.05;24、48 h：P ＜ 0.01）.结论 低氧增强人PDLC 的ALP 活性,上调ALP mRNA 和OCN mRNA 的表达,促进人PDLC 向成骨细胞分化,同时促进成骨细胞的矿化,提示低氧环境对人PDLC 的骨向分化功能产生一定的影响.     

盐酸小檗碱对体外培养人牙周膜细胞生物活性的影响

目的:探讨盐酸小檗碱对原代培养的人牙周膜细胞(PDLC)生物活性的影响。方法:采用细胞培养技术、噻唑蓝(MTT)比色法、考马斯亮蓝法、酶动力学方法观察盐酸小檗碱对PDLC增殖活性、蛋白合成及碱性磷酸酶(ALP)活性的影响。结果:①与对照组比较,作用24h,盐酸小檗碱(0.005～0.030g/L)能明显增强人牙周膜细胞增殖能力(P<0.05);作用48h,盐酸小檗碱(0.005～0.020g/L)能明显增强人牙周膜细胞增殖能力(P<0.05);作用72h,盐酸小檗碱(0.010～0.020g/L)能明显增强人牙周膜细胞增殖能力(P<0.05)。②盐酸小檗碱(0.005～0.020g/L)细胞培养液中蛋白总含量均明显高于对照组(P<0.05)。③盐酸小檗碱(10～20g/L)细胞培养液中ALP活性均明显高于对照组(P<0.05)。结论:盐酸小檗碱在0.010～0.020g/L浓度范围内有促进PDLC的增殖及生物合成作用,能增强牙周膜细胞ALP活性。     

骨形成蛋白-2和碱性成纤维细胞生长因子对牙周膜细胞碱性磷酸酶活性的联合效应

目的 :了解重组人骨形成蛋白 - 2 (rhBMP - 2 )和碱性成纤维细胞生长因子 (bFGF)单独和联合作用对人牙周膜细胞 (PDLC)碱性磷酸酶 (ALP)活性的影响。方法 :体外培养人PDLC ,分别用不同浓度的rhBMP- 2和bFGF单独或联合作用 ,用酶动力学方法检测PDLC的ALP活性。结果 :5 0～ 2 0 0 μg/L浓度的rhBMP - 2可显著增强人PDLC的ALP活性 (P <0 .0 1) ,而 10 μg/L浓度的bFGF可显著抑制人PDLC的ALP活性(P <0 .0 1) ,rhBMP - 2和bFGF联合作用仍可较明显地增强人PDLC的ALP活性 (P <0 .0 5 )。结论 :rhBMP - 2和bFGF联合应用可增强人PDLC的ALP活性     

银杏叶提取物对脂多糖作用下人牙周膜细胞增殖及分泌细胞因子的影响

目的 观察银杏叶提取物(Ginkgo biloba extract,GBE)对脂多糖(lipopolysaccharide,LPS)作用下人牙周膜细胞(periodental ligament cell,PDLC)增殖及分泌白细胞介素(interleukin,IL)6、IL-1β及肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)的影响,为GBE在牙周病防治中的应用提供依据.方法 选取2010年5至7月在遵义医学院附属口腔医院口腔颌面外科门诊因正畸拔除的10例11 ～15岁患者的20颗前磨牙,采用改良组织块培养法体外原代培养人PDLC,实验共分5组:①阴性对照组,含10 ml/L胎牛血清的达尔伯克改良伊格尔培养基(Dulbecco's modified Eagle's medium,DMEM)培养液;②LPS组,含10 ml/L胎牛血清的DMEM培养液+100 mg/L LPS;③LPS+0.1 g/L GBE组,含10 ml/L胎牛血清的DMEM培养液+100 mg/L LPS+0.1g/L GBE;④LPS+0.01 g/L GBE组,含10 ml/L胎牛血清的DMEM培养液+100 mg/L LPS +0.01 g/L GBE;⑤LPS+地塞米松组,含10 ml/L胎牛血清的DMEM培养液+100 mg/L LPS+2 mg/L地塞米松.甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)法测定GBE对LPS作用下人PDLC活性的影响,酶联免疫吸附测定法测定各组IL-6、IL-1β及TNF-α的含量,对所得结果分别进行单因素方差分析,采用LSD-t检验进行组间两两比较,检验水准为双侧α=0.05.结果 实验12、24、48及72 h,LPS+0.1 g/L GBE组的吸光度值(分别为0.30±0.03、0.33±0.02、0.34 ±0.02及0.35 ±0.02)均显著高于LPS组(分别为0.20±0.03、0.20±0.02、0.22±0.04及0.24±0.02)(P＜0.05),PDLC IL-6、IL-1 β及TNF-α的分泌量均显著低于LPS组(P＜0.05);实验12、24、48及72 h,LPS+0.01 g/L GBE组吸光度值(分别为0.27±0.05、0.31 ±0.03、0.33±0.03及0.32 ±0.01)均显著高于LPS组(P＜0.05),PDLC IL-6、IL-1β及TNF-α的分泌量均显著低于LPS组(P＜0.05).结论 GBE对LPS作用下的人PDLC有保护作用.     

富血小板血浆对人牙周组织细胞矿化作用的影响

目的观察富血小板血浆(PRP)对人牙龈成纤维细胞(GF)、牙周膜细胞(PDLC)和牙槽骨成骨细胞(AOB)的矿化诱导作用。方法抽取健康成人的新鲜全血,经过两次离心得到富血小板血浆。培养GF、PDLC和AOB,取第4代细胞用于实验,3种细胞均以4×107个/L接种于6孔培养板内,标准条件下培养96h后换不同的培养液,实验组换用含有矿化液(10nmol/L地塞米松、10mmol/Lβ-甘油磷酸钠和新鲜制备的0·2mmol/L抗坏血酸)和50ml/LPRP的DMEM(100ml/LFBS)培养液,对照组换用含有矿化液的DMEM培养液,空白组换用普通DMEM培养液。隔天换液,培养至第30天进行vonKossa染色,显微镜下观察并记录结果,用图像分析软件半定量分析染色阳性面积比及细胞衰减的面积比。结果对于PDLC和AOB,实验组及对照组的矿化量多于空白组(P<0·05),同时实验组的矿化量高于对照组(P<0·05);对于GF,3个组之间的矿化量差异无统计学意义(P>0·05)。AOB实验组在形成基质矿化时的衰减比对照组和空白组少(P<0·05)。结论PRP可促进PDLC和AOB在体外形成矿化小结,并能减少HAOB在矿化形成时的衰减,提示PRP可提高牙周骨组织的矿化和再生。     

中药黄芩苷对脂多糖介导人牙周膜细胞增殖的时间效应和细胞周期的影响

目的:观察中药黄芩苷对脂多糖(lipopolysaccharide,LPS)介导体外培养的人牙周膜细胞(periodontal ligament cell,PDLC)增殖的时间效应和细胞周期的影响.方法:采用细胞培养技术培养PDLC,应用噻唑盐比色测定法(MTT)和流式细胞仪技术测定黄芩苷对LPS介导PDLC增殖的时间效应和细胞周期.结果:加入0.01 μg/ml黄芩苷3 h后,明显促进人牙周膜细胞的增殖(P＜0.05);加入100 μg/ml LPS 3 h后,即明显抑制人牙周膜细胞的增殖(P＜0.05),抑制作用随时间的延长而增强,而同时加入0.01μg/ml黄芩苷和100μg/ml LPS 3 h后,其抑制作用则显著减弱(P＜0.05).加入0.01μg/ml黄芩苷后12 h,DNA合成前期细胞所占百分比(G1%)较对照组降低,而DNA合成期细胞所占百分比(S%)较对照组增高;加入100 μg/ml LPS后12 h,G1%较对照组增高,而s%则降低;加入0.01μg/ml黄芩苷和100μg/ml LPS后12 h,G1%较LPS组显著降低(P＜0.01),S%则有显著提高(P＜0.01).结论:中药黄芩许能显著促进PDLC增殖分化,对LPS抑制PDLC的活性关系到有保护作用,原理可能足LPS抑制静止态的G1期细胞进入S期,从而抑制细胞的增殖;而黄芩苷则相反促进细胞由G1期细胞进入S期,从而促进细胞的增殖.     

新生大鼠牙乳头细胞的体外培养和矿化特征

目的：观察体外培养的新生大鼠牙乳头细胞的生物学特征。方法：体外培养出生1d大鼠磨牙牙乳头细胞，进行组织学染色，透射扫描电镜观察。结果：体外培养的新生大鼠牙乳头细胞能形成细胞克隆，第3代细胞在含2mmol／L β-甘油磷酸钠、50mg／L抗坏血酸、10^-8moL/L地塞米松的培养液中培养7～8d后可呈现复层生长，培养14～16d后细胞结节中出现矿化，茜素红染色呈阳性反应；透射电镜观察细胞内出现大量含有致密小体的基质小泡样超微结构。结论：发育阶段较早的大鼠牙乳头细胞具有增殖和矿化能力。     

In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in six different media

The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objectives of this study were to evaluate the effectiveness of six different media: culture medium, alpha minimal essential medium (alpha-MEM), milk, Hank's balanced salt solution (HBSS), ViaSpan and conditioned medium (CM) to preserve cultured periodontal ligament fibroblasts (PDLF). Periodontal ligament fibroblasts were obtained from explants of human healthy extracted teeth. Plates with confluent PDLF were soaked in the various media for 2, 8 and 24 h at 4 degrees C. A control group was incubated with culture medium at 37 degrees C. After incubation, cell viability was determined by trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic capacity (by culturing one cell/well). Storage of PDLF up to 24 h decreased their vitality by only 2%-14%. Vitality of the PDLF after 2, 8 and 24 h was highest when stored in milk or HBSS (91%-97%) and lowest when stored in ViaSpan or CM (82%-93%). PDLF stored for 2-8 h in various media had a mitogenic capacity comparable to the control. However, increasing the storage period to 24 h decreased the mitogenicity of the cells by 3%-39%. The highest mitogenicity was found in PDLF stored in milk or HBSS and the lowest in CM or ViaSpan. The clonogenic capacity of the cells dropped by 38%-71% after 24 h and was the best indicator of the deteriorating effect of long storage. Milk and HBSS were the most effective in preserving the clonogenic capacity. Nevertheless, reduction in the viability, mitogenicity or clonogenic capacity was statistically significant in nearly all the tested media only after 24 h of incubation. In conclusion, HBSS and milk were the most effective media for preserving the viability, mitogenicity and clonogenic capacity after storage for up to 24 h at 4 degrees C.     

In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in four media at room temperature

The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objective of this study was to compare the effectiveness of four recommended storage media (Hank's balanced salt solution [HBSS], culture medium, alpha minimal essential medium [alpha-MEM], and ViaSpan) to preserve cultured periodontal ligament fibroblasts (PDLF) at room temperature (22 degrees C). PDLF were obtained from explants of extracted healthy human teeth. Plates with confluent PDLF were soaked in the various media for 2, 8 and 24 h at room temperature. A control group was incubated with culture medium at 37 degrees C. After incubation, viability of the cells was determined by trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic capacity (by culturing one cell/well). Viability of PDLF stored up to 24 h was comparable in all tested media, and the differences were limited to 1%-3%. PDLF stored for up to 24 h in various media had statistically comparable mitogenicity to the control group. After 8 h of storage, the differences were limited to 2%-9%, except for the alpha-MEM group which had 23%-29% lower mitogenic capacity compared to the control group. Increasing the storage time up to 24 h further decreased the mitogenicity of the cells by 22%-47%. The highest mitogenicity after 24 h of storage was found in PDLF stored in culture medium or HBSS, and the lowest in alpha-MEM. PDLF stored for 2-8 h in various media had a comparable clonogenic capacity to the control group. However, after 24 h, the cells' clonogenic ability dropped by 14%-66%. A similar trend of reduction was noted in the mitogenic and clonogenic capacity, although it was statistically significant only in the clonogenic capacity. Culture medium and ViaSpan, followed by HBSS, were the most effective in preserving the clonogenic capacity of PDLF after 24 h of storage. The lowest clonogenic capacity after 24 h of storage was in the alpha-MEM group (66%, P < 0.0025). In conclusion, culture medium, followed by HBSS and ViaSpan, was the most effective media for preserving the viability, mitogenicity and clonogenic capacity of PDLF stored for up to 24 h at room temperature. The lowest functional abilities were found in PDLF stored in alpha-MEM.     

In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in four media at room temperature

Abstract – The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objective of this study was to compare the effectiveness of four recommended storage media (Hank's balanced salt solution [HBSS], culture medium, α minimal essential medium [α‐MEM], and ViaSpan) to preserve cultured periodontal ligament fibroblasts (PDLF) at room temperature (22°C). PDLF were obtained from explants of extracted healthy human teeth. Plates with confluent PDLF were soaked in the various media for 2, 8 and 24 h at room temperature. A control group was incubated with culture medium at 37°C. After incubation, viability of the cells was determined by trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic capacity (by culturing one cell/well). Viability of PDLF stored up to 24 h was comparable in all tested media, and the differences were limited to 1%–3%. PDLF stored for up to 24 h in various media had statistically comparable mitogenicity to the control group. After 8 h of storage, the differences were limited to 2%–9%, except for the α‐MEM group which had 23%–29% lower mitogenic capacity compared to the control group. Increasing the storage time up to 24 h further decreased the mitogenicity of the cells by 22%–47%. The highest mitogenicity after 24 h of storage was found in PDLF stored in culture medium or HBSS, and the lowest in α‐MEM. PDLF stored for 2–8 h in various media had a comparable clonogenic capacity to the control group. However, after 24 h, the cells' clonogenic ability dropped by 14%–66%. A similar trend of reduction was noted in the mitogenic and clonogenic capacity, although it was statistically significant only in the clonogenic capacity. Culture medium and ViaSpan, followed by HBSS, were the most effective in preserving the clonogenic capacity of PDLF after 24 h of storage. The lowest clonogenic capacity after 24 h of storage was in the α‐MEM group (66%, P<0.0025). In conclusion, culture medium, followed by HBSS and ViaSpan, was the most effective media for preserving the viability, mitogenicity and clonogenic capacity of PDLF stored for up to 24 h at room temperature. The lowest functional abilities were found in PDLF stored in α‐MEM.     

In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in six different media

Abstract— The choice of storage medium for preserving traumatic-ally avulsed teeth is important for the success of future replantation. The objectives of this study were to evaluate the effectiveness of six different media: culture medium, α minimal essential medium (αMEM), milk, Hank's balanced salt solution (HBSS), ViaSpan and conditioned medium (CM) to preserve cultured periodontal ligament fibroblasts (PDLF). Periodontal ligament fibroblasts were obtained from explants of human healthy extracted teeth. Plates with confluent PDLF were soaked in the various media for 2, 8 and 24 h at 4°C. A control group was incubated with culture medium at 37°C. After incubation, cell viability was determined by trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic capacity (by culturing one cell/well). Storage of PDLF up to 24 h decreased their vitality by only 2%-14%. Vitality of the PDLF after 2, 8 and 24 h was highest when stored in milk or HBSS (91%-97%) and lowest when stored in ViaSpan or CM (82%-93%). PDLF stored for 2–8 h in various media had a mitogenic capacity comparable to the control. However, increasing the storage period to 24 h decreased the mitogenicity of the cells by 3%-39%. The highest mitogenicity was found in PDLF stored in milk or HBSS and the lowest in CM or ViaSpan. The clonogenic capacity of the cells dropped by 38%-71% after 24 h and was the best indicator of the deteriorating effect of long storage. Milk and HBSS were the most effective in preserving the clonogenic capacity. Nevertheless, reduction in the viability, mitogenicity or clonogenic capacity was statistically significant in nearly all the tested media only after 24 h of incubation. In conclusion, HBSS and milk were the most effective media for preserving the viability, mitogenicity and clonogenic capacity after storage for up to 24 h at 4°C.     

In vitro viability, mitogenicity and clonogenic capacities of periodontal ligament fibroblasts after storage in four media supplemented with growth factors

Abstract – The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objective of this study was to evaluate the effectiveness of growth factors (IGF‐1 and PDGF‐BB) when added to storage media in preserving the functional abilities of cultured periodontal ligament fibroblasts (PDLF). The evaluated storage media were: ViaSpan, Hanks’ balanced salt solution (HBSS), α minimal essential medium (α MEM), and α MEM supplemented with FCS and antibiotic (α MEM‐S). PDLF were obtained from explants of human healthy extracted teeth. Plates with confluent PDLF were soaked in the various media supplemented with IGF‐1 (10 ng/ml) and PDGF‐BB (4 ng/ml) for 2, 8 and 24 h at room temperature (24 °C). The control group was incubated with the examined storage media without growth factors at 24 °C. An additional control group was incubated with culture medium at 37 °C without growth factors. After incubation, the viability of the cells was determined by Trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic (by culturing one cell/well) capacities. Storage of PDLF with growth factors (GF) for 2, 8 and 24 h decreased their vitality by only 3% (not statistically significant). The mitogenicity of PDLF stored for 2, 8 and 24 h in various media with GF was statistically comparable to that of the control group. Generally, the highest mitogenic capacity of PDLF stored with or without GF was found after 8 h of storage. Increasing the storage period to 24 h decreased the mitogenic capacity of the cells stored with GF by only 10–40% compared to the control group. In contrast, the clonogenic capacity of PDLF stored with GF increased with increasing storage periods by 100–300%, and the highest clonogenic capacity was found in most storage media after 24 h of storage with GF. The highest clonogenic and mitogenic capacities were found in cells stored in HBSS followed by α MEM‐S. The mitogenic and clonogenic capacities of PDLF stored in various media supplemented with GF for 2–8 h were generally lower than without GF supplementation. The mitogenic and clonogenic effects of GF‐supplementation was observed only after 24 h of storage. After 24 h of storage with GF, the clonogenic capacity increased by 8–224% and the mitogenicity by 20–37%, except in cells stored in α MEM (‐1%). However, these differences were generally not statistically significant. In conclusion, the mitogenic and clonogenic effects of GF were observed only after 24 h of storage at room temperature. HBSS and α MEM‐S supplemented with GF were the most effective media for preserving the viability, mitogenicity and clonogenic capacity of PDLF stored for 24 h at room temperature. For short periods of storage (2 and 8 h), HBSS and α MEM‐S without GF were preferable.     

Effect of HBSS storage time on human periodontal ligament fibroblast viability

Abstract – Hank’s balanced salt solution (HBSS) is recommended for the storage of avulsed teeth. The objective of this study was to evaluate if the HBSS storage time influences its ability to maintain the viability of human periodontal ligament fibroblasts (PDLF) by the analysis of cell metabolic function using MTT assay. PDLF were kept at 20°C for 3, 6, 24, 48, 72, 96 and 120 h in recently prepared HBSS (HBSS), HBSS stored for 6 months (HBSS 6 M), HBSS stored for 12 months (HBSS 12 M), and in Save‐A‐Tooth system’s HBSS (Save). Minimum essential medium (MEM) at 37°C and tap water at 20°C served as positive and negative controls, respectively. Cell viability was determined by the tetrazolium salt‐based colorimetric (MTT) assay. Data were statistically analyzed by the Kruskal–Wallis and Scheffé tests (α = 5%). Starting with the 6 h time‐point, HBSS was significantly more effective than HBSS 6 M, HBSS 12 M and Save in maintaining cell viability. HBSS 6 M effectiveness was similar to that of HBSS 12 M for up to 48 h, becoming higher at 72 h. In conclusion, the storage time of HBSS had a negative influence on its ability to maintain PDLF viability.     

碱性成纤维细胞生长因子对人牙周膜细胞多配体蛋白聚糖-4基因表达的影响

目的 通过研究碱性成纤维细胞生长因子(bFGF)对人牙周膜细胞(PDLC)多配体蛋白聚糖-4 mRNA水平表达的影响,探讨bFGF在人PDLC迁移中的作用.方法 收集正畸拔除的12～18岁青少年健康前磨牙68颗,体外原代培养人PDLC,用外源性bFGF刺激细胞,培养24、48、72 h后,采用实时荧光定量聚合酶链反应检...     

The influence of storage conditions on the clonogenic capacity of periodontal ligament cells: implications for tooth replantation

Viable periodontal ligament (PL) cells are required for PL healing of avulsed teeth following replantation. If immediate replantation cannot be accomplished, the ability of PL progenitor cells to reproduce (clonogenic capacity) and recolonize the wound may be extended by prevention of desiccation and storage in physiological media. This investigation examined the effects of storage in saliva, milk, Hank's balanced salt solution (HBSS) and Eagle's medium (αMEM) on the clonogenic capacity of human PL progenitor cells at 30 and 60 min extra-alveolar time. Twenty erupted human premolar teeth extracted as atraumatically as possible for orthodontic purposes were used in the present study. Fifteen premolars were placed immediately in freshly collected autologous saliva at room temperature, (+ 23°C) for 15 min. These 15 premolars were next divided into three groups of five and stored in either saliva, milk or HBSS at + 4°C in plastic cups surrounded by ice. The remaining five teeth served as positive controls and were immediately placed in αMEM at + 4°C. PL tissue was scraped from one-half of the root surface with a scalpel at 30 and 60 min total extra-alveolar duration. Cells were released from the tissue sample with a 30 min enzymatic digestion procedure and the cells from the tissue samples analyzed for clonogenic capacity. There was a reduction in clonogenic capacity with time for all protocols. Periodontal ligament cells stored in αMEM showed the least reduction between 30 and 60 min and the greatest reduction was observed for PL cells stored in saliva. The difference in clonogenic capacity following transfer from saliva to milk or HBSS was not significant at 30 min. At 60 min, cells transferred from saliva to HBSS had a statistically higher percentage of clonogenic cells than those transferred to milk (5.9% vs. 3.5%; P < 0.05). We conclude that immediate storage of avulsed teeth in autologous saliva, followed by transfer to chilled milk, preserves the presence of sufficient progenitor cells in the PL to warrant replantation and the possibility of PL healing at 60 min extra-alveolar duration.