骨髓基质细胞膜片对骨质疏松大鼠牙周组织再生的影响

Effect of bone marrow stromal cell sheet on healing of periodontal defects in osteoporosis rats

目的探讨骨髓基质细胞(bone marrow stromal cells,BMSCs)膜片在骨质疏松大鼠牙周组织再生中的作用。方法将30只Sprague-Dawley大鼠随机分为卵巢切除术组(n=24)和假手术组(n=6),再将24只卵巢切除组大鼠随机分为对照组和细胞膜片组各12只。分离培养大鼠BMSCs并构建细胞膜片,将构建好的细胞膜片植入骨质疏松大鼠下颌牙周组织缺损内,分别于术后10和28 d取材制作组织切片行苏木素-伊红(hematoxylin-eosin,HE)和Masson染色,通过组织学观察和测量,评价BMSCs细胞膜片对骨质疏松状态下牙周组织再生的作用。结果成功培养构建大鼠BMSCs细胞膜片,组织染色及细胞死活荧光染色结果显示所构建的细胞膜片具有较好的活性,富含胞外基质。将膜片植入骨质疏松大鼠牙周组织缺损10 d和28 d后,对照组牙周缺损区以纤维结缔组织为主,存在少量的新生牙槽骨,新生的牙周韧带和血管样组织;细胞膜片组牙周缺损部位再生明显,新生牙槽骨样组织填充其中,牙根上可见牙骨质新生,且通过新生牙周韧带附着于新生牙槽骨。植入后10 d和28 d,对照组新生牙周组织面积分别为(294.47±94.41)μm^2和(1 873.96±391.62)μm^2,细胞膜片组分别为(13 496.27±9 831.42)μm^2和(85 967.72±7 632.15)μm^2,细胞膜片组较对照组均显著增加(P<0.001)。结论 BMSCs细胞膜片有望应用于临床实现骨质疏松状态下的牙周组织再生。

Objective To investigate the effects of rat bone marrow stromal cell(BMSCs) sheets on the repair of periodontal defect in osteoporotic rat.Methods Thirty female Sprague Dawley rats were divided into ovariectomy group(OVX group,n=24) and sham operation group(sham group,n=6).Then 24 rats in OVX group were randomly divided into control group(n=12) and BMSCs sheet group(n=12).Rats’ bone marrow stromal cells were isolated and expanded,then we constructed BMSCs sheet.Postmenopausal osteoporosis model was established by ovariectomy.The model of periodontal defect was prepared on the buccal side of the first and second molar teeth.The constructed cell sheets were implanted into periodontal defects.After 10 days and 28 days of surgery,the animals were sacrificed.Specimens were decalcified,embedded in paraffin,making pathological section,HE and Masson stain.The effects on the regeneration of periodontal tissue under postmenopausal osteoporosis(PMO) was evaluated by histological observation and measurement.Results BMSCs sheets were constructed,histologic staining and Live & Dead cell staining showed that the constructed cell sheets had good activity and were rich in extracellular matrix.A periodontal defect model of PMO rat was established and the BMSCs sheets were implanted for 10 days and 28 days.In the control group,the periodontal defect area were dominated by fibrous connective tissue,with a small amount of new alveolar bone,new periodontal ligament and vascular tissue.In the BMSCs sheet group,the periodontal defects were regenerated obviously.The new alveolar bone was filled with the new alveolar bone,and the new cementum was found on the root of the tooth,and attached to the new alveolar bone through the new periodontal ligament.The area of newborn periodontal tissue in the control group were(294.47±94.41) μm^2 and(1 873.96±391.62) μm^2,respectively.The area of newborn periodontal tissue in the BMSCs sheet group were(13 496.27±9 831.42) μm^2 and(85 967.72±7 632.15) μm^2.Compared with the control group,the new formed periodontal tissue in the BMSCs sheet group was significantly increased(P < 0.001).Conclusion BMSCs sheet can regenerate periodontal tissue of rats with osteoporosis.