富血小板血浆对牙周膜成纤维细胞在病变牙根表面胶原形成的影响

Effect of Platelet-rich Plasma Collagen Formation of Periodontal Ligament Fibroblast on the Diseased Root Surfaces

目的：研究不同浓度的富血小板血浆（platelet-rich plasma，PRP）对牙周膜成纤维细胞（peridontal fibro—blasts，PDLFs）在脱矿的病变牙根表面形成胶原的影响。以进一步探讨富血小板血浆在促牙周组织再生中的作用。方法：采用组织块培养法培养原代人牙周膜成纤维细胞，取5～8代细胞用于实验。富血小板血浆的制备采用二步密度梯度离心法，获取PRP待用，取因重度牙周病拔除的病牙，制取牙根片，收集的根片经高压处理后留存待用。观察富血小板血浆对牙周膜成纤维细胞在脱矿的病变牙根表面胶原的形成，用天狼星红组织特染的方法进一步观察病变根片表面胶原形成的情况。结果：MTT结果和天狼星红特异性染色结果：实验组与对照组均有阳性染色，用Histogram功能来分析统计胶原所占面积的百分数，取其平均值，实验组与对照组差异有显著性（P〈0．05）。结论：浓度为20％PRP为促进PDLFs在脱矿处理的病变牙根表面附着的最佳浓度，并能促进其在处理的脱矿的病变牙根表面胶原的形成。推测PRP和根面脱矿处理在牙周再生中起重要作用。

Objective： To study the effect of PRP with various concentrations collagen formation of PDLFs on the diseased root surface of human teeth demineralized with EDTA and to detect the role of PRP in the process of periodontal regeneration. Methods： PDLFs was grown from tissue explants and cells between the 5th and 8th passage in culture were used. Human whole blood obtained from healthy subjects was collected in tubes containing an anticoagulant （acid--citrate--dextrose） was firstly centrifuged （1300 rpm for10 minutes） and the supernatant PPP layer was removed. They were analysed utilizing MTT assay. We studied the effect of PRP collagen formation of PDLFs on the diseased root surface of human teeth demineralize with EDTA. The cultures were incubated for two weeks. Cell morphology, growth pattern and collagen formation were observed by scanning electron microscopy. Furthermore, the collagen formation was observed by Sirius red tissue staining method. Results： Sirius red tissue staining method showed positive both experimental（A） and control（B） groups. The pictures were anlysed experimental（A） and con- trol（B） groups with software of phothshop： A,B groups were 85.32 4.22,70.51 1.64 respectively there were sig- nificant difference between both groups （P〈 0. 05）. Conclusion： PRP could promote the collagen formation of PDLFs on the diseased root surface of human teeth demineralize with EDTA and treated with PRP. But the effect could be reduced when the dose exceeded 20%.