本文選題：富血小板血漿 + 白細(xì)胞　； 參考：《廣西醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的：分別探討不同類型的富血小板血漿(platelet-rich plasma, PRP）支架和復(fù)合牙髓細(xì)胞的PRP支架體內(nèi)誘導(dǎo)牙髓再生的可行性；評(píng)價(jià)不同類型的PRP支架募集內(nèi)源性細(xì)胞歸巢的能力，為PRP支架應(yīng)用于牙髓再生的臨床治療提供實(shí)驗(yàn)基礎(chǔ)和理論依據(jù)。 方法： 1.實(shí)驗(yàn)一：改良的Curasn’s方法制備富白細(xì)胞富血小板血漿（leucocyte-rich plasma,L-PRP）支架，將小型豬乳牙牙根段進(jìn)行化學(xué)預(yù)備，隨機(jī)分為4組，每組6個(gè)樣本，分別為①全血組；②100%L-PRP組；③50%L-PRP組：將100%L-PRP用DMEM培養(yǎng)基稀釋成50%L-PRP（v/v）④空白組，空的牙根段。分組植入裸鼠背部皮下，5周后取出樣本進(jìn)行組織學(xué)觀察。 2.實(shí)驗(yàn)二：二次離心法制備貧白細(xì)胞富血小板血漿（leucocyte-poorplatelet-rich plasma，P-PRP）支架，組織塊法分離培養(yǎng)人牙髓細(xì)胞（humandental plus cells, hDPCs）并進(jìn)行鑒定。將人牙根管段進(jìn)行機(jī)械化學(xué)預(yù)備，隨機(jī)分為3組，，每組8個(gè)樣本,分別為①陰性對(duì)照組：P-PRP支架；②實(shí)驗(yàn)組：P-PRP支架復(fù)合hDPCs；③空白組，空的牙根段。分組植入裸鼠背部皮下，8周后取出樣本進(jìn)行組織學(xué)觀察分析。 結(jié)果： 1.小型豬全血制備的100%L-PRP中血小板和白細(xì)胞的含量分別是全血的6倍和4倍。移植了100%L-PRP的兩只裸鼠出現(xiàn)皮膚損害和炎癥反應(yīng)。 2.50%L-PRP組根管內(nèi)有新的結(jié)締組織形成，沒(méi)有血管生成；100%L-PRP組根管內(nèi)充滿了大量的炎性細(xì)胞，可見(jiàn)少量散在的梭形細(xì)胞。 3.培養(yǎng)的第4代hDPCs礦化誘導(dǎo)后堿性磷酸酶染色呈陽(yáng)性，茜素紅，Von Kossa染色，可觀察到少量鈣結(jié)節(jié)生成。人全血制備的P-PRP中血小板的含量是全血的1.55倍，白細(xì)胞含量遠(yuǎn)遠(yuǎn)低于全血中的含量。單獨(dú)移植P-PRP支架的根管內(nèi)觀察到不規(guī)則的組織形成。 4．P-PRP支架復(fù)合hDPCs的根管內(nèi)觀察到有血管化的牙髓樣組織形成，但是沒(méi)有牙本質(zhì)樣組織生成。根管內(nèi)大部分細(xì)胞抗人線粒體免疫組織化學(xué)染色陽(yáng)性，表明新生組織中細(xì)胞主要是人來(lái)源的細(xì)胞，但在靠近根管開(kāi)口附近有較集中的抗人線粒體免疫組織化學(xué)染色陰性的梭形細(xì)胞，表明有內(nèi)源性宿主細(xì)胞參與組織再生。 結(jié)論： 1.復(fù)合hDPCs的P-PRP支架在體內(nèi)再生牙髓樣組織是可行的。 2.單獨(dú)移植L-PRP支架或者P-PRP支架可以募集內(nèi)源性細(xì)胞參與組織再生，但是募集的細(xì)胞類型和數(shù)量有限。
[Abstract]:Objective: to investigate the feasibility of inducing pulp regeneration in different types of platelet-rich plasma (PRP-rich) stents and composite dental pulp cells, and to evaluate the ability of different types of platelet-rich stents to recruit endogenous cells to homing. To provide experimental and theoretical basis for the application of PRP stent in the clinical treatment of dental pulp regeneration. Methods: 1. Experiment 1: modified Curasnines method was used to prepare leucocyte-rich plasma-rich plasma (L-PRP) stents. The root segments of primary teeth of miniature pigs were chemically prepared and randomly divided into 4 groups, 6 samples in each group: 1 whole blood group (2100g / L PRP group); 350L-PRP group: 100% L-PRP was diluted into 50 L-PRP (v / v) 4 blank group with DMEM medium, empty root segment. After 5 weeks of subcutaneous implantation in nude mice, the samples were taken out for histological observation. 2. Experiment 2: leucocyte-poorplatelet-rich plasma-rich plasma (leucocyte-poorplatelet-rich PPRP) scaffolds were prepared by secondary centrifugation, and (humandental plus cells, hDPCs were isolated and identified by tissue block method. The human root canal segment was prepared by mechanochemistry and was randomly divided into 3 groups with 8 samples in each group: 1 negative control group: 1: P-PRP stent and 2 experimental group: 1 P-PRP stent combined with hDPCs3 blank group, empty root segment. After 8 weeks of subcutaneous implantation of nude mice, the samples were taken out for histological observation and analysis. Results: 1. The contents of platelet and leukocyte in 100% of L-PRP prepared from whole blood of miniature pig were 6 times and 4 times of that of whole blood, respectively. Two nude mice transplanted with 100% L-PRP had skin damage and inflammatory reaction. 2.50 L-PRP group had new connective tissue formation in the root canal, and the root canal in the 100L-PRP group was filled with a large number of inflammatory cells without angiogenesis. A few scattered fusiform cells can be seen. 3. After mineralization induction of the fourth generation of hDPCs, alkaline phosphatase staining was positive, alizarin red Von Kossa staining, a small amount of calcium nodule formation could be observed. The platelet content of P-PRP prepared by human whole blood was 1.55 times of that of the whole blood, and the white blood cell content was much lower than that of the whole blood. Irregular tissue formation was observed in the root canal of P-PRP scaffold alone. 4. The formation of vascularized dental pulp tissue was observed in the root canal of P-PRP scaffold combined with hDPCs, but no dentin tissue formation was observed. Most of the cells in the root canal were positive for anti-human mitochondria immunohistochemical staining, indicating that the cells in the newborn tissue were mainly human derived cells. However, near the root canal opening, there were concentrated spindle cells with negative anti-human mitochondria immunohistochemical staining, indicating that endogenous host cells were involved in tissue regeneration. Conclusion: 1. P-PRP scaffold combined with hDPCs is feasible to regenerate pulp tissue in vivo. 2. 2. L-PRP stents or P-PRP stents alone could recruit endogenous cells to participate in tissue regeneration, but the type and number of cells recruited were limited.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R781.3

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本文編號(hào)：2091859