【摘要】：牙周疾病是引起牙周組織缺損最終導(dǎo)致牙齒缺失的常見病因，如何有效防治牙周病所引起的組織缺損及功能障礙，成為口腔科醫(yī)生亟待解決的問題。組織工程學(xué)理論和技術(shù)的飛速發(fā)展，為牙周組織再生研究提供了新的思路[1]。用組織工程學(xué)方法促進(jìn)牙周再生已成為這一領(lǐng)域的研究熱點(diǎn)之一。如何選擇具有增殖分化潛能的理想種子細(xì)胞及符合組織工程要求的支架材料是組織工程研究的重要內(nèi)容。 目的：以犬牙周膜細(xì)胞（PDLCS）為種子細(xì)胞，殼聚糖-絲素蛋白-磷酸三鈣復(fù)合物作為支架材料，，應(yīng)用于牙周組織工程，探討殼聚糖-絲素蛋白-磷酸三鈣復(fù)合物的優(yōu)越性。 方法：選擇1-2歲的健康雄性雜種犬6只，以每只犬下頜雙側(cè)第2、3、4前磨牙為實(shí)驗(yàn)牙，建立雄性雜種犬的牙周缺損模型；模型建立后，應(yīng)用隨機(jī)化法將其分為空白對(duì)照組、殼聚糖組、殼聚糖-絲素蛋白-磷酸三鈣復(fù)合物組，每組2只犬。體外培養(yǎng)犬牙周膜細(xì)胞，取傳至第4代細(xì)胞接種到支架材料上，并于2 h內(nèi)分別植入犬下頜雙側(cè)第2、3、4前磨牙的牙周缺損處；術(shù)后8周處死動(dòng)物，取實(shí)驗(yàn)處牙槽骨；經(jīng)常規(guī)固定、脫鈣、包埋后，HE染色，光鏡下觀察牙周組織再生情況。MTT法檢測(cè)支架材料對(duì)狗PDLCs增殖的影響。所得各組數(shù)據(jù)使用SPSS17.0軟件進(jìn)行統(tǒng)計(jì)分析。 結(jié)果：HE染色結(jié)果顯示，殼聚糖-絲素蛋白-磷酸三鈣復(fù)合支架材料用于牙周組織工程，其牙周間隙減小、成骨細(xì)胞數(shù)量增多較殼聚糖組和空白對(duì)照組更為明顯。MTT法顯示，殼聚糖/絲素蛋白/磷酸三鈣復(fù)合支架對(duì)狗PDLCs的生長(zhǎng)、增殖與單純殼聚糖作為支架材料對(duì)照相比，差異無統(tǒng)計(jì)學(xué)意義（P 0.05） 結(jié)論：以犬牙周膜細(xì)胞（PDLCs）為種子細(xì)胞，殼聚糖-絲素蛋白-磷酸三鈣復(fù)合支架材料較單純殼聚糖支架材料具有更強(qiáng)的組織相容性和再生能力，具有可行性。
[Abstract]:Periodontal disease is the common cause of periodontal tissue defect and tooth loss. How to effectively prevent and cure the tissue defect and dysfunction caused by periodontal disease has become an urgent problem for stomatologists to solve. The rapid development of tissue engineering theory and technology provides a new idea for the study of periodontal tissue regeneration. Tissue engineering to promote periodontal regeneration has become one of the research hotspots in this field. How to select ideal seed cells with proliferation and differentiation potential and scaffold materials which meet the requirements of tissue engineering is an important part of tissue engineering research. Aim: to investigate the advantages of chitosan-silk fibroin-tricalcium phosphate complex in periodontal tissue engineering by using canine periodontal ligament cell (PDLCS) as seed cell and chitosan-fibroin-tricalcium phosphate complex as scaffold material. Methods: six healthy male hybrid dogs aged 1-2 years were used as experimental teeth to establish periodontal defect model of male hybrid dogs with the 2nd, 3rd and 4th premolars of each dog's mandible as experimental teeth. After the model was established, the dogs were randomly divided into control group, chitosan-silk fibroin-tricalcium phosphate complex group (2 dogs in each group). Canine periodontal ligament cells were cultured in vitro and seeded into the scaffolds in passage 4, then implanted into the periodontal defects of canine mandibular premolars at the 2nd, 3rd and 4th premolars within 2 hours, and the animals were killed 8 weeks after operation and the alveolar bones were taken from the experimental sites. After routine fixation, decalcification, embedding and HE staining, the regeneration of periodontal tissue was observed under light microscope. The effect of scaffolds on the proliferation of dog PDLCs was detected by MTT method. The data were analyzed by SPSS17.0 software. Results: the results of HE staining showed that when chitosan-silk fibroin-tricalcium phosphate composite scaffold was used for periodontal tissue engineering, the periodontal space decreased and the number of osteoblasts increased more obviously than that of chitosan group and blank control group. The growth and proliferation of chitosan / silk fibroin / tricalcium phosphate composite scaffold on dog PDLCs were compared with that of chitosan as scaffold material. There was no significant difference (P 0.05). Conclusion: (PDLCs) is the seed cell of canine periodontal ligament cells. Chitosan-silk fibroin-tricalcium phosphate composite scaffolds have stronger histocompatibility and regeneration ability than pure chitosan scaffolds, so it is feasible to use chitosan-silk protein-tricalcium phosphate composite scaffolds.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R318.08

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