【摘要】：本研究制備了一種新型的復(fù)合支架材料，通過(guò)多肽水凝膠對(duì)納米羥基磷灰石/膠原（nano-Hydroxyapatite/Collagen, nHAC）進(jìn)行表面修飾，未復(fù)合水凝膠的nHAC為對(duì)照組，探討其對(duì)大鼠脂肪干細(xì)胞（Rat adipose-derived stemcells，rADSCs）黏附增殖和骨向分化能力以及體內(nèi)骨再生能力的影響。本實(shí)驗(yàn)將rADSCs接種到兩種支架材料上，使用掃描電子顯微鏡（Field emission scanningelectron microscope, FESEM）和共聚焦顯微鏡（Confocal laser scanningmicroscope, CLSM）進(jìn)行表征，以CCK-8檢測(cè)細(xì)胞毒性， DNA熒光比色法檢測(cè)細(xì)胞增殖情況。以堿性磷酸酶（ALP）活性為早期成骨分化指標(biāo)，并通過(guò)實(shí)時(shí)定量PCR檢測(cè)成骨標(biāo)志基因的表達(dá)，比較rADSCs的骨向分化情況。將支架植入大鼠顱頂骨缺損模型，在植入4周和6周后處死大鼠，觀測(cè)體內(nèi)骨再生情況。結(jié)果顯示多肽水凝膠成納米纖維多孔網(wǎng)狀結(jié)構(gòu)，孔徑約300μm，在nHAC表面形成涂層，，rADSCs在多肽水凝膠復(fù)合支架材料（cnHAC）上黏附、鋪展的更好。CCK-8及DNA熒光比色結(jié)果顯示cnHAC能促進(jìn)rADSCs在支架上的增殖，兩組間差異有統(tǒng)計(jì)學(xué)意義。ALP和實(shí)時(shí)定量PCR結(jié)果表明cnHAC能顯著促進(jìn)rADSCs的骨向分化。大鼠顱頂骨缺損實(shí)驗(yàn)結(jié)果顯示cnHAC能在早期有效地促進(jìn)大鼠新骨形成。本研究結(jié)果表明cnHAC與nHAC相比，能顯著促進(jìn)細(xì)胞的黏附增殖、骨向分化及體內(nèi)骨再生能力。用多肽水凝膠對(duì)nHAC進(jìn)行表面修飾可作為一種簡(jiǎn)單有效的改性方式，提高支架材料的生物相容性和骨引導(dǎo)性。
[Abstract]:In this study, a new composite scaffold material was prepared. The nano-hydroxyapatite / collagen (nHAC) was modified by polypeptide hydrogel, and the nHAC without hydrogel was used as control group. Objective: to investigate the effects of ASCs on the adhesion, proliferation, bone differentiation and bone regeneration of rat adipose stem cells (Rat adipose-derived stem cells). RADSCs was inoculated into two scaffolds and characterized by scanning electron microscope (Field emission scanningelectron microscope, FESEM) and confocal microscope (Confocal laser scanningmicroscope, CLSM). Cytotoxicity was detected by CCK-8 and cell proliferation was detected by DNA fluorescence colorimetry. The activity of alkaline phosphatase (ALP) was used as the early osteogenic differentiation index, and the expression of osteogenic marker gene was detected by real-time quantitative PCR to compare the bone differentiation of rADSCs. The rat model of cranio-parietal bone defect was implanted with scaffold. The rats were killed at 4 and 6 weeks after implantation and bone regeneration in vivo was observed. The results showed that polypeptide hydrogels formed a porous network of nanofibers with pore size of about 300 渭 m, and a coating was formed on the surface of nHAC to adhere to the polypeptide hydrogel composite scaffold (cnHAC). The results of CCK-8 and DNA fluorescence colorimetry showed that cnHAC could promote the proliferation of rADSCs on the scaffold. The difference between the two groups was statistically significant. ALP and real-time quantitative PCR showed that cnHAC could significantly promote the bone differentiation of rADSCs. The experimental results of cranio-parietal bone defect in rats showed that cnHAC could effectively promote the formation of new bone in early stage. The results showed that cnHAC could significantly promote cell adhesion proliferation, bone differentiation and bone regeneration compared with nHAC. The surface modification of nHAC by polypeptide hydrogel can be used as a simple and effective way to improve the biocompatibility and bone guidance of scaffolds.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R783.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 嵇偉平;韓培;蔣W

本文編號(hào)：2162730