Creation and clinical application of the three-dimensional cultured cell transplantation for hard tissue regeneration

三维培养细胞移植硬组织再生技术的创建及临床应用

• 批准号: 17591996
• 负责人: IKEDA Takeshi
• 金额: $ 2.24万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591996/
• 关键词: chitosan; hard tissue regeneration; bone marrow-derived progenitor cells; cDNA microarray; ALP; osteocalcin; BMP-; real-time PCR; 培養骨芽細胞

The present study was designed to investigate histochemically the biodegradation processes of chitin and chitosan implanted in rat alveolar bone. Lysozyme was immunohistochemically detected using postembedding immunogold labeling. The degradation process was ultrastructurally observed using the lectin-colloidal gold technique with electron microscopy. Three groups of chitin were specially prepared according to their degree of deacetylation : 100% deacetylated chitin (DDAC 100) ; 50% (DDAC 50) ; and 0% (DDAC 0). The present immunohistochemical study indicated that lysozyme expression was not detected in the DDAC 100 group. Furthermore, electron microscopy clearly demonstrated that the contour of implanted chitosan changed over time, and that chitosan-like fragments were present in the phagosomes in the DDAC 50 and 100 groups. These findings strongly suggest that phagocytes, such as multinuclear cells, are easily supplied in bone tissue and that the phagocytosis is more effective than en … More zymatic digestion for chitin and chitosan biodegradation in bone tissue. DDAC 100 should be a suitable biomaterial for bone surgery and bone regeneration therapy.The other study was undertaken to evaluate the applicability of chitosan monomer (D-glucosamine hydrochloride) as a pulp capping medicament. Both in vitro and in vivo experiments were carried out to study the cell metabolism and wound healing mechanisms following the application of chitomonosaccharide. After 3 days of osteoblast culture, alkaline phosphatase (ALP) activity significantly increased in the chitosan group. Reverse transcription polymerase chain reaction analysis revealed that chitosan induced an increase in the expression of ALP mRNA after 3 days and bone morphogenetic protein-2 mRNA after 7 days of osteoblast incubation. Inflammatory cytokine, interleukin (IL)-8, synthesis in fibroblasts was strongly suppressed in the medium supplemented with chitosan monomer. Histopathological effects were evaluated in rat experiments. After 1 day, inflammatory cell infiltrations were observed to be weak when compared with the application of chitosan polymer. After 3 days, a remarkable proliferation of fibroblasts was seen near the applied chitosan monomer. The inflammatory cell infiltration had almost completely disappeared. After 5 days, the fibroblastic proliferation progressed, and some odontoblastic cells appeared at the periphery of the proliferated fibroblasts. These findings indicate that the present study is the first report that chitosan monomer acts as a biocompatibly stable medicament even at the initial stage of wound healing in comparison with the application of chitosan polymer. Less

本研究旨在从组织化学上研究植入大鼠肺泡骨的几丁质和壳聚糖的生物降解过程。溶菌酶使用后置免疫金标记在免疫组织化学上检测到。使用带有电子显微镜的讲座 - 胶体金技术观察到了降解过程。根据其脱乙酰基化程度特别制备三组几丁质：100％脱乙酰化几丁质（DDAC 100）； 50％（DDAC 50）;和0％（DDAC 0）。目前的免疫组织化学研究表明，在DDAC 100组中未检测到溶菌酶表达。此外，电子显微镜清楚地表明，植入壳聚糖的轮廓会随着时间的流逝而变化，并且在DDAC 50和100组的吞噬体中存在类似壳聚糖的片段。这些发现强烈表明，吞噬细胞（例如多核细胞）很容易在骨组织中提供，并且吞噬作用比EN更有效……对于骨组织中的几根和壳聚糖生物降解的酶消化更多。 DDAC 100应该是用于骨手术和骨再生疗法的合适生物材料。进行了另一项研究以评估壳聚糖单体（D-葡萄糖盐酸盐）作为纸浆封端药物的适用性。在应用切con糖酸后，进行了体外和体内实验，以研究细胞代谢和伤口愈合机制。在成骨细胞培养3天后，壳聚糖组的醇磷酸酶（ALP）活性显着增加。逆转录聚合酶链反应分析表明，壳聚糖在3天后诱导ALP mRNA的表达增加，成骨育育7天后骨形态发生蛋白-2 mRNA。炎性细胞因子，白介素（IL）-8，成纤维细胞中的合成在补充壳聚糖单体的培养基中被强烈抑制。在大鼠实验中评估了组织病理学效应。 1天后，与壳聚糖聚合物的应用相比，观察到炎症细胞浸润较弱。 3天后，在施加的壳聚糖单体附近看到了成纤维细胞的显着扩散。炎症细胞浸润几乎完全消失了。 5天后，成纤维细胞增殖进行了，一些牙成细胞出现在增殖的成纤维细胞的外围。这些发现表明，本研究是第一个报告，即即使在伤口愈合的初始阶段，与壳聚糖聚合物的应用相比，壳聚糖单体也是一种生物相容性的药物。较少的

项目成果

Early gene expression analyzed by cDNA microarray and real-time PCR in osteoblasts cultured with chitosan monomer

• DOI: 10.1002/jbm.a.31130
• 发表时间: 2007-07-01
• 期刊: JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A
• 影响因子: 4.9
• 作者: Ganno, Tomoko;Yamada, Shizuka;Hayashi, Yoshihiko
• 通讯作者: Hayashi, Yoshihiko

Chitosan monomer promotes tissue regeneration on dental pulp wounds

• DOI: 10.1002/jbm.a.30588
• 发表时间: 2006-03-15
• 期刊: JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A
• 影响因子: 4.9
• 作者: Matsunaga, T;Yanagiguchi, K;Hayashi, Y
• 通讯作者: Hayashi, Y

Chitosan monomer accelerates alkaline phosphatase activity on human osteoblastic cells under hypofunctional conditions

• DOI: 10.1002/jbm.a.31234
• 发表时间: 2007-11-01
• 期刊: JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A
• 影响因子: 4.9
• 作者: Yamada, Shizuka;Ganno, Tomoko;Hayashi, Yoshihiko
• 通讯作者: Hayashi, Yoshihiko

Immunohistochemical and electron microscopic study of the biodegradation processes of chitin and chitosan implanted in rat alveolar bone.

免疫组织化学和电子显微镜研究植入大鼠牙槽骨的几丁质和壳聚糖的生物降解过程。

• 发表时间: 2005
• 期刊: Oral Medicine ＆ Pathology 10(4)
• 作者: T.Matsunaga;K.Yanagiguchi;S.Yamada;N.Ohara;T Ikeda;Y.Hayashi;Tsunenori Matsunaga;Takeshi Ikeda
• 通讯作者: Takeshi Ikeda