関連論文
-
Clumps of MSCs/ECM complexes generated with chondro-induction medium induces bone regeneration
-
Cryopreserved clumps of MSCs/ECM complexes treated with INF-γ retain immunosuppressive capacity and induce rat calvarial bone regeneration
-
Cryopreserved clumps of MSCs/ECM complexes treated with INF-γ retain immunosuppressive capacity and induce rat calvarial bone regeneration
-
Clumps of MSCs/ECM complexes can induce periodontal tissue regeneration via direct differentiation
-
Clumps of MSCs/ECM complexes can induce periodontal tissue regeneration via direct differentiation
参考文献
1.
Pihlstrom, B.L., B.S. Michalowicz, and N.W. Johnson, Periodontal diseases. Lancet,
2005. 366(9499): p. 1809-1820.
2.
Kuo, L.C., A.M. Poison, and T. Kang, Associations between periodontal diseases and
systemic diseases: A review of the inter-relationships and interactions with
diabetes, respiratory diseases, cardiovascular diseases and osteoporosis. Public
Health, 2008. 122(4): p. 417-433.
3.
Bassani, D.G., M.T.A. Olinto, and N. Kreiger, Periodontal disease and perinatal
outcomes: a case-control study. Journal of Clinical Periodontology, 2007. 34(1): p. 3139.
4.
Bartold, P.M., R.I. Marshall, and D.R. Haynes, Periodontitis and rheumatoid
arthritis: A review. Journal of Periodontology, 2005. 76(11): p. 2066-2074.
5.
Scannapieco, F.A., R.B. Bush, and S. Paju, Associations between periodontal disease
and risk for atherosclerosis, cardiovascular disease, and stroke. A systematic
review. Ann Periodontol, 2003. 8(1): p. 38-53.
6.
Yoneda, M., et al., Involvement of a periodontal pathogen, Porphyromonas
gingivalis on the pathogenesis of non-alcoholic fatty liver disease. BMC
Gastroenterol, 2012. 12: p. 16.
7.
Langer, R. and J.P. Vacanti, TISSUE ENGINEERING. Science, 1993. 260(5110): p.
920-926.
8.
Christgau, M., et al., Guided tissue regeneration in intrabony defects using an
experimental bioresorbable polydioxanon (PDS) membrane. A 24-month split-mouth
study. J Clin Periodontol, 2002. 29(8): p. 710-23.
9.
Kitamura, M., et al., Periodontal Tissue Regeneration Using Fibroblast Growth
Factor-2: Randomized Controlled Phase II Clinical Trial. Plos One, 2008. 3(7). doi:
10.1371/journal.pone.0002611.
10.
Takayama, S., et al., Effects of basic fibroblast growth factor on human periodontal
ligament cells. Journal of Periodontal Research, 1997. 32(8): p. 667-675.
11.
Lynch, S.E., et al., A combination of platelet-derived and insulin-like growth factors
enhances periodontal regeneration. J Clin Periodontol, 1989. 16(8): p. 545-8.
12.
Mohammed, S., A.R.C. Pack, and T.B. Kardos, The effect of transforming growth
factor beta one (TGF-beta(1)) on wound healing, with or without barrier
membranes, in a Class II furcation defect in sheep. Journal of Periodontal
Research, 1998. 33(6): p. 335-344.
25
13.
Giannobile, W.V., et al., Recombinant human osteogenic protein-1 (OP-1) stimulates
periodontal wound healing in class III furcation defects. J Periodontol, 1998. 69(2):
p. 129-37.
14.
Wikesjö, U.M., et al., Periodontal repair in dogs: rhBMP-2 significantly enhances
bone formation under provisions for guided tissue regeneration. J Clin Periodontol,
2003. 30(8): p. 705-14.
15.
Blumenthal, N.M., et al., Effect of surgical implantation of recombinant human
bone morphogenetic protein-2 in a bioabsorbable collagen sponge or calcium
phosphate putty carrier in intrabony periodontal defects in the baboon. Journal of
Periodontology, 2002. 73(12): p. 1494-1506.
16.
Takeda, K., et al., Brain-derived neurotrophic factor enhances periodontal tissue
regeneration. Tissue Engineering, 2005. 11(9-10): p. 1618-1629.
17.
Bose, S., M. Roy, and A. Bandyopadhyay, Recent advances in bone tissue
engineering scaffolds. Trends in Biotechnology, 2012. 30(10): p. 546-554.
18.
Treiser, M.D., et al., Cytoskeleton-based forecasting of stem cell lineage fates.
Proceedings of the National Academy of Sciences of the United States of America,
2010. 107(2): p. 610-615.
19.
Thomson, J.A. and V.S. Marshall, Primate embryonic stem cells. Current Topics in
Developmental Biology, Vol 38, 1998. 38: p. 133-165.
20.
Takahashi, K., et al., Induction of pluripotent stem cells from adult human
fibroblasts by defined factors. Cell, 2007. 131(5): p. 861-872.
21.
Takahashi, K. and S. Yamanaka, Induction of pluripotent stem cells from mouse
embryonic and adult fibroblast cultures by defined factors. Cell, 2006. 126(4): p.
663-676.
22.
Pittenger, M.F., et al., Multilineage potential of adult human mesenchymal stem
cells. Science, 1999. 284(5411): p. 143-147.
23.
Colnot, C., Cell Sources for Bone Tissue Engineering: Insights from Basic Science.
Tissue Engineering Part B-Reviews, 2011. 17(6): p. 449-457.
24.
Kawaguchi, H., et al., Enhancement of periodontal tissue regeneration by
transplantation of bone marrow mesenchymal stem cells. Journal of Periodontology,
2004. 75(9): p. 1281-1287.
25.
Hasegawa, N., et al., Behavior of transplanted bone marrow-derived mesenchymal
stem cells in periodontal defects. Journal of Periodontology, 2006. 77(6): p. 10031007.
26.
Nagahara, T., et al., Introduction of a Mixture of beta-Tricalcium Phosphate Into a
Complex of Bone Marrow Mesenchymal Stem Cells and Type I Collagen Can
26
Augment the Volume of Alveolar Bone Without Impairing Cementum Regeneration.
Journal of Periodontology, 2015. 86(3): p. 456-464.
27.
Liechty, K.W., et al., Human mesenchymal stem cells engraft and demonstrate site-
specific differentiation after in utero transplantation in sheep. Nature Medicine,
2000. 6(11): p. 1282-1286.
28.
Hoogendoorn, H.A., et al., LONG-TERM STUDY OF LARGE CERAMIC
IMPLANTS (POROUS HYDROXYAPATITE) IN DOG FEMORA. Clinical
Orthopaedics and Related Research, 1984(187): p. 281-288.
29.
Oreffo, R.O.C. and J.T. Triffitt, Future potentials for using osteogenic stem cells and
biomaterials in orthopedics. Bone, 1999. 25(2): p. 5S-9S.
30.
Kittaka, M., et al., Clumps of a mesenchymal stromal cell/extracellular matrix
complex can be a novel tissue engineering therapy for bone regeneration.
Cytotherapy, 2015. 17(7): p. 860-873.
31.
Takewaki, M., et al., MSC/ECM Cellular Complexes Induce Periodontal Tissue
Regeneration. Journal of Dental Research, 2017. 96(9): p. 984-991.
32.
Takeshita, K., et al., Xenotransplantation of interferon-gamma-pretreated clumps of
a human mesenchymal stem cell/extracellular matrix complex induces mouse
calvarial bone regeneration. Stem Cell Research & Therapy, 2017. 8(1). doi:
10.1186/s13287-017-0550-1.
33.
Motoike, S., et al., Clumps of mesenchymal stem cell/extracellular matrix complexes
generated with xeno-free conditions facilitate bone regeneration via direct and
indirect osteogenesis. International Journal of Molecular Sciences, 2019. 20(16). doi:
10.3390/ijms20163970.
34.
Ernestina, S, et al., Hypoxia in cartilage: HIF-1α is essential for chondrocyte growth
arrest and survival. Genes & Development, 2001. 15(21): p. 2865-2876.
35.
Steck, E., et al., Mesenchymal stem cell differentiation in an experimental cartilage
defect: restriction of hypertrophy to boneclose neocartilage. Stem Cells and
Development, 2009. 18: p. 969-978.
36.
Phuong, N.D., et al., Endochondral Ossification in Critical-Sized Bone Defects via
Readily Implantable Scaffold-Free Stem Cell Constructs. Stem Cells Translational
Medicine, 2017. 6(7): p. 1644-1659.
37.
Bouxsein, M.L., et al., Guidelines for Assessment of Bone Microstructure in Rodents
Using Micro-Computed Tomography. Journal of Bone and Mineral Research, 2010.
25(7): p. 1468-1486.
27
38.
Xin, Z., et al., Chondrocytes transdifferentiate into osteoblasts in endchondral bone
during development, postnatalgrowth and fracture healing in mice. PLOS Genetics,
2014. 10(12). doi: 10.1371/journal.pgen.1004820.
39.
Y Jing., et al.,
Chondrocytes Directly Transform into Bone Cells in Mandibular
Condyle Growth. Journal of Dental Research, 2015. 94(12): p. 1668-1675.
40.
Adam, J.E., et al., Matrix elasticity directs stem cell lineage specification. Cell,
2006. 126(4): p. 677-689.
41.
Komatsu, N., et al., Type I collagen deposition via osteoinduction ameliorates
YAP/TAZ activity in 3D floating culture clumps of mesenchymal stem
cell/extracellular matrix complexes. Stem Cell Research & Therapy, 2018. 9(1). doi:
10.1186/s13287-018-1085-9.
42.
Li, F., X. Wang, and C. Niyibizi, Bone marrow stromal cells contribute to bone
formation following infusion into femoral cavities of a mouse model of osteogenesis
imperfecta. Bone, 2010. 47(3): p. 546-555.
43.
Zhou, Y., et al., Implantation of osteogenic differentiated donor mesenchymal stem
cells causes recruitment of host cells. Journal of Tissue Engineering and
Regenerative Medicine, 2015. 9(2): p. 118-126.
44.
Zhou, Y., et al., Mesenchymal stromal cells regulate the cell mobility and the
immune response during osteogenesis through secretion of vascular endothelial
growth factor A. Journal of Tissue Engineering and Regenerative Medicine, 2018.
12(1): p. E566-E578.
45.
Chelsea, S. B., et al., Stem cell derived endochondral cartilage stimulates bone
healing by tissue transformation. Journal of Bone and Mineral Research, 2014.
29(5): p. 1269-1282.
46.
Motoike, S., et al., Cryopreserved clumps of mesenchymal stem cell/extracellular
matrix complexes retain osteogenic capacity and induce bone regeneration. Stem
Cell Research & Therapy, 2018. 9(1). doi: 10.1186/s13287-018-0826-0.
28
図. 1
Xeno-free/Serum-free の軟骨誘導培地による C-MSCs 作製
(A&B) 5、10、15 日目でサンプルの回収を行い、HE 染色(A)およびサフラニン O 染色(B)
を行った。左は増殖培地で作製した C-MSCs、右は軟骨誘導培地で作製した C-MSCs を示
す。それぞれ左図が弱拡大、右図が強拡大像 (スケールバー=100 µm) を示す。
29
図. 1
Xeno-free/Serum-free の軟骨誘導培地による C-MSCs 作製
(C) 3、5、7、10、13、15 日目で回収を行い、real-time PCR を行った。使用したマーカ
ーは、Sox9, Aggrecan (ACAN), Col 2, Col 10, IHH である。ハウスキーピング遺伝子とし
て 18s を用いて、ΔΔCT 法により目的遺伝子の相対比を産出した。黒線が増殖培地で作製
した C-MSCs、赤線が軟骨誘導培地で作製した C-MSCs を示す。
30
図. 2
Xeno-free/Serum-free の軟骨誘導培地で作製された C-MSCs の SCID マウスの頭
蓋冠骨欠損への移植
(A) C-MSCs 移植時の肉眼所見を示す。上段が移植前、下段が C-MSCs 移植後。写真は軟
骨誘導培地で 10 日間培養した C-MSCs 移植時のもの。(B) 移植 4、8、12 週後の micro
CT 画像。比較として非移植群および増殖培地で 5 日間培養した C-MSCs を用いた。(スケ
ールバー=250 µm) (C) 移植 4、8、12 週後の骨欠損部における再生骨量 (BV) /全体組織量
(TV)。グラフはそれぞれ、左が 4 週、中央が 8 週、右が 12 週目のもの。エラーバーは
S.D.値を示す。**P < 0.01、*P < 0.05
31
32
図. 3
Xeno-free/Serum-free の軟骨誘導培地で培養した C-MSCs の頭蓋冠骨欠損への移
植時の組織像
(A&B) 移植 12 週後の組織像を示す。(A) 厚さ 8 µm で切片を作製し、HE 染色を行っ
た。(スケールバー=250 µm) (B) 厚さ 20 µm で切片を作製し、抗ヒト Vimentin 抗体で蛍
光免疫染色を行った。青色が細胞核、緑色がヒト Vimentin の分布を示す。上段が弱拡大
(スケールバー=250 µm)、下段が強拡大像 (スケールバー=50 µm)。左が非移植群、中央が
通常培地 5 日培養群、右が軟骨誘導培地 5 日培養群。
(C) 移植 4、8、12 週後の組織像を示す。厚さ 8 µm で切片を作製し、HE 染色およびサフ
ラニ O 染色を行った。上図が軟骨誘導培地 10 日培養群、下図が軟骨誘導培地 15 日培養
群を示す。それぞれ上段が HE 染色弱拡大像、下段左図が HE 染色強拡大像、下段右図が
サフラニン O 染色像を示す (スケールバー=250 µm)。左が移植 4 週後、中央移植 8 週
後、右が移植 12 週後。
(D&E) 移植 4、8、12 週後の組織像を示す。 厚さ 20 µm で切片を作製し、抗ヒト
Vimentin 抗体で蛍光免疫染色を行った。軟骨誘導培地 10 日培養群 (D)、軟骨誘導培地 15
日培養群(E)。青色が細胞核、緑色がヒト Vimentin の分布を示す。上段が弱拡大 (スケー
ルバー=250 µm)、下段が強拡大像 (スケールバー=50 µm)。それぞれ左が移植 4 週後、中
央移植 8 週後、右が移植 12 週後。
33
図. 4
Xeno-free/Serum-free の軟骨誘導培地で 10 日間培養した C-MSCs の頭蓋冠骨欠
損への移植時の短期的な観察
移植 3、7、14 日後に観察した。(A) micro CT 画像を示す。(スケールバー=250 µm) (B)上
段が HE 染色像、下段がサフラニン O 染色像を示す。左図が弱拡大、右図が強拡大像。
(スケールバー=500 µm) (C) 抗ヒト Vimentin 抗体で蛍光免疫染色を行った。それぞれ左
図が弱拡大(スケールバー=500 µm)、右図が強拡大像(スケールバー=50 µm)。青色が細胞
核、緑色がヒト Vimentin の分布を示す。
34
表. 1 real-time PCR プライマー
18s
Sox9
ACAN
Col 2
IHH
Col 10
F: GTAACCCGTTGAACCCCATT
R: CCATCCAATCGGTAGTAGCG
F: CATGAGCGAGGTGCACTCC
R: TCGCTTCAGGTCAGCCTTG
F: TGAGGAGGGCTGGAACAAGTACC
R: GGAGGTGGTAATTGCAGGGAACA
F: TTTCCCAGGTCAAGATGGTC
R: CTTCAGCACCTGTCTCACCA
F: AACTCGCTGGCTATCTCGGT
R: GCCCTCATAATGCAGGGACT
F: CCCTTTTTGCTGCTAGTATCC
R: CTGTTGTCCAGGTTTTCCTGGCAC
35
...