Abstract
The aim of this paper was to observe and visualize the changes in osteoblasts by electron microscopy during osteogenesis using tissue engineering technique. We also studied the feasibility of improving tissue vascularization of the engineered bone by using small intestine submucosa (SIS) as the scaffold. Bone mesenchymal stem cells (BMSCs) were isolated by gradient centrifugation method. Bone mesenchymal stem cells were seeded in the SIS, and the scaffold-cell constructs were cultured in vitro for 2 weeks. Small intestine submucosa without BMSCs served as control. Both SIS scaffolds were then implanted subcutaneously in the dorsa of athymic mice. The implants were harvested after in vivo incubation for 4, 8 and 12 weeks. The changes in osteoblasts and vascularization were observed under a transmission electron microscope and a scanning electron microscope. The BMSCs grew quite well, differentiating on the surface of the SIS and secreting a great deal of extracellular matrices. The scaffold-cell constructs formed a lot of bone and blood vessels in vivo. The scaffold degraded after 12 weeks. No osteoblasts, but vascularization and fibroblasts were observed, in the control. The SIS can be used as a scaffold for constructing tissue-engineered bone as it can improve the formation of bone and vessels in vivo.
Similar content being viewed by others
References
Lewandrowski K, Gresser J D, Wise D L, Trantolo D J. Bioresorbable bone graft substitutes of different osteoconductivities: a histo-logic evaluation of osteointegration of poly (propylene glycol-co-fumaric acid)-based cement implants in rats. Biomaterials, 2000, 21(8): 757–764
Breitbart S, Graride D A, Keler R. Tissue engineered bone repair of calvarial defects using cultured periosteal cells. Plast Reconstr Surg, 1998; 101(3): 567–574.
Sittinger M. Tissue engineering:artificial tissue replacment containing vital components. Laryngorhinootologie, 1995, 74(11): 695–699
Menko A S, Boettiger D. Occupation of the extracellular matrixreceptor integrin is a control point for myogenic differentiation. Cell, 1987, 51(1): 51–57
Adams J C, Watt F M. Changes in keratinocyte adhesion during terminal differentiation: reduction in fibronectin binding precedes alpha 5 beta 1 integrin loss from the cell surface. Cell, 1990, 63(2): 425–435
Zhang Kaigang; Zeng Bingfang; Zhang Changqing. Experimental study on the biocompatibility of small intestinal submucosa with bone marrow mesenchymal stem cells. Chin J Orthop Trauma, 2005, 7(4): 344–348
Abraham G A, Murray J, Billiar K, Sullivan S J. Evaluation of the porcine intestinal collagen layer as a biomaterial. J Biomed Mater Res, 2000, 51(3): 442–452
Griffith L G, Naughton G. Tissue engineering-current challenges and expanding opportunities. Science, 2002, 295(3): 1009–1014
Mooney D J, Mikos A G. Growing new organs. Sci Am, 1999, 280(1): 60–65
McPherson T B, Badylak S F. Characterization of fibronectin derived from porcine small intestinal submucosa. Tissue Eng, 1998, 4: 75–81
Voytik-Harbin S L, Brightman A O, Kraine M R, Waisner B, Badylak S F. Identification of extractable growth factors from small intestinal submucosa. J Cell Biochem, 1997, 67(4): 478–491
McDevitt C A, Wildy G M, Cutrone R M. Transforming growth factor-beta1 in a sterilized tissue derived from the pig small intestine submucosa. J Biomed Mater Res, 2003, 67(2): 637–640
Hsu F Y, Chueh S C, Wang Y J. Microspheres of hydroxyapatite/reconstituted collagen as supports for osteoblast cell growth. Biomaterials, 1999, 20(20): 1931–1936
Cook J L, Tomlinson J L, Kreeger J M, Cook C R. Induction of meniscal regeneration in dogs using a novel biomaterial. Am J Sports Med, 1999, 27(5): 658–665
Rabah D M, Spiess P E, Begin L R, Corcos J. Tissue reaction of the rabbit urinary bladder to tension-free vaginal tape and porcine small intestinal submucosa. BJU Int, 2002, 90(6): 601–606
Roeder R A, Lantz G C, Geddes L A. Mechanical remodeling of small-intestine submucosa small-diameter vascular grafts—a preliminary report. Biomed Instrum Technol, 2001, 35(2): 110–120
Dejardin L M, Arnoczky S P, Ewers B J, Haut R C, Clarke R B. Tissue-engineered rotator cuff tendon using porcine small intestine submucosa: histologic and mechanical evaluation in dogs. Am J Sports Med, 2001, 29(2): 175–184
Malekzadeh R, Hollinger J O, Buck D. Isolation of human osteoblastic like cells and in vitro amplification for tissue engineering. J Periodontol, 1998, 69(11): 1256–1263
Kaigler D, Krebsbach P H, Polverini P H, Mooney D J. Role of vascular endothelial growth factor in bone marrow stromal cell modulation of endothelial cells. Tissue Eng, 2003, 9(1): 95–103
Ferrara N. Vascular endothelial growth factor and the regulation of angiogenesis. Recent Prog Horm Res, 2000, 55(1): 15–35
Author information
Authors and Affiliations
Corresponding author
Additional information
Translated from Journal of Shanghai Jiaotong University (Medical Science), 2006, 26(2): 113–116 [译自: 上海交通大学学报 (医学版)]
Rights and permissions
About this article
Cite this article
Zhang, K., Zeng, B. & Zhang, C. Visualization of vascular ultrastructure during osteogenesis by tissue engineering technique. Front. Med. China 1, 181–184 (2007). https://doi.org/10.1007/s11684-007-0034-2
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11684-007-0034-2