Accession Number:

ADA615215

Title:

Manipulation of Human Primary Endothelial Cell and Osteoblast Coculture Ratios to Augment Vasculogenesis and Mineralization

Descriptive Note:

Journal article

Corporate Author:

ARMY INST OF SURGICAL RESEARCH FORT SAM HOUSTON TX

Report Date:

2014-06-01

Pagination or Media Count:

8.0

Abstract:

Tissue-engineering scaffolds are often seeded with a single type of cell, but there has been more focus on cocultures to improve angiogenesis and bone formation for craniofacial applications. Investigation of bone-derived osteoblasts OBs is important because of the use of bone grafts and migration of OBs from native bone into constructs in vivo and therefore, their contribution to bone formation in vivo. The limitation of primary OBs has been their inability to mineralize without osteogenic factors in vitro. Through coculture of OBs and endothelial cells ECs and manipulation of the coculture ratio, mineralization can be achieved without osteogenic media or additional growth factors, thus enhancing their utility for tissue-engineering applications. An optimal ratio of ECOB for vasculogenesis and mineralization has not been determined for human primary cells. Human umbilical vein ECs were cultured with normal human primary OBs in different ECOB ratios, namely, 101, 51, 11, 15, and 110 with EC and OB monocultures as controls. The number of vasculogenic networks in a collagen matrix was highest in ratios of 51 and 11. ECs lined up and formed capillary-like networks by day 10, which was not seen in the other groups. On polystyrene, cells were cocultured with ECs and OBs in direct contact direct coculture or separated by a transwell membrane indirect coculture. At day 21, Alizarin Red staining showed mineralization on the 15 and 110 direct coculture ratios, with 15 having more mineralization nodules present than 110. No mineralization was seen in other direct coculture ratios or in any of the indirect coculture ratios.

Subject Categories:

  • Anatomy and Physiology
  • Medicine and Medical Research
  • Biomedical Instrumentation and Bioengineering

Distribution Statement:

APPROVED FOR PUBLIC RELEASE