Seven million people suffer bone fractures in the U.S. each year. Musculoskeletal conditions cost the U.S. more than $200 billion annually. These numbers are increasing rapidly as the population ages. Human embryonic stem cells (hESCs) offer unlimited supplies of stem cells with a high potential for bone regeneration. However, there has been no report on the use of hESCs for bone tissue engineering via injectable calcium phosphates. While human bone marrow mesenchymal stem cells (hBMSCs) are useful, their harvest requires an invasive procedure, and their proliferation and differentiation potential is lot due to aging and diseases. Therefore, the objectives of this project are to: (1) investigate hESCs in injectable calcium phosphate cement (CPC) for bone engineering, in comparison with human umbilical cord MSCs (hUCMSCs) and hBMSCs;(2) establish the first knowledge base on hESC interactions with CPC scaffolds to guide hESCs for proliferation and osteodifferentiation;(3) design a RGD-grafted CPC scaffold for hESC encapsulation to enhance bone regeneration. While the Arg-Gly-Asp (RGD) peptide has been used in other scaffolds, there has been no report on its use in CPC.
Aim 1 will investigate in vitro the hESC encapsulation and differentiation in injectable and macroporous CPC-RGD constructs, and test these hypotheses: (1) hESC-derived MSCs encapsulated in degradable microbeads and incorporated into CPC will synthesize the most amount of bone matrix, followed by hUCMSCs. The gold-standard hBMSCs will make the least bone matrix;(2) RGD-grafted CPC will greatly enhance cell function and bone matrix synthesis, without compromising the CPC injectability and the mechanical properties, which will match the reported strength of cancellous bone.
Aim 2 will investigate the hESC-CPC-RGD constructs for bone regeneration in animal model, and test these hypotheses: (1) New bone volume, mineral density, and blood vessel density generated by CPC-RGD with hESC-derived MSCs will be the highest, followed by hUCMSCs. Both of them will far exceed those generated by hBMSCs;(2) hESC-CPC- RGD will be completely replaced by new bone across the entire critical-sized cranial defect at six months;(3) hESC-CPC-RGD will induce much more new bone than that without RGD. This project will yield ground- breaking knowledge on hESC encapsulation in CPC, hESC interaction with RGD-CPC and the guidance of hESCs for osteodifferentiation, and bone regeneration in animal model comparing hESCs, hUCMSCs, and hBMSCs side-by-side. The new stem cell paste can be used in minimally-invasive surgeries, fill complex- shaped defects, and be easily shaped for esthetics in dental and craniofacial applications. The novel hESC- CPC-RGD construct is expected to have wide orthopedic and craniofacial applications, with greatly enhanced bone regeneration to improve the health and quality of life for millions of people. If indeed, hESCs delivered via CPC are superior in osteogenesis compared to the gold-standard hBMSCs, which is anticipated to be shown by this project for the first time, the results will broadly impact the regenerative medicine field.

Public Health Relevance

Despite the high promise of human embryonic stem cells (hESCs), there has been no report on hESC seeding with injectable calcium phosphate constructs for bone regeneration. The proposed research will investigate hESCs for bone tissue engineering via novel injectable calcium phosphate scaffolds for the first time, and establish ground-breaking knowledge on hESC guidance via peptide-calcium phosphate cement scaffold to enhance hESC function and bone regeneration in animal model. The novel hESC-calcium phosphate construct is expected to have a wide range of craniofacial and orthopedic applications, with greatly enhanced bone regeneration capability to improve the health and quality of life for millions of people.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21DE022625-02
Application #
8429388
Study Section
Oral, Dental and Craniofacial Sciences Study Section (ODCS)
Program Officer
Lumelsky, Nadya L
Project Start
2012-04-01
Project End
2015-03-31
Budget Start
2013-04-01
Budget End
2015-03-31
Support Year
2
Fiscal Year
2013
Total Cost
$221,040
Indirect Cost
$77,040
Name
University of Maryland Baltimore
Department
None
Type
Schools of Dentistry
DUNS #
188435911
City
Baltimore
State
MD
Country
United States
Zip Code
21201
Wang, Ping; Song, Yang; Weir, Michael D et al. (2016) A self-setting iPSMSC-alginate-calcium phosphate paste for bone tissue engineering. Dent Mater 32:252-63
Wang, Ping; Liu, Xian; Zhao, Liang et al. (2015) Bone tissue engineering via human induced pluripotent, umbilical cord and bone marrow mesenchymal stem cells in rat cranium. Acta Biomater 18:236-48
Lee, Kangwon; Weir, Michael D; Lippens, Evi et al. (2014) Bone regeneration via novel macroporous CPC scaffolds in critical-sized cranial defects in rats. Dent Mater 30:e199-207
Wang, Ping; Zhao, Liang; Liu, Jason et al. (2014) Bone tissue engineering via nanostructured calcium phosphate biomaterials and stem cells. Bone Res 2:14017
Chen, Wenchuan; Thein-Han, WahWah; Weir, Michael D et al. (2014) Prevascularization of biofunctional calcium phosphate cement for dental and craniofacial repairs. Dent Mater 30:535-44
Tang, Minghui; Chen, Wenchuan; Liu, Jun et al. (2014) Human induced pluripotent stem cell-derived mesenchymal stem cell seeding on calcium phosphate scaffold for bone regeneration. Tissue Eng Part A 20:1295-305
Liu, Xian; Wang, Ping; Chen, Wenchuan et al. (2014) Human embryonic stem cells and macroporous calcium phosphate construct for bone regeneration in cranial defects in rats. Acta Biomater 10:4484-93
Wang, P; Zhao, L; Chen, W et al. (2014) Stem Cells and Calcium Phosphate Cement Scaffolds for Bone Regeneration. J Dent Res 93:618-25
Liu, Jun; Chen, Wenchuan; Zhao, Zhihe et al. (2014) Effect of NELL1 gene overexpression in iPSC-MSCs seeded on calcium phosphate cement. Acta Biomater 10:5128-38
TheinHan, Wahwah; Weir, Michael D; Simon, Carl G et al. (2013) Non-rigid calcium phosphate cement containing hydrogel microbeads and absorbable fibres seeded with umbilical cord stem cells for bone engineering. J Tissue Eng Regen Med 7:777-87

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