We are using cookies to implement functions like login, shopping cart or language selection for this website and to create anonymized statistical reports of the usage. You will find more information in our privacy policy. OK, I have understood
International Poster Journal of Dentistry and Oral Medicine
Login:
username:

password:

Plattform:

Forgotten password?

Registration

Int Poster J Dent Oral Med 13 (2011), No. 3     15. Sep. 2011

Int Poster J Dent Oral Med 2011, Vol 13 No 3, Poster 553

Intraindividual comparison of the osteogenic differentiation potential of mesenchymal stem cells derived from adult adipose tissue and bone marrow

Intraindividual comparison of osteogenic differentiation potential of ATSC and BMSC

Language: English
 

Authors:
Dr. Dr. Konstanze Scheller,
Martin-Luther-University Halle-Wittenberg, Universitätsklinikum Kröllwitz, Klinik und Poliklinik für Mund-, Kiefer- und Plastische Gesichtschirurgie, Halle-Wittenberg, Germany
Dr. Dirk Halama,
University of Leipzig, Klinik und Poliklinik für Mund-, Kiefer- und Plastische Gesichtschirurgie, Leipzig, Germany
Alexander Burkhardt, PD. Dr. Frank Gaunitz,
University of Leipzig, Institut für Biochemie, Leipzig, Germany
Prof. Dr. Dr. Bernhard Frerich,
University of Rostock, Klinik und Poliklinik für Mund-, Kiefer- und Plastische Gesichtschirurgie, Rostock, Germany

Date/Event/Venue:
31.05. -02.06.2006
Strategies in Tissue Engineering
Würzburg, Germany
 

Introduction

Adipose tissue, like bone marrow (BMSC), is a promising source to recover multipotent mesenchymal stromal cells (ATSC) for tissue enginnering and regeneration of bone. FACS-analyses even show a specific marker profile for ATSC, like for BMSC. Osteogenic differentiation of BMSC and ATSC is conventionally mediated by ascorbic acid, dihydroxycholecalciferol and dexamethasone/ß-glycerophosphate (conventional conditions) and by the addition of rhBMP-2 in 2D- and 3D-cell culture systems (collagenous scaffolds).
 

Objectives

The lack of sources for autologous bone transplantation has led to the development of different strategies for bone replacement. The long-term objective of our work is the establishment of bone regeneration strategies for the treatment of osseous defects after trauma or ablative tumor surgery by tissue engineering. It is considered, that cure of osseous defects is even ideally approachable to regenerative strategies.
 

Material and Methods

Human adipose tissue (AT) and bone marrow (BM) were obtained from 4 patients and mesenchymal stromal cells were isolated and analysed (FACS, n=14).
Osteogenic differentiation under conventional conditions was compared to the addition of rhBMP-2 (10mg/ml). Histological stainings were prepared of 2D- and 3D-cell culture systems. RNA-expression of osteocalcin (OC), osteopontin (OP), osteonectin (ON) and alkaline phosphatase (AP) was analysed quantitative by RT-PCR (LightCycler) in 2D-cultures.
 

Results

1. Osteogenic differentiation of ATSC in vitro:

Isolated ATSC show the same marker-profile expression (CD29+, CD31-, CD34-, CD44+, CD73+, CD90+, CD105+, CD133-, CD166-) like BMSC in FACS-analyses (Fig.1).
In 2D- and 3D-cell culture systems in vitro (Fig. 2) the osteogenic differentiation of ATSC could be shown by the detection of OC-producing cells (immunhistological staining) in the mineralisation areas (v.Kossa).
3D-cell culture systems in a SCID-mouse model show in vivo (Fig. 3) even a osteogenic differentiation of ATSC in immunhistological stainng of OC and trabecular bone formation.

Fig. 1: FACS-analyses (n=14) Fig. 2: Histological staining of osteogenic differentiated ATSC (A) with mineralisation areas (v.-Kossa, B/C), and osteocalcin-expression (D/E)
Fig. 3a: Imunhistological staining of OC Fig. 3b: Imunhistological staining of HE

2. Comparison of the osteogenic differentiation potential of ATSC/BMSC:

Osteogenic differentiated ATSC and BMSC showed under conventional differentiation conditions a high expression of osteocalcin (OC) in FACS-analysis and RT-PCR (relative: ATSC > BMSC: 15:1, Fig.4).

Fig. 4a-c: RT-PCR and FACS-analysis of OC
 >
Fig. 4a-c: RT-PCR and FACS-analysis of OC  

AP-expression incresed during the whole differentiation period in ATSC, while ON-expression (active remodeling) increased clearly in the 5th week of osteogenic differentaition in BMSC while OPN-expression (tissue integrity) decreased during the differentiation in both (ATSC/BMSC, Fig.5).
By application of rhBMP-2 was only an increase of the ON- and OPN-expression in BMSC during the differentiation rhBMP-2 period in shown (Fig.6).

Fig. 5a-d: RT-PCR of AP-, ON-and OPN-expression
Fig. 5a-d: RT-PCR of AP-, ON-and OPN-expression
Fig. 6a-b: ON-and OPN-expression under rhBMP2-application during differentiation
 

Conclusions

Osteogenic differentiation is possible in ATSC and BMSC (different potential) and showed in ATSC a lower osteogenic potential than in BMSC. The osteogenic differentiation potential of rhBMP-2 in our protocol is less effective
 

Literature

  1. Frerich B, Kurtz-Hoffmann K, Lindemann N, Müller S. (1999) Untersuchungen zum Tissue engineering vaskularisierter knöcherner und weichgewebiger Transplantate. Mund-, Kiefer- und Gesichtschirurgie 4 Suppl 2: 490-5.
  2. Frerich B, Kurtz-Hoffmann K, Lindemann N, Müller S. (1999) Untersuchungen zum Tissue engineering vaskularisierter knöcherner und weichgewebiger Transplantate. Mund-, Kiefer- und Gesichtschirurgie 4 Suppl 2: 490-5.
  3. Frerich, B., Kurtz-Hoffmann, J., Lindemann, J. (2005): Influence of Growth Hormone on Maintenance of Capillary-like Structures in an In Vitro Model of Stromal Vascular Tissue-Results From Morphometric Analysis. Artif Organs 29, 338-341.
  4. Weinzierl, K.; Lazcano, M.; Dammann, B.; Mating, H.; Frerich, B. (2003) Differentiation potential of adipose tissue derived mesenchymal stem cells for in vitro engineering. Int J Artif Organs 26: 850.
  5. Weinzierl, K., Hemprich, A., Frerich, B. Bone engineering with adipose tissue derived stromal cells. J Craniomaxillofac Surg. 34: (2006); 466-471.
  6. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng, 2001, 7: 211-228.
  7. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI,Mizuno H, Alfonso ZC, Fraser JK, Benhaim P, Hedrick MH. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell, 2002, 13: 4279-4295.
     

This Poster was submitted by Dr. Dr. Konstanze Scheller.
 

Correspondence address:
Dr. Dr. Konstanze Scheller
Universität Halle-Wittenberg
Klinik und Poliklinik für Mund-, Kiefer- und Plastische Gesichtschirurgie, Universitätsklinikum Kröllwitz
Ernst-Grube-Strasse 40
06114 Halle
Germany