gms | German Medical Science

German Congress of Orthopedic and Trauma Surgery (DKOU 2018)

23.10. - 26.10.2018, Berlin

Treatment of a critical size segmental bone defects using spheroidal versus monolayer stem cell culture

Meeting Abstract

  • presenting/speaker Julia Bolte - UniversitätsCentrum für Orthopädie und Unfallchirurgie an der Technischen Universität Dresden, Dresden, Germany
  • Lars Müller - UniversitätsCentrum für Orthopädie und Unfallchirurgie an der Technischen Universität Dresden, Dresden, Germany
  • Corina Vater - Zentrum für Translationale Knochen-, Gelenk- und Weichgewebeforschung an der Technischen Universität Dresden, Dresden, Germany
  • Cathleen Petzold - Zentrum für Translationale Knochen-, Gelenk- und Weichgewebeforschung an der Technischen Universität Dresden, Dresden, Germany
  • Christian Männel - UniversitätsCentrum für Orthopädie und Unfallchirurgie an der Technischen Universität Dresden, Dresden, Germany
  • Maik Stiehler - UniversitätsCentrum für Orthopädie und Unfallchirurgie an der Technischen Universität Dresden, Dresden, Germany
  • Michael Gelinsky - Zentrum für Translationale Knochen-, Gelenk- und Weichgewebeforschung an der Technischen Universität Dresden, Dresden, Germany
  • Stefan Zwingenberger - UniversitätsCentrum für Orthopädie und Unfallchirurgie an der Technischen Universität Dresden, Dresden, Germany

Deutscher Kongress für Orthopädie und Unfallchirurgie (DKOU 2018). Berlin, 23.-26.10.2018. Düsseldorf: German Medical Science GMS Publishing House; 2018. DocGF14-766

doi: 10.3205/18dkou474, urn:nbn:de:0183-18dkou4741

Published: November 6, 2018

© 2018 Bolte et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 License. See license information at http://creativecommons.org/licenses/by/4.0/.


Outline

Text

Objectives: Bone loss may be associated with trauma, tumor, infection- and wear-induced periprosthetic osteolysis, or congenital musculoskeletal disorders. Recently was found in vitro, and in a murine calvarial defect model, that spheroidal culture of murine stem cells is associated with a higher osteoinductive potential, compared to monolayer culture. The aim of our study was to investigate the bone regenerative potential of human primary mesenchymal stem cells (MSCs) cultivated by spheroidal versus monolayer culture in a immunodeficient murine critical size femoral bone defect model.

Methods: Bone marrow aspirates of two male and two female healthy donors (aged 70 to 79 years) were collected during total hip arthroplasty procedure. MSCs were isolated by density gradient centrifugation and subsequent plastic adherence. After osteogenic pre-differentiation of monolayer MSCs for 7 days either spheroids containing 5x10^4 cells were formed by centrifugation in suspension plates or cells were further cultured in monolayer in osteogenic medium for additional 2 days. Implants were produced in cylindrical shape (2 mm length and 2 mm diameter) with either 5x10^4 monolayer-cultured cells or one cell spheroid.

Twenty-six 12-week old nude mice were randomized to two equal groups. A 2 mm femoral bone defect model was used. After six weeks µCT-scans and histological analysis were done. Descriptive statistics included means and SD. Unpaired t-tests were used to detect statistical significance. Differences were considered significant if p<0.05.

Results and conclusion: All twenty-six animals survived the observation period. There were no intergroup differences regarding BV in and around the defect area detected (spheroidal MSCs vs. monolayer MSCs; 2,1 ± 0,2 mm³ vs. 2,1 ± 0,2 mm³, p=0,9443). BMD was significantly higher in the spheroidal (917 ± 9 mgHA/mm³) compared with the monolayer group (882 ± 14 mgHA/mm³, p=0,043). Histological analysis showed no difference in the degree of defect healing between the groups (spheroidal vs. monolayer; 5,8 vs. 5,9; p=0,2032).In our experiments osteogenically pre-differentiated spheroidal and monolayer MSCs showed comparable regenerated BV. The reason for the reduced osteoinductive potential compared previous studies might be that we investigated these cells for the first time in a challenging critical size femoral bone defect. However, we found an improved BMD of the defect area, when spheroidal MSCs were used. The application of spheroidal MSC culture might be beneficial for the treatment of site-specific bone regeneration, especially in osteoporotic patients.

Human MSCs cultivated by spheroidal culture showed a significantly increased BMD at the defect site compared to monolayer cultured MSCs of the same donor in a murine critical size femoral bone defect model. No difference was found for the regenerated bone volume and the degree of defect healing.