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Comparative study of the in vitro characteristics of osteoblasts from paralytic and non-paralytic children
K Anselme1, B Noël, D Limosino
1Institut de Recherche sur les Biomatériaux et les Biotechnologies (IR2B), Université du Littoral Côte d'Opale, Berck sur mer, France.
Insights
Children with cerebral palsy show altered bone metabolism. Osteoblast cells from flaccid patients produced more osteocalcin and collagen in vitro, suggesting a potential regulatory factor like leptin.
Area of Science:
- Biochemistry and Molecular Biology
- Cell Biology
- Pediatric Endocrinology
Background:
- Cerebral palsy (CP) is associated with significant bone loss in children and adults.
- Human osteoblastic cell culture is a valuable tool for studying bone metabolism in metabolic bone disorders.
- Limited research exists on the in vitro characteristics of osteoblasts from immobilized or normal bones, particularly differentiating between spastic and flaccid CP subtypes.
Purpose of the Study:
- To characterize mature osteoblasts (OB) and bone marrow-stromal cells (BM) from immobile and normal children.
- To compare in vitro viability, proliferation, and differentiation parameters of cells from paralytic (spastic and flaccid CP) and control groups.
Main Methods:
- Isolation of osteoblasts and bone marrow cells from iliac crests of young control and paralytic patients undergoing pelvic osteotomies.
- In vitro assessment of cell viability, proliferation, and differentiation.
- Comparison of cellular parameters between paralytic and control groups.
Main Results:
- No significant differences in cell proliferation were observed between groups.
- Initial cell viability was lower in the paralytic group compared to controls.
- Osteoblasts from flaccid patients synthesized more osteocalcin (fresh cells) and collagen (thawed cells) than spastic and control groups; BM cultures showed opposite results.
Conclusions:
- A negative feedback mechanism, not conserved in vitro, likely regulates in vivo osteocalcin and collagen synthesis in flaccid paralytic osteoblasts.
- The high fat/lean mass in flaccid patients suggests leptin as a potential regulatory factor in this hypothesized feedback mechanism.
- These findings highlight potential differences in bone cell function related to CP subtypes and suggest novel therapeutic targets.
Study Design:
Cerebral palsy is known to provoke a high loss of bone in children and adults. The potential interest of human osteoblastic cell culture for assessing the osteoblastic function in metabolic bone disorders has been demonstrated by many previous studies. Few studies have attempted to evaluate the capacities of osteoblasts isolated from immobilized or normal bones by in vitro culture methods. Moreover, a few teams did make the distinction between young spastic and flaccid patients.
Objectives:
We attempted to characterize mature osteoblasts (OB) and bone marrow-stromal cells (BM) originating from 56 immobile and normal children. Spastic and flaccid patients formed the paralytic group.
Setting:
France.
Methods:
Osteoblasts and bone marrow cells were isolated from iliac crests obtained during pelvic osteotomies of young control and paralytic patients. The in vitro viability, proliferation and differentiation parameters of the cells from paralytic patients were compared with those of cells coming from normal controls.
Results:
No significant differences in the cell proliferation parameters were observed between the two groups. Only initial cell viability before inoculation was lower for the paralytic group, compared to the control group. On the other hand, contrary to expectations, we found that fresh and thawed OB cells from flaccid patients synthesized more osteocalcin and more collagen respectively than those of the spastic and control groups. Opposite results were obtained from BM cultures.
Conclusion:
A negative feedback mechanism by systemic or local factors, which is not conserved in vitro but controls the in vivo osteocalcin and collagen synthesis of flaccid paralytic OB cells, is hypothesized. Because these flaccid patients are known to have a high fat/lean mass, we suggest that leptin may be the potential regulating factor implicated in the hypothesized negative feedback mechanism.
