Isolation and characterization of osteoblast cultures from normal and osteopenic sheep for biomaterials evaluation

Abstract

Being very useful in the analysis of bone cell differentiation and activity, osteoblast cultures are also used in the in vitro biocompatibility study of new materials. The aim of this work was to evaluate sheep osteoblast cultures derived from normal and ovariectomized animals, and then to assess the in vitro biomaterial behavior on these cultures, taking into account the quality of bone where orthopedic devices are clinically used. For this purpose, we characterized sheep osteoblast cultures, isolated from iliac crest bone of normal (NB osteoblast culture) and osteopenic after ovariectomy (OB osteoblast culture) sheep. Moreover, we studied cell behavior when cultured on different biomaterials (titanium and two biological glasses, RKKP and AP40). Cell characterization at baseline demonstrated that both cultures (NB and OB) showed normal osteoblastic behavior. On the contrary, osteoblasts derived from osteopenic bone and cultivated on AP40 for 6 days revealed a different behavior in terms of both cell morphology and metabolic activity. Statistical analysis (one-way analysis of variance and Scheffé's post hoc multiple-comparison tests) revealed significant differences in Ca level (p<0.0005), MTT test (p<0.0005) and OC production (p<0.05). These in vitro tests demonstrated that sheep osteoblast cultures can be useful when determining biocompatibility and osteointegration of orthopedic materials, and also when evaluating for the presence of osteoporosis.

A new austenitic stainless steel with a negligible amount of nickel: An in vitro study in view of its clinical application in osteoporotic bone

“…As previously observed for Ti6Al4V in both sheep and rat nOB and oOB cultures,19, 20 P558 did not compromise the behavior of osteopenic bone‐derived cells. Osteopenic bone‐derived cells appear to proliferate and differentiate on P558 without differing from normal bone‐derived cells, thus demonstrating that this biomaterial could be a promising option for the implantation in osteoporotic bone in the future.…”

“…The purpose of the present in vitro study was therefore to assess the behavior of this material after a 24‐ and 72‐h culture with primary osteoblasts from healthy and osteopenic rat bone, in order to acquire a better preclinical understanding of the biomaterial behavior in view of its clinical application in osteoporotic and aged bone. P558 was not compared with a control material because in previous studies Ti6Al4V was similarly evaluated in osteopenic bone‐derived osteoblasts 19, 20…”

“…In previous studies, the present authors investigated the use of osteoblasts derived from osteopenic bone to assess whether it could represent a new significant step in the in vitro research on biomaterials 17–20. According to the results, a different reactivity after drug stimulation or contact with biomaterial was observed:17–21 When the same biomaterial was positioned in direct contact with osteoporotic and normal bone‐derived cells, it behaved differently. This behavior was consistent with the in vivo implant results obtained in osteopenic animals with the same biomaterial, thus supporting the reliability of the in vitro pathological model adopted.…”

Biological assessment of the bone–screw interface after insertion of uncoated and hydroxyapatite‐coated pedicular screws in the osteopenic sheep

“…Moreover, the bone formation rates of primates, canines, sheep, and pigs are recognized to be closer to the human bone formation rates in comparison with small‐size animals 12. The sheep is considered a good model for osteoporosis13–18 and in vivo spinal research 19–23. Recent studies of the current authors have demonstrated both the development of osteopenia in the pedicle and vertebral trabecular bone of sheep 24 months after ovariectomy and the significantly improved fixation strength of HA‐coated screws in osteopenic sheep pedicles compared with uncoated screws 16, 24…”

Osteoblasts Cultured from Osteoporotic Bone: A Comparative Investigation on Human and Animal‐Derived Cells

“…Apart from bone biopsies in patients (Battmann et al, 1997;Evans et al, 1990;Kassem et al, 1994;Lomri and Marie, 1990;Marie et al, 1989Marie et al, , 1992Marie et al, , 1993Walsh et al, 2000;Wong et al, 1994), the rat is, to the current authors' knowledge, the animal species most widely used as donor of osteoblasts for in vitro studies on osteoporosis (Egrise et al, 1999;Fini et al, 2001a -c;Modrowski et al, 1992;Torricelli et al, 2000aTorricelli et al, , 2001). In addition, since an increasing use is being made of sheep as a model of osteoporosis (Bellino, 2000;Chavassieux et al, 2001;Fini et al, 2000;Giavaresi et al, 2001a;Martini et al, 2001;Thorndike and Turner, 1998), osteoblasts derived from osteopenic sheep bone also appear to be a promising alternative when conducting research on metabolic bone diseases (Torricelli et al, 2000b).…”