Zoledronic acid at subtoxic dose extends osteoblastic stage span of primary human osteoblasts.

OBJECTIVE: This study aimed to check the effect of zoledronic acid (ZA) at subtoxic dose on human osteoblasts (HOs) in terms of cell viability, apoptosis occurrence, and differentiation induction. ZA belongs to the family of bisphosphonates (BPs), largely used in the clinical practice for the treatment of bone diseases, often associated with jaw osteonecrosis onset. Their pharmacological action consists in the direct block of the osteoclast-mediated bone resorption along with indirect action on osteoblasts. MATERIALS AND METHODS: HOs were treated choosing the highest limit concentration (10(-5) M) which does not induce toxic effects. Live/dead staining, flow cytometry, mitochondrial membrane potential assay, osteocalcin western blotting, gp38 RT-PCR, collagen type I, PGE2, and IL-6 ELISA assays were performed. RESULTS: Similar viability level between control and ZA-treated samples is found along with no significant increase of apoptotic and necrotic cells in ZA-treated sample. To establish if an early apoptotic pathway was triggered, Bax expression and mitochondrial membrane potential were evaluated finding a higher protein expression in control sample and a good integrity of mitochondrial membrane in both experimental points. Type I collagen secretion and alkaline phosphatase (ALP) activity appear increased in ZA-treated sample, osteocalcin expression level is reduced in ZA-treated cells, whereas no modifications of gp38 mRNA level are evidenced. No statistical differences are identified in PGE2 secretion level whereas IL-6 secretion is lower in ZA-treated HOs with respect to control ones. CONCLUSIONS: These results highlight that ZA, delaying the osteoblastic differentiation process versus the osteocytic lineage, strengthens its pharmacological activity enhancing bone density. CLINICAL RELEVANCE: The knowledge of ZA effects on osteoblasts at subtoxic dose allows to improve therapeutic protocols in order to strengthen drug pharmacological activity through a combined action on both osteoclastic and osteoblastic cells.

Equine and porcine bone substitutes in maxillary sinus augmentation: a histological and immunohistochemical analysis of VEGF expression.

The aim of this work was to investigate the morphological structure and the expression of vascular endothelial growth factor (VEGF) after maxillary sinus augmentation through equine and porcine bone substitutes in humans. Ten patients showing edentulous posterior maxilla underwent maxillary sinus augmentation through particulate equine bone substitute and 10 patients through particulate porcine bone substitute. At the moment of implants insertion, 6 months after grafting, bone specimens were withdrawn and processed for morphological and immunohistochemical analyses. Notwithstanding the almost comparable clinical performances of both bone substitutes, histological results showed a better integration when an equine bone substitute was used compared to a porcine one. In particular, evident signs of particles resorption were observed in equine bone substitute group specimens compared to porcine ones. Immunohistochemical analysis showed a statistically significant increase of VEGF expression in equine compared to porcine bone substitute group specimens. These results showed both bone substitutes to achieve comparable clinical performance, indicating their successful use for bone regenerative procedures. However, in the same experimental time, equine group specimens showed evident resorption phenomena, whereas no or little signs of resorption were evident in the porcine group specimens. However, a more rapid and intense vascularization was achieved in equine bone substitute group, as demonstrated by immunohistochemical analysis for VEGF expression. Even if differences in vascularization significantly affect the clinical performance of a heterologous bone substitute, its ability to be resorbed is also very important in influencing long-term integration and long-term predictability of implant-prosthetic rehabilitation in regenerated sites.

Histological evaluation of fresh frozen bone integration at different experimental times.

The aim of this work was to investigate, through histological evaluation, the in vivo behavior of fresh frozen bone (FFB) used as particulate bone substitute in intraoral regenerative procedures. A total of 10 patients (group 1) received particulate FFB graft for bone regeneration in postextractive sockets, and 10 (group 2) underwent maxillary sinus augmentation by using the same bone substitute as filling. Fresh frozen bone was supplied from the Tissue Bank of the Veneto Region, Treviso Section.Healing was uneventful for all the patients and was monitored by periodical radiographs. Patients were scheduled for implant insertion according to the radiographic aspect. However, the mean healing time for group 1 was 45 days, whereas for group 2 patients, it was 100 days. At the moment of implant insertion, bone specimens were collected at the site of implant placement, from both groups and processed for histological analysis.Histological analysis after hematoxylin-eosin staining obtained from group 1 patients showed the presence of newly formed bone tissue, still well distinguishable from the grafted bone substitute. In samples from group 2 patients, a better integration could be recognized associated with active bone remodeling phenomena.These results showed a good integration of the considered FFB graft within the host tissue both at 45 and 100 days after grafting, displaying this biomaterial as suitable for preimplant regenerative procedures.