Regenerating Critical Size Rat Segmental Bone Defects with a Self-Healing Hybrid Nanocomposite Hydrogel: Effect of Bone Condition and BMP-2 Incorporation.

The aim of the current study is to assess the biological performance of self-healing hydrogels based on calcium phosphate (CaP) nanoparticles and bisphosphonate (BP) conjugated hyaluronan (HA) in a critical size segmental femoral bone defect model in rats. Additionally, these hydrogels are loaded with bone morphogenetic protein 2 (BMP-2) and their performance is compared in healthy and osteoporotic bone conditions. Treatment groups comprise internal plate fixation and placement of a PTFE tube containing hydrogel (HABP -CaP) or hydrogel loaded with BMP-2 in two dosages (HABP -CaP-lowBMP2 or HABP -CaP-highBMP2). Twelve weeks after bone defect surgery, bone formation is analyzed by X-ray examination, micro-CT analysis, and histomorphometry. The data show that critical size, segmental femoral bone defects cannot be healed with HABP -CaP gel alone. Loading of the HABP -CaP gel with low dose BMP-2 significantly improve bone formation and resulted in defect bridging in 100% of the defects. Alternatively, high dose BMP-2 loading of the HABP -CaP gel does not improve bone formation within the defect area, but leads to excessive bone formation outside the defect area. Bone defect healing is not affected by osteoporotic bone conditions.

Bioinorganic supplementation of calcium phosphate-based bone substitutes to improve in vivo performance: a systematic review and meta-analysis of animal studies.

Supplementation of CaP-based bone graft substitutes with bioinorganics such as strontium, zinc or silicon is an interesting approach to increase the biological performance in terms of bone regenerative potential of calcium phosphate (CaP)-based bone substitutes. However, the in vivo efficacy of this approach has not been systematically analyzed, yet. Consequently, we performed a systematic review using the available literature regarding the effect of bioinorganic supplementation in CaP-based biomaterials on new bone formation and material degradation in preclinical animal bone defect models and studied this effect quantitatively by performing a meta-analysis. Additional subgroup analyses were used to study the effect of different bioinorganics, animal model, or phase category of CaP-based biomaterial on bone formation or material degradation. Results show that bioinorganic supplementation increases new bone formation (standardized mean difference [SMD]: 1.43 SD, confidence interval [CI]: 1.13-1.73). Additional subgroup analysis showed that strontium, magnesium and silica significantly enhanced bone formation, while zinc did not have any effect. This effect of bioinorganic supplementation on new bone formation was stronger for DCPD or ß-TCP and biphasic CaPs than for HA or α-TCP (p < 0.001). In general, material degradation was slightly hindered by bioinorganic supplementation (mean difference [MD]: 0.84%, CI: 0.01-1.66), with the exception of strontium that significantly enhanced degradation. Overall, bioinorganic supplementation represents an effective approach to enhance the biological performance of CaP-based bone substitutes.

Platinum-loaded, selenium-doped hydroxyapatite nanoparticles selectively reduce proliferation of prostate and breast cancer cells co-cultured in the presence of stem cells.

Chemotherapeutic treatment of patients with bone tumors or bone metastases often leads to severe side effects such as high drug toxicity, lack of tumor specificity and induced drug resistance. A novel strategy to treat early stages of bone metastases involves local co-delivery of multiple chemotherapeutic agents to synergistically improve the curative effect and overcome shortcomings of traditional chemotherapy. Herein we show that selenite-doped hydroxyapatite nanoparticles loaded with a hydroxyapatite-binding anti-tumor platinum complex (PtPP-HASe) selectively reduce proliferation of cancer cells without reducing proliferation of bone marrow stem cells. These PtPP-HASe particles were nanocrystalline with selenium (Se) and platinum (Pt) contents ranging between 0-10 and 1.5-3 wt%, respectively. Release kinetics of Se and Pt from PtPP-HASe nanoparticles resulted in a cumulative release of ∼10 and ∼66 wt% after 7 days, respectively. At a Pt/Se ratio of 8, released Pt and Se species selectively reduced cell number of human prostate (PC3) and human breast cancer cells (MDA-MB-231) by a factor of >10 with limited effects on co-cultured human bone marrow stem cells (hBMSc). These novel nanoparticles demonstrate high anti-cancer selectivity, which offers ample opportunities for the design of novel biomaterials with potent and selective chemotherapeutic efficacy against cancer cells.

Preclinical evaluation of injectable bone substitute materials.

Injectable bone substitutes (IBSs) represent an attractive class of ready-to-use biomaterials, both ceramic- and polymer-based, as they offer the potential benefit of minimally invasive surgery and optimal defect filling. Although in vitro assessments are the first step in the process of development, the safety and efficacy of an IBS strongly depend on validated preclinical research prior to clinical trials. However, the selection of a suitable preclinical model for performance evaluation of an IBS remains a challenge, as no gold standard exists to define the best animal model. In order to succeed in this attempt, we identified three stages of development, including (a) proof-of-principle, (b) predictive validity and (c) general scientific legitimacy, and the respective criteria that should be applied for such selection. The second part of this review provides an overview of commonly used animals for IBSs. Specifically, scientific papers published between January 1996 and March 2012 were retrieved that report the use of preclinical models for the evaluation of IBSs in situations requiring bone healing and bone augmentation. This review is meant not only to describe the currently available preclinical models for IBS application, but also to address critical considerations of such multi-factorial evaluation models (including animal species, strain, age, anatomical site, defect size and type of bone), which can be indicative but in most cases edge away from the human reality. Consequently, the ultimate goal is to guide researchers toward a more careful and meaningful interpretation of the results of experiments using animal models and their clinical applications.

Osteogenic capacity of human BM-MSCs, AT-MSCs and their co-cultures using HUVECs in FBS and PL supplemented media.

Human bone marrow-derived mesenchymal stem cells (BM-MSCs) and human adipose tissue-derived mesenchymal stem cells (AT-MSCs) are the most frequently used stem cells in tissue engineering. Due to major clinical demands, it is necessary to find an optimally safe and efficient way for large-scale expansion of these cells. Considering the nutritional source in the culture medium and method, this study aimed to analyze the effects of FBS- and PL-supplemented media on osteogenesis in stem cell mono- and co-cultures with human umbilical vein endothelial cells (HUVECs). Results showed that cell metabolic activity and proliferation increased in PL- compared to FBS-supplemented media in mono- and co-cultures for both BM-MSCs and AT-MSCs. In addition, calcium deposition was cell type dependent and decreased for BM-MSCs but increased for AT-MSCs in PL-supplemented medium in both mono- and co-cultures. Based on the effects of co-cultures, BM-MSCs/HUVECs enhanced osteogenesis compared to BM-MSCs monocultures in both FBS- and PL-supplemented media whereas AT-MSCs/HUVECs showed similar results compared to AT-MSCs monocultures.

The effect of platelet-rich plasma in vitro on primary cells: rat osteoblast-like cells and human endothelial cells.

The aim of this study was to evaluate the effects of standardized platelet-rich plasma (PRP) concentrates from 10 human donors on cellular behavior. The standardized PRPs used were fivefold average and fivefold maximum baseline values in whole blood. Both these standardized PRPs were characterized by determining platelet numbers and subsequently growth factor concentrations in activated PRPs, called PRP derivatives. Platelet numbers in both types of standardized PRPs were significantly increased compared with whole blood. Further, both PRP derivatives contained significantly higher concentrations of platelet-derived growth factor-AA, platelet-derived growth factor-AB, and transforming growth factor-beta 1. Vascular endothelial growth factor concentrations were significantly elevated in only the most concentrated PRP derivative. Cell culture experiments with osteoblast-like cells showed that both PRP derivatives stimulated cell proliferation without inducing cell differentiation, whereas tube formation in endothelial cell cultures was significantly increased by adding low volume percentages of PRP derivative (2%–8%). Consequently, it can be concluded that there is no direct relationship between the number of platelets and the level of growth factors released from these platelets. PRP derivatives have the potency to stimulate angiogenesis dose dependently, while lacking the capacity to induce osteogenic differentiation. Yet, the proliferation of osteoblast-like cells can significantly be enhanced by supplementation of PRP derivatives.

Hard tissue formation of STRO-1-selected rat dental pulp stem cells in vivo.

The objective of this study was to examine hard tissue formation of STRO-1-selected rat dental pulp-derived stem cells, seeded into a calcium phosphate ceramic scaffold, and implanted subcutaneously in mice. Previously, STRO-1 selection was used to obtain a mesenchymal stem cell progenitor subpopulation from primary dental pulp-derived stem cells. In the current study, these cells were cultured with three different media: "BMP-plus" medium containing dexamethasone and 100 ng/mL of rhBMP-2, "odontogenic" medium containing dexamethasone, and "control" medium without supplements. The cell-scaffold complexes were cultured in these media for 1, 4, or 8 days before implantation. Histological analysis demonstrated that the cultures with BMP-plus and 4 days of culture gave the highest percentage of hard tissue formation per implant (36 +/- 9% of pore area). Real-time PCR confirmed these results. In conclusion, STRO-1-selected dental pulp stem cells show effective hard tissue formation in vivo, and a short in vitro culture period and addition of BMP-2 can enhance this effect.