Injectable bone substitute (IBS) materials are commonly used to fill irregular-shaped bone voids in non-load-bearing areas and can offer greater utility over those which are in prefabricated powder, granule, or block forms. This work investigates the impact of liquid-to-solid ratio (LSR) on the rheology and cytocompatibility of IBSs formulated from bioactive glass particles and β-tricalcium phosphate (β-TCP) in glycerol and poly(ethylene glycol) (PEG). IBS formulations of varying LSR were prepared and packed in 3 cc open-bore syringes and sterilized via gamma irradiation (10 kGy, 25 kGy). Gamma-irradiated formulations with high PEG content required the highest (73 N) mechanical force for injection from syringes. Oscillatory viscosity measurements revealed that the viscosity of samples was directly proportional to glycerol content. PEG and glycerol displayed competing effects on the washout resistance and cohesiveness of samples, which were based on total weight loss in media and Ca2+ ion release, respectively. Cell viability in 24-h extracts of 10 kGy gamma-sterilized and 25 kGy gamma-irradiated samples were 22.94% and 56.53%, respectively. The research highlights the complex interplay of IBS components on IBS rheology and, moreover, the cytotoxicity behaviors of beta-tricalcium phosphate-based injectable bone substitutes by in vitro experiments.

Simple bone cysts (SBCs) and intraosseous lipoma (IOL) of the calcaneus are rare tumor entities that are primarily diagnosed due to unspecific heel pain, incidental findings, or rarely due to pathological fractures. Compared to traditional open tumor resections, endoscopic resection of these benign tumors aims to minimize surgical morbidity and maximize surgical efficiency without compromising safety. Grafting is regularly performed to reduce the risk of recurrence and stimulate osseous consolidation of the lytic lesion. As the incidence is low and treatment strategies are heterogeneous, there is no clear consensus for the treatment of simple cysts or intraosseous lipomas of the calcaneus. The objectives of this study are (a) to present medium to long-term results after endoscopic resection and grafting with allogenic cancellous bone or bioresorbable hydroxyapatite and calcium sulfate cement, and (b) to add further evidence to the discussion of whether calcaneal SBC and IOL are the same entity at different developmental stages. Between 2012 and 2019, a total of 25 benign bone tumors consisting of 17 SBCs and 8 IOLs were treated by A.T. with endoscopic resection and grafting, comprising the largest cohort to date. For grafting, 12 patients received allogenic cancellous bone (group A) and 13 patients received injectable bone substitute (group B). Pre- and postoperative imaging using plain X-rays and MRI was retrospectively analyzed with a mean follow-up time of 24.5 months to assess tumor size, osseous consolidation (modified Neer classification), and tumor recurrence. A retrospective chart analysis focusing on adverse intra- and perioperative events and other complications associated with the surgical procedure was performed using the modified Clavien-Dindo classification (CD1-3). A total of 12/13 cases with allogenic bone grafting showed a Neer Type 1 osseous healing of the tumorous lesion after endoscopic resection, whereas only 5/11 cases with injectable bone substitute showed sufficient healing (types 1 and 2). There were three recurrent cysts (Neer 4) and two persistent cysts (Neer 3) after using injectable bone substitute. Two CD1 complications were observed in group A (prolonged wound drainage, sural neuritis) and eight complications were observed in group B (6× CD1, 2× CD3). At least two IOLs diagnosed preoperatively using MRI were ultimately identified as SBCs upon histopathologic examination. Allogenic cancellous bone grafting after endoscopic resection of calcaneal SBC or IOL showed a very low rate of complications and no tumor recurrence in our series. On the other hand, depending on the material used, injectable bone substitute showed a high rate of \"white-out\" (excessive drainage), resulting in multiple complications such as prolonged wound healing, insufficient permanent defect filling, recurrence, and revision surgery. Over time, calcaneal SBC may transform into IOL, exhibiting distinct features of both entities simultaneously during ossoscopy and histopathological analysis.

The injectable bone substitute (IBS) is a self-setting local drug delivery system that adjusts the shape of the bone gap in the fracture. This study aimed to examine the effectiveness of IBS composites of bovine hydroxyapatite (BHA) and alendronate (Ale) in accelerating bone growth in osteoporotic rats. IBS was made by mixing BHA with gelatin 5%, hydroxypropyl methylcellulose (HPMC) 2%, and Ale 10%. The physical properties of IBS were viscosity, injectability, and density tests. Twenty-four female Wistar rats were divided into four groups. After 8 weeks, 2 mm gap was made in the right femur of all rats and filled with IBS. The healing process was observed after 6 weeks with X-ray imaging and H and E staining. The obtained results showed viscosity, injectability, and density value of IBS from 30.4 to 39.4 dPa.s, 98.22%-98.64%, and 0.6325-0.8409 g/cm3, respectively. X-ray imaging and histology results proved the condition of osteoporosis in rats with ovariectomy. The addition of BHA-Gel-HPMC-Ale significantly affected the number of osteoblasts, osteocytes, and osteoclasts (P < 0.05). After 45 days of observation, the addition of BHA-Gel-HPMC-Ale showed the highest mean number of osteoblasts, osteocytes, and osteoclasts, which were 25.00 ± 3.00, 64.33 ± 11.15, and 5.67 ± 0.58 compared to BHA-Gel-HPMC and positive control groups. The BHA-Gel-HPMC-Ale IBS has the potential to reverse osteoporosis. Nevertheless, the underlying potential of these biomaterials to reverse osteoporosis needs further research.

Demineralized bone matrix (DBM), has been used as a bone-graft material because of its osteoconductivity and osteoinductivity. However, the previous research report that supports the single use of DBM is limited by its rapid resorption caused by the lack of calcium and phosphate. β-Tricalcium phosphate (TCP) is an enriched calcium phosphate material suitable for bone healing with osteoconductive properties. In this study, we have developed injectable bone graft by the loading two kinds of TCP in DBM particles and thermo-sensitive DBM-derived hydrogel (hDBM). TCP powder (pTCP) and TCP granules (gTCP) were loaded into hDBM and DBM, respectively. The bone formation effect was investigated according to the morphological features of TCP. Residual growth factor concentrations were investigated; microstructure and morphology were characterized by SEM. In-vitro studies showed that hDBM/DBM/pTCP and hDBM/DBM/gTCP bone grafts were biocompatible and could promote osteogenesis by up-regulating the expression of Runx2 and OPN, bone-related genes. In-vivo studies using the rabbit-femur defect model revealed that the implanted hDBM/DBM/pTCP bone graft showed similar histology to that of fibrous dysplasia with the expression of CD68, whereas hDBM/DBM/gTCP showed good bone formation. Loading of gTCP in place of pTCP was noticed as an effective way to improve bone regeneration in an injectable hDBM/DBM hydrogel-based bone graft.

In the present study, an injectable bone substitute system which utilized porous bioglass (BG)-derived granules supplemented with hyaluronic acid (Hya), was evaluated. Hya plays ultimate role in wound healing, promoting cell motility. The BG were synthesized by a simple and low sintering temperature process without any foreign phase incorporation. Furthermore, the physical properties in the porous scaffold were optimized to investigate thein vitroandin vivoperformance. The porous BG60 scaffolds system showed excellent bioactivity in anin vitrosimulated body fluid test in which the ions dissolved from the composite materials influenced apatite growth, countered the acidic pH, and increased material degradation. In anin vitrostudy with pre-osteoblasts cells (MC3T3-E1), the porous scaffold supported cell adhesion and proliferation. A post-implantation study conducted in femoral defects showed implant degradation and surprisingly fast bone formation just after 2 weeks of implantation. Initialin vivodegradation of Hya promotes releasing ions which regulates the bone forming cells, clues to tissue repair, and regeneration. On the other hand it also prevent the scattering of BG granule after grafting at implant site. The faster dissolution of the porous BG scaffold increased the resorption of the composite material and hence, facilitated bone tissue regeneration. Our findings suggest that the porous BG scaffold could potentially be used as an injectable bone substitute for fast, early bone regeneration applications.

The purpose of this study was to evaluate the effects of an injectable composite made up of calcium sulfate (CAS), fluvastatin (FS), and atelocollagen on bone augmentation in rats. Porous structures and compressive strength of composites were evaluated. The cumulative release kinetics of FS were determined in vitro by a spectrophotometer. To observe bone regeneration in vivo, five different materials (normal saline; atelocollagen gel only; composite of CAS and atelocollagen; composite containing 0.5% FS; and composite containing 1.0% FS) were injected in extraction sockets and in the crania of rats. Microcomputed tomography and histological evaluation were performed after 2, 4, and 8 weeks of healing time. The composites had high porosity (greater than 55%). FS kept a slow and stable release for >30 days. In vivo results demonstrated that more new bone was formed in the FS groups compared with the other groups, and both bone mass and bone density had prominent increase in maxillae and crania. Resorption of the composite was also observed for cranial tissues. In conclusion, this composite can be applied percutaneously, without any incision. It has excellent properties with replaceability into bone and anabolic effects for bone formation, as well as a drug delivery system for bone formation.

We described a method to produce an injectable bone substitute consisting of a solid and liquid phase, this solid was formed using the coacervation method consisting of a mixture of Hydroxyapatite (HAp) and beta-Tricalcium Phosphate (β-TCP) which the sodium alginate - precursor - was removed during sinterization. The biphasic calcium phosphate microspheres had varying size distributions depending on the flow rate and these microspheres were mixed with a polymeric solution, chitosan and polyethylene glycol, and depending on the ratio of these phases, the injectability results varied. Nonetheless, the force required for complete removal will not disrupt the accuracy of injection into the bone defect while the biomaterial exhibited no cytotoxicity with promising results from in vivo using tibia bone defect in rabbits at 30 and 60 days whereas bone repair was more intense and accentuated with the usage of the biomaterial, and was gradually absorbed during the evaluated periods.

OBJECTIVE: The aim of this in vivo study was to evaluate the utility of bone graft gel containing hydroxyapatite nanoparticles in promoting bone regeneration in a mouse model of vertical bone augmentation.
METHODS: Gel implants with high and low viscosity were compared for their bone regenerating ability. Bone formation at 12 weeks and material reactions were observed radiographically and histologically.
RESULTS: Radiological analysis showed that most bone augmentation area in the graft material occurred in the fourth week after surgery regardless of the viscosity of the gel, and then gradually decreased. The volume of bone augmentation area was greater in the high-viscosity implant group than in the low-viscosity implant group at all time points, the difference was statistically significant at 8 and 12 weeks. Histological evaluation indicated that the new bone area was significantly smaller in the high-viscosity implant group.
CONCLUSIONS: Gelatinous graft materials containing hydroxyapatite nanoparticles were confirmed to be useful in vertical bone augmentation.

Bioglass-calcium phosphate cement (CPC) composite materials have recently received increased attention for bone regeneration purposes, owing to their improved properties in term of biocompatibility and bone ingrowths. In this study, an injectable bone substitute (IBS) system which utilizes bioglass microspheres incorporated into brushite based cement, was evaluated. The microspheres were synthesized with a simple and low sintering temperature process; there was no significant phase difference shown from the powder and good interactivity with cells was obtained. Furthermore, physical properties were optimized in microsphere incorporated brushite cement in order to investigate in vitro and in vivo performance. Accordingly, setting time and compressive strength were hardly altered until a microsphere content of 40% (v/v) was reached. The brushite (BR)/bioglass microsphere (BM) system showed excellent bioactivity to the in-vitro simulated body fluid test: dissolution ions from composite materials influenced apatite growth, countered acidic pH, and increased material degradation. In an in-vitro study with preosteoblasts (MC3T3-E1), BR/BM supported cell adhesion and proliferation, while cell differentiation experiments without osteogenic supplements, demonstrated that BR/BM induced osteogenic differentiation. A post-implantation study conducted in femoral defects showed higher materials degradation and bone formation in BR/BM than in BR. The faster dissolution of bioglass microspheres increased BR/BM composite resorption and hence facilitated bone tissue integration. Our findings suggest that bioglass microspheres incorporated in cement could potentially be used as an injectable bone substitute for bone regeneration applications.

The present work is to investigate the efficiency of hyaluronic acid (HyA) supplemented biphasic calcium phosphate (BCP) injectable granule to promote the bone regeneration. The effect of adding HyA to the multichannel BCP granule (MCG-HyA) was studied in terms of morphology, chemical structure, porosity, in-vitro and in-vivo biocompatibility, RT-PCR, western blot and compared with MCG. The addition of HyA to MCG successfully made the granules injectable type. In-vivo studies in rabbit model showed an enhancement in bone formation after 4 weeks of implantation and better handling characteristics for MCG-HyA than MCG. RT-PCR and Western Blotting studies revealed that MCG-HyA significantly unregulated the osteogenic gene and protein expressions respectively. Our results indicated that MCG-HyA could be used as a promising injectable bone substitute in clinical applications.