Biphasic Calcium Phosphate Research Articles

Enhanced osteogenesis and angiogenesis of biphasic calcium phosphate scaffold by synergistic effect of silk fibroin coating and zinc doping.

Biphasic calcium phosphate (BCP) scaffold has been widely applied to bone regeneration because of its good biocompatibility and bone conduction property. However, the low mechanical strength and the lack of angiogenic and osteogenic induction properties have restricted its application in bone tissue regeneration. In this study, we combined zinc (Zn2+) doping and silk fibroin (SF) coating with expectation to enhance compressive strength, osteogenesis and angiogenesis of BCP scaffolds. The phase composition, morphology, porosity, compressive strength, in vitro degradation and cell behaviors were investigated systematically. Results showed that the scaffold coated with SF exhibited almost 3 times of compressive strength without compromising its porosity compared with the uncoated scaffold. Zn2+ doping and SF coating synergistically enhanced the alkaline phosphatase activity and osteogenesis-related genes expression of mouse bone mesenchymal stem cells (mBMSCs). Furthermore, SF coating notably improved the proliferation, cell viability and in vitro angiogenesis of human umbilical vein endothelial cells (HUVECs). This work provides a novel way to modify BCP scaffolds simultaneously with enhancing mechanical strength and biological properties.

Leukemia Inhibitory Factor Facilitates Self-Renewal and Differentiation and Attenuates Oxidative Stress of BMSCs by Activating PI3K/AKT Signaling.

Objective Transplantation of bone marrow-derived mesenchymal stem cells (BMSCs) remains a hopeful therapeutic approach for bone defect reconstruction. Herein, we investigated the effects and mechanisms of leukemia inhibitory factor (LIF) in the function and viability of hypoxic BMSCs as well as bone defect repair. Methods The effects of LIF on apoptosis (flow cytometry, TUNEL staining), mitochondrial activity (JC-1 staining), proliferation (colony formation, EdU staining), and differentiation (CD105, CD90, and CD29 via flow sorting) were examined in hypoxic BMSCs. LIF, LIFR, gp130, Keap1, Nrf2, antioxidant enzymes (SOD1, catalase, GPx-3), bone-specific matrix proteins (ALP, BSP, OCN), PI3K, and Akt were detected via immunoblotting or immunofluorescent staining. BMSCs combined with biphasic calcium phosphate scaffolds were implanted into calvarial bone defect mice, and the therapeutic effect of LIF on bone defect was investigated. Results Hypoxic BMSCs had increased apoptosis and oxidative stress and reduced mitochondrial activity. Additionally, LIF, LIFR, and gp130 were upregulated and PI3K/Akt activity was depressed in hypoxic BMSCs. Upregulated LIF alleviated apoptosis and oxidative stress and heightened mitochondrial activity and PI3K/Akt signaling in hypoxic BMSCs. Additionally, LIF overexpression promoted self-renewal and osteogenic differentiation of BMSCs with hypoxic condition. Mechanically, LIF facilitated self-renewal and differentiation as well as attenuated oxidative stress of BMSCs through enhancing PI3K/AKT signaling activity. Implantation of LIF-overexpressed BMSC-loaded BCP scaffolds promoted osteogenesis as well as alleviated oxidative stress and apoptosis through PI3K/Akt signaling. Conclusion Our findings demonstrate that LIF facilitates self-renewal and differentiation and attenuates oxidative stress of BMSCs by PI3K/AKT signaling.

Regenerative potential of biphasic calcium phosphate and enamel matrix derivatives in the treatment of isolated interproximal intrabony defects: a randomized controlled trial.

BackgroundThe combined use of biomaterials for regeneration may have great biological relevance. This study aimed to compare the regenerative potential of biphasic calcium phosphate (BCP) alone and with growth factor enamel matrix derivatives (EMDs) for the regeneration of intrabony defects at 1 year.MethodsThis randomized controlled trial included 40 sites in 29 patients with stage II/III periodontitis and 2/3 wall intrabony defects that were treated with BCP alone (control group) or a combination of BCP and EMD (test group). BCP alloplastic bone grafts provide better bio-absorbability and accelerate bone formation. EMDs are commercially available amelogenins. Mean values and standard deviations were calculated for the following parameters: plaque index (PI), papillary bleeding index (PBI), vertical probing pocket depth (V-PPD), vertical clinical attachment level (V-CAL), and radiographic defect depth (RDD). Student paired and unpaired t-tests were used to compare the data from baseline to 12 months for each group and between the groups, respectively. The results were considered statistically significant at p<0.05.ResultsAt 12 months, the PI and PBI scores of the control and test groups were not significantly different (p>0.05). The mean V-PPD difference, V-CAL gain, and RDD difference were statistically significant in both groups at 12 months (p<0.001 for all parameters). Intergroup comparisons showed that the mean V-PPD reduction (2.13±1.35 mm), V-CAL gain (2.53±1.2 mm), and RDD fill (1.33±1.0 mm) were statistically significant between the groups at 12 months (p<0.001 for all parameters).ConclusionBCP and EMDs combination is a promising modality for the regeneration of intrabony defects.

Sr and Mg Doped Bi-Phasic Calcium Phosphate Macroporous Bone Graft Substitutes Fabricated by Robocasting: A Structural and Cytocompatibility Assessment.

Bi-phasic calcium phosphates (BCPs) are considered prominent candidate materials for the fabrication of bone graft substitutes. Currently, supplemental cation-doping is suggested as a powerful path to boost biofunctionality, however, there is still a lack of knowledge on the structural role of such substituents in BCPs, which in turn, could influence the intensity and extent of the biological effects. In this work, pure and Mg- and Sr-doped BCP scaffolds were fabricated by robocasting from hydrothermally synthesized powders, and then preliminarily tested in vitro and thoroughly investigated physically and chemically. Collectively, the osteoblast cell culture assays indicated that all types of BCP scaffolds (pure, Sr- or Sr–Mg-doped) delivered in vitro performances similar to the biological control, with emphasis on the Sr–Mg-doped ones. An important result was that double Mg–Sr doping obtained the ceramic with the highest β-tricalcium phosphate (β-TCP)/hydroxyapatite mass concentration ratio of ~1.8. Remarkably, Mg and Sr were found to be predominantly incorporated in the β-TCP lattice. These findings could be important for the future development of BCP-based bone graft substitutes since the higher dissolution rate of β-TCP enables an easier release of the therapeutic ions. This may pave the road toward medical devices with more predictable in vivo performance.

SURGICAL MANAGEMENT OF GOLDENHAR SYNDROME PATIENT WITH ORTHOGNATHIC SURGERY AND HEMIMANDIBULAR PROSTHESIS: A CASE REPORT

Goldenhar syndrome, also known as hemifacial microsomia (HFM), is a rare congenital disease associated with facial asymmetry and hypoplasia of the jaw. We describe the clinical case of a 13-year-old boy with HFM treated with orthognathic surgery and a customized biphasic calcium phosphate (BCP) bioceramic hemimandibular prosthesis. Clinical and radiographic examination revealed facial asymmetry, right hemifacial microsomia, hypoplasia of the mandible, retrognathism, hypomentonism, canted occlusal plan, and epibulbar dermoid. The primary treatment consisted of bimaxillary orthognathic surgery for occlusal plane correction. Five years later, the patient underwent surgery for hemimandibular reconstruction using BCP customized prosthesis for correction of asymmetry. An improvement of esthetics was noted after surgery.

Effect of Photobiomodulation Therapy Associated With Biphasic Phosphate Calcium on Bone Repair: A Histomorphometric Study in Rats.

Introduction: This study aimed to investigate the effects of photobiomodulation (PBM) therapy associated with biphasic calcium phosphate on calvaria critical defects in rats. Methods: Forty-eight (90 days old) adult male rats (Rattus norvegicus, Albinus variation, Wistar) received critical defects of 5 mm in diameter, which were made on their skull, and they were randomly assigned into the following groups: C-blood clot, B-biphasic calcium phosphate, L-photobiomodulation therapy, and B + L-biphasic calcium phosphate + photobiomodulation therapy. A low-level a gallium aluminum arsenide (GaAlAs) laser was applied in a single dose during surgery, in a wavelength of 660 nm and total energy density of 45 J/cm2. On 30th and 60th days, the animals from each group were euthanized. Histological and histomorphometric analyses were performed. Results:In 30 days, almost all specimens (C, L, B and B + L) showed bone neoformation areas in regions near the borders of the surgical defect. In 60 days, in many specimens (C, L, B, B + L), it was possible to see a narrow neoformed bone structure along almost the whole extension of the surgical defect, though it was thinner than the original calvary bone. Data were recorded as mean ± standard deviation, and after normality was tested, a suitable statistical test was applied (α = 5%). On day 60, there was a statistically significant difference when comparing the proportion of neoformation area between group L (0.52%±0.13) and group B+L (0.20%±0.08). Group L showed a difference compared with all the groups when we compared the remaining distance between the edges of neoformed bone (C×L, P=0.0431; B × L, P=0.0386; L×B+L, P=0.0352), demonstrating a great defect closure. Conclusion: Our findings suggest that although biphasic calcium phosphate exerts some osteogenic activity during bone repair, PBM therapy is not able to modulate this process.

Enhanced reparative dentinogenesis of biphasic calcium phosphate ceramics containing calcium-deficient hydroxyapatite (CDHA) and strontium-incorporated CDHA in direct pulp capping

Despite that dental materials and therapeutic strategies have made progress, reserving dental pulp vitality remains a great challenge during clinical practices of dentistry. Biphasic calcium phosphate (BCP) ceramic is a promising biomaterial for bone regeneration and dentin restoration, whose biological activities need to be further improved to meet the needs of regenerative medicine. Calcium-deficient hydroxyapatite (CDHA), which is more similar in composition to natural bone and tooth minerals, imparts superior bioactivity to BCP ceramics. Besides, owing to the positive roles of strontium in stimulating bone formation, the strontium-incorporated CDHA has excellent biocompatibility and further promotes bone regeneration. However, the effect of CDHA and strontium-incorporated CDHA in dental pulp capping has been rarely investigated. Herein, we presented an effective method to integrate CDHA and strontium-incorporated CDHA into BCP ceramics and investigated their biological properties as a potential strategy for dental pulp-capping and dentin regeneration. The characterization of phase composition and crystal structure confirmed that novel BCP ceramics with high CDHA content (BCP-A) and strontium-incorporated CDHA (BCP-A/Sr) have been successfully synthesized. In vitro, BCP-C (conventional BCP), BCP-A, and BCP-A/Sr provided an appropriate microenvironment and significantly promoted the proliferation of human dental pulp cells (hDPCs). BCP-A did not affect the migration of hDPCs, yet BCP-C and BCP-A/Sr displayed a suppressing trend. In vivo, BCP-A significantly reduced the inflammation response and improved the formation of reparative dentin bridges. Compared with mineral trioxide aggregate (MTA), a clinical direct pulp capping material, the texture of reparative dentin bridges in BCP-A showed more uniformity. Moreover, RNA-seq analysis further clarified that direct co-culture of hDPCs with BCP-A mainly up-regulated the cell cycle signaling pathway and CDC20 protein. These findings indicate that the novel BCP-A is a promising biomaterial for direct dental pulp capping due to its unique intrinsic biological activities, which warrants further investigation.

A Novel Hollow‐Tube‐Biphasic‐Whisker‐Modified Calcium Phosphate Ceramics with Simultaneously Enhanced Mechanical Strength and Osteogenic Activity

AbstractDue to the inherent brittleness and low mechanical strength, it is still a challenge for calcium phosphate (Ca‐P) ceramics to be used in load‐bearing bone defect repair. To achieve a good balance between mechanical strength and osteogenic activity, hollow‐tube‐whisker‐modified biphasic calcium phosphate (BCP) ceramics (BCP‐HW) are successfully fabricated by an in situ growth process in the present study. Compared to the initial BCP ceramics (BCP‐C) and those with solid whiskers, BCP‐HW exhibits larger specific surface area (3.9 times vs BCP‐C) and higher mechanical strength (3.4 times vs BCP‐C), endowing it with stronger stimulation on adhesion, proliferation, and osteogenic differentiation of bone marrow mesenchymal stem cells. In an intramuscular implantation model of canine, BCP‐HW shows excellent osteoinductivity and promotes the maturation of new bone, and the resultant compressive strength of the implant increases to ≈12 MPa at 3 months postoperatively. In another critical‐sized segmental bone defect model of rabbit femur, BCP‐HW has the best repairing effect. After implantation for 6 months, much more new bone ingrowth and higher bending load are observed in BCP‐HW than BCP‐C. Collectively, these findings suggest that the in situ hollow‐tube whisker construction possesses immense potential in expanding the applications of Ca‐P ceramics to load‐bearing bone defect repair.

Conversion of borate glass microspheres to hollow biphasic calcium phosphate ceramic microspheres for bone defects

AbstractHollow biphasic calcium phosphate (BCP) microspheres with different fractions of nanocrystalline hydroxyapatite (HA) and β‐tricalcium phosphate (β‐TCP) were prepared, and their in vitro and in vivo reactivity were assessed. The microspheres were prepared by reacting soluble borate glass microspheres in phosphate solutions (0.1‐ or 0.25‐M K2HPO4) with different values of pH (7–12) to produce hollow calcium‐deficient hydroxyapatite (CDHA) microspheres with Ca/P ratios that increased with solution pH. Heating the hollow CDHA microspheres to 800°C produced hollow, nanoporous BCP microspheres with ratios of nanocrystalline HA and β‐TCP that correspond to the Ca/P ratio of the CDHA precursors. Hollow BCP microspheres with a greater fraction of β‐TCP released calcium ions most quickly in a K‐acetate acidic buffer solution, consistent with their greater bioactivity in the in vivo tests. CDHA and BCP microspheres converted to nanocrystalline carbonated HA when immersed in 37°C simulated body fluid. Hollow CDHA and BCP microspheres were implanted in rat calvarial defects, and after 8 weeks, the BCP microspheres with a β‐TCP/HA ratio of 70/30 (by weight) promoted a greater fraction of new bone formation (33% ± 7%) compared to BCP microspheres with a β‐TCP/HA ratio of 44/56 (20% ± 9%) and compared to the respective CDHA precursor microspheres (17% ± 4%, 17% ± 5%). Bone formation was not observed in the subcutaneous implants of any types of these hollow microspheres, after 8 and 12 weeks, and the more reactive BCP microspheres (β‐TCP/HA = 70/30) were associated with significantly more blood vessel formation in the subcutaneous implants. These results indicate that the properties of hollow BCP microspheres can be tailored by their processing conditions and are useful as scaffold materials for tissue regeneration.

Synthesis and Thermal Behaviour of Calcium Alkyl Phosphates as Bioceramic Precursors

Powders of alkyl phosphoric acids and calcium alkyl phosphates with various alkyl chains (butyl, octyl, and dodecyl) have been synthesized. The resulting powders were characterized by X-ray phase analysis, electron microscopy, and thermal analysis. It was shown that the calcium alkyl phosphates correspond to the composition of acid salts of calcium alkyl phosphates Ca(RPO4H)2, data on which are not presented in the literature. The thermal behaviour of calcium alkyl phosphates can be described as a complex phase transformation into biphasic calcium phosphate mixture (of Ca2P2O7 and Ca3(PO4)2) with the increase of the Ca to P ratio in comparison to initial materials. The powders thermally treated in the range of 400–600 °C could be recommended as single precursors of biphasic bioceramics.

Powder 3D Printing of Bone Scaffolds with Uniform and Gradient Pore Sizes Using Cuttlebone-Derived Calcium Phosphate and Glass-Ceramic.

The pore geometry of bone scaffolds has a major impact on their cellular response; for this reason, 3D printing is an attractive technology for bone tissue engineering, as it allows for the full control and design of the porosity. Calcium phosphate materials synthesized from natural sources have recently attracted a certain interest because of their similarity to natural bone, and they were found to show better bioactivity than synthetic compounds. Nevertheless, these materials are very challenging to be processed by 3D printing due to technological issues related to their nanometric size. In this work, bone scaffolds with different pore geometries, with a uniform size or with a size gradient, were fabricated by binder jetting 3D printing using a biphasic calcium phosphate (BCP) nanopowder derived from cuttlebones. To do so, the nanopowder was mixed with a glass-ceramic powder with a larger particle size (45–100 µm) in 1:10 weight proportions. Pure AP40mod scaffolds were also printed. The sintered scaffolds were shown to be composed mainly by hydroxyapatite (HA) and wollastonite, with the amount of HA being larger when the nanopowder was added because BCP transforms into HA during sintering at 1150 °C. The addition of bio-derived powder increases the porosity from 60% to 70%, with this indicating that the nanoparticles slow down the glass-ceramic densification. Human mesenchymal stem cells were seeded on the scaffolds to test the bioactivity in vitro. The cells’ number and metabolic activity were analyzed after 3, 5 and 10 days of culturing. The cellular behavior was found to be very similar for samples with different pore geometries and compositions. However, while the cell number was constantly increasing, the metabolic activity on the scaffolds with gradient pores and cuttlebone-derived powder decreased over time, which might be a sign of cell differentiation. Generally, all scaffolds promoted fast cell adhesion and proliferation, which were found to penetrate and colonize the 3D porous structure.

Guided Bone Regeneration Prior to Implant Therapy in the Esthetic Zone: A Case Report

Background:Implant placement on the anterior maxilla requires a comprehensive concern of adequate alveolar bone and gingival biotype. Insufficient support for hard and soft tissues becomes one of the main causes of implant complications. To avoid this problem, pre-surgical treatment with Guided Bone Regeneration (GBR) is suggested with respect to the alveolar ridge defect.Objective:In this study, we reported the predictability of a sandwich technique of GBR prior to implant placement.Case Presentation:Case: Thirty-four-year-old male was referred with a chief complaint of missing teeth on area #21 accompanied by labial bone dehiscence due to trauma. Bone thickness measurement displayed was approximately 3/4/5 mm (coronal/middle third/apical third of the alveolar ridge). This case was diagnosed as soft and hard tissue deformity on area #21 prior to implant. Case management: GBR and implant placement were performed in two stages of surgery. GBR was executed by using a combination of Biphasic Calcium Phosphate (BCP) (consisting of 40% Beta-Tri Calcium Phosphate (β-TCP) and 60% Hydroxyapatite (HA)) and Demineralized Freeze-Dried Bone Allografts (DFDBA) bone graft materials. Alveolar ridge re-measurement five months after GBR showed significant bone augmentation, 6/6/6 mm (coronal/middle third/apical third of the alveolar ridge). Furthermore, a 3.5 mm (diameter) x 10 mm (length) bone-level implant was inserted accordingly. The two-year follow-up exhibited gingival stability and no sign of either recession or bone resorption.Conclusion:Herein, we demonstrated the effectiveness of BCP and DFDBA bone substitutes to obtain proper conditions for the long-termed stability and predictability of implant.

Microstructures and mechanical properties of biphasic calcium phosphate bioceramics fabricated by SLA 3D printing

Porous biphasic calcium phosphate (BCP) bioceramics are considered to be the most promising bone repair materials in clinical medicine. Stereolithography (SLA) 3D printing can precisely fabricate bioceramic scaffolds with complex porous structure. During this process, how to obtain a suitable sintering procedure for BCP bioceramics by SLA 3D printing is the key to determine the microstructures and mechanical properties, but this is absent. In this present study, we prepared BCP bioceramics with superior densification and mechanical properties by SLA 3D printing, and mainly investigated the effects of sintering temperature and condition on the microstructures and mechanical properties of SLA 3D printed BCP bioceramics for the first time. At the optimized procedure of sintering temperature 1250 °C for 2 h, the 3D printed BCP bioceramics showed uniform shrinkage in all directions, and especially the mechanical properties were close to that of human cortical bone. Furthermore, complex porous BCP scaffolds with high porosity (51.49 %) and compressive strength (8.14 MPa) were successfully obtained. BCP bioceramics greatly promoted the proliferation of MC3T3-E1 cells by the release of Ca and P ion and presented excellent bioactivity. This work proves that SLA 3D printing technology can prepare BCP bioceramics with high mechanical and functional properties, and provides a new route for manufacturing high performance BCP bioceramic scaffolds with complex structure by SLA 3D printing for repairing bone defect in clinical.

HA/β-TCP Biphasic Calcium Phosphate Ceramics Derived From Butterfish Bones Loaded With Bone Marrow Mesenchymal Stem Cells Promote Osteogenesis

Biphasic calcium phosphate (BCP) containing hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP) is considered the gold standard for bone repair. However, the complex synthesis of BCP restricts the wide application of BCP. In the preliminary work, we proved that the HA/β-TCP BCP ceramic material extracted from fishery waste had a good prospect as a bone replacement graft material for filling bone defects. This study aimed to explore the osteogenesis effect of BCP material derived from butterfish bones in vivo and in vitro. After loading human bone marrow mesenchymal stem cells (hBMSCs) with materials, we used scanning electron microscopy to observe cell adhesion and survival. Western blot analysis was used to detect osteogenic expression in vitro. Micro-computed tomography and hematoxylin–eosin staining were used to detect bone regeneration and material degradation rate in the rat femoral defect model. The results showed that hBMSCs grew well and adhered closely to the material. In vitro, the expression levels of bone formation–related markers ALP and Runx-2 of butterfish bones calcined at 900°C were generally higher than those in the other groups. Notably, in vivo, the osteogenesis ability of butterfish bones calcined at 900°C was almost comparable to that of the most commonly used Bio-Oss in clinical practice and was significantly better than that in the other groups. In summary, the BCP ceramic material derived from butterfish bones had good biocompatibility, osteoconductivity, and osteoinductivity, and had a good application prospect in the field of bone graft substitutes.

Preparation and characterization of two novel osteoinductive fishbone-derived biphasic calcium phosphate bone graft substitutes.

Many studies have reported on the conversion of natural resources into xenografts with hydroxyapatite (HA) as major component, but the extraction of biphasic calcium phosphate (HA/β-TCP) from animal bones and transformation into bone graft substitutes are rarely reported. In this research, two kinds of fish bones were made into granular porous biphasic calcium phosphate bone graft substitutes with particle sizes between 500 to 1000μm through a series of preparation procedures (Salmo salar calcined at 900°C named Sa900 and Anoplopoma fimbria calcined at 800°C named An800). The chemical composition was characterized by X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). The morphology and porous structure of the scaffolds were comparatively analyzed by scanning electron microscopy (SEM) and mercury porosimeter. The specific surface area of materials was measured by the nitrogen adsorption technique based on BET theory. Cytotoxicity and ectopic osteogenesis were also carried out to investigate the biocompatibility and osteoinductive potential of these materials. The results showed that both fishbone-derived scaffolds were composed of HA and β-TCP with different proportions, and numerous interconnected pores with different sizes were observed at the surface of materials. An800 had higher total porosity reaching 74.8% with higher interconnectivity and micropores mostly distributed at 0.27μm and 0.12μm, while Sa900 had a higher specific surface area and higher intraparticle porosity with nanopores mostly distributed at 0.07μm. CCK-8 assays and Live/dead staining demonstrated excellent biocompatibility. Material-induced osteoid formation were observed on the interface of both internal pores and periphery of materials after implantation in muscle pouch of Wistar rats for 8weeks which indicated some extent of osteoinductive potential of materials. The possible mechanism of material-induced osteogenesis and the effects of chemical composition, surface topography, and spatial structure on osteogenesis were also discussed in this paper.

Biological evaluation of polydopamine and chitosan composite coatings on the 3D printed porous biphasic calcium phosphate scaffold

A current, gradually growing trend is to use bioactive ceramics to develop new bone scaffolds to replace traditional bone implants. Biphasic calcium phosphate (BCP), a mixed material of hydroxyapatite and tricalcium phosphate, has limited osseointegration ability. Therefore, improving the bioactivity and osteoinductive ability of BCP scaffolds through various surface modification methods is an important issue. Polydopamine (PDA) coating, chitosan (CS) coating, and their composite coatings have been previously reported to individually possess a good ability to improve scaffold bioactivity, but few comparisons between them and the optimal coating sequence of PDA and CS have been reported. Herein, we fabricated a BCP scaffold by using a self-made 3D printer, sintered the scaffold, and then prepared PDA, CS, PDA/CS and CS/PDA coatings by solution immersion. Afterwards, the surface morphology, element composition and proportion, compressive strength and surface contact angle of the five groups of scaffolds were characterized. Finally, the adhesion, proliferation and osteogenesis-related gene expression levels of MC3T3-E1 on the five groups in vitro were evaluated in detail. The results showed that the five types of scaffolds were fabricated successfully and that the PDA/CS and CS/PDA coatings had different surface morphologies. The compressive strengths of the composite-coating groups were higher than those of the single-coating groups and the BCP group. The CCK-8 experiment revealed that the composite coatings can promote the initial adhesion and proliferation of cells. The detection results of three osteogenesis-related genes (OCN, COL-1 and Runx-2) proved that the composite coatings had a synergistic effect in promoting osteodifferentiation and that the CS/PDA coating had the greatest effect in improving cell viability and promoting osteogenic differentiation.

In Vitro Biocompatibility of a Novel Semi-Rigid Shell Barrier System: As a New Application for Guided Bone Regeneration.

This study evaluated the in vitro biocompatibility of a novel, semi-rigid shell barrier system for guided bone regeneration (GBR) based on polycaprolactone and biphasic calcium phosphate membranes and consisting of a semi-rigid shell (SR) and two semi-resorbable barrier membranes, i.e., a buffered (BF) and an airdried (AD) membrane. In vitro biocompatibility, cell cytotoxicity, cell proliferation and differentiation were evaluated with osteoblast (MC3T3-E1) and fibroblast (L929) cells compared to the d-PTFE membrane (Cytoplast®, CP). The osteoblasts and fibroblasts were well attached and proliferated on all materials from days 1, 3, and 7 without cell cytotoxicity. All groups showed that osteoblast and fibroblast cell proliferation increased from day 1 to day 14–17 and decreased on day 21. On day 21, the CP membrane presented significantly higher osteoblast cell numbers than the BF membrane and the SR shell (p = 0.000). The CP membrane presented a significantly higher amount of fibroblast cells than the other groups (p = 0.000). The SR shell and the BF membrane demonstrated higher osteoblast cell differentiation markers including ALP activity, osteocalcin, and mineral secretion than the CP and the AD membrane. The semi-rigid shell barrier system demonstrated good in vitro biocompatibility and supported osteogenic cell proliferation and differentiation better than the d-PTFE membrane.

Three-dimensionally printed biphasic calcium phosphate blocks with different pore diameters for regeneration in rabbit calvarial defects

BackgroundBiphasic calcium phosphate (BCP) is the most frequently used synthetic bone substitutes, which comprises a combination of hydroxyapatite (HA) and beta-tricalcium phosphate (b-TCP). Thanks to the recent advances in digital dentistry and three-dimensional (3D) printing technology, synthetic block bone substitutes can be customized to fit individual defect morphologies. The diameter of the pores can influence the rate of bone formation and material resorption. The aim of this study was to compare three-dimensionally printed biphasic calcium phosphate (BCP) block bone substitutes with different pore diameters (0.8-, 1.0-, and 1.2- mm) for use in the regeneration of rabbit calvarial defects.MethodsFour circular defects were formed on the calvaria of ten rabbits. Each defect was randomly allocated to one of the following study groups: (i) control group, (ii) 0.8-mm group, (iii) 1.0-mm group, and (iv) 1.2-mm group. All specimens were postoperatively harvested at 2 and 8 weeks, and radiographic and histomorphometric analyses were performed on the samples.ResultsHistologically, the BCP blocks remained unresorbed up to 8 weeks, and new bone formation occurred within the porous structures of the blocks. After the short healing period of 2 weeks, histomorphometric analysis indicated that new bone formation was significantly greater in the BCP groups compared with the control (p < 0.05). However, there were no significant differences between the groups with different pore diameters (p > 0.05). At 8 weeks, only the 1.0-mm group (3.42 ± 0.48 mm2, mean ± standard deviation) presented a significantly larger area of new bone compared with the control (2.26 ± 0.59 mm2) (p < 0.05). Among the BCP groups, the 1.0- and 1.2-mm groups exhibited significantly larger areas of new bone compared with the 0.8-mm group (3.42 ± 0.48 and 3.04 ± 0.66 vs 1.60 ± 0.70 mm2, respectively).ConclusionsWithin the limitations of this study, the BCP block bone substitutes can be applied to bone defects for successful bone regeneration. Future studies should investigate more-challenging defect configurations prior to considering clinical applications.

Digital light processing (DLP) of nano biphasic calcium phosphate bioceramic for making bone tissue engineering scaffolds

Nano-sized ceramic particles are beneficial for the properties of 3D printed structures via digital light processing (DLP). However, achieving optimal conditions for the preparation and processing of ceramic nanoparticle-based slurry is a significant challenge. In this work, a stable and well dispersed slurry with nano-sized biphasic calcium phosphate (BCP) powders was developed and porous BCP bioceramic scaffolds with good bioactivity were accurately fabricated via the DLP additive manufacturing technology. Effects of dispersant, photoinitiator and solid loading on rheological properties and curing abilities of BCP slurries were systematically studied. The slurry having 65 wt % of nano-sized BCP powders showed a relatively low viscosity of 400 mPa s at the 50 s−1 shear rate. It was found that the optimal photoinitiator concentration was tightly associated with the energy dosage in DLP. Furthermore, the effects of sintering temperature (1100 °C, 1200 °C and 1300 °C) on the surface morphology, shrinkage, phase constitution, hardness, compressive properties, and in vitro biological performance of DLP-formed bioceramics were comprehensively investigated. The results showed that DLP-formed BCP ceramics sintered at 1300 °C had the best mechanical properties, in vitro cytocompatibility, and homogeneous bone-like apatite formation ability. Finally, DLP-formed 3D scaffolds with different pore sizes ranging from 300 μm to 1 mm were successfully fabricated, which exhibited high fidelity and accuracy. The current study has demonstrated the great potential of using DLP technology to construct functional complex BCP bioceramic scaffolds for bone tissue regeneration.

A Comparative In Vitro and In Vivo Study of Osteogenicity by Using Two Biomaterials and Two Human Mesenchymal Stem Cell Subtypes.

Background: Bone repair induced by stem cells and biomaterials may represent an alternative to autologous bone grafting. Mesenchymal stromal/stem cells (MSCs), easily accessible in every human, are prototypical cells that can be tested, alone or with a biomaterial, for creating new osteoblasts. The aim of this study was to compare the efficiency of two biomaterials—biphasic calcium phosphate (BCP) and bioactive glass (BG)—when loaded with either adult bone marrow mesenchymal stem cells (BMMSCs) or newborn nasal ecto-mesenchymal stem cells (NE-MSCs), the latter being collected for further repair of lip cleft-associated bone loss. Materials and Methods: BMMSCs were collected from two adults and NE-MSCs from two newborn infants. An in vitro study was performed in order to determine the best experimental conditions for adhesion, viability, proliferation and osteoblastic differentiation on BCP or BG granules. Bone-associated morphological changes and gene expression modifications were quantified using histological and molecular techniques. The in vivo study was based on the subcutaneous implantation in nude mice of the biomaterials, loaded or not with one of the two cell types. Eight weeks after, bone formation was assessed using histological and electron microscopy techniques. Results: Both cell types—BMMSC and NE-MSC—display the typical stem cell surface markers—CD73+, CD90+, CD105+, nestin - and exhibit the MSC-associated osteogenic, chondrogenic and adipogenic multipotency. NE-MSCs produce less collagen and alkaline phosphatase than BMMSCs. At the transcript level, NE-MSCs express more abundantly three genes coding for bone sialoprotein, osteocalcin and osteopontin while BMMSCs produce extra copies of RunX2. BMMSCs and NE-MSCs adhere and survive on BCP and BG. In vivo experiments reveal that bone formation is only observed with BMMSCs transplanted on BCP biomaterial. Conclusion: Although belonging to the same superfamily of mesenchymal stem cells, BMMSCs and NE-MSCs exhibit striking differences, in vitro and in vivo. For future clinical applications, the association of BMMSCs with BCP biomaterial seems to be the most promising.