Biphasic Calcium Phosphate Scaffolds Research Articles

Biphasic Calcium Phosphate Ceramic Scaffold Composed of Zinc Doped β-Tricalcium Phosphate and Silicon Doped Hydroxyapatite for Bone Tissue Engineering.

The rapid repair of bone defects remains a significant clinical challenge to this day. To address this issue, a 3D-printed biphasic calcium phosphate (BCP) scaffold consisting of 40 wt % hydroxyapatite (HA) and 60 wt % β-tricalcium phosphate (β-TCP) was created. Silicon and zinc were incorporated into HA and β-TCP, respectively, to enhance the angiogenic and osteogenic properties of the BCP scaffold. The physicochemical properties, in vitro cell responses, and bone defect repair efficacy of the modified BCP scaffold were comprehensively investigated. Results showed that the fabricated scaffold possessed a 3D interconnected pore structure. Zinc doping enhanced the sintering of the BCP scaffold, increased its density and strength, but decreased its degradation rate. Conversely, silicon doping had the opposite effect. The modified scaffold was capable of a gradual release of zinc/silicon ions, which promoted the proliferation and differentiation of cells. Specifically, the scaffold doped with zinc significantly promoted the osteogenic differentiation of stem cells. Moreover, co-doping with silicon and zinc synergistically promoted in vitro angiogenesis, with BCP-3 (doped with 2.5 mol % zinc and 4 mol % silicon) exhibiting the best pro-angiogenic activity. BCP-3 significantly induced regeneration of blood vessels and bone tissue in vivo, indicating its potential to accelerate the process of bone defect repair.

Ank-mediated pyrophosphate regulates shear stress-induced small extracellular vesicle production in 3D-cultured osteocytes

ABSTRACT Osteocytes are located in the lacunae of fluid-filled bone and communicate with neighboring or distant cells by secreting small extracellular vesicles (sEVs) and growth factors as well as via dendrite–dendrite direct connections. However, the mechanism regulating sEV production in osteocytes is yet to be elucidated. In this study, we investigated sEV production and its underlying mechanism in osteocytes cultured on a three dimensional (3D) scaffold. We employed a perfusion system to apply shear stress stimulation to MLO-Y4 cells cultured on a 3D biphasic calcium phosphate (BCP) scaffold and analyzed sEV production and gene expression using RNA sequencing. We found that the expression of genes associated with sEV biogenesis and the secretory pathway were enhanced by fluid shear stress in MLO-Y4 cells cultured on a 3D BCP scaffold. In particular, fluid shear stress induced the expression of Ank, a pyrophosphate transporter, in 3D-cultured MLO-Y4 cells. The role of Ank in sEV production was further examined. Probenecid, an Ank inhibitor, significantly suppressed shear stress-induced sEV production, whereas Ank cDNA overexpression stimulated it. The inhibition of shear stress-induced sEV production by probenecid was recovered by the exogenous addition of pyrophosphate to MLO-Y4 cells. These findings suggest that shear stress-mediated sEV production in 3D-cultured osteocytes is regulated by extracellular pyrophosphate transported by Ank.

A 3D in vitro model of biphasic calcium phosphate (BCP) scaffold combined with human osteoblasts, osteoclasts, and endothelial cells as a platform to mimic the oral microenvironment for tissue regeneration

Objectives: This study aimed to develop an innovative 3D in vitro model based on the biphasic calcium phosphate (BCP) scaffold combined with human osteoblasts (hOBs), osteoclasts (hOCs), and endothelial cells to evaluate its effects on bone and vascular cells behavior. Methods: To this end, an optimized mixture of hydroxyapatite (HA) and β-tricalcium phosphate (TCP) with a weight ratio of 30/70 was employed to develop a BCP scaffold using the computer-aided design (CAD) approach. The BCP scaffold was combined with primary cultures of hOBs, hOCs and human umbilical vein endothelial cells (HUVECs). Results: Morphometric analyses using scanning electron microscopy (SEM) and X-ray micro-computed tomography, along with biomechanical testing, revealed that BCP scaffold exhibited a regular 3D structure with large interconnected internal pores (700 µm) and high mechanical strength. In terms of biological behavior, after 14 days of tri-culture with hOBs, hMCs and HUVECs, SEM, immunofluorescence, and histological analyses showed that all cell types were viable and adhered well to the entire surface of the scaffold. Interestingly, SEM and energy-dispersive X-ray spectroscopy analyses also revealed on the BCP scaffold the presence of mineralized matrix crystals of Ca, P, O and C within a tissue-like cell layer produced by the interaction of the three cell types. Conclusions: Data confirmed the high performance of the BCP scaffold through biomechanical studies. Notably, for the first time, this study demonstrated the feasibility of combining BCP scaffold with hOBs, hOCs, and HUVEC, which remained viable and maintained their native phenotypes, creating also tissue-like cell layer. Clinical significanceAlthough further investigation is needed, these results underscore the potential to develop a 3D in vitro model that mimics the oral microenvironment, which could be valuable for BTE approaches in in vivo studies.

Role of Wnt5a in modulation of osteoporotic adipose-derived stem cells and osteogenesis.

Osteoporosis, a condition marked by the deterioration of bone microarchitecture and increased facture risk, arises from a disruption in bone metabolism, with osteoclasts surpassing osteoblasts in bone resorption versus formation. The Wnt signalling pathway, a key regulator of bone maintenance, remains partially understood in osteoporosis. Our research delves into the role of Wnt-related molecules in this disease. In osteoporotic adipose-derived stem cells (OP-ASCs), we detected a significant decrease in Ctnnb1 and Frizzled-6 (Fzd6), contrasted by an increase in Gsk-3β and Wnt5a. Activation of the Wnt pathway by LiCl resulted in elevated Ctnnb1 and Fzd6, but decreased Gsk-3β and Wnt5a levels, promoting OP-ASCs' bone-formation capacity. In contrast, inhibition of this pathway by DKK-1 led to diminished Ctnnb1 and Fzd6, and increased Gsk-3β and Wnt5a, adversely affecting osteogenesis. Furthermore, our findings show that overexpressing Wnt5a impedes, while silencing it enhances the bone-forming capability of OP-ASCs. In a cranial bone defect model, the implantation of Wnt5a-silenced OP-ASCs with biphasic calcium phosphate scaffolds significantly promoted new bone formation. These observations indicated a repression of the canonical Wnt pathway and a stimulation of the non-canonical pathway in OP-ASCs. Silencing Wnt5a increased the osteogenic and regenerative abilities of OP-ASCs. Our study suggests targeting Wnt5a could be a promising strategy for enhancing bone regeneration in post-menopausal osteoporosis.

3D printing of strontium-enriched biphasic calcium phosphate scaffolds for bone regeneration

Calcium phosphate (CaP) scaffolds doping with therapeutic ions are one of the focuses of recent bone tissue engineering research. Among the therapeutic ions, strontium stands out for its role in bone remodeling. This work reports a simple method to produce Sr-doped 3D-printed CaP scaffolds, using Sr-doping to induce partial phase transformation from β-tricalcium phosphate (β-TCP) to hydroxyapatite (HA), resulting in a doped biphasic calcium phosphate (BCP) scaffold. Strontium carbonate (SrCO3) was incorporated in the formulation of the 3D-printing ink, studying β-TCP:SrO mass ratios of 100:0, 95:5, and 90:10 (named as β-TCP, β-TCP/5-Sr, and β-TCP/10-Sr, respectively). Adding SrCO3 in the 3D-printing ink led to a slight increase in viscosity but did not affect its printability, resulting in scaffolds with a high printing fidelity compared to the computational design. Interestingly, Sr was incorporated into the lattice structure of the scaffolds, forming hydroxyapatite (HA). No residual SrO or SrCO3 were observed in the XRD patterns of any composition, and HA was the majority phase of the β-TCP/10-Sr scaffolds. The addition of Sr increased the compression strength of the scaffolds, with both β-TCP/5-Sr and β-TCP/10-Sr performing better than the β-TCP. Overall, β-TCP/5-Sr presented higher mineralized nodules and mechanical strength, while β-TCP scaffolds presented superior cell viability. The incorporation of SrCO3 in the ink formulation is a viable method to obtain Sr-BCP scaffolds. Thus, this approach could be explored with other CaP scaffolds aiming to optimize their performance and the addition of alternative therapeutic ions.

Three-Dimensionally Printed Biphasic Calcium Phosphate Ceramic Substrates as the Sole Inducer of Osteogenic Differentiation in Stromal Vascular Fraction Cells.

The stromal vascular fraction (SVF) is a derivate of fat tissue comprising both adipose-derived mesenchymal stem cells and endothelial cells and serves as a promising cell source for engineering vascularized bone tissues. Its combination with osteoconductive biphasic calcium phosphate (BCP) ceramic may represent a point-of-care agent for bone reconstruction. Here we assessed the proliferation and osteogenic differentiation capacities of SVF on 3D printed BCP implants, in comparison with isolated adipose-derived mesenchymal stem cells (AD-MSCs). AD-MSCs and SVF isolated from human donors were seeded on plastic or 3D printed BCP ceramics with sinusoidal or gyroid macrotopography and cultured in the presence or absence of osteogenic factors. Vascular, hematopoietic and MSC surface markers were assessed by flow cytometry whereas osteogenic activity was investigated through alizarin red staining and alkaline phosphatase activity. Osteogenic factors were necessary to trigger osteogenic activity when cells were cultured on plastic, without significant difference observed between the two cell populations. Interestingly, osteogenic activity was observed on BCP implants in the absence of differentiation factors, without significant difference in level activity between the two cell populations and macrotopography. This study offers supportive data for the use of combined BCP scaffolds with SVF in a perspective of a one-step surgical procedure for bone regeneration.

Optimizing silicon doping levels for enhanced osteogenic and angiogenic properties of 3D-printed biphasic calcium phosphate scaffolds: An in vitro screening and in vivo validation study

Biphasic calcium phosphate (BCP) ceramics are valued for their osteoconductive properties but have limited osteogenic and angiogenic activities, which restricts their clinical utility in bone defect repair. Silicon doping has emerged as an effective strategy to enhance these biological functions of BCP. However, the biological impact of BCP is influenced by the level of silicon doping, necessitating determination of the optimal concentration to maximize efficacy in bone repair. This study investigated the effects of silicon doping on both the physicochemical and biological properties of BCP, with a specific focus on osteogenic and angiogenic potentials. Results indicated that silicon doping exceeding 4 mol.% led to the formation of α-TCP, accelerating BCP degradation, enhancing silicon ion release, and promoting mineralization product formation. Simultaneously, silicon doping increased the porosity of BCP scaffolds, which typically reduces their compressive strength. Nevertheless, scaffolds doped with ≤4 mol.% silicon maintained compressive strengths exceeding 2 MPa. In vitro biological experiments indicated that higher levels of silicon doping (≥6 mol.%) partially inhibited the successful differentiation of stem cells and the vascularization of endothelial cells. Optimal conditions for promoting osteogenic differentiation and angiogenesis were identified between 2 and 4 mol.% silicon doping, with an optimal level of approximately 4 mol.%. Subsequent in vivo experiments confirmed that BCP scaffolds doped with 4 mol.% silicon effectively promoted vascularization and new bone formation, highlighting their potential for clinical bone defect repair.

Evaluating osteogenic potential of a 3D-printed bioceramic-based scaffold for critical-sized defect treatment: an in vivo and in vitro investigation.

The integration of precision medicine principles into bone tissue engineering has ignited a wave of research focused on customizing intricate scaffolds through advanced 3D printing techniques. Bioceramics, known for their exceptional biocompatibility and osteoconductivity, have emerged as a promising material in this field. This article aims to evaluate the regenerative capabilities of a composite scaffold composed of 3D-printed gelatin combined with hydroxyapatite/tricalcium phosphate bioceramics (G/HA/TCP), incorporating human dental pulp-derived stem cells (hDPSCs). Using 3D powder printing, we created cross-shaped biphasic calcium phosphate scaffolds with a gelatin layer. The bone-regenerating potential of these scaffolds, along with hDPSCs, was assessed through in vitro analyses and in vivo studies with 60 rats and critical-sized calvarial defects. The assessment included analyzing cellular proliferation, differentiation, and alkaline phosphatase activity (ALP), and concluded with a detailed histological evaluation of bone regeneration. Our study revealed a highly favorable scenario, displaying not only desirable cellular attachment and proliferation on the scaffolds but also a notable enhancement in the ALP activity of hDPSCs, underscoring their pivotal role in bone regeneration. However, the histological examination of calvarial defects at the 12-wk mark yielded a rather modest level of bone regeneration across all experimental groups. The test and cell group exhibited significant bone formation compared to all other groups except the control and cell group. This underscores the complexity of the regenerative process and paves the way for further in-depth investigations aimed at improving the potential of the composite scaffolds.

Effect of Citric Acid as a Porogenic Agent for Fabrication of Macroporous Biphasic Calcium Phosphate Scaffolds

Effect of Citric Acid as a Porogenic Agent for Fabrication of Macroporous Biphasic Calcium Phosphate Scaffolds

3D-printed Biphasic Calcium Phosphate Scaffold to augment cytocompatibility evaluation for load-bearing implant applications

In this work, we developed and analyzed a biphasic calcium phosphate (BCP) bioceramic for bone regeneration using stereolithography (SLA). The SLA method is a promising additive manufacturing (AM) technique capable of creating BCp parts with high accuracy and efficiency. However, the ceramic suspension used in SLA exhibits significantly higher viscosity and is not environmentally friendly. Therefore, adequate preparation of a suspension with low viscosity and high solid loading is essential. In this paper, we optimized the effects of surfactant doses and solid loading on the BCp slurry, and initially examined the process parameters of photocuring, debinding, and sintering. The utilization of 9 wt % Disperbyk (BYK) with a 40 vol % loading of BCp bioceramics exhibited a reasonably low viscosity of 8.9 mPa·s at a shear level of 46.5 s−1. Functional and structural analyses confirmed that BCp was retained after photocuring and subsequent treatment, which were incorporated into the BYK dispersion. The 3D printed objects with different sintered temperatures, specifically at 1100 °C, 1200 °C, and 1300 °C, were further optimized. Additionally, the surface roughness, porosity, and mechanical properties of BCp green parts were systematically investigated. Most importantly, in vitro analysis of cell attachment, differentiation, and red alizarin analysis could support the application of bone regeneration.

Controlling sintering temperature for biphasic calcium phosphate scaffolds using submicron hydroxyapatite slurries for LCD 3D printing

Liquid crystal display (LCD)-based 3D printing, a precise and surface-smooth technology, has found widespread use in dentistry due to its exceptional precision and surface quality. This research endeavors to create a scaffold through 3D printing using bone graft materials, specifically addressing the challenge of molding bone graft powders to fit defect areas in dental surgery. Our approach involved developing biphasic calcium phosphate (BCP) scaffolds by regulating the sintering temperature of an HA-based green body. Initially, by using wet ball mill technology, HA slurries with 30 %, 40 %, and 50 % HA content by weight were prepared and exhibited viscosities below 0.15 Pa s. Subsequently, these slurries were 3D printed via an LCD-based VP printer. The utilization of submicron-sized HA particles played a critical role in maintaining slurry stability and preventing sedimentation during the printing process. We investigated sintering temperatures of 1100 °C, 1200 °C, and 1300 °C, discovering that submicron-sized HA particles enabled precise alignment of the decomposition temperature with the typical HA sintering range. Consequently, we achieved the production of BCP scaffolds with adjustable properties by tuning the sintering temperature. Subsequent analysis delved into scaffold shrinkage, mechanical characteristics, degradation behavior, and biological responses. Among the sintered scaffolds, the 50 % HA slurry printed and sintered at 1200 °C exhibited the highest compressive strength. The cell viabilities of these sintered scaffolds consistently met the ISO 10993-5 standard, affirming their biocompatibility. Moreover, the biological responses of MG63 cells demonstrated that the sintered scaffolds not only exhibited cytocompatibility but also facilitated strong cell attachment and proliferation. Furthermore, scaffolds sintered at 1300 °C containing TCP showed the formation of a layer of HA with plate-like structures due to ion-induced precipitation, indicating their bioactivity. In conclusion, this study highlights the promising potential of LCD-based VP 3D printing technology for ceramic slurries, with clear implications for clinical applications in bone tissue engineering.

Cytotoxicity and mechanical properties of biphasic calcium phosphate scaffold from Tegilarca granosa due to its composition

Background: Biphasic calcium phosphate (BCP) is graft material contained HA and TCP. Tegilarca granosa shell is a natural source that may converted into BCP. This study aims to determine the composition and cytotoxicity of BCP synthesized from Tegilarca granosa shell used various hydrothermal hours and to evaluate the mechanical properties of BCP scaffold. Methods: Tegilarca granosa shell was converted into BCP using hydrothermal method at 200˚C for 6h (Group 1); 9h (Group 2); and 12h (Group 3). The composition was determined by XRD and the cell viability were evaluated using MTT Assays. Each group was added with 20% gelatin ratio 50:50 (w/v) and freeze-dried to form scaffold. Scaffolds (Ø6mm x 4mm) were prepared for diametral tensile strength (DTS) test (n=6) and scaffolds (Ø7mm x 11mm) were used for compressive strength (CS) test (n=6). All data were analyzed using Kruskall-Wallis followed by Mann-Whitney test. Results: The composition of BCP (HA/ TCP) at Group 1, Group 2, and Group 3 were 81.80%/14,10%; 87%/6%; and 72%/21%. The cell viabilities were good for all groups. The DTS and CS test showed there was a significant difference between Group 1 and Group 3 scaffold, meanwhile there was no significant differences between Group 2 and Group 3 scaffold. Group 3 scaffold showed the highest DTS and CS, 6.921 MPa and 1,233 MPa. Conclusion: The BCP composition were depent on hydrothermal hours. Although all scaffold groups were non-toxic, but BCP scaffold synthesized from Tegilarca granosa shell using hydrothermal for 12 hours showed the highest mechanical properties.

The role of integrin αvβ3 in biphasic calcium phosphate ceramics mediated M2 Macrophage polarization and the resultant osteoinduction

Calcium phosphate ceramics-based biomaterials were reported to have good biocompatibility and osteoinductivity and have been widely applied for bone defect repair and regeneration. However, the mechanism of their osteoinductivity is still unclear. In our study, we established an ectopic bone formation in vivo model and an in vitro macrophage cell co-culture system with calcium phosphate ceramics to investigate the effect of biphasic calcium phosphate on osteogenesis via regulating macrophage M1/M2 polarization. Our micro-CT data suggested that biphasic calcium phosphate had significant osteoinductivity, and the fluorescence co-localization detection found increased F4/80+/integrin αvβ3+ macrophages surrounding the biphasic calcium phosphate scaffolds. Besides, our study also revealed that biphasic calcium phosphate promoted M2 polarization of macrophages via upregulating integrin αvβ3 expression compared to tricalcium phosphate, and the increased M2 macrophages could subsequently augment the osteogenic differentiation of MSCs in a TGFβ mediated manner. In conclusion, we demonstrated that macrophages subjected to biphasic calcium phosphate could polarize toward M2 phenotype via triggering integrin αvβ3 and secrete TGFβ to increase the osteogenesis of MSCs, which subsequently enhances bone regeneration.

3D gel-printing of hierarchically porous BCP scaffolds for bone tissue engineering

In this study, a conventional technique of porous preparation was used to improve the constructive capability of direct ink writing on microstructures, and the hierarchically porous scaffolds were successfully prepared by 3D gel printing (3DGP). Micron-sized hydroxyapatite (HA) was coated with tricalcium phosphate (TCP) nanopowders synthesized by chemical co-precipitation to form biphasic calcium phosphate (BCP). The random structure of concave micropores was achieved by filling the BCP slurry with PMMA microspheres while successfully controlling the internal porosity of printed filaments. The results showed that the three-stage porous structure was successfully constructed, i.e., macroscopic pores of 1.50-2.00 mm, spherical micropores of 100-200 μm, and inter-powder interstices of 1.00-10.00 μm. Nano-TCP coated micron-HA powders improved the sintering activity of BCP particles. The compressive strength and porosity of the scaffolds sintered at 1400 °C were 2.78 MPa and 84.98%. The hierarchically porous BCP scaffolds had bright applications in bone tissue engineering.

In Vitro Evaluation of Biphasic Calcium Phosphate Scaffolds Derived from Cuttlefish Bone Coated with Poly(ester urea) for Bone Tissue Regeneration.

This study investigates the osteogenic differentiation of umbilical-cord-derived human mesenchymal stromal cells (hUC-MSCs) on biphasic calcium phosphate (BCP) scaffolds derived from cuttlefish bone doped with metal ions and coated with polymers. First, the in vitro cytocompatibility of the undoped and ion-doped (Sr2+, Mg2+ and/or Zn2+) BCP scaffolds was evaluated for 72 h using Live/Dead staining and viability assays. From these tests, the most promising composition was found to be the BCP scaffold doped with strontium (Sr2+), magnesium (Mg2+) and zinc (Zn2+) (BCP-6Sr2Mg2Zn). Then, samples from the BCP-6Sr2Mg2Zn were coated with poly(ԑ-caprolactone) (PCL) or poly(ester urea) (PEU). The results showed that hUC-MSCs can differentiate into osteoblasts, and hUC-MSCs seeded on the PEU-coated scaffolds proliferated well, adhered to the scaffold surfaces, and enhanced their differentiation capabilities without negative effects on cell proliferation under in vitro conditions. Overall, these results suggest that PEU-coated scaffolds are an alternative to PCL for use in bone regeneration, providing a suitable environment to maximally induce osteogenesis.

Mangiferin-Enriched Mn-Hydroxyapatite Coupled with β-TCP Scaffolds Simultaneously Exhibit Osteogenicity and Anti-Bacterial Efficacy.

Biphasic calcium phosphate (BCP) containing β-tricalcium phosphate and manganese (Mn)-substituted hydroxyapatite (HAP) was synthesized. Biomedical scaffolds were prepared using this synthesized powder on a sacrificial polyurethane sponge template after the incorporation of mangiferin (MAN). Mn was substituted at a concentration of 5% and 10% in HAP to examine the efficacy of Mn at various concentrations. The phase analysis of the as-formed BCP scaffold was carried out by X-ray diffraction analysis, while the qualitative observation of morphology and the osteoblast cell differentiation were carried out by scanning electron microscopy and confocal laser scanning microscopy techniques. Gene expressions of osteocalcin, collagen 1, and RUNX2 were carried out using qRT-PCR analyses. Significantly higher (p < 0.05) levels of ALP activity were observed with extended osteoblast induction on the mangiferin-incorporated BCP scaffolds. After characterization of the specimens, it was found that the scaffolds with 10% Mn-incorporated BCP with mangiferin showed better osteogenicity and simultaneously the same scaffolds exhibited higher anti-bacterial properties as observed from the bacterial viability test. This study was carried out to evaluate the efficacy of Mn and MAN in BCP for osteogenicity and antibacterial action.

Regenerative Potential of Bone Morphogenetic Protein 7-Engineered Mesenchymal Stem Cells in Ligature-Induced Periodontitis.

Periodontitis is an oral disease caused by bacterial infection that has stages according to the severity of tissue destruction. The advanced stage of periodontitis presents irreversible destruction of soft and hard tissues, which finally results in loss of teeth. When conventional treatment modalities show limited results, tissue regeneration therapy is required in patients with advanced periodontitis. In the present study, we aimed to evaluate the effect of bone marrow-derived mesenchymal stem cells (BM-MSCs) delivering bone morphogenetic protein 7 (BMP7) on tissue regeneration in a periodontitis model. BMP7 is a member of the BMP family that shows bone-forming ability; however, BMPs rapid clearing and degradation and unproven efficacy make it difficult to apply it in clinical dentistry. To overcome this, we established BMP7-expressing engineered BM-MSCs (BMP7-eBMSCs) that showed superior osteogenic differentiation potential when subcutaneously transplanted with a biphasic calcium phosphate scaffold into immunocompromised mice. Furthermore, the efficacy of BMP7-eBMSC transplantation for periodontal tissue regeneration was evaluated in a rat ligature-induced periodontitis model. Upon measuring two-dimensional and three-dimensional amounts of regenerated alveolar bone using microcomputed tomography, the amounts were found to be significantly higher in the BMP7-eBMSC transplantation group than in the eBMSC transplantation group. Most importantly, fibrous periodontal ligament (PDL) tissue regeneration was also achieved upon BMP7-eBMSC transplantation, which was evaluated by calculating the modified relative connective tissue attachment. The amount of connective tissue attachment in the BMP7-eBMSC transplantation group was significantly higher than that in the ligature-induced periodontitis group, although the increase was comparable between the BMP7-eBMSC and human PDL stem cell transplantation groups. Taken together, our results suggested that sustainable release of BMP7 induces periodontal tissue regeneration and that transplantation of BMP7-eBMSCs is a feasible treatment option for periodontal regeneration. Impact Statement Periodontitis is the second most common human dental disease affecting chronic systemic diseases. Despite the tremendous efforts trying to cure the damaged periodontal tissues using tissue engineering technologies, a definitive regenerative method has not been in consensus. Researchers are seeking more feasible and abundant source of mesenchymal stem cells (MSCs), and furthermore, how to use reliable growth factors under more efficient control are the issues to be solved. In this study, we aimed to evaluate the effect of bone morphogenetic protein 7 (BMP7) gene delivering bone marrow-derived MSCs on periodontal tissue regeneration to evaluate the efficacy of BMP7 and engineered BMSCs for periodontal tissue regeneration.

Enhancing osteoinduction and bone regeneration of biphasic calcium phosphate scaffold thought modulating the balance between pro-osteogenesis and anti-osteoclastogenesis by zinc doping

Biphasic calcium phosphate (BCP) is a promising bone regeneration material with an adjustable degradation rate, and ions doping is an effective method to improve its bone repair effect. However, few studies have focused on the role of ions doping-mediated osteogenesis and osteoclastogenesis balance in bone regeneration. In order to improve the bone repair effect of BCP and emphasize the role of osteogenesis and osteoclastogenesis balance regulated by ions doping to promote bone regeneration. Here, zinc (Zn), an essential trace element with pro-osteogenesis and anti-osteoclastogenesis, was dopped into BCP scaffolds, and its effects on the physicochemical properties, in vitro cytological responses, in vivo osteoinduction, and bone defect regeneration of BCP were systematically investigated. Results showed that Zn was successfully introduced into the crystal lattice of BCP. The differentiation of both osteoblasts and osteoclasts showed a doping content-dependent behavior. By increasing the doping content (0–5 mol.%), the anti-osteoclastogenesis effect of BCP gradually increased, while its pro-osteogenesis performance first increased and then decreased. ThoughBCP with a doping content of 5 mol.% could promote osteogenic differentiation of stem cells and reduced fibrous capsule encapsulation, it could not enhance ectopic bone formation or bone regeneration due to its excessive anti-osteoclastogenesis. BCP with a doping content of 2.5 mol.% remarkably enhanced the osteoinduction activity and accelerated new bone formation, which were associated with its highest pro-osteogenesis and appropriate anti-osteoclastogenesis activity. This study highlights on regulating the balance between pro-osteogenesis and anti-osteoclastogenesis by Zn doping and paves the way for the development of a more osteoinductive calcium phosphate-based material for bone regeneration.

Adjusting physicochemical and cytological properties of biphasic calcium phosphate by magnesium substitution: An in vitro study

Biphasic calcium phosphate (BCP) is a highly study bone defect repair material with adjustable degradation, perfect osteoconduction and good osteoinduction. As one of the essential trace elements, magnesium (Mg) possesses the abilities of pro-osteogenesis and pro-angiogenesis. Therefore, Mg doping may further expand the application of BCP in bone defect repair, but few studies focus on promoting the osteogenesis and angiogenesis of BCP simultaneously by Mg doping, and the optimal doping amount of Mg remains to be explored. In this study, the physicochemical and biological properties of BCP scaffold affected by Mg doping were systematically study. Results showed that Mg doping enhanced the sintering of BCP scaffold, resulting in the decrease of degradation rate at the initial soaking period. However, the introduction of Mg damaged the lattice stability of BCP, leading to the increase of BCP degradation rate at the later soaking period. BCP scaffolds with Mg doping content ≥3 mol.% could achieve a long-term sustained release of Mg. The ion microenvironment created by Mg-doped scaffolds was simultaneously conducive to the osteogenic differentiation of stem cells and the enhanced angiogenic activity of endothelial cells. The scaffold doped with 5 mol.% of Mg (Mg5–S) showed the highest efficiency in promoting osteogenic differentiation. Mg-doped BCP scaffolds with a doping content ≥3 mol.%, especially Mg5–S, significantly improved the proliferation and angiogenic differentiation of endothelial cells. Based on these, we believe that the optimal doping content of Mg in BCP is 5 mol.%, and Mg5–S has great application potential in bone defect repair.

Combined Application of Dentin Noncollagenous Proteins and Odontogenic Biphasic Calcium Phosphate in Rabbit Maxillary Sinus Lifting.

Teeth can be used as a raw material for preparing bone substitutes due to their similar chemical composition to bone. The objective of our study was to evaluate the effect of odontogenic biphasic calcium phosphate (BCP) incorporating dentin noncollagenous proteins (DNCPs) on osteogenesis and stability in maxillary sinus augmentation. The composition, structure and morphology of the odontogenic BCP were tested by X-ray powder diffraction (XRD), Brunauer-Emmett-Teller, and scanning electron microscopy methods. The biocompatibility and osteoinduction of DNCPs and materials were examined in vitro and their bone regeneration capacity was verified in vivo. The results showed that the cells adhered and proliferated well on the DNCP-loaded BCP scaffold. The odontogenic BCP and DNCPs promoted osteogenic differentiation of cells, The new bone formation in the BCP groups and DNCP subgroups was significantly higher than the new bone formation in the control, and the new bone quality was better. The bone regeneration effect of odontogenic BCP was similar to the effect of deproteinized bovine bone mineral, but β-TCP did not maintain the height and volume of bone reconstruction. In conclusion, the combined application of DNCPs and odontogenic BCP is an effective strategy for tissue engineering osteogenesis in the maxillary sinus region. The biomimetic strategy could provide a new approach for patients requiring maxillary sinus lifting.