Craniofacial Bone Tissue Engineering Research Articles

Enhancing craniofacial bone tissue engineering strategy: integrating rapid wet chemically synthesised bioactive glass with photopolymerized resins

BackgroundCraniofacial bone regeneration represents a dynamic area within tissue engineering and regenerative medicine. Central to this field, is the continual exploration of new methodologies for template fabrication, leveraging established bio ceramic materials, with the objective of restoring bone integrity and facilitating successful implant placements.MethodsPhotopolymerized templates were prepared using three distinct bio ceramic materials, specifically a wet chemically synthesized bioactive glass and two commercially sourced hydroxyapatite variants. These templates underwent comprehensive characterization to assess their physicochemical and mechanical attributes, employing techniques including Fourier transform infrared spectroscopy, scanning electron microscopy, and nano-computed tomography. Evaluation of their biocompatibility was conducted through interaction with primary human osteoblasts (hOB) and subsequent examination using scanning electron microscopy.ResultsThe results demonstrated that composite showed intramolecular hydrogen bonding interactions with the photopolymer, while computerized tomography unveiled the porous morphology and distribution within the templates. A relatively higher porosity percentage (31.55 ± 8.70%) and compressive strength (1.53 ± 0.11 MPa) was noted for bioactive glass templates. Human osteoblast cultured on bioactive glass showed higher viability compared to other specimens. Scanning micrographs of human osteoblast on templated showed cellular adhesion and the presence of filopodia and lamellipodia.ConclusionIn summary these templates have the potential to be used for alveolar bone regeneration in critical size defect. Photopolymerization of bioceramics may be an interesting technique for scaffolds fabrication for bone tissue engineering application but needs more optimization to overcome existing issues like the ideal ratio of the photopolymer to bioceramics.

Retraction: Photoencapsulated-BMP2 in visible light-cured thiol-acrylate hydrogels for craniofacial bone tissue engineering.

Retraction: Photoencapsulated-BMP2 in visible light-cured thiol-acrylate hydrogels for craniofacial bone tissue engineering.

Surface Modification of Polylactic Acid Bioscaffold Fabricated via 3D Printing for Craniofacial Bone Tissue Engineering.

Bone tissue engineering is a promising solution for advanced bone defect reconstruction after severe trauma. In bone tissue engineering, scaffolds in three-dimensional (3D) structures are crucial components for cell growth, migration, and infiltration. The three-dimensional printing technique is well suited to manufacturing scaffolds since it can fabricate scaffolds with highly complex designs under good internal structural control. In the current study, the 3D printing technique was utilized to produce polylactic acid (PLA) scaffolds. BMSCs were seeded onto selected scaffolds, either hydrogel-mixed or not, and cultivated in vitro to investigate the osteogenic potential in each group. After osteogenic incubation in vitro, BMSC-seeded scaffolds were implanted onto rat cranium defects, and bone regeneration was observed after 12 weeks. Our results demonstrated that BMSCs were able to seed onto 3D-printed PLA scaffolds under high-resolution observation. Real-time PCR analysis showed their osteogenic ability, which could be further improved after BMSCs were mixed with hydrogel. The in vivo study showed significantly increased bone regeneration when rats' cranium defects were implanted with a hydrogel-mixed BMSC-seeded scaffold compared to the control and those without cell or hydrogel groups. This study showed that 3D-printed PLA scaffolds are a feasible option for BMSC cultivation and osteogenic differentiation. After mixing with hydrogel, BMSC-seeded 3D-printed scaffolds can facilitate bone regeneration.

Advances and Prospects in Materials for Craniofacial Bone Reconstruction.

The craniofacial region is composed of 23 bones, which provide crucial function in keeping the normal position of brain and eyeballs, aesthetics of the craniofacial complex, facial movements, and visual function. Given the complex geometry and architecture, craniofacial bone defects not only affect the normal craniofacial structure but also may result in severe craniofacial dysfunction. Therefore, the exploration of rapid, precise, and effective reconstruction of craniofacial bone defects is urgent. Recently, developments in advanced bone tissue engineering bring new hope for the ideal reconstruction of the craniofacial bone defects. This report, presenting a first-time comprehensive review of recent advances of biomaterials in craniofacial bone tissue engineering, overviews the modification of traditional biomaterials and development of advanced biomaterials applying to craniofacial reconstruction. Challenges and perspectives of biomaterial development in craniofacial fields are discussed in the end.

Hydrogels provide microenvironments to mesenchymal stem cells for craniofacial bone regeneration: Review.

The anatomical and physiological architecture of the craniofacial bone is intricate. Hence, the exact management of osteogenesis is necessary for the regeneration of the deficiencies that present in this area. Stem-based tissue engineering approaches, as opposed to conventional surgical intervention, induce bone growth with minimal postoperative risk and expense. Mesenchymal stem/stromal cells (MSC)'s pluripotent differentiation potential, anti-inflammatory and immunomodulatory properties underpin its versatility as a therapeutic agent in bone tissues. Inspired by the native stem cell niche, hydrogels are preferred choices to mediate cells and adapt to 3-D environment because of their outstanding swelling capabilities and similarity to natural extracellular matrices (ECMs). Due to their remarkable biocompatibility and capacity for stimulating bone regeneration, bone regeneration hydrogels have also received a great deal of interest. This review explores the opportunities of MSC based regenerative skeletal therapies, introduces the application of hydrogel scaffolds as artificial bone microenvironments for stem cells to explore its usage in craniofacial bone tissue engineering.

Bioactive 3D-printed chitosan-based scaffolds for personalized craniofacial bone tissue engineering

• 3D-printing of chitosan-based hydrogel • Increase in pore size and porosity was observed in the 3D-printed scaffolds containing hydroxyapatite. • Hydroxyapatite incorporation increased apatite formation and decreased the size of formed apatite crystals. • No considerable toxicity on MSCs was observed in 3D-printed scaffolds. Regeneration of craniofacial bone defects is a key issue in the bone regeneration field. Hence, novel treatment strategies, such as tissue engineering using porous scaffolds, have been developed. An ideal tissue-engineered scaffold for bone tissue regeneration should possess pores to facilitate nutrients transmission and support reparative tissue ingrowth, bioactivity for osteoconduction and osseointegration, and biocompatibility to improve cell attachment, proliferation, and extracellular matrix formation. In the present study, we manufactured chitosan-based hydrogels substituted with alginate with optimized properties by extrusion-based three-dimensional (3D) printing. 3D printing of the scaffolds enables the designing and developing of complex architectures for craniofacial reconstruction using computer-aided design (CAD). Different ratios (2.5, 5, and 10%) of hydroxyapatite were added to the hydrogel, printed, and subsequently lyophilized to augment the physical and biological characteristics of the scaffolds. Hydroxyapatite incorporation into the chitosan-based scaffolds increased the porosity and pore size of the printed scaffolds. In addition, the presence of hydroxyapatite amplified apatite formation and decreased the size of formed apatite crystals. All the scaffold samples showed biocompatible properties and did not have toxicity toward rat bone marrow mesenchymal stem cells. Furthermore, the scaffolds containing 5% w/V hydroxyapatite exhibited significant growth in cell viability compared to the control. Overall, it is concluded that chitosan-based scaffolds adorned with hydroxyapatite are considerable for regenerating craniofacial bone defects. Graphical Abstract:

State-of-the-art techniques for imaging the vascular microenvironment in craniofacial bone tissue engineering applications.

Vascularization is a crucial step during musculoskeletal tissue regeneration via bioengineered constructs or grafts. Functional vasculature provides oxygen and nutrients to the graft microenvironment, facilitates wound healing, enhances graft integration with host tissue, and ensures the long-term survival of regenerating tissue. Therefore, imaging de novo vascularization (i.e., angiogenesis), changes in microvascular morphology, and the establishment and maintenance of perfusion within the graft site (i.e., vascular microenvironment or VME) can provide essential insights into engraftment, wound healing, as well as inform the design of tissue engineering (TE) constructs. In this review, we focus on state-of-the-art imaging approaches for monitoring the VME in craniofacial TE applications, as well as future advances in this field. We describe how cutting-edge in vivo and ex vivo imaging methods can yield invaluable information regarding VME parameters that can help characterize the effectiveness of different TE constructs and iteratively inform their design for enhanced craniofacial bone regeneration. Finally, we explicate how the integration of novel TE constructs, preclinical model systems, imaging techniques, and systems biology approaches could usher in an era of "image-based tissue engineering."

Innovations in craniofacial bone and periodontal tissue engineering – from electrospinning to converged biofabrication

ABSTRACT From a materials perspective, the pillars for the development of clinically translatable scaffold-based strategies for craniomaxillofacial (CMF) bone and periodontal regeneration have included electrospinning and 3D printing (biofabrication) technologies. Here, we offer a detailed analysis of the latest innovations in 3D (bio)printing strategies for CMF bone and periodontal regeneration and provide future directions envisioning the development of advanced 3D architectures for successful clinical translation. First, the principles of electrospinning applied to the generation of biodegradable scaffolds are discussed. Next, we present on extrusion-based 3D printing technologies with a focus on creating scaffolds with improved regenerative capacity. In addition, we offer a critical appraisal on 3D (bio)printing and multitechnology convergence to enable the reconstruction of CMF bones and periodontal tissues. As a future outlook, we highlight future directions associated with the utilisation of complementary biomaterials and (bio)fabrication technologies for effective translation of personalised and functional scaffolds into the clinics.

Microarray illustrates enhanced mechanistic action towards osteogenesis for magnesium aluminate spinel ceramic-based polyphasic composite scaffold with mesenchymal stem cells and bone morphogenetic protein 2.

Spinel (magnesium aluminate MgAl2 O4 ) ceramic-based polyphasic composite scaffold has been recently reported for craniofacial bone tissue engineering. Improving the osteogenic effects of such composite scaffolds with bone morphogenetic proteins (BMP2) is an intensely researched area. This study investigated the gene interactions of this scaffold with BMP2 and mesenchymal stem cells (MSCs). Human bone marrow MSCs were cultured in 3 groups: Group 1-Control (BMSCs), Group 2-BMSC with BMP2, and Group 3-BMSC with scaffold and BMP2. After RNA isolation, gene expression analysis was done by microarray. Differentially expressed genes (DEGs) (-1.0 > fold changes>1 and p value <.05) were studied for their function and gene ontologies using Database for Annotation, Visualization and Integrated Discovery (DAVID). They were further studied by protein-protein interaction network analysis using STRING and MCODE Cytoscape plugin database. Group 3 showed up regulation of 3222 genes against 2158 of Group 2. Group 3 had five annotation clusters with enrichment scores from 2.08 to 3.93. Group 2 had only one cluster. Group 3 showed activation of all major osteogenic pathways: TGF, BMP2, WNT, SMAD, and Notch gene signaling with effects of calcium and magnesium released from the scaffold. Downstream effect of all these caused significant activation of RUNX2, the key transcriptional regulator of osteogenesis in Group 3. STRING and MCODE Cytoscape plugin demonstrated the interactions. The enhanced MSC differentiation for osteogenesis with the addition of BMP2 to the polyphasic composite scaffold proposed promising clinical applications for bone tissue engineering.

Illuminating the Regenerative Microenvironment: Emerging Quantitative Imaging Technologies for Craniofacial Bone Tissue Engineering.

Tissue engineering has the potential to revolutionize treatments for patients suffering from critical-sized craniofacial bone defects, but it has yet to make a substantial impact in clinical practice. One of the barriers to improving the design of tissue-engineered bone grafts (TEBGs) is the lack of adequate techniques to study how transplanted cells, host cells, and biomaterials interact to facilitate the dynamic healing process. In this perspective, we discuss recent advances in quantitative imaging that may be adapted to provide high spatiotemporal resolution of the 3D tissue microenvironment during cranial bone regeneration. The adoption and application of these imaging technologies will provide a more rigorous framework for evaluating TEBG performance and enable the development of next-generation TEBGs for craniofacial repair.

Craniofacial Bone Tissue Engineering: Current Approaches and Potential Therapy.

Craniofacial bone defects can result from various disorders, including congenital malformations, tumor resection, infection, severe trauma, and accidents. Successfully regenerating cranial defects is an integral step to restore craniofacial function. However, challenges managing and controlling new bone tissue formation remain. Current advances in tissue engineering and regenerative medicine use innovative techniques to address these challenges. The use of biomaterials, stromal cells, and growth factors have demonstrated promising outcomes in vitro and in vivo. Natural and synthetic bone grafts combined with Mesenchymal Stromal Cells (MSCs) and growth factors have shown encouraging results in regenerating critical-size cranial defects. One of prevalent growth factors is Bone Morphogenetic Protein-2 (BMP-2). BMP-2 is defined as a gold standard growth factor that enhances new bone formation in vitro and in vivo. Recently, emerging evidence suggested that Megakaryocytes (MKs), induced by Thrombopoietin (TPO), show an increase in osteoblast proliferation in vitro and bone mass in vivo. Furthermore, a co-culture study shows mature MKs enhance MSC survival rate while maintaining their phenotype. Therefore, MKs can provide an insight as a potential therapy offering a safe and effective approach to regenerating critical-size cranial defects.

Effect of Electrospinning Parameters on the Fiber Diameter and Morphology of PLGA Nanofibers.

Poly lactic-co-glycolic acid (PLGA) has been widely investigated for various biomedical applications, such as craniofacial bone regeneration, wound dressing and tissue engineering. Electrospinning is a versatile technology used to produce micro/nanoscale fibers with large specific surface area and high porosity. The aim of the current study is to prepare PLGA nanofibers using electrospinning for guided tissue regeneration/guided bone regeneration applications. The objective of this study is to determine the appropriate electrospinning parameters such as applied voltage, flow rate, spinneret-collector distance and polymer solution concentration for preparation of PLGA fibrous membrane and their effect on the mean fiber diameter of the electrospun fibers. PLGA pellets were dissolved in Hexafluoroisopropanol (HFIP) in various concentrations overnight using a bench rocker. The resulting PLGA solution was then loaded into a syringe and electrospinning was done by maintaining the other parameters constant. Similarly, various fibrous mats were collected by altering the specific electrospinning parameter inputs such as applied voltage, flow rate and spinneret-collector distance. The morphology of the fibrous mats was characterized using Scanning Electron Microscope. The mean fiber diameter was assessed using ImageJ software and the results were compared using one-way ANOVA. We obtained bead-free uniform fibers with various tested solution concentrations. One-way ANOVA analysis demonstrated significant variation in mean fiber diameter of the electrospun fibers with altering applied voltage, solution concentration, flow rate and spinneret-collector distance. The above-mentioned electrospinning parameters and solution concentration influence the mean fiber diameter of electrospun PLGA nanofibers.

The Potential of FGF-2 in Craniofacial Bone Tissue Engineering: A Review.

Bone is a hard-vascularized tissue, which renews itself continuously to adapt to the mechanical and metabolic demands of the body. The craniofacial area is prone to trauma and pathologies that often result in large bone damage, these leading to both aesthetic and functional complications for patients. The “gold standard” for treating these large defects is autologous bone grafting, which has some drawbacks including the requirement for a second surgical site with quantity of bone limitations, pain and other surgical complications. Indeed, tissue engineering combining a biomaterial with the appropriate cells and molecules of interest would allow a new therapeutic approach to treat large bone defects while avoiding complications associated with a second surgical site. This review first outlines the current knowledge of bone remodeling and the different signaling pathways involved seeking to improve our understanding of the roles of each to be able to stimulate or inhibit them. Secondly, it highlights the interesting characteristics of one growth factor in particular, FGF-2, and its role in bone homeostasis, before then analyzing its potential usefulness in craniofacial bone tissue engineering because of its proliferative, pro-angiogenic and pro-osteogenic effects depending on its spatial-temporal use, dose and mode of administration.

In vitro evaluation of magnesium aluminate [MgAl2O4] spinel ceramic based polyphasic composite scaffold for craniofacial bone tissue engineering

Craniofacial bone regeneration is crucial for restoring the form and function of underlying vital structures. However, the unique physicochemical and biologic requirements of it make this task challenging. Two types of polyphasic composite scaffolds were therefore developed by polymeric template replication process from magnesium aluminate (MgAl2O4) spinel, β-tricalcium phosphate (β-TCP) and polymers; poly lactic-co-glycolic acid (PLGA) or poly caprolactone (PCL). The scaffolds: spinel/β-TCP/PLGA and spinel/β-TCP/PCL were physicochemically characterized at each stage of development followed by evaluation of compressive strength, porosity, in vitro degradation and ion release. Their morphology was then assessed for trabecular parameters: trabecular number, thickness, spacing, structural model index and degree of anisotropy. Their interactions with mesenchymal stem cells were also studied. Both the scaffolds showed compressive strength and porosity of craniofacial bones. However, trabecular parameters, degradation, ion release, cell viability / proliferation / deposition inside the scaffold were better for spinel/β- TCP/PLGA. Further, its internal morphology was similar to cancellous bone, optimum for cellular and vascular permeation. The scaffold also underwent significant degradation in four months releasing calcium, magnesium and phosphorus. Up regulation of genes pertaining to bone morphogenesis, development, differentiation, mineralization, ossification, remodeling were found along with transmembrane and notch receptor genes. All these led to the activation of major osteogenic pathways mediated by TGF/BMP/WNT/integrin families and notch signaling, with significant contributions from Mg ions. The genes with specific role in osteogenesis were also validated. Overall, magnesium aluminate based polyphasic composite scaffold: spinel/β-TCP/PLGA demonstrated physical and biologic characteristics essential for in vitro craniofacial bone tissue engineering.

Comparative Craniofacial Bone Regeneration Capacities of Mesenchymal Stem Cells Derived from Human Neural Crest Stem Cells and Bone Marrow.

Most craniofacial bones are derived from the ectodermal germ layer via neural crest stem cells, which are distinct from mesoderm-derived long bones. However, current craniofacial bone tissue engineering approaches do not account for this difference and utilize mesoderm-derived bone marrow mesenchymal stem cells (BM-MSCs) as a paradigm cell source. The effect of the embryonic origin (ontogeny) of an MSC population on its osteogenic differentiation potential and regenerative ability still remains unresolved. To clarify the effects of MSC ontogeny on bone regenerative ability, we directly compared the craniofacial bone regenerative abilities of an ecto-mesenchymal stem cell (eMSC) population, which is derived from human embryonic stem cells via a neural crest intermediate, with mesodermal adult BM-MSCs. eMSCs showed comparable osteogenic and chondrogenic ability to BM-MSCs in 2-D in vitro culture, but lower adipogenic ability. They exhibited greater proliferation than BM-MSCs and comparable construct mineralization in a well-established 3-D polycaprolactone-tricalcium phosphate (PCL-TCP) scaffold system in vitro. eMSC-derived 3D osteogenic constructs were maintained for longer in a proliferative osteoblast state and exhibited differential levels of genes related to fibroblast growth factor (FGF) signaling compared to BM-MSCs. Although both eMSC and BM-MSC-seeded scaffold constructs could promote bone regeneration in a rat calvarial defect model, eMSC-derived osseous constructs had significantly higher cellularity due to increased number of proliferative (Ki67+) cells than those seeded with BM-MSCs, and exhibited enhanced new bone formation in the defect area as compared to untreated controls. Overall, our study demonstrates the potential of human eMSCs for future clinical use in craniofacial regeneration applications and indicates the importance of considering MSC origin when selecting an MSC source for regenerative applications.

Skeletal Stem Cells-A Paradigm Shift in the Field of Craniofacial Bone Tissue Engineering.

Defects of the craniofacial skeleton arise as a direct result of trauma, diseases, oncological resection, or congenital anomalies. Current treatment options are limited, highlighting the importance for developing new strategies to restore form, function, and aesthetics of missing or damaged bone in the face and the cranium. For optimal reconstruction, the goal is to replace “like with like.” With the inherent challenges of existing options, there is a clear need to develop alternative strategies to reconstruct the craniofacial skeleton. The success of mesenchymal stem cell-based approaches has been hampered by high heterogeneity of transplanted cell populations with inconsistent preclinical and clinical trial outcomes. Here, we discuss the novel characterization and isolation of mouse skeletal stem cell (SSC) populations and their response to injury, systemic disease, and how their re-activation in vivo can contribute to tissue regeneration. These studies led to the characterization of human SSCs which are able to self-renew, give rise to increasingly fate restricted progenitors, and differentiate into bone, cartilage, and bone marrow stroma, all on the clonal level in vivo without prior in vitro culture. SSCs hold great potential for implementation in craniofacial bone tissue engineering and regenerative medicine. As we begin to better understand the diversity and the nature of skeletal stem and progenitor cells, there is a tangible future whereby a subset of human adult SSCs can be readily purified from bone or activated in situ with broad potential applications in craniofacial tissue engineering.

Three-Dimensional Printing for Craniofacial Bone Tissue Engineering.

The basic concepts from the fields of biology and engineering are integrated into tissue engineering to develop constructs for the repair of damaged and/or absent tissues, respectively. The field has grown substantially over the past two decades, with particular interest in bone tissue engineering (BTE). Clinically, there are circumstances in which the quantity of bone that is necessary to restore form and function either exceeds the patient's healing capacity or bone's intrinsic regenerative capabilities. Vascularized osseous or osteocutaneous free flaps are the standard of care with autologous bone remaining the gold standard, but is commonly associated with donor site morbidity, graft resorption, increased operating time, and cost. Regardless of the size of a craniofacial defect, from trauma, pathology, and osteonecrosis, surgeons and engineers involved with reconstruction need to consider the complex three-dimensional (3D) geometry of the defect and its relationship to local structures. Three-dimensional printing has garnered significant attention and presents opportunities to use craniofacial BTE as a technology that offers a personalized approach to bony reconstruction. Clinicians and engineers are able to work together to produce patient-specific space-maintaining scaffolds tailored to site-specific defects, which are osteogenic, osseoconductive, osseoinductive, encourage angiogenesis/vasculogenesis, and mechanically stable upon implantation to prevent immediate failure. In this work, we review biological and engineering principles important in applying 3D printing technology to BTE for craniofacial reconstruction as well as present recent translational advancements in 3D printed bioactive ceramic scaffold technology.

Photoencapsulated-BMP2 in visible light-cured thiol-acrylate hydrogels for craniofacial bone tissue engineering.

Aim: The study aimedto examine the impact of crosslinking BMP2 in biodegradable visible light-cured thiol-acrylate hydrogels. Materials & methods: BMP2 wasphotoencapsulated in 10wt% PEG-diacrylatehydrogels with or without immortalized mouse bone marrow stromal cells (BMSC). Results & conclusion: Photoencapsulated-BMSC with BMP2 (BMBMP2) showeda significantly (p < 0.05) increased level in metabolic activity, by 54.61%, compared with photoencapsulated-BMSCat day 3. Furthermore, BMBMP2 groups showed significantly increased levels in ALPactivity compared with BMSC at days, 1, 3, 7 (p < 0.01) and 10 (p < 0.05). This study shows promising results photoencapsulating BMP2 in thiol-acrylate hydrogels for craniofacial bone tissue engineering applications.

An engineered cell-laden adhesive hydrogel promotes craniofacial bone tissue regeneration in rats.

Cell-laden hydrogels are widely used in tissue engineering and regenerative medicine. However, many of these hydrogels are not optimized for use in the oral environment, where they are exposed to blood and saliva. To address these challenges, we engineered an alginate-based adhesive, photocrosslinkable, and osteoconductive hydrogel biomaterial (AdhHG) with tunable mechanical properties. The engineered hydrogel was used as an injectable mesenchymal stem cell (MSC) delivery vehicle for craniofacial bone tissue engineering applications. Subcutaneous implantation in mice confirmed the biodegradability, biocompatibility, and osteoconductivity of the hydrogel. In a well-established rat peri-implantitis model, application of the adhesive hydrogel encapsulating gingival mesenchymal stem cells (GMSCs) resulted in complete bone regeneration around ailing dental implants with peri-implant bone loss. Together, we have developed a distinct bioinspired adhesive hydrogel with tunable mechanical properties and biodegradability that effectively delivers patient-derived dental-derived MSCs. The hydrogel is photocrosslinkable and, due to the presence of MSC aggregates and hydroxyapatite microparticles, promotes bone regeneration for craniofacial tissue engineering applications.

Decellularized pulp matrix as scaffold for mesenchymal stem cell mediated bone regeneration

Scaffolds that are used for bone repair should provide an adequate environment for biomineralization by mesenchymal stem cells (MSCs). Recently, decellularized pulp matrices (DPM) have been utilized in endodontics for their high regenerative potential. Inspired by the dystrophic calcification on the pulp matrix known as pulp stone, we developed acellular pulp bioscaffolds and examined their potential in facilitating MSCs mineralization for bone defect repair. Pulp was decellularized, then retention of its structural integrity was confirmed by histological, mechanical, and biochemical evaluations. MSCs were seeded and proliferation, osteogenic gene expression, and biomineralization were assessed to verify DPM’s osteogenic effects in vitro. MicroCT, energy-dispersive X-ray (EDX), and histological analyses were used to confirm that DPM seeded with MSCs result in greater mineralization on rat critical-sized defects than that without MSCs. Overall, our study proves DPM’s potential to serve as a scaffolding material for MSC-mediated bone regeneration for future craniofacial bone tissue engineering.