Maxillofacial Bone Regeneration Research Articles

Nanobioactive blood-derived shear-thinning biomaterial for tissue engineering applications

The conventional technique for successful bone grafts, involving the use of a patient's own tissue (autografts), is challenged by limited availability and donor site morbidity. While allografts and xenografts offer alternatives, they come with the risk of rejection. This underscores the pressing need for tailor-made artificial bone graft materials. In this context, injectable hydrogels are emerging as a promising solution for bone regeneration, especially in complex maxillofacial reconstruction cases. These hydrogels can seamlessly adapt to irregular shapes and conservatively fill defects. Our study introduces a shear-thinning biomaterial by blending silicate nanoplatelets (SNs) enriched with human blood-derived plasma rich in growth factors (PRGF) for personalized applications. Notably, our investigations unveil that injectable hydrogel formulations comprising 7.5 % PRGF yield sustained protein and growth factor release, affording precise control over critical growth factors essential for tissue regeneration. Moreover, our hydrogel exhibits exceptional biocompatibility in vitro and in vivo and demonstrates hemostatic properties. The hydrogel also presents a robust angiogenic potential and an inherent capacity to promote bone differentiation, proven through Alizarin Red staining, gene expression, and immunostaining assessments of bone-related biomarkers. Given these impressive attributes, our hydrogel stands out as a leading candidate for maxillofacial bone regeneration application. Beyond this, our findings hold immense potential in revolutionizing the field of regenerative medicine, offering an influential platform for crafting precise and effective therapeutic strategies.

Wireless Electric Cues Mediate Autologous DPSC-Loaded Conductive Hydrogel Microspheres to Engineer the Immuno-Angiogenic Niche for Homologous Maxillofacial Bone Regeneration.

Stem cell therapy serves as an effective treatment for bone regeneration. Nevertheless, stem cells from bone marrow and peripheral blood are still lacking homologous properties. Dental pulp stem cells (DPSCs) are derived from neural crest, in coincidence with maxillofacial tissues, thus attracting great interest in in situ maxillofacial regenerative medicine. However, insufficient number and heterogenous alteration of seed cells retard further exploration of DPSC-based tissue engineering. Electric stimulation has recently attracted great interest in tissue regeneration. In this study, a novel DPSC-loaded conductive hydrogel microspheres integrated with wireless electric generator is fabricated. Application of exogenous electric cues can promote stemness maintaining and heterogeneity suppression for unpredictable differentiation of encapsulated DPSCs. Further investigations observe that electric signal fine-tunes regenerative niche by improvement on DPSC-mediated paracrine pattern, evidenced by enhanced angiogenic behavior and upregulated anti-inflammatory macrophage polarization. By wireless electric stimulation on implanted conductive hydrogel microspheres, loaded DPSCs facilitates the construction of immuno-angiogenic niche at early stage of tissue repair, and further contributes to advanced autologous mandibular bone defect regeneration. This novel strategy of DPSC-based tissue engineering exhibits promising translational and therapeutic potential for autologous maxillofacial tissue regeneration.

Effects of rhBMP-2 with various carriers on maxillofacial bone regeneration through computed tomography evaluation

BackgroundrhBMP-2 is regarded as the most potent osteoinductive growth factor, and it has been used in the oral cavity with different carriers. The purpose of this study is to evaluate the bone-regenerative effect of rhBMP-2 delivered with different carrier systems through three-dimensional cone beam computed tomography analysis.MethodA total of 112 patients underwent oral surgery with rhBMP-2 application (Group 1, n = 53) or without rhBMP-2 application (Group 2, n = 59). Group 1 was divided into 3 groups according to carriers, rhBMP-2 with allograft (Group 1–1, n = 34), rhBMP-2 with xenograft (Group 1–2, n = 5), and rhBMP-2 with absorbable collagen sponge (Group 1–3, n = 14). Cone beam computed tomography scans were taken before surgery (T0) 6 months after surgery (T1). The volume of defects was measured through the three-dimensional image analysis tool.ResultsThe average bone regeneration rate of Group 1 was significantly greater than that of Group 2. Within Group 1, the group that used allograft as a carrier (Group 1–1) showed significantly higher bone regeneration rates than the group that used absorbable collagen sponge as a carrier (Group 1–3).ConclusionThe use of rhBMP-2 after oral surgery results in a superior bone regeneration rate compared to not using rhBMP-2, and its efficacy depends on the carriers it is used with. Allograft affects bone regeneration more than absorbable collagen sponge when it is carried with rhBMP-2. Therefore, the appropriate use of rhBMP-2 with suitable bone grafting materials is useful for promoting postoperative bone regeneration in oral surgery.

3D‐printed degradable hydroxyapatite bioactive ceramics for skull regeneration

AbstractHydroxyapatite (HA) bioceramics have been extensively employed as bone tissue scaffolds owing to their biodegradability and osteoinductivity. In our work, HA, a significant component of natural bone tissue used as the raw material to produce porous scaffolds employing three‐dimensional (3D)‐printing technology. Physical and chemical properties, porosity, and compression resistance of the scaffolds were investigated in vitro. The scaffold was confirmed to have a large number of interconnected pore structures on the surface and inside HA scaffolds showed good cell compatibility and cell adhesion in cell text. To analyze the effect of the scaffold on bone repair and regeneration in vivo, the large‐size defect of beagle skull was repaired with a 3D printing group and an autologous bone group (ABG) for 8 months. Images and histological analysis of the 3D printing group indicated better integration with adjacent tissues. However, there were obvious gaps in the ABG, which indicates weak bone regeneration ability of this group due to unmatched implant dimension. Immunohistochemistry and immunofluorescence results showed that 3D‐printed scaffolds had a highly vascularized structure. This study indicates that 3D‐printed bioceramics scaffolds that are osteoinductivity and biodegradable have great potential in maxillofacial bone regeneration.

Tissue buccal fat pad–stromal vascular fraction as a safe source in maxillofacial bone regeneration: A clinical pilot study

The purpose of this study was to examine the biological properties of the buccal fat pad (BFP)-derived tissue stromal vascular fraction (tSVF) in vitro and compare them with BFP-derived cellular SVF (cSVF). Furthermore, a clinical pilot study assessed the safety of using BFP-derived tSVF for maxillofacial bone regeneration. This study was performed in two sections: 1) experimental section: BFP tissue was harvested from three healthy donors, and then cSVF and tSVF were isolated by enzymatic and mechanical methods to assess their biological properties and 2) clinical section: Ten patients with maxillofacial bone defects were enrolled according to eligibility criteria and offered two options for surgery, including autologous BFP-tSVF (n=5) and autologous bone grafting (n=5), to evaluate safety after a year of follow-up. The BFP-tSVF exhibited high cell viability and various cell surface markers, including CD45, CD31, and CD34. There was no population-doubling time and multilineage differentiation capacity compared with BFP-cSVF. BFP-tSVF is safe because of the lack of intervention-related adverse events reported in donor and surgery sites during a one-year period. In addition, cell therapy was feasible because it can be performed during surgery and requires little preparation time.Patients in the ABG group experienced pain and tenderness in the iliac crest, leading to dissatisfaction and complications. The experimental results confirmed that the cells isolated from BFP-tSVF have stemness properties similar to BFP-cSVF. Clinical evaluation also indicated that this cellular product could be used safely to regenerate maxillofacial bone defects.

Major clinical outcomes of cellular and molecular regulation in maxillofacial bone regeneration: a systematic review

Introduction: The importance of knowing the constitution of this epithelium is because these hair cells play a fundamental role in the physiology of the maxillary sinus. When a dental element is lost in the posterior region of the maxilla, there is natural reabsorption of the alveolar process and, at the same time, pneumatization of the maxillary sinus will occur. Objective: It was to demonstrate, through a systematic review, the main clinical outcomes of cellular and molecular regulation in maxillofacial bone regeneration. Methods: The systematic review rules of the PRISMA Platform were followed. The research was carried out from September to October 2022 in Scopus, PubMed, Science Direct, Scielo, and Google Scholar databases. The quality of the studies was based on the GRADE instrument and the risk of bias was analyzed according to the Cochrane instrument. A total of 110 articles were found, 41 articles were evaluated and 31 were included in this systematic review. Considering the Cochrane tool for risk of bias, the overall assessment resulted in 31 studies at high risk of bias and 25 studies that did not meet the GRADE. Most studies showed homogeneity in their results, with I2 =98.9% >50%. Results and Conclusion: It was found that the search for a solution to large bone defects guided the studies for regeneration therapy tissue or bone regeneration. These studies can promote the use of fillers and epithelial barriers that help in the treatment as an adjuvant to bone graft techniques, favoring greater predictability in alveolar and peri-implant reconstructions and with a good prognosis. The main filler biomaterials can be fibrin-rich plasma (FRP), Bio-Oss®. It is necessary to understand the chemical, physical and biological processes of both the biological material and the biological niche of the host. Crossing compatible information between microenvironments allows cell recognition and signaling cascades for neovascularization and regeneration and bone filling.

Optimization of a concentrated growth factor/mesoporous bioactive glass composite scaffold and its application in rabbit mandible defect regeneration.

Maxillofacial bone defect repair and regeneration remains a tremendous challenge in the field of stomatology. However, the limited osteoinductivity of artificial materials and the high cost of bioactive agents restrain their clinical translation. This study aimed to construct an economical and efficient concentrated growth factor/mesoporous bioactive glass (CGF/MBG) composite scaffold for bone regeneration. The biochemical composition and biological effects of different forms of CGFs were systematically compared, and the results showed that CGF-conditioned medium effectively promoted proliferation, migration and osteogenesis of allogenic BMSCs. Gel phase CGF (gpCGF) exhibited superior bioactivity and osteoinductivity to liquid phase CGF (lpCGF) and liquid/gel mixed phase CGF (lgpCGF), and was further applied to construct CGF/MBG scaffolds. In vitro studies demonstrated that co-culture with gpCGF-conditioned medium further enhanced the biocompatibility of MBG, increasing cell adhesion and proliferation on the scaffold. On this basis, two compositing approaches to construct the scaffold by fibrin gel formation (CGF/FG/MBG) and freeze-drying (fdCGF/MBG) were applied, and the biological efficacy of CGFs was compared in vivo. In a rabbit mandibular defect model, higher osteogenic efficiency in in situ bone regeneration of CGF/FG/MBG composite scaffolds was proved, compared with fdCGF/MBG. Taken together, the CGF/FG/MBG composite scaffold is expected to be an efficient bone repairing therapy for clinical translation, and the CGF-composited scaffold using gpCGF and the fibrin gel formation method is a promising way to enhance the bioactivity and osteoinductivity of current clinical bone repairing materials, providing new thoughts on the development of future orthopedic biomaterials.

Periodontal ligament stem cell-based bioactive constructs for bone tissue engineering.

Objectives: Stem cell-based tissue engineering approaches are promising for bone repair and regeneration. Periodontal ligament stem cells (PDLSCs) are a promising cell source for tissue engineering, especially for maxillofacial bone and periodontal regeneration. Many studies have shown potent results via PDLSCs in bone regeneration. In this review, we describe recent cutting-edge researches on PDLSC-based bone regeneration and periodontal tissue regeneration. Data and sources: An extensive search of the literature for papers related to PDLSCs-based bioactive constructs for bone tissue engineering was made on the databases of PubMed, Medline and Google Scholar. The papers were selected by three independent calibrated reviewers. Results: Multiple types of materials and scaffolds have been combined with PDLSCs, involving xeno genic bone graft, calcium phosphate materials and polymers. These PDLSC-based constructs exhibit the potential for bone and periodontal tissue regeneration. In addition, various osteo inductive agents and strategies have been applied with PDLSCs, including drugs, biologics, gene therapy, physical stimulation, scaffold modification, cell sheets and co-culture. Conclusoin: This review article demonstrates the great potential of PDLSCs-based bioactive constructs as a promising approach for bone and periodontal tissue regeneration.

Carrier systems for bone morphogenetic proteins: An overview of biomaterials used for dentoalveolar and maxillofacial bone regeneration.

Different types of biomaterials have been used to fabricate carriers to deliver bone morphogenetic proteins (BMPs) in both dentoalveolar and maxillofacial bone regeneration procedures. Despite that absorbable collagen sponge (ACS) is considered the gold standard for BMP delivery, there is still some concerns regarding its use mainly due to its poor mechanical properties. To overcome this, novel systems are being developed, however, due to the wide variety of biomaterial combination, the heterogeneous assessment of newly formed tissue, and the intended clinical applications, there is still no consensus regarding which is more efficient in a particular clinical scenario. The combination of two or more biomaterials in different topological configurations has allowed specific controlled-release patterns for BMPs, improving their biological and mechanical properties compared with classical single-material carriers. However, more basic research is needed. Since the BMPs can be used in multiple clinical scenarios having different biological and mechanical needs, novel carriers should be developed in a context-specific manner. Thus, the purpose of this review is to gather current knowledge about biomaterials used to fabricate delivery systems for BMPs in both dentoalveolar and maxillofacial contexts. Aspects related with the biological, physical and mechanical characteristics of each biomaterial are also presented and discussed.

A novel lineage of osteoprogenitor cells with dual epithelial and mesenchymal properties govern maxillofacial bone homeostasis and regeneration after MSFL

Bone regeneration originates from proliferation and differentiation of osteoprogenitors via either endochondral or intramembranous ossification; and the regeneration capacities decline with age and estrogen loss. Maxillary sinus floor lifting (MSFL) is a commonly used surgical procedure for guiding bone regeneration in maxilla. Radiographic analysis of 1210 clinical cases of maxilla bone regeneration after MSFL revealed that the intrasinus osteogenic efficacy was independent of age and gender, however; and this might be related to the Schneiderian membrane that lines the sinus cavity. In view of the particularity of this biological process, our present study aimed to elucidate the underlying mechanism of MSFL-induced bone regeneration. We first established a murine model to simulate the clinical MSFL. By single-cell RNA-sequencing and flow cytometry-based bulk RNA-sequencing, we identified a novel Krt14+Ctsk+ subset of cells that display both epithelial and mesenchymal properties and the transcriptomic feature of osteoprogenitors. Dual recombinases-mediated lineage tracing and loss-of-function analyses showed that these Krt14+Ctsk+ progenitors contribute to both MSFL-induced osteogenesis and physiological bone homeostasis by differentiating into Krt14–Ctsk+ descendants which show robust osteogenic capacity. In addition, we detected a similar population of Krt14+Ctsk+ cells in human samples of Schneiderian membrane, which show a highly similar osteogenic potential and transcriptomic feature to the corresponding cells in mice. The identification of this Krt14+Ctsk+ population, featured by osteoprogenitor characteristics and dual epithelial–mesenchymal properties, provides new insight into the understanding of bone regeneration and may open more possibilities for clinical applications.

Hydroxyapatite/tricalcium phosphate (HA/beta-TCP) scaffold combined with bone and endothelial cells as a potential candidate for oral and maxillofacial bone regeneration.

Cell-based scaffolds have been proposed as useful for tissue repair and bone regeneration in oral and maxillofacial tissue engineering. However, vascularization, a key player in bone regeneration, is often inadequate. To improve this, the use of biphasic calcium phosphate biomaterial (30% hydroxyapatite (HA)-70% tricalcium phosphate (TCP) mixture) HA/beta-TCP-scaffold with defined micron-channel sizes and pore sizes capable of driving angiogenesis, may prove helpful. Therefore, our study aimed to combine the HA/beta-TCP scaffold with in vitro differentiated cells, especially endothelial cells, well-known players in angiogenesis. The HA/beta-TCP-scaffold was combined with primary cultures of human osteoblasts (hOBs), osteoclasts (hOCs), and endothelial cells (Human Umbilical Vein Endothelial Cells, HUVECs). Cells were co-cultured with HA/beta-TCP-scaffold in normoxia (20% oxygen tension) for 14 days. Then, we evaluated: cell adhesion/morphology (Scanning Electron Microscopy and Confocal Microscopy); cell viability (Calcein AM/Propidium Iodide Assay); the presence of multinucleated hOCs (Tartrate Resistant- Acid Phosphatase assay); expression of specific markers of both osteo- and vasculogenic differentiation (Immunohistochemistry). All cell types were viable and well adhered to the entire scaffold surface. Interestingly, bone matrix formation and newly formed vessel-like structures were both detected. Although further investigation is needed, our data proved that differentiated cells combined in vitro with HA/beta-TCP-scaffold prior to implantation could represent a useful strategy for future applications in bone regeneration. Especially, the use of customized micro-vascularized cell-based scaffold would fall within the innovative "personalized medicine” approach.

Bone Tissue Engineering Using Osteogenic Cells: From the Bench to the Clinical Application.

The use of tissue engineering to restore and to build new bone tissue is under active research at present. The following review summarizes the latest studies and clinical trials related to the use of osteogenic cells, biomaterials, and scaffolds to regenerate bone defects in the human jaws. Bone tissue engineering (BTE) combined with scaffolds have provided a range of advantages not only to transport the target cells to their desired destination but also to support the early phases of the mineralization process. The mechanical, chemical, and physical properties of scaffolds have been evaluated as they affect the quantity of bone regeneration, particularly in the oral cavity. This review also highlighted the mechanisms underlying bone homeostasis, including the key transcription factors and signaling pathways responsible for regulating the differentiation of osteoblast lineage. Furthering understanding of the mechanisms of cellular signaling in skeletal remodeling with the use of mesenchymal stem cells and the proper scaffold properties are key-factors to enable the incorporation of new and effective treatment methods into clinical practice for bone tissue regeneration using BTE. Impact Statement The use of mesenchymal stem cells able to differentiate in osteoblast lineage for bone tissue engineering (BTE) remains a major challenge. Viable cells and signaling pathways play an essential role in bone repair and regeneration of critical size defects. Recent advances in scaffolds and biological factors such as growth factors (e.g., cytokines and hormones) controlling the osteogenic signaling cascade are now becoming new players affecting the osteogenic potential of cells. Such techniques will significantly impact the maxillofacial bone tissue replacement, repair, and regeneration for patients without having to rely on donor banks or other surgical sites.

Cell viability of novel composite hydrogels loaded with hydroxyapatite for oral and maxillofacial bone regeneration.

The development of hydrogels for maxillofacial bone regeneration holds vast potential. However, some challenges need to be addressed to further their application in clinical settings. One challenge is optimizing cell viability. To improve mechanical strength, various materials have been investigated; however, incorporation of these materials within the hydrogel network may affect cell viability. The purpose of this study was to evaluate the cell viability of novel gelatin-alginate composite hydrogels loaded with hydroxyapatite (HA) and nano-hydroxyapatite (n-HA) for maxillofacial bone regeneration. Nine different hydrogels were prepared: three loaded with 0.5%, 1%, and 3% w/v HA; three loaded with 0.25%, 0.5%, and 1% w/v n-HA; one not loaded as a control and two HA and n-HA hydrogels with a lower concentration of the EDC crosslinker. Cell viability of human osteoblasts exposed to the hydrogels as affected by the HA type, size, and concentration, as well as to the crosslinker concentration, was investigated. An Alamar Blue assay was used to evaluate cell viability in the presence of hydrogel extracts and in aqueous solutions (without the hydrogel). A qualitative model was developed for explaining cell viability and growth. Higher percentages of cell viability were observed in the hydrogels loaded with hydroxyapatite as compared with the control. The effect of HA-related parameters, i.e., particle size and concentration, was found to increase the cytotoxic effect, as expressed in lower cell viability. The most favorable composites were the n-HA hydrogels. The incorporation of n-HA in the hydrogel to form a composite seems to be a very promising approach for maxillofacial bone regeneration applications.

Polydopamine–Ag composite surface guides HBMSCs adhesion and proliferation

Human bone marrow mesenchymal stem cells (HBMSCs) are regarded as an important resource in the field of maxillofacial bone regeneration because of their favorable properties when compared with other stem cells. Hence, finding suitable materials that could extend the application of HBMSCs has become an emerging medical topic and socioeconomic problem. In this work, polydopamine (PDA)–Ag surface was fabricated by PDA assisted photoreduction method, and the obtained PDA–Ag composite surface significantly promoted HBMSCs adhesion and proliferation. This effect is highly related to the amount of Ag nanoparticles (Ag NPs) present on the PDA surface. The behavior of HBMSCs on PDA–Ag surface could be spatially manipulated by controlling the distribution of Ag NPs on PDA surface (by controlling UV light). The general adhesion property allows the PDA–Ag surface to be fabricated on various substrates, making it a simple, general and controllable method for the fabrication of bioactive surface for HBMSCs.

Considerations for Clinical Use of Concentrated Growth Factor in Maxillofacial Regenerative Medicine

Different strategies have been utilized to facilitate mineralized/soft tissues. Concentrated growth factor (CGF) emerges as a promising biomaterial for regenerative therapy due to high levels of platelets, growth factors, and nucleated cells entrapped in the fibrin scaffold. This review aimed to collect extensive studies on CGF used in maxillofacial regenerative medicine, discussing current obstacles, and expressing some considerations of CGF use. Articles were retrieved systematically without time limitation. In total, 27 human studies were included and separated accordingly. In general, CGF has been most evaluated in implant related therapy and maxillofacial bone regeneration where a majority of articles have revealed favorable outcomes. Little studies have supported the effects on improving probing periodontal depth reductions and clinical attachment level gains for intrabony and furcation defect regeneration. Very little data with high-level evidence was available directly to investigate its effects on the soft tissue regeneration and postoperative complications reduction. The evidence supporting the clinical efficacy of CGF in maxillofacial regenerative medicine is limited. Some conflicting results are worrisome. In addition, the characterization of CGF preparation protocols and CGF's components was not performed in most studies. Further studies should make a consensus on standardized end-product of CGF including characterization of protocols and optimal ratio of CGF' components. Defined algorithms and evidence-based protocols for the clinical use of CGF should be also available.

Extracellular vesicles derived from human dental pulp stem cells promote osteogenesis of adipose-derived stem cells via the MAPK pathway

Recent studies have shown that co-culture systems play an important role in bone tissue engineering. In this study, human dental pulp stem cells (hDPSCs) were co-cultured with human adipose-derived stem cells (hADSCs), and osteoblastic phenotypes were found to be enhanced in co-cultures compared with monocultures of hDPSCs or hADSCs. Furthermore, GW4869, an inhibitor of extracellular vesicle (EV) formation, suppressed the mineralization of co-cultured cells. Studies indicate that the therapeutic potential of DPSCs is realized through paracrine action, in which EVs play an important role. To study their role, we successfully obtained and identified hDPSC-derived extracellular vesicles (hDPSC-EVs), and further investigated their effects on hADSCs and the underlying mechanism. hADSCs were stimulated with hDPSC-EVs, which were found to promote the migration and mineralization of hADSCs. Moreover, hDPSC-EVs promoted osteogenic differentiation by enhancing the phosphorylation of ERK 1/2 and JNK in hADSCs. To investigate the specific proteins in EVs that might play a role in hADSC osteogenic differentiation, we performed proteomic analysis of hDPSC-EVs. We determined the top 30 enriched pathways, which notably included the insulin signaling pathway. The number of genes enriched in the insulin signaling pathway was the largest, in addition to the “protein processing in endoplasmic reticulum” term. The MAPK cascade is a typical downstream pathway mediating insulin signaling. To further study the effects of hDPSC-EVs on maxillofacial bone regeneration, we used hDPSC-EVs as a cell-free biomaterial in a model of mandibular defects in rats. To assess the therapeutic potential of EVs, we analyzed their proteome. Animal experiments demonstrated that hDPSC-EVs promoted the regeneration of bone defects. Overall, these results highlight the potential of hDPSC-EVs to induce lineage specific differentiation of hADSCs. The results also indicated the importance of considering hDPSC-EVs as biomimetic materials for clinical translation of treatments for oral maxillofacial defects.

Platelet-Rich Fibrin as a Bone Graft Material in Oral and Maxillofacial Bone Regeneration: Classification and Summary for Better Application.

Platelet-rich fibrin (PRF) is an autologous platelet concentrate that consists of cytokines, platelets, leukocytes, and circulating stem cells. It has been considered to be effective in bone regeneration and is mainly used for oral and maxillofacial bone. Although currently the use of PRF is thought to support alveolar ridge preservation, there is a lack of evidence regarding the application of PRF in osteogenesis. In this paper, we will provide examples of PRF application, and we will also summarize different measures to improve the properties of PRF for achieving better osteogenesis. The effect of PRF as a bone graft material on osteogenesis based on laboratory investigations, animal tests, and clinical evaluations is first reviewed here. In vitro, PRF was able to stimulate cell proliferation, differentiation, migration, mineralization, and osteogenesis-related gene expression. Preclinical and clinical trials suggested that PRF alone may have a limited effect. To enlighten researchers, modified PRF graft materials are further reviewed, including PRF combined with other bone graft materials, PRF combined with drugs, and a new-type PRF. Finally, we will summarize the common shortcomings in the application of PRF that probably lead to application failure. Future scientists should avoid or solve these problems to achieve better regeneration.

Hydroxyapatite-collagen augments osteogenic differentiation of dental pulp stem cells.

The objectives of this study were to isolate and culture dental pulp stem cells (DPSCs) and to investigate their proliferation and osteogenic differentiation on hydroxyapatite-collagen (HA-Col) scaffold. DPSCs were characterized by fluorescence-activated cell sorting (FACS). Cultured cells were CD73+, CD90+, CD105+ and CD31-, CD45-. A commercially available HA-Col scaffold was used for culture of DPSCs. Cell attachment and viability of DPSCs cultured on scaffold was studied by sulforhodamine assay. Osteoblast differentiation capacity was studied by alkaline phosphatase assay and the effects of growth factors such as PDGF, IGF1 and FGF2 were further studied. Scanning electron microscopy (SEM) of cell seeded scaffolds was also performed. We found that DPSCs cultured exhibited the characteristic mesenchymal stem cells (MSCs) morphology and differentiation properties. Scaffold was found to be non-cytotoxic and had good biocompatibility in vitro. Osteoblast differentiation ability was found to increase at higher concentration of scaffold and additive effects were observed with the use of growth factors. In SEM, cells appeared to cover the entire surface of the scaffold forming continuous cell layer and extending filopodial extensions. HA-Col scaffold is apt for MSCs attachment and proliferation in vitro. Their unique self-renewal and multilineage differential potential make them ideal for use in regenerative medicine. The limitations of currently available bone graft materials have led to the emergence of tissue engineering using mesenchymal stem cells (MSCs). Since, HA-Col scaffold potentiated the proliferation and osteogenic differentiation of DPSCs, this biomimetic material may be an ideal one for maxillofacial and alveolar bone regeneration.

MiRNA-21 promotes osteogenesis via the PTEN/PI3K/Akt/HIF-1\u03b1 pathway and enhances bone regeneration in critical size defects

BackgroundFunctional reconstruction of maxillofacial bone defects is a considerable clinical challenge. Many studies have emphasized the osteogenic and angiopoietic abilities of stem cells for tissue regeneration. We previously showed that microRNA-21 (miRNA-21) can promote angiogenesis in human umbilical cord blood-derived mesenchymal stem cells (UCBMSCs). In the present study, the role of miRNA-21 in osteogenic differentiation of bone marrow-derived stem cells (BMSCs) was investigated.MethodsWestern blotting and qPCR were performed to investigate the influences of miRNA-21 on osteogenic differentiation of BMSCs. The effects of miRNA-21 on PTEN/PI3K/Akt/HIF-1α pathway were also assessed using western blotting. To further evaluate the roles of miRNA-21 in osteogenesis in vivo, we conducted animal experiments in rat and canine. New bone formation was assessed using micro-CT and histological methods.ResultsIn the present study, we found that miRNA-21 promotes the migration and osteogenic differentiation of bone marrow-derived stem cells (BMSCs) in vitro. Using gain- and loss-of-function studies, we found that miRNA-21 promoted the osteogenic ability of BMSCs by increasing P-Akt and HIF-1α activation. Finally, we verified the essential role of miRNA-21 in osteogenesis by implanting a miRNA-21-modified BMSCs/β-tricalcium phosphate (β-TCP) composite into critical size defects. Radiography, micro-CT, and histology revealed significantly greater volume of new bone formation in the miRNA-21 group than in the control group.ConclusionIn conclusion, our study demonstrated an essential role of miRNA-21 in promoting maxillofacial bone regeneration via the PTEN/PI3K/Akt/HIF-1α pathway.

A standardized rat burr hole defect model to study maxillofacial bone regeneration

With high incidence rate and unique regeneration features, maxillofacial burr hole bone defects require a specially designed bone defect animal model for the evaluation of related bone regenerative approaches. Although some burr hole defect models have been developed in long bones or calvarial bones, the mandible has unique tissue development origins and regenerative environments. This suggests that the defect model should be prepared in the maxillofacial bone area. After dissecting the anatomic structures of rat mandibles, we found that creating defects in the anterior tooth area avoided damaging important organs and improved animal welfare. Furthermore, the available bone volume at the anterior tooth area was superior to that of the posterior tooth and ascending ramus areas. We then managed to standardize the model by controlling the age, weight and gender of the animal, creating standardized measurement instruments and reducing the variations derived from various operators. We also succeeded in deterring the self-rehabilitation of the proposed model by increasing the defect size. The 6 × 2 mm and 8 × 2 mm defects were found to meet the requirements of bone regenerative studies. This study provided a step-by-step standardized burr hole bone defect model with minimal tissue damage in small animals. The evaluations resulting from this model testify to the in vitro outcomes of the proposed regenerative approaches and provide preliminary screening data for further large animal and clinical trials. Therefore, the inclusion of this model may optimize the evaluation systems for maxillofacial burr hole bone defect regenerative approaches. Statement of SignificanceUnremitting effort has been devoted to the development of bone regenerative materials to restore maxillofacial burr hole bone defects because of their high clinical incidence rate. In the development of these biomaterials, in vivo testing in small animals is necessary to evaluate the effects of candidate biomaterials. However, little has been done to develop such defect models in small animals. In this study, we developed a standardized rat mandible burr hole bone defect model with minimal injury to the animals. A detailed description and supplementary video were provided to guide the preparation. The development of this model optimizes the maxillofacial bone regenerative approach evaluation system.