Clinical outcomes and risk factor analysis of dental implants inserted with lateral maxillary sinus floor augmentation: A 3- to 8-year retrospective study.

AIM: To evaluate the 3- to 8-year outcomes of dental implants placed with lateral sinus floor augmentation (LSFA) and to identify factors affecting implant survival. MATERIALS AND METHODS: This retrospective study was performed by screening all implants placed with LSFA procedures, which were conducted between January 2012 and December 2016. Subantral bone gain (SABG) and apical bone height (ABH) were assessed using panoramic radiographs. The cumulative survival rate of implants was analysed using life-table analysis and Kaplan-Meier survival curves. The influential risk factors affecting survival were assessed using univariate log-rank tests and multivariable mixture cure rate model. Implant complications were recorded. RESULTS: Based on the established criteria, a total of 449 patients (760 implants) were included in this study. In the 3- to 8-year follow-up (mean ± SD, 5.81 ± 1.33 years), 15 implants in 14 patients failed, with a CRS of 96.81% on an implant basis and 95.07% on a patient basis. A history of periodontitis and poor compliance with supportive periodontal treatment was associated with a significantly higher risk of implant failure at both implant and patient levels. Significant decreases in ABH occurred during each yearly interval except for 3 years. A similar trend has been observed for SABG at 1, 2, 6 and 8 years. The total complication rate was 31.84% on implant basis, with peri-implant mucositis (21.58%) being the most frequent biologic complication and porcelain cracking (5.00%) being the most common technical complication. CONCLUSIONS: Implant with LSFA is a reliable treatment option in atrophic maxilla. A history of periodontitis without regular supportive periodontal treatment was identified as a predictor for implant failure. Slight but significant shrinkage of vertically augmented bone can be observed after implant placement.

The biofilm removal effect and osteogenic potential on the titanium surface by electrolytic cleaning: An in vitro comparison of electrolytic parameters and five techniques.

OBJECTIVES: To determine the optimal current and time of electrolytic cleaning (EC), compare its biofilm removal effect with generic treatments and evaluate the influence of EC to surface characteristics and osteogenic potential of SLA titanium (Ti) discs. MATERIALS AND METHODS: The six-species biofilm-covered Ti discs were placed as cathodes in physiologic saline and subjected to various current and time treatments. The residual biofilms were evaluated to determine the optimal parameters. The contaminated Ti discs were randomized and treated by rotating Ti brush; ultrasonic-scaling with metal tips; ultrasonic-scaling with PEEK tips; air-polishing and EC. The residual biofilms were compared using a lipopolysaccharide kit (LPS), scanning electron microscope (SEM), confocal laser scanning microscopy and colony-forming unit counting. Non-contaminated Ti discs were treated and characterized. The bone marrow mesenchymal stem cells (BMSCs) were cultured on treated non-contaminated Ti discs. The adhesion, proliferation, alkaline phosphatase (ALP) activity and osteocalcin level of BMSCs were assessed. RESULTS: The parameters at 0.6A5min were considered optimal. For LPS and SEM, EC promoted a significantly greater biofilm removal than the other groups. There were no changes in the Ti discs' colour, topography, roughness and chemical elements after EC, and the electrolysis-treated Ti discs obtained a super-hydrophilic surface. EC positively impacted the proliferation and ALP activity of BMSCs, surpassing the efficacy of alternative treatments. CONCLUSIONS: EC achieves a near-complete eradication of contaminants on the SLA surface, causes no surface damage with improved hydrophilicity, and promotes the early osteogenic response of BMSCs, which makes it a promising treatment for peri-implantitis.

Metabolic profiling of peri-implant crevicular fluid in peri-implantitis.

OBJECTS: This study aims to explore the etiology of peri-implantitis by comparing the metabolic profiles in peri-implant crevicular fluid (PICF) from patients with healthy implants (PH) and those with peri-implantitis (PI). MATERIALS AND METHODS: Fifty-six patients were enrolled in this cross-sectional study. PICF samples were collected and analyzed using both non-targeted and targeted metabolomics approaches. The relationship between metabolites and clinical indices including probing depth (PD), bleeding on probing (BOP), and marginal bone loss (MBL) was examined. Additionally, submucosal microbiota was collected and analyzed using 16S rRNA gene sequencing to elucidate the association between the metabolites and microbial communities. RESULTS: Significant differences in metabolic profiles were observed between the PH and PI groups, with 179 distinct metabolites identified. In the PI group, specific amino acids and fatty acids were significantly elevated compared to the PH group. Organic acids including succinic acid, fructose-6-phosphate, and glucose-6-phosphate were markedly higher in the PI group, showing positive correlations with mean PD, BOP, and MBL. Metabolites that increased in the PI group positively correlated with the presence of Porphyromonas and Treponema and negatively with Streptococcus and Haemophilus. CONCLUSIONS: This study establishes a clear association between metabolic compositions and peri-implant condition, highlighting enhanced metabolite activity in peri-implantitis. These findings open avenues for further research into metabolic mechanisms of peri-implantitis and their potential therapeutic implications.

Risk factors associated with loss of variable-thread tapered implants: A retrospective observational study of 1-5 years.

OBJECTIVES: This study aimed to evaluate the survival rate of variable-thread tapered implants (VTTIs) and identify risk factors for early/late implant loss. MATERIALS AND METHODS: From January 2016 to December 2019, patients who received VTTIs were included in this study. The cumulative survival rates (CSRs) at implant/patient levels were calculated by the life table method and presented via Kaplan-Meier survival curves. The relation between investigated variables and early/late implant loss was analyzed by the multivariate generalized estimating equation (GEE) regression model on the implant level. RESULTS: A total of 1528 patients with 2998 VTTIs were included. 95 implants from 76 patients were lost at the end of observation. At the implant level, the CSRs at 1, 3, and 5 years were 98.77%, 96.97%, and 95.39%, respectively, whereas they were 97.84%, 95.31%, and 92.96% at the patient level, respectively. The multivariate analysis revealed that non-submerged implant healing (OR = 4.63, p = .037) was associated with the early loss of VTTIs. Besides, male gender (OR = 2.48, p = .002), periodontitis (OR = 3.25, p = .007), implant length <10 mm (OR = 2.63, p = .028), and overdenture (OR = 9.30, p = .004) could significantly increase the risk of late implant loss. CONCLUSION: Variable-thread tapered implants could reach an acceptable survival rate in clinical practice. Non-submerged implant healing was associated with early implant loss; male gender, periodontitis, implant length <10 mm, and overdenture would significantly increase the risk of late implant loss.

The Influence of Storage in Saline or Irradiation by Ultraviolet on Surface Hydrophilicity of Implant and Osseointegration: An Experimental Study in Rabbits.

Storage in aqueous solution or ultraviolet (UV) irradiation can retain or regain the hydrophilicity of titanium implant surface. In this study, 3 types of commercial titanium implants were used: ZBL (ZDI Bone Level), CEL (C-tech Esthetic Line), and modSLA (Straumann SLActive). ZBL and CEL implants were treated with UV irradiation for 4 hours. Surface characterization of the 4 groups (ZBL, ZBL-UV, CEL-UV, and modSLA) was evaluated by scanning electron microscopy and contact angle measurements. The in vivo bone response was evaluated by removal torque (RTQ) tests and histomorphometric analysis at 3, 6, and 12 weeks postimplantation. A total of 144 implants and 36 rabbits were used for experiments according to a previously established randomization sequence. The ZBL-UV, CEL-UV, and modSLA groups were hydrophilic, and nanostructures were observed on the modSLA implant surface. ModSLA achieved better RTQ value than ZBL at 12 weeks (P < .05). For histomorphometric analysis, ZBL-UV and CEL-UV implants showed higher bone area values in the cancellous bone zone at 6 weeks than did modSLA and ZBL implants (P < .05). In the cortical bone zone, all groups showed comparable bone-to-implant contact at all healing time points (P > .05). Both storage in saline and UV irradiation could retain or provoke hydrophilic surfaces and improve osseointegration. Compared with storage in saline, UV irradiation displayed slight advantages in promoting new bone formation in cancellous bone zone at an early stage.

IRAK4 inhibition: an effective strategy for immunomodulating peri-implant osseointegration via reciprocally-shifted polarization in the monocyte-macrophage lineage cells.

BACKGROUND: The biomaterial integration depends on its interaction with the host immune system. Monocyte-macrophage lineage cells are immediately recruited to the implant site, polarized into different phenotypes, and fused into multinucleated cells, thus playing roles in tissue regeneration. IL-1R-associated kinase 4 (IRAK4) inhibition was reported to antagonize inflammatory osteolysis and regulate osteoclasts and foreign body giant cells (FBGCs), which may be a potential target in implant osseointegration. METHODS: In in-vitro experiments, we established simulated physiological and inflammatory circumstances in which bone-marrow-derived macrophages were cultured on sand-blasted and acid-etched (SLA) titanium surfaces to evaluate the induced macrophage polarization, multinucleated cells formation, and biological behaviors in the presence or absence of IRAK4i. Then, bone marrow stromal stem cells (BMSCs) were cultured in the conditioned media collected from the aforementioned induced osteoclasts or FBGCs cultures to clarify the indirect coupling effect of multinucleated cells on BMSCs. We further established a rat implantation model, which integrates IRAK4i treatment with implant placement, to verify the positive effect of IRAK4 inhibition on the macrophage polarization, osteoclast differentiation, and ultimately the early peri-implant osseointegration in vivo. RESULTS: Under inflammatory conditions, by transforming the monocyte-macrophage lineage cells from M1 to M2, IRAK4i treatment could down-regulate the formation and activity of osteoclast and relieve the inhibition of FBGC generation, thus promoting osteogenic differentiation in BMSCs and improve the osseointegration. CONCLUSION: This study may improve our understanding of the function of multinucleated cells and offer IRAK4i as a therapeutic strategy to improve early implant osseointegration and help to eliminate the initial implant failure.

Strontium-incorporated titanium implant surface treated by hydrothermal reactions promotes early bone osseointegration in osteoporotic rabbits.

OBJECTIVES: The aim of the present study was to investigate the effects of strontium-modified implant surfaces on promoting early bone osseointegration in osteoporotic rabbits. MATERIALS AND METHODS: The surface topographies, chemical elements, contact angles and ionic releases of the SLA and SLA-Sr samples were analysed by special instruments separately. Sixteen ovariectomized New Zealand rabbits received glucocorticoid administration, and sixteen SHAM rabbits were used as controls. After generating a successful osteoporosis-induced model, SLA and SLA-Sr implants were randomly inserted into the tibia and femur metaphysis of each animal. The rabbits were sacrificed after 3 and 6 weeks of bone healing, and then, removal torque values (RTVs), percentage of bone area (BA%) and percentage of bone-to-implant contact (BIC%) were analysed for the SLA-Sr and SLA implants. RESULTS: Multiple nanostructures were found on the Sr-incorporated titanium surface, and appropriate amounts of strontium ions from the SLA-Sr surface were released into the surrounding tissue within 21 days. In vivo, SLA-Sr implants displayed much more new bone around their surfaces than the SLA implants. Significantly higher RTVs and BIC% were observed for the SLA-Sr implants than for the SLA implants in both osteoporotic (p < 0.01) and healthy animals (p < 0.01) at 3 and 6 weeks. The SLA-Sr implants exhibited higher BA% in cortical bone (p < 0.01) and in cancellous bone (p < 0.05) than the SLA implants in osteoporotic rabbits at 3 weeks. CONCLUSIONS: It is suggested that Sr-incorporated surfaces treated through hydrothermal reactions have positive effects on promoting early osseointegration in both osteoporotic and non-osteoporotic rabbits.

Titanium surface interacting with blood clot enhanced migration and osteogenic differentiation of bone marrow mesenchymal stem cells.

Introduction: Blood clot formation is the initial phase upon implantation, and the feature of blood clot orchestrates the following complement system activation, coagulation cascade, and bone marrow mesenchymal stromal cells (BMSCs) recruitment. This study aimed to investigate the effect of implant surface on blood-material interactions and subsequent BMSC cellular behaviors. Methods: This study was established to imitate the physiological process of implantation in vivo and in vitro. Whole blood was incubated with polished titanium (PT) surfaces and sandblasted and double acid-etching (SLA) surfaces for 10 min or 2 h, then seeded with BMSCs. The adhesion, proliferation, migration, and differentiation of cells were studied at specific time points. Titanium implants were implanted into the tibia in vivo and were screwed out after implantation. The activation of the coagulation cascade, platelets, complement system, and clot networks were assessed and further quantitatively analyzed. Results: Compared with the PT surface, the SLA surface induced the earlier and stronger blood coagulation cascade and formed a more stratified clots network with fibrinogen, platelets, and CD14 positive cell. The adhesion, proliferation, and migration of BMSCs were enhanced by pre-incubated surfaces. The higher levels of the osteogenic-related genes, ALP activity, and calcium nodule formation were showed on SLA surfaces with blood incubation. Conclusion: SLA titanium surfaces play a role in influencing the formation of blood clots and coordinating surface-blood interactions and cell biological processes. These findings provide the idea of modifying the blood clots formed on the implant surface by biomaterials modification and thus has implications for the development of better osteogenic biomaterials.

Clinical and patient-centered outcomes following rehabilitation of atrophic edentulous maxilla using six implants placed simultaneously with bilateral maxillary sinus augmentation: A retrospective case series.

OBJECTIVES: To evaluate treatment success, patient satisfaction, and oral health-related quality of life (OHRQoL) after rehabilitation of atrophic edentulous maxilla (AEM) with six implants placed simultaneously with bilateral maxillary sinus floor augmentation (MSFA). MATERIALS AND METHODS: The selected patients were fully edentulous with atrophic maxillary posterior residual ridges and rehabilitated with six implants placed simultaneously with bilateral MSFA and immediate All-on-4 provisional fixed prosthesis (PFP). After 7-12 months of implant surgery, all patients have received an All-on-6 definitive fixed prosthesis (DFP). After at least one year of function with DFP, clinical and radiographic examinations were performed. Patient satisfaction was assessed through a visual analog scale (VAS). The OHRQoL before treatment (T0), during provisional (T1), and after definitive prosthesis (T2) was evaluated using OHI-14. RESULTS: 20 maxillary edentulous patients were rehabilitated with 120 implants, 20 immediate All-on-4 PFP, and 20 All-on-6 DFP. Of those, 12 patients have passed at least a year follow-up after DFP insertion and were eligible to be included in the assessment of treatment success. After a mean of 20 (12-36) months follow-up, the implant and prosthesis survival rates were 100%. The success rate at the implant level was 98.6%. The mean marginal bone loss (MBL) was 0.09 ± 0.06 mm. No prosthetic or postoperative complications, and the mean general satisfaction was (91.75 ± 7.06). There was a significant improvement in all OHIP-14 domains during the final All-on-6 prosthesis (T2) (P < 0.01). CONCLUSIONS: Rehabilitation of atrophic edentulous maxilla using six implants with simultaneous bilateral MSFA and immediate All-on-4 PFP is a successful treatment approach associated with minimal postoperative complications and significant improvement in OHRQoL.

Remodeling of the osteoimmune microenvironment after biomaterials implantation in murine tibia: Single-cell transcriptome analysis.

Osseointegration seems to be a foreign body reaction equilibrium due to the complicated interactions between the immune and skeletal systems. The heterogeneity of the osteoimmune microenvironment in the osseointegration of implant materials remains elusive. Here, a single-cell study involving 40043 cells is conducted, and a total of 10 distinct cell clusters are identified from five different groups. A preliminary description of the osteoimmune microenvironment revealed the diverse cellular heterogeneity and dynamic changes modulated by implant properties. The increased immature neutrophils, Ly6C + CCR2hi monocytes, and S100a8hi macrophages induce an aggressive inflammatory response and eventually lead to the formation of fibrous capsule around the stainless steel implant. The enrichment of mature neutrophils, FcgR1hi and differentiated immunomodulatory macrophages around the titanium implant indicates favorable osseointegration under moderate immune response. Neutrophil-depletion mice are conducted to explore the role of neutrophils in osseointegration. Neutrophils may improve bone formation by enhancing the recruitment of BMSCs via the CXCL12/CXCR3 signal axis. These findings contribute to a better knowledge of osteoimmunology and are valuable for the design and modification of 'osteoimmune-smart' biomaterials in the bone regeneration field.

Radiographic assessment of deproteinized bovine bone mineral (DBBM) and collagen-stabilized DBBM for transalveolar sinus floor elevation: A 2-year retrospective cohort study.

BACKGROUND: Theoretically, collagen-stabilized deproteinized bovine bone mineral (DBBM-C) has better operability compared with DBBM. DBBM-C avoids dispersing during the transalveolar sinus floor elevation (TSFE) because of its block shape. PURPOSE: To evaluate radiological changes of using DBBM-C in TSFE. MATERIALS AND METHODS: Patients who received TSFE using DBBM (Bio-Oss®) or DBBM-C (Bio-Oss® collagen) with simultaneous implantation were recruited. Graft bone height apically (aGH), endo-sinus bone gain (ESBG), and crest bone level (CBL) were assessed through panoramic radiograph and cone beam computed tomography (CBCT). RESULTS: A total of 138 patients (138 implants) were retrospectively enrolled. After 2 years of implantation, the incidence of postoperative complications was 4.2% (95% CI: 0.9%-11.7%) and 4.5% (95% CI: 0.9%-12.7%) for DBBM and DBBM-C groups, respectively. Measured in panoramic radiograph, ΔaGH of DBBM-C (1.8 mm, SD: 1.4, 95% CI: 1.2-2.4, P = 0.044) group was significantly higher than that of DBBM (1.2 mm, SD: 1.4, 95% CI: 0.7-1.7) after 24 months. No significant differences for ΔCBL were noted during the entire observation period. Measured through CBCT, ESBG was 5.0 (SD: 1.8, 95% CI: 4.1-5.8) mm in DBBM group and 4.6 (SD: 1.6, 95% CI: 3.9-5.3) mm in DBBM-C group 24 months after implantation. The aGH value of DBBM-C group was significantly higher compared with DBBM in CBCT (OR = 1.4, 95% CI: 1.1-1.9, P = 0.020). CONCLUSIONS: DBBM-C could achieve similar bone generation as DBBM in TSFE. Both materials could maintain aGH, ESBG, and CBL relatively stable 2 years after implantation.

The Effect of Initial Biologic Width on Marginal Bone Loss: A Retrospective Study.

PURPOSE: To evaluate the short-term effect of dental implant placement, mucosa thickness, and their combined effects (initial biologic width) on marginal bone loss. MATERIALS AND METHODS: This was a retrospective study on patients who received implant surgery in the posterior region without bone augmentation surgery between 2012 and 2016, and implants had been loaded for more than 12 months. Each patient received radiographic examination before and after implant surgery, before the stage-two surgery, and during the 1- to 5-year follow-up. The thickness of mucosa, depth of dental implant placement, and crestal bone loss were evaluated on digital radiographs. The interaction was discussed by defining the combination of initial mucosal thickness and implantation depth as the initial biologic width. The implants were divided into four study groups based on the quartile of the initial biologic width. RESULTS: This study included 266 patients (94 male and 172 female, 22 to 85 years of age, mean age: 51.43 years), with 413 dental implants placed including 239 Straumann implants and 174 Ankylos implants. The average follow-up was 21.50 months. After 1 to 5 years, the median crestal bone loss around implants was 0.35 mm (0.30 mm for Straumann BL and 0.40 mm for Ankylos). The implants were divided into four groups: group A (≤ 2.85 mm), group B (2.85 to 3.40 mm), group C (3.40 to 3.97 mm), and group D (> 3.97 mm). Group B showed significantly less crestal bone loss than group A (0.38 mm vs 0.25 mm; P < .05) and group C (0.25 mm vs 0.40 mm; P < .05) during the follow-up. Significantly more crestal bone loss around implants was observed in the thin mucosa group than in the thick mucosa group (0.50 mm vs 0.30 mm; P < .001), while implants placed beneath the bone level displayed a significantly higher amount of marginal bone loss than implants placed even with the bone crest (0.50 mm vs 0.10 mm; P < .001). CONCLUSION: The initial biologic width has an effect on crestal bone loss. When the initial biologic width was between 2.85 and 3.40 mm, the marginal bone loss was lowest. Based on radiographic evaluation, implants placed in thick gingiva and even with the bone level showed less alveolar marginal bone loss compared with implants placed in thin gingiva and below the crestal bone level.

Modulating autophagy by strontium-doped micro/nano rough titanium surface for promotion of osteogenesis and inhibition of osteoclastogenesis.

Although it has been demonstrated that implant surfaces treated with strontium (Sr) promote osseointegration, the underlying intracellular mechanism remains unknown. Autophagy is a vital intracellular degradation mechanism that plays an essential role in maintaining bone homeostasis. Therefore, while designing implant biomaterials, it is critical to consider the autophagy mechanism. In this study, we fabricated Sr-doped micro/nano rough titanium implant surface by hydrothermal treatment (SLA+Sr). The in vitro results revealed that the SLA+Sr surface promoted osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) via autophagy activation. The SLA+Sr surface, on the other hand, inhibited osteoclast differentiation by downregulating autophagy. Additionally, in vivo, the SLA+Sr implant improved osseointegration, inhibited osteoclastogenesis, and upregulated autophagy levels in surrounding bone tissue cells. Our findings established a novel centralized mechanism by which SLA+Sr regulated osteogenesis and osteoclastogenesis during the osseointegration process through autophagy regulation. Moreover, endowing implants with the ability to modulate autophagy may be a promising strategy for enhancing implant osseointegration in the future translational medicine field.

Zinc- and strontium- co-incorporated nanorods on titanium surfaces with favorable material property, osteogenesis, and enhanced antibacterial activity.

Early infection and peri-implantitis after implant restoration are major reasons for dental implant failure. Implant-associated infections are majorly attributed to biofilm formation. In this study, co-incorporated zinc- (Zn-) and strontium- (Sr-) nanorod coating on sandblasted and acid-etched (SLA) titanium (SLA-Zn/Sr) was fabricated by hydrothermal synthesis. It was aimed at promoting osteogenesis while inhibiting biofilm formation. The nanorod-like particles (φ 30-50 nm) were found to be evenly formed on SLA-Zn/Sr (Zn: 1.49 ± 0.16 wt%; Sr: 21.69 ± 2.74 wt%) that was composed of well-crystallized ZnTiO3 and SrTiO3 phases. With a sufficient interface bonding strength (42.00 ± 3.00 MPa), SLA-Zn/Sr enhanced the corrosion resistance property of titanium. Besides, SLA-Zn/Sr promoted the cellular initial adhesion, proliferation and osteogenic differentiation of rBMSCs in vitro while inhibiting the adhesion of Staphylococcus aureus and Porphyromonas gingivalis . In addition, through down-regulating icaA gene expression, this novel surface reduced the secretion of polysaccharide intercellular adhesion (reduced by 87.9% compared to SLActive) to suppress the S. aureus biofilm formation. We, therefore, propose a new chemical modification on titanium for multifunctional implant material development. Due to the Zn/Sr co-doping in coating, material properties, early osteogenic effect and antibacterial ability of titanium can be simultaneously enhanced, which has the potential to be applied in dental implantation in the future.

Effects of strontium-incorporated micro/nano rough titanium surfaces on osseointegration via modulating polarization of macrophages.

Macrophages perform multiple functions in both inflammation and wound healing, and are one of the fore front cells during implant osseointegration that influence subsequent process. Essential trace element modification may effectively modulate titanium implant surface biological properties. In this work, strontium (Sr) incorporated micro/nano rough titanium surfaces (Sr-SLA) was fabricated by hydrothermal treatment, and immunoreaction of macrophages was further investigated. In vitro results revealed that Sr doping inhibited inflammatory response of macrophages, further attenuated the inhibitory effect on following bone marrow derived cells (BMSCs) osteogenic differentiation. The regulation of macrophages by Sr-SLA likely involved ERK signaling pathway. Consistently, the in vivo study showed that compared with titanium surface sand-blasted with large grit and double acid-etched (SLA) implants, Sr-SLA implants could enhance new bone formation accompanied with more alternatively activated M2 macrophages infiltration and less classically activated M1 macrophages infiltration. These results reveal the immunomodulatory ability of Sr-SLA of adjusting the functional status of macrophages through inhibiting M1 polarization while promoting M2 polarization.

The optimization of sintering treatment on bovine-derived bone grafts for bone regeneration: in vitro and in vivo evaluation.

Modifications of sintering temperature and treatment time of bovine-derived bone grafts affect their physicochemical properties and further influence biological activity. Three different temperature sintered bovine-derived bone grafts: group I (300 °C 3 h), group II (300 °C 3 h plus 530 °C 6 h), and group III (300 °C 3 h plus 1000 °C 2 h) and Bio-Oss® were characterized and then compared in vitro for their effects on bone marrow stromal cells (BMSCs) migration, proliferation, and differentiation as estimated by cell migration assay, Alkaline phosphatase (ALP) activity assay, and Alizarin red staining. Further, the four bone grafts were implanted into the calvarial defects of rabbits to evaluate bone regeneration and graft degradation. The four deproteinized bovine-derived bone grafts displayed different surface topography. Group II displayed the highest potential of attracting cells. Both groups I and II markedly promote BMSCs differentiation. After 6 and 12 weeks, defects grafted with groups I and II displayed a significant higher bone fraction than defects grafted with group III and Bio-Oss®. Bone graft remnants remained in all four groups. Taken together, sintering at 300 °C for 3 h and sintering at 300 °C for 3 h with an addition of 530 °C for 6 h of bovine-dervied bone grafts displayed potential use in bone regeneration. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 108B:272-281, 2020.

Comparison Study of Three Hydroxyapatite-Based Bone Substitutes in a Calvarial Defect Model in Rabbits.

PURPOSE: The aim of this study was to investigate the in vivo performance, including bone regeneration and biomaterial degradation, of three hydroxyapatite-based bone substitutes at surgically created defects in the cranium of rabbits. MATERIALS AND METHODS: A bovine-derived hydroxyapatite (BHA), a synthetic hydroxyapatite (SHA), and a nano-hydroxyapatite with collagen I (nHAC) composite were characterized and then placed in calvarial defects in rabbits. After surgery at 6 weeks and 12 weeks, the animals were sacrificed for further microcomputed tomography, histologic, and histomorphometric analyses. The amounts of newly formed bone, residual grafting materials, connective tissues, and tartrate-resistant acid phosphatase staining positive multinucleate cells were evaluated. RESULTS: The three hydroxyapatite-based bone substitutes displayed different surface topography. BHA had the highest amount of newly formed bone at both 6 weeks and 12 weeks, yet no significant difference was found between the three groups at 6 weeks. The amount of residual materials was significantly higher in the SHA group than the other groups at both time points. The amount of connective tissue was highest in BHA. More multinucleated cells were observed in the nHAC group. CONCLUSION: The three tested bone substitutes are suitable for grafting defects related to early bone healing. BHA had a better impact on bone formation with the prolongation of time. An integration of collagen type I and nano-hydroxyapatite may not improve bone regeneration of pure synthetic hydroxyapatite but helps to modify the degradation rate of pure hydroxyapatite.

Physiochemical characterization and biological effect of anorganic bovine bone matrix and organic-containing bovine bone matrix in comparison with Bio-Oss in rabbits.

Bovine origin matrix has been widely used in clinical applications and investigated by various research institutions. However, the potential factors that influence bone regeneration are still not thoroughly understood and need further investigations. In this study, bone regeneration properties of anorganic bovine bone matrix (ABBM), organic-containing bovine bone matrix (OBBM), and widely acknowledged anorganic bovine bone matrix (Bio-Oss) were compared. Besides, the correlations between physiochemical characterizations and bone regeneration properties of the three xenografts were also investigated. Physiochemical characterizations were measured by special instrumentations. In animal studies, the three xenografts were implanted into 8-mm-diameter cranial defects of 16 New Zealand white rabbits. The biological effects were evaluated by micro-computed tomography and histomorphometric analysis after 6 and 12 weeks of implantation. The physical characterizations showed that anorganic bovine bone matrix and Bio-Oss had more nanostructures, larger surface area, bigger pore volume, and bigger pore size than that of organic-containing bovine bone matrix. The chemical characterizations showed that anorganic bovine bone matrix and Bio-Oss had higher crystallinity than that of organic-containing bovine bone matrix, and organic-containing bovine bone matrix contained organic nitrogen (N) component. In vivo, anorganic bovine bone matrix and Bio-Oss possessed better bone regeneration properties than that of organic-containing bovine bone matrix. Taken together, nanostructures, larger surface area, bigger pore volume, and bigger pore size of xenografts played an active role in new bone formation. Besides, lower crystallinity and organic N element of xenografts produced a positive effect on graft degradation. The abovementioned findings could provide theoretical basis for better choice in clinical applications and better manufacturing hydroxyapatite-derived bone graft in the future.

Optimized Bone Regeneration in Calvarial Bone Defect Based on Biodegradation-Tailoring Dual-shell Biphasic Bioactive Ceramic Microspheres.

Bioceramic particulates capable of filling bone defects have gained considerable interest over the last decade. Herein, dual-shell bioceramic microspheres (CaP@CaSi@CaP, CaSi@CaP@CaSi) with adjustable beta-tricalcium phosphate (CaP) and beta-calcium silicate (CaSi) distribution were fabricated using a co-concentric capillary system enabling bone repair via a tailorable biodegradation process. The in vitro results showed the optimal concentration (1/16 of 200 mg/ml) of extracts of dual-shell microspheres could promote bone marrow mesenchymal cell (BMSC) proliferation and enhance the level of ALP activity and Alizarin Red staining. The in vivo bone repair and microsphere biodegradation in calvarial bone defects were compared using micro-computed tomography and histological evaluations. The results indicated the pure CaP microspheres were minimally resorbed at 18 weeks post-operatively and new bone tissue was limited; however, the dual-shell microspheres were appreciably biodegraded with time in accordance with the priority from CaSi to CaP in specific layers. The CaSi@CaP@CaSi group showed a significantly higher ability to promote bone regeneration than the CaP@CaSi@CaP group. This study indicates that the biphasic microspheres with adjustable composition distribution are promising for tailoring material degradation and bone regeneration rate, and such versatile design strategy is thought to fabricate various advanced biomaterials with tailorable biological performances for bone reconstruction.

The effects of Sr-incorporated micro/nano rough titanium surface on rBMSC migration and osteogenic differentiation for rapid osteointegration.

Recruitment of endogenous bone marrow-derived mesenchymal stem cells (BMSCs) has been widely discussed as an alternative strategy for bone regeneration. Strontium (Sr) is known to direct the BMSCs' commitment to the bone lineage and encourage bone formation; however, the underlying mechanisms remain elusive. In this study, an Sr-incorporated micro/nano rough titanium surface (MNT-Sr) was fabricated by hydrothermal treatment in an attempt to facilitate BMSCs' recruitment and their osteogenic differentiation to enhance rapid osseointegration. Micro rough titanium (MT) was set as the control biomaterial. In vitro, MNT-Sr and its extracts promoted the migration and osteogenic differentiation of BMSCs. In animal studies, green fluorescent protein (GFP)-labeled BMSCs were intravenously injected into wild-type rats for tracing before tibial implantation surgery. The GFP+BMSC recruitment to the implantation site was successfully triggered by MNT-Sr implantation. A trend for increased bone area (BA%), bone-implant contact (BIC%) and removal torque values (RTVs) was observed for the MNT-Sr implant compared to that observed for MT at 2 weeks. Advanced mechanism analysis indicated that Sr2+ enhanced the SDF-1α/CXCR4 signaling pathway both in vitro and in vivo. Taken together, these findings suggest that MNT-Sr has promising therapeutic potential for future use in dental implants by homing endogenous stem cells to stimulate bone regeneration.