Residual Biomaterial Research Articles

Dynamics of CSBD Healing after Implementation of Dentin and Xenogeneic Bone Biomaterial

Autologous dentin is frequently used in guided bone regeneration due to its osteoinductive properties, which come from its similarity to native bone. On the other hand, the xenogeneic bone biomaterial Cerabone® serves as a biocompatible, but hardly resorbed biomaterial. During bone healing, an inflammatory, vascular, and osteogenic response occurs in which cytokines such as tumor necrosis factor-alpha (TNF-α), vascular endothelial growth factor (VEGF), and osteopontin (OPN) are released locally and systemically. The aim was to follow up the dynamics (on days 3, 7, 15, 21, and 30) of critical-sized bone defect (CSBD) healing after the implantation of bovine devitalized dentin, rat dentin, and xenogeneic bone biomaterial. For this purpose, histological and histomorphometric methods were employed. Additionally, serum concentrations of TNF-α, VEGF, and OPN were monitored in parallel to better understand the biomaterial-dependent systemic response in rats. At the last time interval, the results showed that the bone defect was bridged over in all three groups of biomaterials. The rat dentin group had the highest percentage of bone volume (BV/TV) and the least percentage of residual biomaterial (RB), which makes it the most optimal biomaterial for bone regeneration. Serum concentrations of the TNF-α, VEGF, and OPN refer to systemic response, which could be linked to intense bone remodeling between days 15 and 21 of the bone healing.

Prospective Clinical Study with New Materials for Tissue Regeneration: A Study in Humans.

This study was performed to evaluate the clinical, radiographic, and histomorphometric outcomes of novel bone grafting materials and dental membranes and to compare the results with current data from the literature. New synthetic bone substitutes, consisting of biphasic calcium phosphate in the ratio of 60% hydroxyapatite and 40% β-tricalcium phosphate, were applied in bony defects and covered by either a novel synthetic poly(lactic-co-glycolic) acid (PLGA) or porcine collagen membrane. A sample of 51 biomaterials was placed in a total of 20 patients during different surgical protocols. Implants were simultaneously inserted, and in the case of sinus floor elevations 6 months later. Pre- and postoperative cone-beam computed tomographies were taken. Bone biopsies were harvested from augmented sides and processed for histomorphometric evaluation. Averages and ranges were calculated for the percentage of newly formed bone, residual biomaterial, and connective tissue. Data were submitted to analyze the radiological mean differences in length, width, and density. Paired t-tests were deployed for the analysis of differences within each group between the baseline (preoperative) and the final (postoperative) measurements. The mean bone gain in length and width were 0.96 ± 3.33 mm (+27.59%) and 1.22 ± 1.87 mm (+30.48%), respectively. The bone density was increased by a factor of 4, reaching an average of 387.47 ± 328.86 HU. Histomorphometric evaluations revealed new bone formation of 41.44 ± 5.37%, residual biomaterial of 24.91 ± 7.31%, and connective tissue of 33.64 ± 4.81%. The mean healing period was 8.32 ± 3.00 months. Data from this study confirmed the suitability of the tested materials in dental surgery. The biomaterials may be recommended for various clinical procedures. A satisfactory level of increase of new bone was reported in augmented sides. No significant differences were observed between the tested membranes. PLGA membranes might be superior to collagen membranes for their easier handling.

Periosteal Flaps Enhance Prefabricated Engineered Bone Reparative Potential

The clinical translation of bone tissue engineering for reconstructing large bone defects has not advanced without hurdles. The in vivo bioreactor (IVB) concept may therefore bridge between bone tissue engineering and reconstructive surgery by employing the patient body for prefabricating new prevascularized tissues. Ideally, IVB should minimize the need for exogenous growth factors/cells. Periosteal tissues are promising for IVB approaches to prefabricate tissue-engineered bone (TEB) flaps. However, the significance of preserving the periosteal vascular supply has not been adequately investigated. This study assessed muscle IVB with and without periosteal/pericranial grafts and flaps for prefabricating TEB flaps to reconstruct mandibular defects in sheep. The sheep (n = 14) were allocated into 4 groups: muscle IVB (M group; nM = 3), muscle + periosteal graft (MP group; nMP = 4), muscle + periosteal flap (MVP group; nMVP = 4), and control group (nControl = 3). In the first surgery, alloplastic bone blocks were implanted in the brachiocephalic muscle (M) with a periosteal graft (MP) or with a vascularized periosteal flap (MVP). After 9 wk, the prefabricated TEB flaps were transplanted to reconstruct a mandibular angle defect. In the control group, the defects were reconstructed by non-prevascularized bone blocks. Computed tomography (CT) scans were performed after 13 wk and after 23 wk at termination, followed by micro-CT (µCT) and histological analyses. Both CT and µCT analysis revealed enhanced new bone formation and decreased residual biomaterial volume in the MVP group compared with control and MP groups, while the M group showed less new bone formation and more residual biomaterial. The histological analysis showed that most of the newly formed bone emerged from defect edges, but larger areas of new bone islands were found in MP and MVP groups. The MVP group showed enhanced vascularization and higher biomaterial remodeling rates. The periosteal flaps boosted the reconstructive potential of the prefabricated TEB flaps. The regenerative potential of the periosteum was manifested after the transplantation into the mechanically stimulated bony defect microenvironment.

CSBD Healing in Rats after Application of Bovine Xenogeneic Biomaterial Enriched with Magnesium Alloy.

Xenogeneic biomaterials Cerbone® and OsteoBiol® are widely used in oral implantology. In dental practice, xenogeneic biomaterial is usually combined with autologous bone to provide bone volume stability needed for long-term dental implants. Magnesium alloy implants dissolve and form mineral corrosion layer that is directly in contact with bone tissue, allowing deposition of the newly formed bone. CSBD heals by intramembranous ossification and therefore is a convenient model for analyses of ostoconductive and osteoinductive properties of different type of biomaterials. Magnesium alloy-enriched biomaterials have not yet been applied in oral implantology. Therefore, the aim of the current study was to investigate biological properties of potentially new bovine xenogeneic biomaterial enriched with magnesium alloy in a 5 mm CSBD model. Osteoconductive properties of Cerabone®, Cerabone® + Al. bone, and OsteoBiol® were also analyzed. Dynamics of bone healing was followed up on the days 3, 7, 15, 21, and 30. Calvary bone samples were analyzed by micro-CT, and values of the bone morphometric parameters were assessed. Bone samples were further processed for histological and immunohistochemical analyses. Histological observation revealed CSBD closure at day 30 of the given xenogeneic biomaterial groups, with the exception of the control group. TNF-α showed high intensity of expression at the sites of MSC clusters that underwent ossification. Osx was expressed in pre-osteoblasts, which were differentiated into mature osteoblasts and osteocytes. Results of the micro-CT analyses showed linear increase in bone volume of all xenogeneic biomaterial groups and also in the control. The highest average values of bone volume were found for the Cerabone® + Mg group. In addition, less residual biomaterial was estimated in the Cerabone® + Mg group than in the Cerabone® group, indicating its better biodegradation during CSBD healing. Overall, the magnesium alloy xenogeneic biomaterial demonstrated key properties of osteoinduction and biodegradidibility during CSBD healing, which is the reason why it should be recommended for application in clinical practice of oral implantology.

A retrospective preliminary histomorphometric and clinical investigation on sinus augmentation using enzyme-deantigenic, collagen-preserving equine bone granules and plasma rich in growth factors

BackgroundEnzyme-deantigenic equine bone (EDEB) is a substitute of autogenous bone. Mixing it with plasma rich in growth factors (PRGF) seems a viable option to achieve enhanced bone formation in alveolar bone augmentation surgeries. This retrospective study aims to first report the histomorphometric and clinical outcomes achieved when using the EDEB/PRGF mixture for performing sinus augmentation procedures followed by delayed implant placement.Materials and methodsRecords of 11 patients who underwent 14 sinus augmentation surgeries using EDEB/PRGF followed by delayed implant placement were retrospectively collected and analyzed to assess histomorphometric data concerning newly formed bone (NFB) and residual biomaterial (RB) recorded at implant placement, marginal bone loss (MBL) values of implants placed in the augmented sinuses, and implant and prosthetic success and survival rates.ResultsAt 5.6 ± 1.1 months after grafting, NFB and RB were 34.0 ± 9.1% and 11.3 ± 2.2% respectively, and no histologic signs of inflammation or immune reaction were observed in any of the 34 bone biopsies being collected. Further, 86.5 ± 4.3 months after implant placement, MBL was 0.40 ± 0.07 mm. No implant or prosthesis failed, and the implant success and survival rates were 100%ConclusionsWithin the limitations of the present study, grafting EDEB/PRGF for lateral sinus augmentation and delayed implant placement seems to be safe. Compared to published data concerning EDEB alone, results of the present study do not suggest that the EDEB/PRGF combination may provide a histomorphometric or medium-/long-term clinical advantage.

The effect of synthetic bone graft substitutes on bone formation in rabbit calvarial defects

The aim of this study was to evaluate the influence of the intensity of the biomimetic hydroxyapatite (HA) coating of α-tricalcium phosphate (α-TCP) on biomaterial degradation and bone formation. Twenty-four female NZW rabbits of approximately 12 weeks of age were used. Critical size defects were randomly treated with 3%:97% HA:α-TCP (BBCP1), 12%:88% HA:α-TCP (BBCP2), and 23%:77% HA:α-TCP (BBCP3), respectively or sham. All defects were covered with a resorbable collagen membrane. Animals were euthanized after 3 and 12 weeks of healing and samples were investigated by micro-CT and histologic analysis. Ingrowth of newly formed woven bone from the original bone at 3-week healing period was observed in all samples. At the 12-week healing period, the new bone in the peripheral area was mainly lamellar and in the central region composed of both woven and lamellar bone. New bony tissue was found on the surface of all three types of granules and at the interior of the BBCP1 granules. Samples with 3% HA showed significantly less residual biomaterial in comparison to the other two groups. Furthermore, BBCP1 significantly promoted new bone area as compared to other three groups and more bone volume as compared to the control. Within its limitations, this study indicated the highest degradation rate in case of BBCP1 concomitant with the highest rate of bone formation. Hence, formation of new bone can be affected by the level of biomimetic HA coating of α-TCP.

Maxillary sinus augmentation with three different biomaterials: Histological, histomorphometric, clinical, and patient-reported outcomes from a randomized controlled trial.

Lateral maxillary sinus augmentation (MSA) is a predictable bone regeneration technique in case of atrophy of the posterior-upper maxilla. Aimed at obtaining quantity and quality of bone suitable for receiving osseointegrated implants, its success is largely due to the skill of the surgeon, but also to the characteristics of the biomaterial used. Twenty-four patients needing MSA were included in the study. The patients were randomly allocated to three different groups: anorganic bovine bone mineral as control, tricalcium phosphate with or without hyaluronic acid (HA) as test groups. Nine months after MSA, bone biopsies were harvested for the histomorphometric analysis. Secondary outcomes were mean bone gain, intraoperative and postoperative complications, implant insertion torque, implant failure, and patient-reported outcome measures. Although the percentage of new bone was not statistically different between the three groups (P = .191), the percentages of residual biomaterial was significantly higher (P < .000) and nonmineralized tissue significantly lower (P < .000) in the control than in the test groups. Test groups did not differ significantly from each other for all histomorphometric parameters. The implant insertion torque was significantly higher in the control group (P < .0005). The rest of the secondary outcomes were not significantly different between the groups. MSA is a safe and predictable procedure in terms of histological, clinical, and PROAMs, regardless of the biomaterial used. The addition of HA did not influence the outcomes.

Ridge Preservation Using a Novel Enzyme-Treated Xenograft. A Preliminary Retrospective Histomorphometric Investigation

The use of xenografts to preserve the post-extraction alveolar ridge is an established and effective procedure. Recently, a novel freeze-dried, enzyme-deantigenic equine bone (EDEB) particulate combined with a hydrogel carrier (Exur®) containing ascorbic acid has been developed (EDEBEX). The aim of this study was to preliminarily investigate histomorphometric and early implant survival outcomes following the graft of EDEBEX in post-extractive sockets. Records of patients who underwent ridge preservation using EDEBEX followed by two-step implant placement were retrospectively collected and analyzed. Newly Formed Bone (NFB) and Residual Biomaterial (RB) at the implant placement site were measured through histomorphometric analysis, and early Marginal Bone Loss (MBL) for implants was calculated at the final follow-up. Records concerned 13 patients (nine women and four men, average age 54.1 ± 9.5 years). The 13 sockets were considered healed 4.5 ± 2.6 months (mean ± SD) after grafting, with NFB and RB values of 43.2 ± 22.1% and 8.8 ± 5.9%, respectively. 8.4 ± 5.8 months after implant placement, the median MBL was 0.20 [0.00–0.45] mm. No correlation was observed between MBL and NFB. EDEBEX grafted in post-extractive sockets for ridge preservation seems to allow for new bone formation with satisfactory implant outcomes. Future prospective studies are necessary to confirm these preliminary findings.

Chemical, Clinical and Histomorphometric Comparison between Equine Bone Manufactured through Enzymatic Antigen-Elimination and Bovine Bone Made Non-Antigenic Using a High-Temperature Process in Post-Extractive Socket Grafting. A Comparative Retrospective Clinical Study.

Enzyme-deantigenic equine bone (EDEB) and anorganic bovine bone (ABB) are two xenografts made non-antigenic through different processing methods. This study aimed to characterize them for the presence of native bone collagen and other proteins and to compare their histomorphometric outcome when they were used to graft post-extractive sockets. The records of 46 patients treated with EDEB (n = 22) or ABB (n = 24) and followed-up for at least four months after delayed implant placement, were retrospectively collected. Samples of EDEB and ABB were analyzed using Attenuated Total Reflection Fourier Transform Infrared and Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis for the presence of collagen and other proteins. For histomorphometric analysis on bone specimens, newly formed bone and residual biomaterial percentages were calculated. Results of the present study show that EDEB contains type I bone collagen in its native conformation, while no proteins were detected in ABB. Grafting EDEB resulted in a significantly greater quantity of newly formed bone and less residual biomaterial. Our findings suggest that the manufacturing process can greatly affect the graft behavior and a process preserving collagen in its native form may favor bone tissue regeneration.

Utilization of agroindustrial wastes for the production of laccase by Achromobacter xylosoxidans HWN16 and Bordetella bronchiseptica HSO16

Agroindustrial residual lignocellulosic biomaterial provides an economical and renewable natural bioresource for the large-scale, gainful biofuel production, as well as the production of fine bulk chemicals, which may include industrial biocatalysts. To this end, the laccase-inducing aptitude of some agroindustrial, lignocellulosic residues were appraised in submerged fermentation batch culture of two woodland betaproteobacteria (Hb9c; Achromobacter xylosoxidans HWN16, Hb16c; Bordetella bronchiseptica HSO16). Significant fermentation factors for laccase production were identified following a one-variable-at-a-time: OVAT screening method, levels of significant factors were optimized using response surface methodology: RSM. Mandarin peelings: MP and wheat bran: WB were suitable substrates for laccase production in Hb9c; 29.4 U/mL and Hb16c; 28.2 U/mL, respectively. However, the numerical optimization of significant factors for laccase production in both isolates presented an overall maximum laccase output encountered throughout the study (Hb9c; 169.39 U/mL, Hb16c; 45.22 U/mL), albeit the simulated conditions of the statistical model were outside the design space of the algorithm such as pH 5, 0.5 g MP, 100 rpm, 0.25 g NaNO3 for Hb9c and pH 3, 2.5 g WB, 50 rpm, 0.05 g yeast extract for Hb16c. Furthermore, a record 17.5 and 15.54 fold increase in laccase turnover depicts the astuteness of the statistical method in the valorization of these lignocellulosic residues for enhanced laccase production, hence, the incorporation of these outcomes at industrial scales might yield tremendous outputs.

Morphological and Structural Study of a Novel Porous Nurse's A Ceramic with Osteoconductive Properties for Tissue Engineering.

The characterization process of a new porous Nurse’s A ceramic and the physico chemical nature of the remodeled interface between the implant and the surrounding bone were studied after in vivo implantation. Scaffolds were prepared by a solid-state reaction and implanted in New Zealand rabbits. Animals were sacrificed on days 15, 30, and 60. The porous biomaterial displayed biocompatible, bioresorbable, and osteoconductive capacity. The degradation processes of implants also encouraged osseous tissue ingrowths into the material’s pores, and drastically changed the macro- and microstructure of the implants. After 60 healing days, the resorption rates were 52.62% ± 1.12% for the ceramic and 47.38% ± 1.24% for the residual biomaterial. The elemental analysis showed a gradual diffusion of the Ca and Si ions from the materials into the newly forming bone during the biomaterial’s resorption process. The energy dispersive spectroscopy (EDS) analysis of the residual ceramic revealed some particle categories with different mean Ca/P ratios according to size, and indicated various resorption process stages. Since osteoconductive capacity was indicated for this material and bone ingrowth was possible, it could be applied to progressively substitute an implant.

Novel Nanostructured Zn-substituted Monetite Based Biomaterial for Bone Regeneration

A bone regeneration biomaterial must be biocompatible, osteoconductive and osteoinductive, and be gradually replaced by newly formed bone in the shorter time possible. Nanostructured materials, emulating bone composition and morphology, have shown great potential in bone repair because of their higher reactivity, faster reabsorption and improved biological behavior over microstructured materials. This work was aimed at developing a new biomaterial that meets the requirements for effective bone regeneration. Combination of elements with different solubility such as monetite, hydroxyapatite, amorphous calcium phosphate and silica gel, provided with the means to modulate the rate of material reabsorption of this novel biomaterial were Zn was found to be present as a partial substitution of Ca in the monetite lattice. The biomaterial was obtained by a hydraulic cementing reaction and was characterized by XRD, FTIR, NMR, chemical analysis, N2 adsorption porosimetry, Hg porosimetry, picnometry, SEM, TEM and evaluated in vitro and in vivo. Biomaterial showed a nanometric structure with a very high specific surface area (≈80 m2/g), high surface roughness and high intragranular porosity (50%) ranging from macro to nanopores. All of these are key features for a bone regeneration material. Solubility studies demonstrate the different solubility of its components and the release of Ca, P, Si and Zn. In vivo evaluation showed the effectiveness of the material to regenerate and to maintain 89 ± 9% of the volume of a critical size bone defect in sheep at 16 weeks. In the restored volume residual biomaterial was found to occupy 9 ± 5% of the space while newly formed trabeculae occupied 32 ± 6% of the space. The newly formed bone showed abundant vascularization and osteogenic activity. Biomaterial was estimated to have resorbed by 86 ± 7% with a reduction of 67 ± 8% in diameter and clear signs of cell-mediated resorption.

Ultrastructural study by backscattered electron imaging and elemental microanalysis of biomaterial-to-bone interface and mineral degradation of bovine xenografts in maxillary sinus floor elevation.

The aim of this study was to perform an ultrastructural study of the bone-to-biomaterial interface and biomaterial mineral degradation in retrieved bone biopsies following maxillary sinus augmentation using bovine xenografts (Endobon(®)) in 15 clinical cases. Nine months after sinus lifting, bone cores were harvested from the maxillary sinus. The specimens were processed for observation under a scanning electron microscope with backscattered electron imaging (SEM-BSE). In addition, chemical analysis and elemental mapping of the mineral composition were generated using a microanalytical system based on energy-dispersive X-ray spectrometry (EDX). No clinical complications occurred during surgery. Scanning electron microscopy revealed that newly formed bone was closely attached to the xenograft. Elemental analysis showed a significantly high Ca/P ratio in the residual biomaterials (3.031 ± 0.104) compared with the interface (2.908 ± 0.115) and new bone (2.889 ± 0.113), which suggests that there may be a gradual diffusion of Ca ions from the biomaterial into the newly forming bone at the interface as part of the biomaterial's resorption process. EDX analysis of the residual bovine biomaterial showed particle categories with different mean Ca/P ratios according to size, pointing to different stages of the resorption process. The biomaterial proved to be biocompatible and osteoconductive when used as a bone substitute for maxillary sinus elevation. The grafting material used is not a completely resorbable material over the time period covered by this study.

Histological evaluation of healing after transalveolar maxillary sinus augmentation with bioglass and autogenous bone

The aim was to evaluate histologically the outcome of a bioglass and autogenous bone (at 1 : 1 ratio) composite implantation for transalveolar sinus augmentation. In 31 patients, during implant installation ca. 4 months after sinus augmentation, biopsies were harvested through the transalveolar osteotomy by means of a trephine bur and non-decalcified sections through the long axis of the cylinder were produced. After a strict selection process, taking into account the presurgical residual bone height and biopsy length, 8 and 15 biopsies representing the new tissues formed inside the sinus and the transalveolar osteotomy, respectively, qualified for analysis. The tissue fractions occupied by newly formed bone (mineralized tissue+bone marrow), soft connective tissue, residual biomaterial+empty spaces, and debris inside the sinus cavity or the transalveolar osteotomy were estimated. Bone and connective tissue fraction in the newly formed tissues inside the sinus cavity averaged 23.4 ± 13.2% and 54.1 ± 23.5%, respectively. Residual biomaterial, empty spaces, and debris averaged 1.9 ± 3.5%, 10.5 ± 6.3%, and 8.4 ± 14.5%, respectively. In the transalveolar osteotomy, bone and connective tissue fraction averaged 41.6 ± 14.3% and 46.1 ± 13%, respectively, while the amount of residual biomaterial, empty spaces, and debris was 2.8 ± 5%, 4.7 ± 1.9%, and 3.2 ± 2.6%, respectively. Statistically significant differences between the sinus cavity and the transalveolar osteotomy were found only for bone and empty spaces' values (P=0.02 and 0.04, respectively). Sinus augmentation with a bioglass and autogenous bone composite is compatible with bone formation that, in a short distance from the floor of the sinus, shows similar density as that reported previously for other commonly used bone substitutes. New bone fraction inside the transalveolar osteotomy was almost twice as much as in the sinus cavity, while the amount of residual biomaterial was much less than that inside the sinus.

Bone Formation Following Implantation of Bone Biomaterials Into Extraction Sites

Adequate bone volume is imperative for the osseointegration of endosseous implants, but postextraction resorption and remodeling may challenge implant placement. The use of bone biomaterials has been advocated to fill extraction sites and to enhance primary implant stability during osseointegration. The objective of the case series was to evaluate bone formation histologically and biomechanically in extraction sites following implantation of three commercially available bone biomaterials to compare their ability to allow guided bone regeneration. Thirty-six periodontally involved teeth were extracted from eight healthy non-smoking subjects. At least two bone biomaterials, a synthetic sponge based on polylactic-polyglycolic acid technology (FIS), bovine porous bone mineral (BPBM), or a natural coral derivative physically and chemically transformed into a calcium carbonate ceramic (COR), and one non-grafted control were applied to the extraction sockets within each subject and were covered by an expanded polytetrafluoroethylene device. The devices were removed after 2 months, and trephine biopsies were obtained from each site 4 months later. At that time, endosseous implants were placed in 25 of the sites, and healing abutments were placed; measurements were taken 4 to 6 months later with an electronic mobility testing device. The percentage of residual biomaterial was 5.6% +/- 8.9% for FIS (P <0.001), 20.2% +/- 17.0% for BPBM (P <0.05), and 12.0% +/- 16.4% for COR (P <0.001). The amount of residual biomaterial after 6 months showed a significant relationship with the insertion torque measurements during the first third of implant insertion (P <0.05) and with values of the electronic mobility testing device at the abutment connection (P = 0.05). Histologically, new bone apposition was seen on BPBM particles. FIS sites showed similar ingrowth of blood vessels and osteocytes as empty controls. All sites revealed good primary stability at implant insertion and proper implant rigidity at abutment placement, indicating that early implant osseointegration was not influenced by the application of bone biomaterials used in this study.