Efficacy and Safety of Antibiotic Impregnated Microporous Nanohydroxyapatite Beads for Chronic Osteomyelitis Treatment: A Multicenter, Open-Label, Prospective Cohort Study.

Chronic osteomyelitis is still a serious health problem that causes disabling conditions and has an impact on the quality of life. The objective of this study was to determine the clinical efficacy and safety of localized antibiotics delivery via impregnated microporous nanohydroxyapatite (nHA-ATB) beads for chronic osteomyelitis treatment. A total of 62 patients were enrolled in this study. After radical surgical debridement, the bone defect was filled with three types of antibiotics (vancomycin or gentamicin or fosfomycin) impregnated HA beads. The follow-up period was 48 weeks. It was found that the success rate was approximately 98% with a re-infection in only one patient. Quality of life of all patients after treatment improved significantly over time. Systemic exposure to vancomycin and gentamicin after beads implantation was limited and high local antibiotics concentrations were found in wound drainage fluid at 24, 48 and 72 h. Blood biochemistry measurements did not show any nephrotoxic or hepatotoxic effects. 20 adverse events were reported, but 90% of the events were resolved without having to remove the beads and the patients recovered. Satisfactory outcomes were observed in terms of success rate, quality of life and adverse effect. nHA-ATB beads impregnated by vancomycin or gentamicin or fosfomycin could potentially be employed as an alternative product of choice for localized antibiotics delivery in chronic osteomyelitis treatment.

Customized 3D printed nanohydroxyapatite bone block grafts for implant sites: A case series.

PATIENTS: A case series of 12 patients (mean age, 53.5 years) with horizontal ridge deficiencies had augmentations with customized 3D printed nanohydroxyapatite (3DHA) block grafts prior to implant placement. 3DHA graft materials were fabricated to fit the individual patient defects using DICOMs from CBCT images obtained from each patient. The CBCT images were then converted into the STL file format and 3DHA was reconstructed by 3D printing. Surgical bone augmentation consisted of 3DHA incorporating concentrated growth factors (CGFs) and platelet-rich fibrin (PRF) membrane. At 6 months, a bone biopsy and implantation were performed. The primary outcome was horizontal bone gain after 6 months. The secondary outcomes included information on the clinical outcomes, dimensions, and histomorphometric results. DISCUSSION: The 3DHA block graft was successful in 10 of 12 patients. Graft adjustment was not required. All 3DHA adapted and fit well at all defect sites. Maximum mean horizontal bone gains were 3.06 ± 1.02 and 3.56 ± 0.23 mm from the DICOMs and STL data sets, respectively. The volume gain was 229.8 ± 82.96 mm3. A low pain score after surgery was reported of 1.41 ± 0.51, while the healing index score increased with a maximum mean of 4.7 ± 0.67. Thirteen implants were placed with good primary stability (ISQ = 65 ± 4.08), without additional guided bone regeneration. Histomorphometric analysis revealed that new bone formation, bone tissue, residual grafts, and connective tissue were 28.6 ± 1.88, 30.48 ± 4.81, 19.82 ± 4.07, and 20.81 ± 4.41%, respectively. CONCLUSIONS: A customized 3DHA block graft is a viable treatment option for primary implant-site augmentation.

Osteogenic differentiation and proliferation potentials of human bone marrow and umbilical cord-derived mesenchymal stem cells on the 3D-printed hydroxyapatite scaffolds.

Mesenchymal stem cells (MSCs) are a promising candidate for bone repair. However, the maintenance of MSCs injected into the bone injury site remains inefficient. A potential approach is to develop a bone-liked platform that incorporates MSCs into a biocompatible 3D scaffold to facilitate bone grafting into the desired location. Bone tissue engineering is a multistep process that requires optimizing several variables, including the source of cells, osteogenic stimulation factors, and scaffold properties. This study aims to evaluate the proliferation and osteogenic differentiation potentials of MSCs cultured on 2 types of 3D-printed hydroxyapatite, including a 3D-printed HA and biomimetic calcium phosphate-coated 3D-printed HA. MSCs from bone marrow (BM-MSCs) and umbilical cord (UC-MSCs) were cultured on the 3D-printed HA and coated 3D-printed HA. Scanning electron microscopy and immunofluorescence staining were used to examine the characteristics and the attachment of MSCs to the scaffolds. Additionally, the cell proliferation was monitored, and the ability of cells to differentiate into osteoblast was assessed using alkaline phosphatase (ALP) activity and osteogenic gene expression. The BM-MSCs and UC-MSCs attached to a plastic culture plate with a spindle-shaped morphology exhibited an immunophenotype consistent with the characteristics of MSCs. Both MSC types could attach and survive on the 3D-printed HA and coated 3D-printed HA scaffolds. The MSCs cultured on these scaffolds displayed sufficient osteoblastic differentiation capacity, as evidenced by increased ALP activity and the expression of osteogenic genes and proteins compared to the control. Interestingly, MSCs grown on coated 3D-printed HA exhibited a higher ALP activity and osteogenic gene expression than those cultured on the 3D-printed HA. The finding indicated that BM-MSCs and UC-MSCs cultured on the 3D-printed HA and coated 3D-printed HA scaffolds could proliferate and differentiate into osteoblasts. Thus, the HA scaffolds could provide a suitable and favorable environment for the 3D culture of MSCs in bone tissue engineering. Additionally, biomimetic coating with octacalcium phosphate may improve the biocompatibility of the bone regeneration scaffold.

Clinical evaluation of 3D printed nano-porous hydroxyapatite bone graft for alveolar ridge preservation: A randomized controlled trial.

BACKGROUND/PURPOSE: Ridge resorption after tooth extraction may result in inadequate bone volume and unfavorable ridge architecture for ideal implant placement. The use of bone substitutes has been advocated to fill extraction sites and to enhance primary implant stability. This study was made to evaluate the clinical efficacy of novel 3D printed nano-porous hydroxyapatite (3DP HA, test group) in comparison to nano-crystalline bone graft (NanoBone®, control group) in alveolar ridge preservation prior to implant placement. MATERIALS AND METHODS: Thirty patients were randomized into two groups following tooth extraction. All extracted sockets were filled with 3DP HA or NanoBone® and covered with a non-resorbable membrane. After four months, cone-beam computed tomography (CBCT) and intraoral scanner were used to measure dimensional changes of bone and soft tissue surface. Bone core specimens were harvested for histological analysis during implant osteotomy. Implant stability was assessed using a modified damping capacity analysis. RESULTS: At four months postoperatively, dimensional changes in soft tissue surface resorption were less in the test group than in the control group; however, alveolar bone resorption was the same in both groups. Histological analysis revealed new bone formation, residual graft and fibrous connective tissue in both groups. The average primary implant stability (IST) value for both groups was approximately 70. There was no statistically significant difference in all parameters between two groups (p > 0.05). CONCLUSION: 3DP HA could potentially be used as an alternative bone graft material for alveolar ridge preservation.

In vitro resorbability and granular characteristics of 3D-printed hydroxyapatite granules versus allograft, xenograft, and alloplast for alveolar cleft surgery applications.

Three-dimensionally printed hydroxyapatite (3DP HA) was investigated in regards to its functional properties supporting bone regeneration and tooth movement in alveolar cleft applications. Commercially available bovine xenograft (BXG), biphasic calcium phosphate alloplast (BCP), and two types of freeze-dried bone allograft granules (FDBA and FDBA-CMC) were employed as control samples. Degradability was studied by submerging the samples in pH 7.4 buffered solution at 37°C for 28 days and determining subsequent weight loss percentage. The wicking property and granular agglomeration were evaluated by putting the granules in contact with deionized water, blood, and phosphate-buffered saline (PBS). Both of FDBA and FDBA-CMC showed the greatest weight loss at 28 days followed by 3DP HA. In contrast, 3DP HA showed significantly greater wicking ability than other samples for all liquid types. FDBA-CMC exhibited the greatest granular agglomeration for all liquid types followed by 3DP HA. 3DP HA was found to be a favorable candidate for bone grafting in alveolar cleft treatment.

Performance evaluation of bilayer oxidized regenerated cellulose/poly ε-caprolactone knitted fabric-reinforced composites for dural substitution.

Ideally, alloplastic dural substitute should have functional properties resembling human dura mater and retain a watertight closure to prevent cerebrospinal leakage. Therefore, functional properties for successful dural closure application of newly developed bilayer oxidized regenerated cellulose knitted fabric/poly ε-caprolactone knitted fabric-reinforced composites were studied and compared with human cadaveric dura mater and three commercial dural substitutes including two collagen matrices and one synthetic poly-L-lactide patch. It was found that oxidized regenerated cellulose knitted fabric/poly ε-caprolactone knitted fabric-reinforced composites uniquely contained a bilayer structure consisting of micropores distributed within the relatively dense microstructure. Density, tensile properties and stitch tear strength of oxidized regenerated cellulose knitted fabric/poly ε-caprolactone knitted fabric-reinforced composites were found to be closed to human cadaveric dura mater than those of dense-type and porous-type dural substitutes. Water tightness performance in both sutured and non-sutured forms of oxidized regenerated cellulose knitted fabric/poly ε-caprolactone knitted fabric-reinforced composites was slightly inferior to human cadaveric dura mater, but still better than those of commercial dural substitutes. This study revealed that oxidized regenerated cellulose knitted fabric/poly ε-caprolactone knitted fabric-reinforced composite showed better functional properties than typical dural substitutes and was found to be a good candidate for being employed as a dural substitute. The role and relationship of both microstructure and the type of materials on the functional properties and water tightness of the dural substitutes were also elucidated.

Enhancement of mechanical properties of 3D printed hydroxyapatite by combined low and high molecular weight polycaprolactone sequential infiltration.

A new infiltration technique using a combination of low and high molecular weight polycaprolactone (PCL) in sequence was developed as a mean to improve the mechanical properties of three dimensional printed hydroxyapatite (HA). It was observed that using either high (M n~80,000) or low (M n~10,000) molecular weight infiltration could only increase the flexural modulus compared to non-infiltrated HA, but did not affect strength, strain at break and energy at break. In contrast, a combination of low and high molecular infiltration in sequence increased the flexural modulus, strength and energy at break compared to those of non-infiltrated HA or infiltrated by high or low molecular weight PCL alone. This overall enhancement was found to be attributed to the densification of low molecular weight PCL and the reinforcement of high molecular PCL concurrently. The combined low and high molecular weight infiltration in sequence also maintained high osteoblast proliferation and differentiation of the composites at the similar level of the HA. Densification was a dominant mechanism for the change in modulus with porosity and density of the infiltrated HA/PCL composites. However, both densification and the reinforcing performance of the infiltration phase were crucial for strength and toughening enhancement of the composites possibly by the defect healing and stress shielding mechanisms. The sequence of using low molecular weight infiltration and followed by high molecular infiltration was seen to provide the greatest flexural properties and highest cells proliferation and differentiation capabilities.

Influence of process parameters on the content of biomimetic calcium phosphate coating on titanium: a Taguchi analysis.

In this study, a statistical design of experimental methodology based on Taguchi orthogonal design has been used to study the effect of various processing parameters on the amount of calcium phosphate coating produced by such technique. Seven control factors with three levels each including sodium hydroxide concentration, pretreatment temperature, pretreatment time, cleaning method, coating time, coating temperature and surface area to solution volume ratio were studied. X-ray diffraction revealed that all the coatings consisted of the mixture of octacalcium phosphate (OCP) and hydroxyapatite (HA) and the presence of each phase depended on the process conditions used. Various content and size (-1-100 µm) of isolated spheroid particles with nanosized plate-like morphology deposited on the titanium surface or a continuous layer of plate-like nanocrystals having the plate thickness in the range of -100-300 nm and the plate width in the range of 3-8 µm were formed depending on the process conditions employed. The optimum condition of using sodium hydroxide concentration of 1 M, pretreatment temperature of 70 degrees C, pretreatment time of 24 h, cleaning by ultrasonic, coating time of 6 h, coating temperature of 50 degrees C and surface area to solution volume ratio of 32.74 for producing the greatest amount of the coating formed on the titanium surface was predicted and validated. In addition, coating temperature was found to be the dominant factor with the greatest contribution to the coating formation while coating time and cleaning method were significant factors. Other factors had negligible effects on the coating performance.

Preparation and characterization of nanosized silver phosphate loaded hydroxyapatite by single step co-conversion process.

The preparation and characterization of silver phosphate nanoparticles loaded hydroxyapatite aiming to enhance the bactericidal performance by a single step co-conversion technique using low temperature phosphorization in the presence of various silver nitrate concentration (AgNO3, ranging 0.001-0.1M) was performed. Characterization by using X-ray diffraction, infrared spectroscopy and transmission electron microscopy showed that hydroxyapatite and silver phosphate were the main phases in all converted samples and the microstructure comprised the distribution of spherical-shaped silver phosphate nanoparticles within the cluster of hydroxyapatite nanocrystals. Total silver content (ranging 0.09-5.6%) in the converted samples was found to increase with increasing silver nitrate content. Flexural modulus and strength of converted samples remained unchanged for samples using silver nitrate between 0.001 and 0.01M, but decreased at greater silver nitrate concentration. Antibacterial activity of two selected samples (0.001 and 0.005M AgNO3) against two bacterial strains (Pseudomonas aeruginosa and Staphylococcus aureus) was observed since 100% reduction of viable cells after 24h contact was detected. Cytotoxic potential by MTT assay of sample using 0.001M AgNO3 was only observed at 24h extraction, but was seen at all extraction periods (24-72h) for sample using 0.005M AgNO3.