Pull-out strength of multifunctional bioabsorbable ciprofloxacin-releasing polylactide-polyglycolide 80/20 tacks: an experimental study allograft cranial bone.

In this study, the pull-out forces of recently developed multifunctional bioabsorbable self-reinforced ciprofloxacin-releasing polylactide/polyglycolide (SR-PLGA+AB) 80/20 tacks were compared with plain SR-PLGA 80/20 tacks in human cadaver parietal bones. Pieces of parietal bone (approximately 6 x 20 cm) were harvested from 5 human cadavers (all were male, 29-77 years old). Fifty plain SR-PLGA tacks (diameter, 2.0 mm; length, 6.0 mm) and 50 ciprofloxacin-releasing SR-PLGA tacks of similar dimensions were applied to drill holes using a special tack shooter without tapping the drill holes. The force needed to pull the tacks from human parietal cadaver bones was measured using a universal tensile-testing machine, by tack pull-out speed of 10 mm/min. Means and SDs were calculated and analyzed using Student's t-test (SPSS version 10.0 for Windows). The pull-out forces of the ciprofloxacin-releasing and plain tacks were 147.0 +/- 5.94 and 141.4 +/- 8.97 N respectively (P = 0.14, statistically insignificant difference between the 2 groups). The cause of failure in all cases was barb breakage. Ciprofloxacin-releasing SR-PLGA 80/20 tacks seem to have a similar holding power to cranial bone as plain SR-PLGA (80/20) tacks but have additional advantage of ciprofloxacin release.

Sustained release of ciprofloxacin from an osteoconductive poly(DL)-lactide implant.

BACKGROUND AND PURPOSE: Antibiotic-releasing bioresorbable implants are used for local treatment of bone infections, but most drug delivery systems release antibiotic for too short a time. METHODS: We used pellets (0.9 x 1.0 mm) made of bioabsorbable poly(DL) lactic acid matrix, ciprofloxacin (7.3 +/- 0.4 wt%), and bioactive glass microspheres of 90-125 microm (29.3 +/- 0.2 wt%). The ciprofloxacin release and antibacterial activity was measured in elution tests in vitro and local tissue concentrations were measured in rabbits. RESULTS: In elution tests in vitro, the therapeutic level (> 2 microg/mL) of ciprofloxacin was achieved within 6 h of the start of the test, and it was maintained for up to 300 days. The antibacterial activity of the antibiotic released from sterilized composites was similar to that of the unprocessed ciprofloxacin. In vivo measurements showed high local tissue concentrations (16-86 micrg/g of bone tissue) for 3 months. Compared to previous experiments on two-component polymeric matrices (PLGA or PDLLA) with ciprofloxacin alone, adding bioactive glass microspheres into the composite resulted in morphological changes that facilitated fluid intrusion into the microstructure and quickened ciprofloxacin release. INTERPRETATION: This type of composition of implant may fulfill the requirements of bone infection therapy, for sustained local release of the selected antibiotic over several months.

In vitro and in vivo testing of bioabsorbable antibiotic containing bone filler for osteomyelitis treatment.

The use of local antibiotics from a biodegradable implant is appealing concept for treatment of chronic osteomyelitis. Our aim was to develop a new drug delivery system based on controlled ciprofloxacin release from poly(D/L-lactide). Cylindrical composite pellets (1.0 x 0.9 mm) were manufactured from bioabsorbable poly(D/L-lactide) matrix and ciprofloxacin (7.4 wt %). In vitro studies were carried out to delineate the release profile of the antibiotic and to verify its antimicrobial activity by means of MIC testing. A long-term study in rabbits was performed to validate the release of ciprofloxacin from the composite in vivo. Therapeutic level of ciprofloxacin (>2 microg/mL) was maintained between 60 and 300 days and the concentration remained below the potentially detrimental level of 20 microg/mL in vitro. The released ciprofloxacin had retained its antimicrobial properties against common pathogens. In an exploratory long-term in vivo study with three rabbits, ciprofloxacin could not be detected from the serum after moderate filling (160 mg) of the tibia (follow-up 168 days), whereas after high dosing (a total dose of 1,000 mg in both tibias) ciprofloxacin was found temporarily at low serum concentrations (14-34 ng/mL) during the follow-up of 300 days. The bone concentrations of ciprofloxacin could be measured in all samples at 168 and 300 days. The tested copolylactide matrix seems to be a promising option in selection of resorbable carriers for sustained release of antibiotics, but the composite needs modifications to promote ciprofloxacin release during the first 60 days of implantation.

Ciprofloxacin-releasing bioabsorbable polymer is superior to titanium in preventing Staphylococcus epidermidis attachment and biofilm formation in vitro.

Antibiotic coating systems have been successfully used to prevent bacterial attachment and biofilm formation. Our purpose was to evaluate whether bioabsorbable polylactide-co-glycolide (PLGA) 80/20 on its own, and PLGA together with ciprofloxacin (PLGA+C) have any advantages over titanium in preventing Staphylococcus epidermidis attachment and biofilm formation in vitro. Cylindrical specimens of titanium, PLGA, and PLGA+C in triplicate were examined for S. epidermidis ATCC 35989 attachment and biofilm formation after incubation with a bacterial suspension of about 10(5) cfu/mL for 1, 3, 7, 14, and 21 days, using scanning electron microscopy. Growth inhibition properties of PLGA and PLGA+C cylinders were tested on agar plates. On days 1, 3, and 21, no bacterial attachment was seen in 19.5, 9.2, and 41.4% of the titanium specimens; in 18.4, 28.7, and 34.5% of the PLGA specimens; and in 57.5, 62.1, and 57.5% of the PLGA+C specimens, respectively. During the whole study period, no biofilm was observed on 74-93% of the titanium specimens, 58-78% of the PLGA specimens, and 93-100% of the PLGA+C specimens. PLGA+C showed clear bacterial growth inhibition on agar plates, while PLGA and titanium did not show any inhibition. PLGA+C bioabsorbable material was superior to titanium in preventing bacterial attachment and biofilm formation and may have clinical applicability, for example, in prevention of infection in trauma surgery or in the treatment of chronic osteomyelitis.

Osteointegration of PLGA implants with nanostructured or microsized ß-TCP particles in a minipig model.

Bioresorbable suture anchors and interference screws have certain benefits over equivalent titanium-alloy implants. However, there is a need for compositional improvement of currently used bioresorbable implants. We hypothesized that implants made of poly(l-lactide-co-glycolide) (PLGA) compounded with nanostructured particles of beta-tricalcium phosphate (ß-TCP) would induce stronger osteointegration than implants made of PLGA compounded with microsized ß-TCP particles. The experimental nanostructured self-reinforced PLGA (85L:15G)/ß-TCP composite was made by high-energy ball-milling. Self-reinforced microsized PLGA (95L:5G)/ß-TCP composite was prepared by melt-compounding. The composites were characterized by gas chromatography, Ubbelohde viscometry, scanning electron microscopy, laser diffractometry, and standard mechanical tests. Four groups of implants were prepared for the controlled laboratory study employing a minipig animal model. Implants in the first two groups were prepared from nanostructured and microsized PLGA/ß-TCP composites respectively. Microroughened titanium-alloy (Ti6Al4V) implants served as positive intra-animal control, and pure PLGA implants as negative control. Cone-shaped implants were inserted in a random order unilaterally in the anterior cortex of the femoral shaft. Eight weeks after surgery, the mechanical strength of osteointegration of the implants was measured by a push-out test. The quality of new bone surrounding the implant was assessed by microcomputed tomography and histology. Implants made of nanostructured PLGA/ß-TCP composite did not show improved mechanical osteointegration compared with the implants made of microsized PLGA/ß-TCP composite. In the intra-animal comparison, the push-out force of two PLGA/ß-TCP composites was 35-60% of that obtained with Ti6Al4V implants. The implant materials did not result in distinct differences in quality of new bone surrounding the implant.

Osteoconductive properties of poly(96L/4D-lactide)/beta-tricalcium phosphate in long term animal model.

The objective of this study was to determine the effect of calcium phosphate mineral content on the bone in-growth at the expense of composite of co-polylactide polymer charged with 2 different ratios of ß-TCP granules (10 and 24 w-% of ß-TCP). The evaluation was realized in a long term rabbit bone model. After 24, 48 and 76 weeks, the implants were examined by micro CT, scanning electron microscopy (SEM) using backscattered electron (BSE) and light microscopy (polarized and blue light microscopy). No foreign body reaction was detected during the 76 weeks follow-up in any of the test samples. Polymer hydrolysis began at approximately 24 weeks, by 76 weeks, the pure polymer implant had begun to release P(96L/4D)LA particles and show signs of peripheral localized bone resorption. A decrease in the amount of CaP was noticed between 24 and 76 weeks in both 10 wt-% and 24 wt-% ß-TCP/P(96L/4D)LA composites. The study showed that the highest bone in-growth was with 24 wt-% ß-TCP/P(96L/4D)LA composite. Bone in-growth and mineralization were evident for the composites associated with specific peripheral bone architecture. Fluorescent labelling demonstrated high bone in-growth and remodeling at the interface, while for pure co-polymer no bone remodeling or bone activity was maintained after 48 weeks. The study demonstrated the positive effect of calcium phosphate content into P(96L/4D)LA. This kind of composite is a suitable resorbable osteoconductive matrix, which provides long term stability required for ligament fixation device.

Release of diclofenac sodium from polylactide-co-glycolide 80/20 rods.

Due to inflammatory reactions complicating bioabsorbable devices, the aim of this study was to develop and characterize bioabsorbable implants with anti-inflammatory drug releasing properties. Polylactide-co- glycolide (PLGA) 80/20 was compounded with diclofenac sodium (DS) to produce rods. Thermal properties were analyzed using differential scanning calorimetry (DSC). Inherent viscosity (eta(inh)) was measured to evaluate the drug effect on the extrude polymer. Drug release measurements were performed using UV-spectrophotometer. Five parallel samples from each type of rods were examined, first at 6 hour intervals, then on daily basis, and later twice a week. DS was released in 110 days from thinner rods and in 150 days from thicker rods. Drug release comprised a starting peak, slow release phase, then a high release phase, and a burst release phase. DSC analysis showed that DS containing rods had crystallinity in their structure. In conclusions, it is feasible to combine PLGA 80/20 and DS by using melt extrusion. Released DS concentrations reached local therapeutic levels, but the release profile was complex and therapeutic levels were not reached all the time.

Tissue reactions to bioabsorbable ciprofloxacin-releasing polylactide-polyglycolide 80/20 screws in rabbits' cranial bone.

The aim of this study was to assess tissue reactions to bioabsorbable self-reinforced ciprofloxacin-releasing polylactide/polyglycolide (SR-PLGA) 80/20 screws in rabbits' cranial bone. Two screws were implanted in each rabbit, one screw on either side of the sagittal suture (n = 28 rabbits). Animals were sacrificed after 2, 4, 8, 16, 24, 54 and 72 weeks, four animals per group. On histological examination the number of macrophages, giant cells, active osteoblasts and fibrous tissue layers were assessed and degradation of the screws was evaluated. At 2 weeks, the highest number of macrophages and giant cells were seen near the heads of the screws. After 4 and 8 weeks, the number of giant cells decreased but that of macrophages decreased from 16 weeks and on. Screws were surrounded by fibrous tissue capsule that progressively was growing in thickness by time. Active osteoblasts were seen around the shaft of the screws with the highest number seen at 4 weeks postoperatively. At 16 weeks, compact fragmentation of the screw heads was seen with macrophages seen inside the screw matrices. After 24 weeks, no polarization of the screws was seen. After one year, PLGA screws had been replaced by adipose tissue, fibrous tissue and "foamy macrophages" which had PLGA particles inside them. After 1(1/2) years, the amount of biomaterial remaining had decreased remarkably. The particles of biomaterial were inside "foamy macrophages." Ciprofloxacin-releasing SR-PLGA 80/20 screws elicited a mild inflammatory reaction but did not interfere with osteoblast activity. No complications were seen when implanted in cranial bone of rabbit.

Self-reinforced ciprofloxacin-releasing polylactide-co-glycolide 80/20 inhibits attachment and biofilm formation by Staphylococcus epidermidis: an in vitro study.

We have observed the efficiency of antibiotic-releasing polylactide-co-glycolide (PLGA) 80/20 in preventing Staphylococcus epidermidis attachment and biofilm formation in vitro. The aim of the present study was to evaluate the effect of self-reinforced (SR) implants with enhanced antibiotic release on bacterial attachment and biofilm formation rates, and also on growth inhibition of Staphylococcus epidermidis. Cylindrical SR-PLGA+AB specimens (length 30 mm, diameter 3 mm) were examined by scanning electron microscopy (SEM) for attachment of S. epidermidis ATCC 35989 on biomaterial surface and formation of biofilm, after incubating with bacterial suspension of ca. 10 cfu/mL for 1, 3, 7, 14 and 21 days. SR-PLGA and SR-PLGA+AB implants were tested on agar plates by measuring the inhibition distance around implants. On the surface of SR-PLGA+AB, at days 1, 3, 7, 14 and 21, the percentage of areas with not a single bacteria attached, was 88.6%, 71.1%, 73.7%, 73.7%, and 68.4%, respectively. On the areas where bacteria were detected, the number of bacterial cells remained low during whole study period, and no significant increase by time was seen. There was no biofilm observed on 97-99% of the examined areas during the whole study period on SR-PLGA+AB. In agar plates, the SR-PLGA+AB showed inhibition of bacterial growth, with (mean) 53.2 mm diameter of inhibition area with peeled implants and 50.5 mm with non-peeled implants. There was no inhibition seen around implants without ciprofloxacin. Bioabsorbable ciprofloxacin-releasing self-reinforced PLGA (SR-PLGA+AB) was superior to plain SR-PLGA in preventing bacterial attachment, biofilm formation, and also the growth of Staphylococcus epidermidis.

Efficacy of ciprofloxacin-releasing bioabsorbable osteoconductive bone defect filler for treatment of experimental osteomyelitis due to Staphylococcus aureus.

The concept of local antibiotic delivery via biodegradable bone defect fillers with multifunctional properties for the treatment of bone infections is highly appealing. Fillers can be used to obliterate surgical dead space and to provide targeted local bactericidal concentrations in tissue for extended periods. Eventually, the osteoconductive component of the filler could guide the healing of the bone defect. The present experimental study was carried out to test this concept in a localized Staphylococcus aureus osteomyelitis model in the rabbit (n = 31). A metaphyseal defect of the tibia was filled with a block of bone cement, followed by insertion of a bacterial inoculum. After removal of the bone cement and surgical debridement at 2 weeks, the defect was filled with a ciprofloxacin-containing (7.6% +/- 0.1%, by weight) composite (treated-infection group) or with a composite without antibiotic (sham-treated group). Both a positive control group (untreated-infection group) and a negative control group were also produced. The treatment response, monitored by positron emission tomography (PET) with fluorine-18-labeled fluorodeoxyglucose ([18F]FDG) at 3 and 6 weeks, showed rapidly decreasing amounts of [18F]FDG uptake in the treated-infection group (P = 0.001 compared with the results for the untreated-infection group at 6 weeks). The bacteriological analysis confirmed the eradication of the bone pathogen in the treated-infection group. However, three animals had culture-positive soft tissue infections. All animals in the sham-treated and untreated-infection groups had culture-positive bone infections with typical radiographic changes of osteomyelitis. Histomorphometry, peripheral quantitative computed tomography, and backscattered electron imaging of scanning electron microscopy images verified the osteoconductive properties of the bioactive glass microspheres within the composite. The median bone ciprofloxacin concentrations were 1.2 and 2.1 microg/g at two anatomic locations of the tibia. This is the first report to show the value of [18F]FDG PET for quantitative monitoring of the treatment response in bone infections. The collaborative results of bacteriologic and [18F-FDG] PET studies showed that use of the multifunctional composite was successful for eradication of the S. aureus pathogen from bone.

Comparison of the pull-out forces of bioabsorbable polylactide/glycolide screws (Biosorb and Lactosorb) and tacks: a study on the stability of fixation in human cadaver parietal bones.

The aim of this study was to compare the pull-out forces of bioabsorbable polylactide/glycolide (PLGA) tacks and screws in human cadaver parietal bones. Parietal bone pieces (c. 6 cm x 20 cm) were collected from five human male cadavers (age range: 47-75 years). Forty-nine BioSorbPDX (self-reinforced [SR] PLGA 80/20) tacks (1.5-mm diameter, 4.0-mm length), 47 BioSorbPDX (SR-PLGA 80/20) screws (1.5-mm diameter, 4.0-mm length), and 46 LactoSorb (PLGA 82/18) screws (1.5-mm diameter, 4.0-mm length) were applied. The tacks were applied to drill holes using a special applicator gun (no tapping or tightening). The screws were applied to drill holes in the traditional way using tapping and tightening with a screwdriver. A tensile testing machine was used. All the implants were tested thus: the head of the implant was held by an aluminum jig, and the jig was pulled with wire until implant failure. The testing pull speed was 10 mm/min. Means and SDs were calculated, and the data were analyzed using ANOVA. The pull-out force of the tacks was 115.9 +/- 8.3 N, that of Lactosorb screws was 112.9 +/- 12.1 N, and that of Biosorb screws was 110.4 +/- 8.9 N (statistically insignificant difference between the three groups). The most common reason for failure in the case of tacks was barb breakage (55.1%); it was thread breakage in the case of BioSorb screws (66%) and stem split in the case of Lactosorb screws (56%). Tacks seem to have a similar, perhaps even a little better, holding power to cranial bone as screws and can hence be recommended for clinical application, as the procedure saves time and, consequently, costs.

Holding power of bioabsorbable ciprofloxacin-containing self-reinforced poly-L/DL-lactide 70/30 bioactive glass 13 miniscrews in human cadaver bone.

Antibiotics-plus bioactive glass-containing bioabsorbable self-reinforced (SR) polylactide screws have been developed for antibacterial osteoconductive bone fixation. The aim of the present study was to test the pullout properties of these recently developed miniscrews. Ciprofloxacin-plus bioactive glass-containing SR-polylactide miniscrews (BC) were compared with miniscrews made of neat SR-polylactide (A), SR-polylactide with bioactive glass (B), and ciprofloxacin-containing SR-polylactide (C). BC miniscrews and their controls (A, B, C) (all of length 6.0 mm, core diameter 1.45 mm, thread diameter 2.0 mm) were applied to one pair of cadaveric fibulae. Pullout force was measured using a materials testing machine. We carried out 49-50 pullout tests for each implant type. The Mann-Whitney test and Student's t-test were used for statistical evaluation. The pullout force for BC miniscrews was 114.9 +/- 34.0 (SD) N. Pullout forces for control miniscrews were 162.7 +/- 37.8 N (A), 99.1 +/- 16.2 N (B), and 142.9 +/- 26.9 N (C). Differences between the four groups were statistically significant (p < 0.001). Ciprofloxacin-plus bioactive glass-containing polylactide miniscrews have good holding power to human cadaver fibulae. However, adding bioactive glass and ciprofloxacin components to neat SR-polylactide results in lower pullout values.

Bioabsorbable ciprofloxacin-containing and plain self-reinforced polylactide-polyglycolide 80/20 screws: pullout strength properties in human cadaver parietal bones.

The aim of this study was to compare the pullout forces of recently developed bioabsorbable ciprofloxacin-containing and plain self-reinforced polylactide/polyglycolide (SR-PLGA) miniscrews in human cadaver parietal bones. Parietal bone pieces (approximately 6 x 20 cm) were collected from five human male cadavers (44-75 years of age). Fifty plain SR-PLGA 80/20 miniscrews (diameter = 1.5 mm, length = 4.0 mm) and 50 ciprofloxacin-containing SR-PLGA 80/20 miniscrews (diameter = 1.5 mm, length = 4.0 mm) were used in this study. The force needed to pull the screws from human parietal cadaver bones was measured using a tensile strength-testing machine. The screw pullout speed was 10 mm/min. Means and SDs were calculated and analyzed using the Student t test (SPSS version 10.0 for Windows). The pullout forces of the ciprofloxacin-containing and plain miniscrews were 66.8 +/- 4.9 N and 96.3 +/- 9.3 N (significant difference, P < 0.001), respectively. The most common cause of failure was screw-shaft breakage (60% in the case of ciprofloxacin-containing screws and 52% in the case of plain SR-PLGA screws). Scanning electron microscopy showed that the fibrillar strip-like microstructure of plain SR-PLGA miniscrews turns into a coarse, uniaxial, platelet-like morphology in antibiotic SR-PLGA miniscrews as a result of the addition of ciprofloxacin. Ciprofloxacin-containing SR-PLGA screws consequently have a lower pullout strength than corresponding plain conventional SR-PLGA screws. Nevertheless, it is evident that the ciprofloxacin-containing screws can be applied in craniomaxillofacial surgery in nonload-bearing or slightly load-bearing applications.