In vitro elution of ofloxacin from a bioabsorbable polymer

A drawback with antibiotic-loaded polymethylmethacrylate (PMMA) beads is the second surgical procedure that may be required to remove the beads. We explored alternatives for the local delivery of antimicrobials and studied the release characteristics of ofloxacin (10 mg) when incorporated into either 30 or 50 mg beads of two different molar ratios of a bioabsorbable lactide: glycolide polymer. An elution method was employed over a 60-day period at 37°C. After 60 days, about half of the ofloxacin had been released from the composites containing 50 mg and 30 mg of the 85:15 polymer, whereas nearly all of the ofloxacin had been released from both sizes of the 50:50 polymer composites.     

The in vitro elution characteristics of vancomycin and tobramycin from calcium sulfate beads

The purpose of this study was to determine the elution characteristics of vancomycin and tobramycin when mixed with calcium sulfate to form antibiotic beads. Calcium sulfate was combined with vancomycin and tobramycin separately to form 2 types of antibiotic beads, which were packaged and labeled separately. The packaged calcium sulfate beads with vancomycin and tobramycin were then gas sterilized. The beads were placed in phosphate-buffered saline and kept at 36 degrees C for 6 weeks. Two separate series of assays were run simultaneously for both types of beads. In one assay, a bead containing vancomycin was placed in a fresh vial of phosphate buffered saline after each assay. The same was done with beads containing tobramycin. In the second series of assays, 9 vials of phosphate buffered saline each containing 1 vancomycin bead and 9 vials of phosphate buffered saline each containing 1 tobramycin bead was arranged. The phosphate-buffered saline was then assayed at predetermined times for both the vancomycin bead series and the tobramycin bead series. The amount of vancomycin and tobramycin assayed nearly equaled the calculated amount of antibiotic per bead measured before bead construction. Also, the elution of antibiotic from the calcium sulfate was complete within 72 hours. In conclusion, the construction and gas sterilization of calcium sulfate beads containing vancomycin and tobramycin does not destroy vancomycin and tobramycin. Also, the complete elution of available vancomycin and tobramycin in calcium sulfate beads occurs within 72 hours.     

In vitro elution of ciprofloxacin from polymethylmethacrylate cement beads

The use of antibiotic-impregnated polymethylmethacrylate (PMMA) cement beads for the local delivery of antibiotics in the treatment of chronic osteomyelitis has become a standard orthopaedic practice. The increasing resistance to antibiotics of organisms associated with orthopaedic infections has led to interest in the incorporation of more effective antibiotics into PMMA cement. Ciprofloxacin, a synthetic fluoroquinolone, is potent against a broad spectrum of bacteria associated with osteomyelitis. In this study, strands of ciprofloxacin-impregnated PMMA cement beads were prepared with 0.2, 0.5, or 1.0 g of ciprofloxacin per 40 g of PMMA. The elution concentration of ciprofloxacin was at least 1–2 mcg/ml for 7 days (0.2 g), 30 days (0.5 g), and 42 days (1.0 g). This concentration is equivalent to the minimum inhibitory concentration for the common pathogens associated with osteomyelitis. Concurrent systemic and local ciprofloxacin therapy appears to be a method for the treatment of chronic osteomyelitis.     

Vancomycin vs tobramycin elution from polymethylmethacrylate: an in vitro study

We fabricated batches of cement containing 0.5 gm, 1.0 gm, and 2.0 gm vancomycin and one with 1.0 gm tobramycin and shaped them into cylinders. They were immersed into 0.5 L of normal saline and the fluid volume was changed daily. Samples of fluid were obtained on days 1, 2, 3, 5, 7, 14, and 28. All fluid samples had antibiotic assays performed to quantitate the amount of elution for vancomycin or tobramycin. Vancomycin elution from PMMA occurred under our study conditions in similar quantities to that measured for tobramycin controls. Vancomycin-loaded PMMA cement may have a clinical role in the treatment of musculoskeletal sepsis caused by gram-positive bacteria, particularly if organisms resistant to the usual antibiotic agents are identified.     

In vitro elution characteristics of commercially and noncommercially prepared antibiotic PMMA beads.

The successful treatment of osteomyelitis with commercially prepared gentamicin-polymethylmethacrylate (PMMA) (Septopal) beads and surgical debridement has led to the use of this technique in the United States. However, commercially prepared gentamicin-PMMA beads are not currently available to orthopedic surgeons in the United States. Therefore, these surgeons commonly manufacture their own antibiotic-containing cement beads in the operating room at the time of surgery. There is little data that compare the antibiotic elution characteristics of such preparations to commercially prepared gentamicin-PMMA beads. This study compares the measured amount of antibiotic elution of either gentamicin or tobramycin from laboratory manufactured Zimmer, Simplex, or Palacos beads to commercially prepared gentamicin-PMMA (Septopal) beads. During a 30-day study period, commercially prepared gentamicin-PMMA beads eluted more total antibiotic and maintain higher concentrations than did antibiotic acrylic composites manufactured in the authors' laboratory.     

In vitro elution of vancomycin from calcium phosphate cement

Antibiotic-impregnated bone cement beads have become popular for the treatment of osteomyelitis and/or prosthesis infection. However, bone cement has the disadvantage of heating up during polymerization of cement. Recently, calcium phosphate cement (CPC) has been used as a bone replacement and augmentation, and it does not heat up during polymerization. First, we measured the release rate of vancomycin (VCM) from bone cement and CPC impregnated with VCM for 2 weeks in vitro. The mean concentration of VCM for CPC was 62.6 times at 7 days (258 ± 29 vs 4.12 ± 1.0) and 6.7 times at 13 days (15.5 ± 5.5 vs 2.3 ± 0.7). Second, we were successful in treating 2 cases of osteomyelitis and prosthesis infection with VCM-impregnated CPC. From this study, we concluded that VCM-impregnated CPC might be an effective material for the treatment of osteomyelitis and/or prosthesis infection.     

Elution characteristics of tobramycin from polycaprolactone in a rabbit model.

This study investigated the elution characteristics of tobramycin from polycaprolactone, a bioabsorbable polymer, in a rabbit model. Sixty rabbits were divided into two groups. Group 1 had polycaprolactone rods impregnated with 6% tobramycin surgically implanted into the proximal femoral intramedullary canal. Group 2 received polymethylmethacrylate rods of like size, shape, and antibiotic concentration. Serum and urine samples were obtained, and tobramycin levels were determined via fluorescent immunosorbent assay. Rabbits were sacrificed as long as 56 days after surgery. Local bone tobramycin concentration was determined using the agar diffusion method. Polycaprolactone delivered a significantly higher peak bone concentration of tobramycin (22.4 microg/mL) than did polymethylmethacrylate (13.59 microg/mL). Polycaprolactone also had a more gradual decrease in local tobramycin concentration than did polymethylmethacrylate. Neither polycaprolactone nor polymethylmethacrylate yielded consistently detectable (> 0.1 microg/mL) serum tobramycin levels. Urine concentrations mirrored those seen in bone, with polycaprolactone achieving significantly higher tobramycin concentrations than did polymethylmethacrylate. Polycaprolactone had superior elution characteristics compared with polymethylmethacrylate in this lapine model, suggesting that polycaprolactone might be a promising local antibiotic delivery vehicle for the treatment of osteomyelitis.     

The effect of sampling method on the elution of tobramycin from calcium sulfate

Release rate is a critical property of all drug delivery vehicles, including antibiotic-laden bioerodibles. In vitro elution studies, used to evaluate release rates, use different sampling methods, including changing the entire amount of buffer and partial exchanges each day. Two groups of 10% calcium sulfate-tobramycin pellets were eluted in 20 mL of buffer for 30 days. Group I had 5 mL of buffer withdrawn and replaced daily whereas Group II had the entire 20 mL of buffer changed daily. The results show that the complete exchange method caused a significantly faster release of antibiotic than the partial exchange method. In the complete exchange group, greater than 50% of the tobramycin was released by 24 hours, whereas in the partial exchange group, 50% of the antibiotic was not released until Day 6. The two methods of sampling used to evaluate this bioerodible material provide data that allow the user to anticipate how the material will function in relatively inert and volatile environments. The method used to sample the elution of antibiotics from bioerodible materials affects the amount of antibiotic eluted. It therefore is important to know the method of sampling when making a decision to use a bioerodible material to deliver antibiotics locally.     

In vitro elution of antibiotic from antibiotic-impregnated biodegradable calcium alginate wound dressing.

OBJECTIVE: The authors investigated the calcium alginate dressing as a drug-delivery system for the treatment of various surgical infections. METHODS: Cytotoxicity of the calcium alginate dressing to fibroblasts and HeLa cells was evaluated by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MITT) colorimetric assay. The calcium alginate dressing was mixed with vancomycin, and lyophilized or not lyophilized to form two types of antibiotic dressings. The antibiotic dressings were placed in 2 mL of phosphate buffered saline (PBS) or in PBS containing 0.01% calcium ions, and incubated at 37 degrees C. The PBS was changed daily, and the removed solutions were stored at -70 degrees C until the antibiotic concentration in each sample was determined by high performance liquid chromatography assay. RESULTS: The results suggested that the antibiotic dressings present no obvious toxic risk to their use as a drug-delivery system. The concentration of vancomycin in each sample was well above the breakpoint sensitivity concentration (the antibiotic concentration at the transition point between bacterial kill. ing and resistance to the antibiotic) for more than 14 days. The release was most marked during the first 48 hours. The concentration of calcium ions in PBS and the lyophilization of the manufacture process of antibiotic dressings prolonged the antibiotic diffusion duration. The diameter of the sample inhibition zone ranged from 10 to 11 mm, and the relative activity of vancomycin ranged from 62.88% to 92.18%. CONCLUSION: All antibiotic dressings released bactericidal concentrations of the antibiotics in vitro for the period of time needed to treat surgical infections. This study offers a convenient method to meet the specific antibiotic requirement for different patients.     

In vitro comparison of elution characteristics of vancomycin from calcium phosphate cement and polymethylmethacrylate

Background

Calcium phosphate cement [CPC (Biopex®)] has been used as the drug delivery system of choice for treatment of infected joint replacement because of its good elution efficiency. The influence of CPC polymerization on the bactericidal activity of vancomycin (VCM) impregnated into CPC has not been investigated. We compared VCM concentration, bactericidal activity, and profile of eluates between CPC and polymethylmethacrylate (PMMA; Cemex RX®).

Methods

Test specimens consisted of a powder composite of CPC or PMMA, VCM and solvent (10:0.25:3.3 g). Each test specimen was immersed in sterile phosphate-buffered saline. Eluates obtained on days 1, 3, 7, and 14 and weeks 4, 8, and 12 were evaluated by high performance liquid chromatography (HPLC) and by microbiological assay (MBA).

Results

The elution level of VCM from CPC/VCM on day 1 was 8.1 fold greater than that from PMMA/VCM. The detection periods of VCM from CPC/VCM and PMMA/VCM were 8 weeks and 14 days, respectively. The values of eluates from CPC/VCM and PMMA/VCM obtained by HPLC were comparable to those obtained by MBA. HPLC chromatogram showed that the elution profiles of VCM from CPC/VCM and PMMA/VCM on day 1 were very close to those of standard solutions.

Conclusions

CPC could release more VCM over a longer period than PMMA. The polymerization of CPC and PMMA did not alter the inhibitory activity of VCM and did not denature VCM.     

The elution of colistimethate sodium from polymethylmethacrylate and calcium phosphate cement beads

Gram-negative bacilli resistance to all antibiotics, except for colistimethate sodium (CMS), is an emerging healthcare concern. Incorporating CMS into orthopedic cement to treat bone and soft-tissue infections due to these bacteria is attractive, but the data regarding the elution of CMS from cement are conflicting. The in vitro analysis of the elution of CMS from polymethylmethacrylate (PMMA) and calcium phosphate (CP) cement beads is reported. PMMA and CP beads containing CMS were incubated in phosphate-buffered saline and the eluate sampled at sequential time points. The inhibition of the growth of a strain of Acinetobacter baumannii complex by the eluate was measured by disk diffusion and microbroth dilution assays, and the presence of CMS in the eluate was measured by mass spectroscopy. Bacterial growth was inhibited by the eluate from both PMMA and CP beads. Mass spectroscopy demonstrated greater elution of CMS from CP beads than PMMA beads. The dose of CMS in PMMA beads was limited by failure of bead integrity. CMS elutes from both CP and PMMA beads in amounts sufficient to inhibit bacterial growth in vitro. The clinical implications of these findings require further study.     

Antibiotic elution from acrylic bone cement loaded with high doses of tobramycin and vancomycin

Two‐stage revision treatment of prosthetic joint infection (PJI) frequently employs the use of a temporary bone cement spacer loaded with multiple antibiotic types. Tobramycin and vancomycin are commonly used antibiotics in cement spacers, however, there is no consensus on the relative concentrations and combinations that should be used. Therefore, the purpose of this study was to investigate the influence of dual antibiotic loading on the total antibiotic elution and compressive mechanical properties of acrylic bone cement. Varying concentrations of tobramycin (0–3 g) and vancomycin (0–3 g) were added either alone or in combination to acrylic cement (Palacos R), resulting in 12 experimental groups. Samples were submerged in 37°C saline for 28 d and sampled at specific time points. The collected eluent was analyzed to determine the cumulative antibiotic release. In addition, the cement's compressive mechanical properties and porosity were characterized. Interestingly, the cement with the highest concentration of antibiotics did not possess the best elution properties. Cement samples containing both 3 g of tobramycin and 2 g vancomycin demonstrated the highest cumulative antibiotic release after 28 d, which was coupled with a significant decrease in the mechanical properties and an increased porosity. The collected data also suggests that tobramycin elutes more effectively than vancomycin from cement. In conclusion, this study demonstrates that high antibiotic loading in cement does not necessarily lead to enhanced antibiotic elution. Clinically this information may be used to optimize cement spacer antibiotic loading so that both duration and amount of antibiotics eluted are optimized. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1078–1085, 2018.

     

美罗培南骨水泥体外生物力学及洗脱特性研究

目的 研究不同含量的美罗培南骨水泥的体外生物力学特性和药物洗脱特性.方法 40 g骨水泥(Palacos LV,40 g聚合物粉剂+20 ml单体液剂)中分别加入2、4、6 g注射用美罗培南和2、4 g注射用盐酸万古霉素复合,制备成A2、A4、A6、B2、B4和不含抗菌药物的对照组共6组样品.对6组样品分别进行能承受的...     

In vitro release of antibiotics from commercial PMMA beads and articulating hip spacers

The efficacy and benefits of high-dose antibiotic cement spacers compared with beads in the treatment of an infected prosthesis have been shown. However, in clinical practice, commercial, low-dose antibiotic bone cement is often used. This study investigated the in vitro antibiotic release of hip spacers made from Refobacin-Palacos-R or Antibiotic-Simplex-P cement compared with Septopal beads. Antibiotic concentrations were measured during 6 weeks. All carriers showed a burst release, but spacers showed little additional release after the first week. Cumulative release was 27.5 +/- 2.3 mg for Palacos, 23.8 +/- 0.2 mg for Simplex, and 188.3 +/- 9.3 mg for Septopal (P < .001). Despite the efficacy of high-dose antibiotic bone cement spacers, we believe one should be cautious toward using low-dose antibiotic bone cement for spacers because this could result in an unsuccessful eradication of infection.     

Rapid release of gentamicin from collagen sponge. In vitro comparison with plastic beads

The gentamicin-containing collagen sponge is a new product intended for local application in bone and soft-tissue infections. The release of gentamicin from the collagen sponges was compared in vitro to that from polymethyl-methacrylate (PMMA) beads. A static and kinetic experimental design was used. In the static model, pieces of collagen sponge or PMMA beads were added to 20 mL of distilled water, and during the following hours the gentamicin concentrations in the water were repeatedly measured. This simple model was extended to the kinetic model as the released gentamicin was removed from the water exponentially by means of an infusion-withdrawal pump. The gentamicin was released from the carrier substances with increasing half lives. During the first 4 hours, the half life increased from 0.2 to 1.5 hours for the collagen sponge and from 3 to 78 hours for the PMMA beads. After 1.5 hours, 95 percent of the gentamicin was released from the sponges, whereas only 8 percent was released from the beads.     

In vitro and in vivo evaluation of antibiotic diffusion from antibiotic-impregnated polymethylmethacrylate beads.

The elution of antibiotics from antibiotic-impregnated polymethylmethacrylate (PMMA) beads was measured in mongrel dogs. The antibiotics, used in mixture with Simplex cement, included cefazolin (Ancef; 4.5 g/40 g cement powder), ciprofloxacin (Cipro; 6 g/40 g powder), clindamycin (Cleocin; 6 g/40 g powder), ticarcillin (Ticar; 12 g/40 g powder), tobramycin (Nebcin; 9.8 g/40 g powder), and vancomycin (Vancocin; 4 g/40 g powder). After a pneumatic drill was used to dredge a trough in the tibia, five beads were implanted. During the next 28 days, seroma samples and serum samples were taken for antibiotic measurements. On Day 28, the dogs were killed, beads removed, and the seroma, serum, bone, and granulation tissue sampled. The results of the study showed that clindamycin, vancomycin, and tobramycin exhibited good elution characteristics and had consistently high levels in bone and granulation tissue.     

Rapid release of gentamicin from collagen sponge: In vitro comparison with plastic beads

The gentamicin-containing collagen sponge is a new product intended for local application in bone and soft-tissue infections. The release of gentamicin from the collagen sponges was compared in vitro to that from polymethyl-methacrylate (PMMA) beads. A static and kinetic experimental design was used. In the static model, pieces of collagen sponge or PMMA beads were added to 20 mL of distilled water, and during the following hours the gentamicin concentrations in the water were repeatedly measured. This simple model was extended to the kinetic model as the released gentamicin was removed from the water exponentially by means of an infusion-withdrawal pump. The gentamicin was released from the carrier substances with increasing half lives. During the first 4 hours, the half life increased from 0.2 to 1.5 hours for the collagen sponge and from 3 to 78 hours for the PMMA beads. After 1.5 hours, 95 percent of the gentamicin was released from the sponges, whereas only 8 percent was released from the beads.     

In vitro culture of human dermal fibroblasts on electrospun polycaprolactone collagen nanofibrous membrane

Novel cost-effective electrospun nanofibrous membrane is established for wound dressing and allogeneic cultured dermal substitute through the cultivation of human dermal fibroblast for skin defects. Synthetic polymers are generally used for tissue engineering and drug delivery applications because of their remarkable mechanical stability and slow degradation. Polycaprolactone (PCL) is used as a bioresorbable polymer in numerous medical devices as well as for tissue engineering applications. The large surface area of the polymer nanofibers with specific modifications facilitates cell adhesion and control of their cellular functions. The objectives of this study was to fabricate electrospun nanofibrous membrane from biodegradable PCL for wound dressing and collagen-blended nanofibrous membrane, and to examine fibroblast attachment, cell proliferation, and morphology of cell matrix interaction. Results of the present investigation prove that the porous nanofibrous membrane is suitable for wound dressing and modified PCL-blended collagen nanofibrous membrane is suitable for the attachment and proliferation of fibroblast, and might have the potential to be applied in tissue engineering as a dermal substitute for the treatment of skin defects and burn wounds.     

The in vitro elution characteristics of vancomycin from tendons

Background  

Infection after ACL reconstruction is uncommon but catastrophic. Prophylactic graft saturation in vancomycin reportedly reduces infection rates.