Topical vancomycin and its effect on survival and migration of osteoblasts, fibroblasts, and myoblasts: An in vitro study

Liu, James X.; Bravo, Dalibel; Buza, John A.; Kirsch, Thorsten; Kennedy, Oran D.; Rokito, Andrew S.; Zuckerman, Joseph D.

doi:10.1016/j.jor.2018.01.032

Cited by 22 publications

(25 citation statements)

References 25 publications

(39 reference statements)

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“…They found that as CHX concentration increased there was a significant (p<0.001) difference in fibroblast proliferation - even at their lowest tested concentration (1%), exposure to CHX resulted in in vitro fibroblast survival of 51.7% compared to no exposure 33 . The dose-dependent cytotoxicity of CHX on fibroblasts, myoblasts, and osteoblasts is not a unique property of CHX; prior in vitro studies have demonstrated that dilute povidone-iodine and topical vancomycin powder, also used for prevention of surgical site infections, have similarly specific dose-dependent and time-dependent cytotoxicity profiles on fibroblasts, myoblasts, and osteoblasts, as measured via the scratch test and Cell Counting Kit-8 cell survival assay 34 , 35 .…”

Section: Discussionmentioning

confidence: 99%

Cytotoxicity evaluation of chlorhexidine gluconate on human fibroblasts, myoblasts, and osteoblasts

Liu

Werner

Kirsch

et al. 2018

J. Bone Joint Infect.

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113

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Introduction: Chlorhexidine gluconate (CHX) is widely used as a preoperative surgical skin-preparation solution and intra-wound irrigation agent, with excellent efficacy against wide variety of bacteria. The cytotoxic effect of CHX on local proliferating cells following orthopaedic procedures is largely undescribed. Our aim was to investigate the in vitro effects of CHX on primary fibroblasts, myoblasts, and osteoblasts.Methods: Cells were exposed to CHX dilutions (0%, 0.002%, 0.02%, 0.2%, and 2%) for either a 1, 2, or 3-minute duration. Cell survival was measured using a cytotoxicity assay (Cell Counting Kit-8). Cell migration was measured using a scratch assay: a “scratch” was made in a cell monolayer following CHX exposure, and time to closure of the scratch was measured.Results: All cells exposed to CHX dilutions of ≥ 0.02% for any exposure duration had cell survival rates of less than 6% relative to untreated controls (p < 0.001). Cells exposed to CHX dilution of 0.002% all had significantly lower survival rates relative to control (p < 0.01) with the exception of 1-minute exposure to fibroblasts, which showed 96.4% cell survival (p = 0.78). Scratch defect closure was seen in < 24 hours in all control conditions. However, cells exposed to CHX dilutions ≥ 0.02% had scratch defects that remained open indefinitely.Conclusions: The clinically used concentration of CHX (2%) permanently halts cell migration and significantly reduces survival of in vitro fibroblasts, myoblasts, and osteoblasts. Further in vivo studies are required to examine and optimize CHX safety and efficacy when applied near open incisions or intra-wound application.

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Section: Discussionmentioning

confidence: 99%

Cytotoxicity evaluation of chlorhexidine gluconate on human fibroblasts, myoblasts, and osteoblasts

Liu

Werner

Kirsch

et al. 2018

J. Bone Joint Infect.

Self Cite

113

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“…The authors found that 1 h vancomycin exposure reduced osteoblast and myoblast survival and migration only at the highest dosage (12 mg/cm 2 ). Conversely, a prolonged vancomycin exposure significantly impaired both survival and migration in all cell types at all concentrations tested [ 34 ]. According to our results, it may be reasonable to reduce vancomycin concentration for graft soaking under 5 mg/mL to reduce the risk of graft damage without affecting antimicrobial efficacy, as the MIC for most bacteria involved in post-ACLR septic arthritis is approximately 2 mg/mL [ 23 ].…”

Section: Discussionmentioning

confidence: 99%

Does Vancomycin Wrapping in Anterior Cruciate Ligament Reconstruction Affect Tenocyte Activity In Vitro?

et al. 2021

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Knee septic arthritis is a devastating complication following anterior cruciate ligament (ACL) reconstruction. To prevent this issue, intraoperative soaking of ACL grafts with vancomycin is often performed before implantation. Although vancomycin cytotoxicity has been reported several times, little is known about its biological effect on tenocytes. The aim of this study was to evaluate the in vitro effects of vancomycin on human primary tenocytes (hTCs). hTCs were isolated from hamstring grafts of four patients undergoing ACL reconstruction. After expansion, hTCs were treated with different concentrations of vancomycin (0, 2.5, 5, 10, 25, 50 and 100 mg/mL) for 10, 15, 30 and 60 min. In vitro cytotoxicity was evaluated measuring metabolic activity, cell toxicity, and apoptosis. hTC metabolic activity was affected starting from 10 mg/mL vancomycin and decreased markedly at 100 mg/mL. Cell viability remained unaffected only at a concentration of 2.5 mg/mL vancomycin. Vancomycin cytotoxicity was detected from 10 mg/mL after 15 min and at all higher concentrations. Cells died when treated with concentrations higher than 5 mg/mL. The use of this antibiotic on tendons to prevent infections could be useful and safe for resident cells if used at a concentration of 2.5 mg/mL for up to 1 h of treatment.

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“…Previous in vitro studies on cellular toxicity of vancomycin have been conducted on relevant cell types within a joint. 1,3,9,19 Bariteau et al 3 found that exposing human mesenchymal stromal cells (hMSCs) to 5000 µg/mL vancomycin for 3 days resulted in significantly decreased cell viability, proliferation, and mineralization. Chu et al 9 established a dose-dependent increase in cell death for hMSCs exposed to vancomycin for 24 hours, starting at 400 µg/mL.…”

Section: Discussionmentioning

confidence: 99%

“…This finding suggests that various cell types respond differently to antibiotic exposure. Liu et al 19 exposed osteoblasts, fibroblasts, and myoblasts to vancomycin for a short duration (1 hour) or continuous duration (48 hours). They found that for short durations, osteoblast and myoblast survival and migration were not affected by vancomycin, except for at the highest concentration of 12 mg/mL.…”

Section: Discussionmentioning

confidence: 99%

Soaking of Autologous Tendon Grafts in Vancomycin Before Implantation Does Not Lead to Tenocyte Cytotoxicity

et al. 2020

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Background: Surgical site infections (SSIs) after anterior cruciate ligament (ACL) reconstruction procedures are an unfortunate complication. Soaking grafts in vancomycin before implantation has been reported to reduce the incidence of postoperative SSI after ACL reconstruction. There is potential for vancomycin to compromise graft integrity because of tenocyte toxicity. Purpose: To examine the in vitro toxicity of varying doses of vancomycin on human tenocytes. Study Design: Controlled laboratory study. Methods: Human patellar tenocytes were isolated and expanded in vitro. Tenocytes in culture were exposed to vancomycin at 5 different concentrations (400, 1600, 3200, 6400, and 12,800 μg/mL) and 3 time intervals (2, 6, and 24 hours). The control for all series was tenocyte exposure to only culture medium for each time interval. After treatment, a 10% Cell Counting Kit-8 solution in cellular growth medium was applied to the cells to examine cytotoxicity. A live/dead assay was used to assess tenocyte viability through fluorescence microscopy and flow cytometry. Results were analyzed statistically using multivariable logistic regression models with Tukey honest significant difference post hoc tests. Results: Vancomycin did not cause significant changes in tenocyte viability after 2 and 6 hours of incubation at any concentration between 0 and 12,800 µg/mL. Incubation with vancomycin for 24 hours led to a significant decrease in cell viability at higher concentrations. Conclusion: Tenocytes derived from human patellar tendons exposed to relatively high concentrations of vancomycin for short periods of time do not demonstrate significant cell death and toxicity. Clinical Relevance: Exposing tendons to vancomycin for a short period of time, such as before ACL reconstruction, is not likely to cause tenocyte toxicity because of vancomycin administration.

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Topical vancomycin and its effect on survival and migration of osteoblasts, fibroblasts, and myoblasts: An in vitro study

Cited by 22 publications

References 25 publications

Cytotoxicity evaluation of chlorhexidine gluconate on human fibroblasts, myoblasts, and osteoblasts

Cytotoxicity evaluation of chlorhexidine gluconate on human fibroblasts, myoblasts, and osteoblasts

Does Vancomycin Wrapping in Anterior Cruciate Ligament Reconstruction Affect Tenocyte Activity In Vitro?

Soaking of Autologous Tendon Grafts in Vancomycin Before Implantation Does Not Lead to Tenocyte Cytotoxicity

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