Effects of high-concentration of vancomycin and meropenem on viability and osteogenic differentiation of rat bone marrow mesenchymal stem cells_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

HU Junjie , WANG Fei , LI Yichang , LI Yicheng , ZHU Guangzhao , MENG Xingbu , LIU Bo ,  CAO Li

Department of Orthopedics, the First Affiliated Hospital of Xinjiang Medical University, Urumqi Xinjiang, 830054, P. R. China;

Corresponding?author：

CAO Li, Email: xjbone@sina.com

Keywords：

Periprosthetic joint infection; vancomycin; meropenem; bone marrow mesenchymal stem cells; osteogenic differentiation; rat

DOI：

10.7507/1002-1892.202601040

Video：

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Objective To investigate the effects of high-concentration of vancomycin and meropenem on the viability and osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs), and to provide experimental evidence for local antibiotic treatment of periprosthetic joint infection (PJI). Methods Rat BMSCs were cultured with vancomycin and meropenem at different concentrations (10-10 000 μg/mL). A cell counting kit 8 assay was used to assess the effects of the antibiotics on cell viability. The antibiotic concentration at which cell viability began to decrease by more than 50% compared to the control group (cells cultured without antibiotics) after 7 days was selected as the upper concentration limit for subsequent experiments. The lowest concentration at which cell viability significantly decreased compared to the control group after 7 days (P<0.05) was selected as the lower concentration limit. The cells were then further cultured in osteogenic induction medium. After 7 days, the proportion of late apoptotic cells was measured by flow cytometry, and the protein expression of Runt-related transcription factor 2 (Runx2), osteopontin (OPN), and Osterix (Osx) was detected by Western blot. After 21 days, Alizarin red staining was performed to evaluate the effects of the antibiotics on BMSCs mineralization and osteogenic differentiation. Results The inhibitory effects of vancomycin and meropenem on BMSCs viability gradually increased with prolonged intervention. After 7 days, the threshold concentration for a significant decrease in BMSCs viability was 500 μg/mL for both antibiotics (P<0.05). Concentrations of 500, 1 000, and 3 000 μg/mL were selected for subsequent experiments. After 7 days of osteogenic induction, flow cytometry showed that the proportions of late apoptotic cells were significantly higher in the all antibiotics groups than in the control group (P<0.05). Western blot showed that low-concentration vancomycin (500 μg/mL) promoted Runx2 expression but inhibited OPN and Osx expressions (P<0.05), whereas high-concentration vancomycin (3 000 μg/mL) inhibited Runx2, OPN, and Osx expressions (P<0.05). Low-concentration meropenem (500 μg/mL) promoted Runx2, OPN, and Osx expressions (P<0.05), whereas high-concentration meropenem (3 000 μg/mL) inhibited the expressions of these osteogenesis-related proteins (P<0.05). After 21 days, the percentage of mineralized area in the 3 000 μg/mL vancomycin and meropenem groups was significantly lower than that in the control group (P<0.05). Conclusion High-concentration (3 000 μg/mL) of vancomycin and meropenem administered for more than 7 days may inhibit the viability and osteogenic differentiation of rat BMSCs.

Citation： HU Junjie, WANG Fei, LI Yichang, LI Yicheng, ZHU Guangzhao, MENG Xingbu, LIU Bo, CAO Li. Effects of high-concentration of vancomycin and meropenem on viability and osteogenic differentiation of rat bone marrow mesenchymal stem cells. Chinese Journal of Reparative and Reconstructive Surgery, 2026, 40(7): 1121-1127. doi: 10.7507/1002-1892.202601040 Copy

1.	Zimmerli W, Trampuz A, Peter E, et al. Prosthetic-joint infections. N Engl J Med, 2004, 351: 1645-3654.
2.	Kapadia B, Berg R, Daley J, et al. Periprosthetic joint infection. Lancet, 2015, 387: 386-394.
3.	Bruyninckx S, Metsemakers WJ, Depypere M, et al. Local antibiotic delivery via intra-articular catheter infusion for the treatment of periprosthetic joint infection: a systematic review. Archives of Orthopaedic & Trauma Surgery, 2024, 144(12): 5177-5189.
4.	Yao L, Ma Y, Hu Q, et al. Impact of tranexamic acid on the efficacy of vancomycin and meropenem against periprosthetic joint infections: an in vitro analysis. Bone Joint Res, 2025, 14(7): 601-608.
5.	Li YC, Wuermanbieke S, Wang F, et al. Efficacy and safety of intra-articular-only meropenem after one-stage revision for treating Escherichia coli-induced periprosthetic joint infection in a rat model. Bone Joint Res, 2024, 13(10): 546-558.
6.	Berberich CE. Current concepts of local antibiotic delivery in bone and joint infections—A narrative review of techniques and clinical experiences. Microorganisms, 2025, 13(10): 2276. doi: 10.3390/microorganisms13102276.
7.	Yamamoto Y, Fukui T, Sawauchi K, et al. Effects of high antibiotic concentrations applied to continuous local antibiotic perfusion on human bone tissue-derived cells. Bone Joint Res, 2024, 13(3): 91-100.
8.	Zou C, Guo WT, Mu WB, et al. Synovial vancomycin and meropenem concentrations in periprosthetic joint infection treated by single-stage revision combined with intra-articular infusion. Bone & Joint Research, 2024, 13(10): 535-545.
9.	Azzam K, Parvizi J, Jungkind D, et al. Microbiological, clinical, and surgical features of fungal prosthetic joint infections: a multi-institutional experience. J Bone Joint Surg (Am), 2009, 91 Suppl 6: 142-149.
10.	Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science, 1999, 284(5418): 1318-1322.
11.	Hsu YH, Hu CC, Hsieh PH, et al. Vancomycin and ceftazidime in bone cement as a potentially effective treatment for knee periprosthetic joint infection. J Bone Joint Surg (Am), 2017, 99(3): 223-231.
12.	van Hal SJ, Paterson DL, Lodise TP. Systematic review and meta-analysis of vancomycin-induced nephrotoxicity associated with dosing schedules that maintain troughs between 15 and 20 milligrams per liter. Antimicrob Agents Chemother, 2013, 57(2): 734-744.
13.	He J, Wang J, Cao L, et al. Determination of vancomycin and meropenem in serum and synovial fluid of patients with prosthetic joint infections using UPLC-MS/MS. Journal of Mass Spectrometry, 2024, 59(6): e5041. doi: 10.1002/jms.5041.
14.	Okae Y, Nishitani K, Sakamoto A, et al. Estimation of minimum biofilm eradication concentration (MBEC) on in vivo biofilm on orthopedic implants in a rodent femoral infection model. Frontiers in Cellular and Infection Microbiology, 2022, 12: 896978. doi: 10.3389/fcimb.2022.896978.
15.	Rathbone CR, Cross JD, Brown KV, et al. Effect of various concentrations of antibiotics on osteogenic cell viability and activity. J Orthop Res, 2011, 29(7): 1070-1074.
16.	Hanson K, Isder C, Shogren K, et al. The inhibitory effects of vancomycin on rat bone marrow-derived mesenchymal stem cell differentiation. J Neurosurg Spine, 2021, 34(6): 931-935.
17.	Arimura Y, Yano T, Hirano M, et al. Mitochondrial superoxide production contributes to vancomycin-induced renal tubular cell apoptosis. Free Radic Biol Med, 2012, 52(9): 1865-1873.
18.	Sakamoto Y, Yano T, Hanada Y, et al. Vancomycin induces reactive oxygen species-dependent apoptosis via mitochondrial cardiolipin peroxidation in renal tubular epithelial cells. Eur J Pharmacol, 2017, 800: 48-56.
19.	Yao L, Ma Y, Liu R, et, al. Ptential interactions between vancomycin and meropenem in culture-negative periprosthetic joint infection: an in vitro study. BMC Microbiology, 2025, 25(1): 197. doi: 10.1186/s12866-025-03918-4.
20.	Okurowska K, Monk PN, Karunakaran E. Increased tolerance to commonly used antibiotics in a Pseudomonas aeruginosa ex vivo porcine keratitis model. Microbiology (Reading), 2024, 170(5): 001459. doi: 10.1099/mic.0.001459.
21.	Bush K, Bradford PA. β-lactams and β-lactamase inhibitors: An overview. Cold Spring Harbor Perspectives in Medicine, 2016, 6(8): a025247. doi: 10.1101/cshperspect.a025247.
22.	Wan J, He Z, Peng R, et al. Injectable photocrosslinking spherical hydrogel-encapsulated targeting peptide-modified engineered exosomes for osteoarthritis therapy. Journal of Nanobiotechnology, 2023, 21(1): 284. doi: 10.1186/s12951-023-02050-7.

1. Zimmerli W, Trampuz A, Peter E, et al. Prosthetic-joint infections. N Engl J Med, 2004, 351: 1645-3654.
2. Kapadia B, Berg R, Daley J, et al. Periprosthetic joint infection. Lancet, 2015, 387: 386-394.
3. Bruyninckx S, Metsemakers WJ, Depypere M, et al. Local antibiotic delivery via intra-articular catheter infusion for the treatment of periprosthetic joint infection: a systematic review. Archives of Orthopaedic & Trauma Surgery, 2024, 144(12): 5177-5189.
4. Yao L, Ma Y, Hu Q, et al. Impact of tranexamic acid on the efficacy of vancomycin and meropenem against periprosthetic joint infections: an in vitro analysis. Bone Joint Res, 2025, 14(7): 601-608.
5. Li YC, Wuermanbieke S, Wang F, et al. Efficacy and safety of intra-articular-only meropenem after one-stage revision for treating Escherichia coli-induced periprosthetic joint infection in a rat model. Bone Joint Res, 2024, 13(10): 546-558.
6. Berberich CE. Current concepts of local antibiotic delivery in bone and joint infections—A narrative review of techniques and clinical experiences. Microorganisms, 2025, 13(10): 2276. doi: 10.3390/microorganisms13102276.
7. Yamamoto Y, Fukui T, Sawauchi K, et al. Effects of high antibiotic concentrations applied to continuous local antibiotic perfusion on human bone tissue-derived cells. Bone Joint Res, 2024, 13(3): 91-100.
8. Zou C, Guo WT, Mu WB, et al. Synovial vancomycin and meropenem concentrations in periprosthetic joint infection treated by single-stage revision combined with intra-articular infusion. Bone & Joint Research, 2024, 13(10): 535-545.
9. Azzam K, Parvizi J, Jungkind D, et al. Microbiological, clinical, and surgical features of fungal prosthetic joint infections: a multi-institutional experience. J Bone Joint Surg (Am), 2009, 91 Suppl 6: 142-149.
10. Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science, 1999, 284(5418): 1318-1322.
11. Hsu YH, Hu CC, Hsieh PH, et al. Vancomycin and ceftazidime in bone cement as a potentially effective treatment for knee periprosthetic joint infection. J Bone Joint Surg (Am), 2017, 99(3): 223-231.
12. van Hal SJ, Paterson DL, Lodise TP. Systematic review and meta-analysis of vancomycin-induced nephrotoxicity associated with dosing schedules that maintain troughs between 15 and 20 milligrams per liter. Antimicrob Agents Chemother, 2013, 57(2): 734-744.
13. He J, Wang J, Cao L, et al. Determination of vancomycin and meropenem in serum and synovial fluid of patients with prosthetic joint infections using UPLC-MS/MS. Journal of Mass Spectrometry, 2024, 59(6): e5041. doi: 10.1002/jms.5041.
14. Okae Y, Nishitani K, Sakamoto A, et al. Estimation of minimum biofilm eradication concentration (MBEC) on in vivo biofilm on orthopedic implants in a rodent femoral infection model. Frontiers in Cellular and Infection Microbiology, 2022, 12: 896978. doi: 10.3389/fcimb.2022.896978.
15. Rathbone CR, Cross JD, Brown KV, et al. Effect of various concentrations of antibiotics on osteogenic cell viability and activity. J Orthop Res, 2011, 29(7): 1070-1074.
16. Hanson K, Isder C, Shogren K, et al. The inhibitory effects of vancomycin on rat bone marrow-derived mesenchymal stem cell differentiation. J Neurosurg Spine, 2021, 34(6): 931-935.
17. Arimura Y, Yano T, Hirano M, et al. Mitochondrial superoxide production contributes to vancomycin-induced renal tubular cell apoptosis. Free Radic Biol Med, 2012, 52(9): 1865-1873.
18. Sakamoto Y, Yano T, Hanada Y, et al. Vancomycin induces reactive oxygen species-dependent apoptosis via mitochondrial cardiolipin peroxidation in renal tubular epithelial cells. Eur J Pharmacol, 2017, 800: 48-56.
19. Yao L, Ma Y, Liu R, et, al. Ptential interactions between vancomycin and meropenem in culture-negative periprosthetic joint infection: an in vitro study. BMC Microbiology, 2025, 25(1): 197. doi: 10.1186/s12866-025-03918-4.
20. Okurowska K, Monk PN, Karunakaran E. Increased tolerance to commonly used antibiotics in a Pseudomonas aeruginosa ex vivo porcine keratitis model. Microbiology (Reading), 2024, 170(5): 001459. doi: 10.1099/mic.0.001459.
21. Bush K, Bradford PA. β-lactams and β-lactamase inhibitors: An overview. Cold Spring Harbor Perspectives in Medicine, 2016, 6(8): a025247. doi: 10.1101/cshperspect.a025247.
22. Wan J, He Z, Peng R, et al. Injectable photocrosslinking spherical hydrogel-encapsulated targeting peptide-modified engineered exosomes for osteoarthritis therapy. Journal of Nanobiotechnology, 2023, 21(1): 284. doi: 10.1186/s12951-023-02050-7.

Chinese Journal of Reparative and Reconstructive Surgery

Effects of high-concentration of vancomycin and meropenem on viability and osteogenic differentiation of rat bone marrow mesenchymal stem cells

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