Ex Vivo Murine Skin Model for B. burgdorferi Biofilm
Abstract
:1. Introduction
2. Results
2.1. Establishment of an Ex Vivo Skin Model System for a B. burgdorferi Biofilm
2.2. Analyses of Infected Murine Skin Biopsies Inoculated with 5 × 106 Spirochetes
2.3. Analyses of Infected Murine Skin Biopsies Inoculated with 1 × 107 Spirochetes
2.4. Reverse-Transcriptase PCR (RT-PCR) Analysis on Infected Murine Skin Biopsies
2.5. Confocal Microscopy Analysis of Various Morphological Forms of B. burgdorferi in Infected Murine Skin Biopsies
2.6. Atomic Force Microscopy Analysis of B. burgdorferi Biofilm in Infected Murine Skin Biopsies
3. Discussion
4. Material and Methods
4.1. Bacterial Culture
4.2. Mouse Skin Biopsy Inoculation, Fixation and Processing
4.3. Immunohistochemistry (IHC)
4.4. RNA Extraction and Reverse Transcriptase—PCR
4.5. Confocal Microscopy
4.6. Atomic Force Microscopy
4.7. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
- Cook, M.J. Lyme borreliosis: A review of data on transmission time after tick attachment. Int. J. Gen. Med. 2014, 8, 1–8. [Google Scholar] [CrossRef] [PubMed]
- Feder, H.M.; Abeles, M.; Bernstein, M.; Whitaker-Worth, D.; Grant-Kels, J.M. Diagnosis, treatment, and prognosis of erythema migrans and Lyme arthritis. Clin. Dermatol. 2006, 6, 509–520. [Google Scholar] [CrossRef] [PubMed]
- Müllegger, R.R.; Glatz, M. Skin manifestations of Lyme Borreliosis. Am. J. Clin. Dermatol. 2008, 9, 355–368. [Google Scholar] [CrossRef] [PubMed]
- Eisendle, K.; Grabner, T.; Zelger, B. Focus floating microscopy: “Gold Standard” for cutaneous Borreliosis? Am. J. Clin. Pathol. 2007, 127, 213–222. [Google Scholar] [CrossRef]
- Duray, P.H.; Steere, A.C. Clinical pathologic correlations of Lyme disease by stage. Ann. N. Y. Acad. Sci. 1988, 539, 65–79. [Google Scholar] [CrossRef]
- Yeung, C.; Baranchuk, A. Diagnosis and treatment of Lyme carditis. J. Am. Coll. Cardiol. 2019, 73, 717–726. [Google Scholar] [CrossRef]
- Coyle, P.K. Neurologic aspects of Lyme disease. Med. Clin. N. Am. 2002, 86, 261–284. [Google Scholar] [CrossRef]
- Donta, S.T. Tetracycline therapy for chronic Lyme disease. Clin. Infect. Dis. 1997, 25, S52–S56. [Google Scholar] [CrossRef]
- Wormser, G.P.; Nadelman, R.B.; Dattwyler, R.J.; Dennis, D.T.; Shapiro, E.D.; Steere, A.C.; Rush, T.J.; Rahn, D.W.; Coyle, P.K.; Persing, D.H.; et al. Practice guidelines for the treatment of Lyme disease. Clin. Infect. Dis. 2000, 31, S1–S14. [Google Scholar] [CrossRef] [Green Version]
- Dattwyler, R.J.; Wormser, G.P.; Rush, T.J.; Finkel, M.F.; Choen, R.T.; Grunwaldt, E.; Franklin, M.; Hilton, E.; Bryant, G.L.; Agger, W.A.; et al. A comparison of two treatment regimens of ceftriaxone in late Lyme disease. Wien. Klin. Wochenschr. 2005, 117, 393–397. [Google Scholar] [CrossRef]
- Eppes, S.C.; Childs, J.A. Comparative study of cefuroxime axetil versus amoxicillin in children with early Lyme disease. Pediatrics 2002, 109, 1173–1177. [Google Scholar] [CrossRef] [PubMed]
- Preac-Mursic, V.; Weber, K.; Pfister, H.W.; Wilske, B.; Gross, B.; Baumann, A.; Prokop, J. Survival of Borrelia burgdorferi in antibiotically treated patients with Lyme borreliosis. Infection 1989, 17, 355–359. [Google Scholar] [CrossRef] [PubMed]
- Liegner, K.B.; Rosenkilde, C.E.; Campbell, G.L.; Quan, T.J.; Dennis, D.T. Culture-confirmed treatment failure of cefotaxime and minocycline in a case of Lyme meningoencephalomyelitis in the United States. In Program and Abstracts of the Fifth International Conference on Lyme Borreliosis, Arlington, VA, USA, 30 May–2 June 1992; Federation of American Societies for Experimental Biology: Bethesda, MD, USA, 1992; p. A11. [Google Scholar]
- Liegner, K.B.; Shapiro, J.R.; Ramsay, D.; Halperin, A.J.; Hogrefe, W.; Kong, L. Recurrent erythema migrans despite extended antibiotic treatment with minocycline in a patient with persisting Borrelia burgdorferi infection. J. Am. Acad. Dermatol. 1993, 28, 312–314. [Google Scholar] [CrossRef]
- Steere, A.C.; Angelis, S.M. Therapy for Lyme arthritis: Strategies for the treatment of antibiotic-refractory arthritis. Arthritis Rheumatol. 2006, 54, 3079–3086. [Google Scholar] [CrossRef]
- Klempner, M.S.; Baker, P.J.; Shapiro, E.D.; Marques, A.; Dattwyler, R.J.; Halperin, J.J.; Wormser, G.P. Treatment trials for post-lyme disease symptoms revisited. Am. J. Med. 2013, 126, 665–669. [Google Scholar] [CrossRef] [Green Version]
- Post-Treatment Lyme Disease Syndrome. Centre of Disease Control and Prevention; Lyme Disease; 2018. Available online: https://www.cdc.gov/lyme/postlds/index.html (accessed on 22 May 2020).
- Straubinger, R.K.; Summers, B.A.; Chang, Y.F.; Appel, M.J. Persistence of Borrelia burgdorferi in experimentally infected dogs after antibiotic treatment. J. Clin. Microbiol. 1997, 35, 111–116. [Google Scholar] [CrossRef] [Green Version]
- Hodzic, E.; Feng, S.; Holden, K.; Freet, K.J.; Barthold, S.W. Persistence of Borrelia burgdorferi following antibiotic treatment in mice. Antimicrob. Agents Chemother. 2008, 52, 1728–1736. [Google Scholar] [CrossRef] [Green Version]
- Barthold, S.W.; Hodzic, E.; Imai, D.M.; Feng, S.; Yang, X.; Luft, B.J. Ineffectiveness of tigecycline against persistent Borrelia burgdorferi. Antimicrob. Agents. Chemother. 2010, 54, 643–651. [Google Scholar] [CrossRef] [Green Version]
- Embers, M.E.; Barthold, S.W.; Borda, J.T.; Bowers, L.; Doyle, L.; Hodzic, E.; Jacobs, M.B.; Hasenkampf, N.R.; Martin, D.S.; Narasimhan, S.; et al. Persistence of Borrelia burgdorferi in Rhesus Macaques following antibiotic treatment of disseminated infection. PLoS ONE 2012, 7, e29914. [Google Scholar] [CrossRef]
- Hodzic, E.; Imai, D.; Feng, S.; Barthold, S.W. Resurgence of persisting non-cultivable Borrelia burgdorferi following antibiotic treatment in mice. PLoS ONE 2014, 9, e86907. [Google Scholar] [CrossRef] [Green Version]
- Embers, M.E.; Hasenkampf, N.R.; Jacobs, M.B.; Tardo, A.C.; Doyle-Meyers, L.A.; Philipp, M.T.; Hodzic, E. Variable manifestations, diverse seroreactivity and post-treatment persistence in non-human primates exposed to Borrelia burgdorferi by tick feeding. PLoS ONE 2017, 12, e0189071. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Middelveen, M.J.; Sapi, E.; Burke, J.; Filush, K.R.; Franco, A.; Fesler, M.C.; Stricker, R.B. Persistent Borrelia infection in patients with ongoing symptoms of Lyme disease. Healthcare 2018, 6, 33. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Berndtson, K. Review of evidence for immune evasion and persistent infection in Lyme disease. Int. J. Gen. Med. 2013, 6, 291–306. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bockenstedt, L.K.; Gonzalez, D.G.; Haberman, A.M.; Belperron, A.A. Spirochete antigens persist near cartilage after murine Lyme borreliosis therapy. J. Clin. Investig. 2012, 122, 2652–2660. [Google Scholar] [CrossRef] [PubMed]
- Jutras, B.L.; Lochhead, R.B.; Kloos, Z.A.; Biboy, J.; Strle, K.; Booth, C.J.; Govers, S.K.; Gray, J.; Schumann, P.; Vollmer, W.; et al. Borrelia burgdorferi peptidoglycan is a persistent antigen in patients with Lyme arthritis. Proc. Natl. Acad. Sci. USA 2019, 116, 13498–13507. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Feng, J.; Shi, W.; Zhang, S.; Zhang, Y. Persister mechanisms in Borrelia burgdorferi: Implications for improved intervention. Emerg. Microbes. Infect. 2015, 4, e51. [Google Scholar] [CrossRef]
- Brorson, O.; Brorson, S.H. Transformation of cystic forms of Borrelia burgdorferi to normal, mobile spirochetes. Infection 1997, 25, 240–246. [Google Scholar] [CrossRef]
- Murgia, R.; Cinco, M. Induction of cystic forms by different stress conditions in Borrelia burgdorferi. APMIS 2004, 112, 57–62. [Google Scholar] [CrossRef]
- MacDonald, A.B. Spirochetal cyst forms in neurodegenerative disorders, hiding in plain sight. J. Med. Hypothesis 2006, 67, 819–832. [Google Scholar] [CrossRef]
- Miklossy, J.; Kasas, S.; Zurn, A.D.; McCall, S.; Yu, S.; McGeer, P.L. Persisting atypical and cystic forms of Borrelia burgdorferi and local inflammation in Lyme neuroborreliosis. J. Neuroinflam. 2008, 5, 40. [Google Scholar] [CrossRef] [Green Version]
- Brorson, Ø.; Brorson, S.H.; Scythes, J.; MacAllister, J.; Wier, A.; Margulis, L. Destruction of spirochete Borrelia burgdorferi round-body propagules (RBs) by the antibiotic tigecycline. Proc. Natl. Acad. Sci. USA 2009, 106, 18656–18661. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Vancová, M.; Rudenko, N.; Vaněček, J.; Golovchenko, M.; Stmad, M.; Rego, R.O.M.; Tichá, L.; Grubhoffer, L.; Nebesářová, J. Pleomorphism and viability of the Lyme disease pathogen Borrelia burgdorferi exposed to physiological stress conditions: A correlative cryo-fluorescence and cryo-scanning electron microscopy study. Front. Microbiol. 2017, 8, 596. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Rudenko, N.; Golovchenko, M.; Kybicova, K.; Vancova, M. Metamorphoses of Lyme disease spirochetes: Phenomenon of Borrelia persisters. Parasit. Vectors 2019, 12, 237. [Google Scholar] [CrossRef] [PubMed]
- Sapi, E.; Bastian, S.L.; Mpoy, C.M.; Scott, S.; Rattelle, A.; Pabbati, N.; Poruri, A.; Burugu, D.; Theophilus, P.A.S.; Pham, T.V.; et al. Characterization of biofilm formation by Borrelia burgdorferi in vitro. PLoS ONE 2012, 7, e48277. [Google Scholar] [CrossRef] [PubMed]
- Timmaraju, V.A.; Theophilus, P.A.S.; Balasubramanian, K.; Shakih, S.; Luecke, D.F.; Sapi, E. Biofilm formation by Borrelia sensu lato. FEMS Microbiol. Lett. 2015, 362, fnv120. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sapi, E.; Kaur, N.; Anyanwu, S.; Leucke, D.F.; Data, A.; Patel, S.; Rossi, M.; Stricker, R.B. Evaluation of in-vitro antibiotic susceptibility of different morphological forms of Borrelia burgdorferi. Infect. Drug Resist. 2011, 4, 97–113. [Google Scholar]
- Sapi, E.; Balasubramanian, K.; Poruri, A.; Maghsoudlou, J.S.; Socarras, K.M.; Timmaraju, A.V.; Filush, K.R.; Gupta, K.; Shaikh, S.; Theophilus, P.A.S.; et al. Evidence of in vivo existence of Borrelia biofilm in Borrelial lymphocytomas. Eur. J. Microbiol. Immunol. 2016, 6, 9–24. [Google Scholar] [CrossRef] [Green Version]
- Sapi, E.; Kasliwala, R.S.; Ismail, H.; Torres, J.P.; Oldakowski, M.; Markland, S.; Gaur, G.; Melillo, A.; Eisendle, K.; Liegner, K.B.; et al. The long-term persistence of Borrelia burgdorferi antigens and DNA in the tissues of a patient with Lyme Disease. Antibiotics 2019, 8, 183. [Google Scholar] [CrossRef] [Green Version]
- Costerton, J.W.; Stewart, P.S.; Greenberg, E.P. Bacterial biofilms: A common cause of persistent infections. Science 1999, 284, 1318–1322. [Google Scholar] [CrossRef] [Green Version]
- Sutherland, I. Biofilm exopolysaccharides: A strong and sticky framework. Microbiology 2001, 147, 3–9. [Google Scholar] [CrossRef] [Green Version]
- Römling, U.; Balsalobre, C. Biofilm infections, their resilience to therapy and innovative treatment strategies. J. Intern. Med. 2012, 272, 541–561. [Google Scholar] [CrossRef]
- Sun, F.; Qu, F.; Ling, Y.; Mao, P.; Xia, P.; Chen, H.; Zhou, D. Biofilm-associated infections: Antibiotic resistance and novel therapeutic strategies. Fut. Microbiol. 2013, 8, 877–886. [Google Scholar] [CrossRef]
- Hall, C.W.; Mah, T.F. Molecular mechanisms of biofilm-based antibiotic resistance and tolerance in pathogenic bacteria. FEMS Microbiol. Rev. 2017, 41, 276–301. [Google Scholar] [CrossRef]
- Algburi, A.; Comito, N.; Kashtanov, D.; Leon, M.T.; Dicks, L.M.T.; Chikindas, M.L. Control of biofilm formation: Antibiotics and beyond. App. Environ. Microbiol. 2017, 83, e02508-16. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ciofu, O.; Tolker-Nielsen, T. Tolerance and resistance of Pseudomonas aeruginosa biofilms to antimicrobial agents—How, P. aeruginosa can escape antibiotics. Front. Microbiol. 2019, 10, 913. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Coenye, T.; Nelis, H.J. In vitro and in vivo model systems to study microbial biofilm formation. J. Microbiol. Methods 2010, 83, 89–105. [Google Scholar] [CrossRef] [PubMed]
- Lebeaux, D.; Chauhan, A.; Rendueles, O.; Beloin, C. From in vitro to in vivo models of bacterial biofilm-related infections. Pathogens 2013, 2, 288–356. [Google Scholar] [CrossRef] [Green Version]
- Junka, A.; Szymczyk, P.; Ziółkowski, G.; Karuga-Kuzniewska, E.; Smutnicka, D.; Bil-Lula, I.; Bartoszewicz, M.; Mahabady, S.; Sedghizadeh, P.P. Bad to the bone: On in vitro and ex vivo microbial biofilm ability to directly destroy colonized bone surfaces without participation of host immunity or osteoclastogenesis. PLoS ONE 2017, 12, e0169565. [Google Scholar] [CrossRef]
- Harrison, F.; Muruli, A.; Higgins, S.; Diggle, S.P. Development of an ex vivo porcine lung model for studying growth, virulence and signaling of Pseudomonas aeruginosa. Infect. Immun. 2014, 82, 3312–3323. [Google Scholar] [CrossRef] [Green Version]
- Rubinchik, E.; Pasetka, C. Ex Vivo skin infection model. In Antimicrobial Peptides: Methods and Protocols; Giuliani, A., Rinaldi, A.C., Eds.; Humana Press: Totowa, NJ, USA, 2010; pp. 359–369. [Google Scholar]
- Steinstraesser, L.; Sorkin, M.; Niederbichler, A.D.; Becerikli, M.; Stupka, J.; Daigeler, A.; Kesting, M.R.; Stricker, I.; Jacobsen, F.; Schulte, M. A novel human skin chamber model to study wound infection ex vivo. Arch. Dermatol. Res. 2010, 302, 357–365. [Google Scholar] [CrossRef] [Green Version]
- Mason, L.M.; Wagemakers, A.; van ‘t Veer, C.; Oei, A.; van der Pot, W.J.; Ahmed, K.; van der Poll, T.; Geijtenbeek, T.B.H.; Hovius, J.W.R. Borrelia burgdorferi induces TLR2-mediated migration of activated dendritic cells in an ex vivo human skin model. PLoS ONE 2016, 11, e0164040. [Google Scholar] [CrossRef]
- Duray, P.H.; Yin, S.R.; Ito, Y.; Bezrukov, L.; Cox, C.; Cho, M.S.; Fitzgerald, W.; Dorward, D.; Zimmerberg, J.; Margolis, L. Invasion of human tissue ex vivo by Borrelia burgdorferi. J. Infect. Dis. 2005, 191, 1747–1754. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ding, Z.; Ma, M.; Tao, L.; Peng, Y.; Han, Y.; Sun, L.; Dai, X.; Ji, Z.; Bai, R.; Jian, M.; et al. Rhesus brain transcriptomic landscape in an ex vivo model of the interaction of live Borrelia burgdorferi with frontal cortex tissue explants. Front. Neurosci. 2019, 13, 651. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Grillon, A.; Westermann, B.; Cantero, P.; Jaulhac, B.; Voorduow, M.J.; Kapps, D.; Collin, E.; Barthel, C.; Ehret-Sabatier, L.; Boulanger, N. Identification of Borrelia protein candidates in mouse skin for potential diagnosis of disseminated Lyme borreliosis. Sci. Rep. 2017, 7, 16719. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Høiby, N.; Ciofu, O.; Bjarnsholt, T. Pseudomonas aeruginosa biofilms in cystic fibrosis. Future Microbiol. 2010, 5, 1663–1674. [Google Scholar] [CrossRef]
- Clementi, F. Alginate production by Azotobacter vinelandii. Crit. Rev. Biotechnol. 1997, 17, 327–361. [Google Scholar] [CrossRef]
- Ristow, P.; Bourhy, P.; Kerneis, S.; Schmitt, C.; Prevost, M.C.; Lilenbaum, W.; Picardeau, M. Biofilm formation by saprophytic and pathogenic leptospires. Microbiology 2008, 154, 1309–1317. [Google Scholar] [CrossRef] [Green Version]
- Schaudinn, C.; Dittmann, C.; Jurisch, J.; Laue, M.; Günday-Türeli, N.; Blume-Peytavi, U.; Vogt, A.; Rancan, F. Development, standardization and testing of a bacterial wound infection model based on ex vivo human skin. PLoS ONE 2017, 12, e0186946. [Google Scholar] [CrossRef] [Green Version]
- Popov, L.; Kovalski, J.; Grandi, G.; Bagnoli, F.; Amieva, M.R. Three-dimensional human skin models to understand Staphylococcus aureus skin colonization and infection. Front. Immunol. 2014, 5, 41. [Google Scholar] [CrossRef] [Green Version]
- Corzo-León, D.E.; Munro, C.A.; MacCallum, D.M. An ex vivo human skin model to study superficial fungal infections. Front. Microbiol. 2019, 10, 1172. [Google Scholar] [CrossRef] [Green Version]
- Secor, P.R.; James, G.A.; Fleckman, P.; Olerud, J.E.; McInnerney, K.; Stewart, P.S. Staphylococcus aureus biofilm and planktonic cultures differentially impact gene expression, mapk phosphorylation, and cytokine production in human keratinocytes. BMC Microbiol. 2011, 11, 143. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Elias, S.; Banin, E. Multi-species biofilms: Living with friendly neighbors. FEMS Microbiol. Rev. 2012, 36, 990–1004. [Google Scholar] [CrossRef] [PubMed]
- Omar, A.; Wright, J.B.; Schultz, G.; Burrell, R.; Nadworny, P. Microbial biofilms and chronic wounds. Microorganisms 2017, 5, 9. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lopes, S.P.; Ceri, H.; Azevedo, N.F.; Pereira, M.O. Antibiotic resistance of mixed biofilms in cystic fibrosis: Impact of emerging microorganisms on treatment of infection. Int. J. Antimicrob. Agents 2012, 40, 260–263. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sapi, E.; Gupta, K.; Wawrzeniak, K.; Gauri, G.; Torres, J.; Filush, K.R.; Melillo, A.; Zelger, B. Borrelia and Chlamydia can form mixed biofilms in infected human skin tissues. Europ. J. Microbiol. Immunol. 2019, 9, 46–55. [Google Scholar] [CrossRef]
- Middelveen, M.J.; Filush, K.R.; Stricker, R.B.; Sapi, E. Mixed Borrelia and Helicobacter pylori biofilms in Morgellons disease dermatological specimens. Healthcare 2019, 7, 70. [Google Scholar] [CrossRef] [Green Version]
- Feng, J.; Wang, T.; Shi, W.; Zhang, S.; Sullivan, D.; Auwaerter, P.G.; Zhang, Y. Identification of novel activity against Borrelia burgdorferi persisters using an FDA approved drug library. Emerg. Microbes. Infect. 2014, 3, e49. [Google Scholar] [CrossRef]
- Feng, J.; Shi, W.; Zhang, S.; Zhang, Y. Identification of new compounds with high activity against stationary phase Borrelia burgdorferi from the NCI compound collection. Emerg. Microbes. Infect. 2015, 4, e31. [Google Scholar] [CrossRef]
- Feng, J.; Auwaerter, P.G.; Zhang, Y. Drug combinations against Borrelia burgdorferi persisters in vitro: Eradication achieved by using daptomycin, cefoperazone and doxycycline. PLoS ONE 2015, 10, e0117207. [Google Scholar] [CrossRef] [Green Version]
- Theophilus, P.A.S.; Victoria, M.J.; Socarras, K.M.; Filush, K.R.; Gupta, K.; Luecke, D.F.; Sapi, E. Effectiveness of Stevia rebaudiana whole leaf extract against the various morphological forms of Borrelia burgdorferi in vitro. Eur. J. Microbiol. Immunol. 2015, 5, 268–280. [Google Scholar] [CrossRef] [Green Version]
- Feng, J.; Weitner, M.; Shi, W.; Zhang, S.; Zhang, Y. Eradication of biofilm-like microcolony structures of Borrelia burgdorferi by daunomycin and daptomycin but not mitomycin C in combination with doxycycline and cefuroxime. Front. Microbiol. 2016, 7, 62. [Google Scholar] [CrossRef] [Green Version]
- Fearnley, A.; Gupta, K.; Freeman, P.R.; Horowitz, R.I. Effect of dapsone and its antimicrobial combinations on Borrelia burgdorferi biofilms. In Proceedings of the 16th Annual Meeting of International Lyme and Associated Diseases Society, Boston, MA, USA, 31 October–3 November 2019. [Google Scholar]
- Soccaras, K.M.; Theophilus, P.A.S.; Torres, J.P.; Gupta, K.; Sapi, E. Antimicrobial activity of bee venom and mellittin against Borrelia. Antibiotics 2017, 6, 31. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pothineni, V.; Wagh, D.; Babar, M.M.; Inayathullah, M.; Watts, R.E.; Kim, K.-M.; Parekh, M.B.; Gurjarpadhye, A.A.; Solow-Cordero, D.; Tayebi, L.; et al. Screening of NCI-DTP library to identify new drug candidates for Borrelia burgdorferi. J. Antibiot. 2017, 70, 308–312. [Google Scholar] [CrossRef]
- Feng, J.; Zhang, S.; Shi, W.; Zhang, Y. Activity of sulfa drugs and their combinations against stationary phase B. burgdorferi in vitro. Antibiotics 2017, 6, 10. [Google Scholar] [CrossRef]
- Pothineni, V.R.; Potula, H.S.K.; Ambati, A.; Mallajosyual, V.V.A.; Sridharan, B.; Inayathullah, M.; Ahmed, M.S.; Rajadas, J. Azlocillin can be the potential drug candidate against drug-tolerant Borrelia burgdorferi sensu stricto JLB31. Sci. Rep. 2020, 10, 3798. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Feng, J.; Li, T.; Yuan, Y.; Yee, R.; Bai, C.; Cai, M.; Shi, W.; Embers, M.; Brayton, C.; Saeki, H.; et al. Stationary phase persister/biofilm microcolony of Borrelia burgdorferi causes more severe disease in a mouse model of Lyme arthritis: Implications for understanding persistence, Post-treatment Lyme Disease Syndrome (PTLDS), and treatment failure. Discov. Med. 2019, 148, 125–138. [Google Scholar]
- Theilacker, C.; Coleman, F.T.; Mueschenborn, S.; Llosa, N.; Grout, M.; Pier, G.B. Construction and characterization of a Pseudomonas aeruginosa mucoid exopolysaccharide-alginate conjugate vaccine. Infect. Immun. 2003, 71, 3875–3884. [Google Scholar] [CrossRef] [Green Version]
Culture Media | Concentration (cells/mL) | Positive Spirochete Slides | # of Spirochetes/mm2 |
---|---|---|---|
BSK-H + 6% RS | 5 × 106 | 30/30 | >500 |
1 × 107 | 30/30 | >500 | |
DMEM + 10% CS | 5 × 106 | 30/30 | >500 |
1 × 107 | 30/30 | >500 | |
Uninfected (Both media) | 0 | 0 | 0 |
Culture Media | Concentration (cells/mL) | Positive Biofilm Slides | # of Biofilms/mm2 | Size of Biofilms |
---|---|---|---|---|
BSK-H + 6% RS | 5 × 106 | 0/30 | 0 | 0 |
1 × 107 | 12/30 | (1–2) | 50–300 μm | |
DMEM + 10% CS | 5 × 106 | 0/30 | 0 | 0 |
1 × 107 | 8/30 | 1 | 20–200 μm | |
Uninfected (Both media) | 0 | 0 | 0 | 0 |
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Torres, J.P.; Senejani, A.G.; Gaur, G.; Oldakowski, M.; Murali, K.; Sapi, E. Ex Vivo Murine Skin Model for B. burgdorferi Biofilm. Antibiotics 2020, 9, 528. https://doi.org/10.3390/antibiotics9090528
Torres JP, Senejani AG, Gaur G, Oldakowski M, Murali K, Sapi E. Ex Vivo Murine Skin Model for B. burgdorferi Biofilm. Antibiotics. 2020; 9(9):528. https://doi.org/10.3390/antibiotics9090528
Chicago/Turabian StyleTorres, Jason P., Alireza G. Senejani, Gauri Gaur, Michael Oldakowski, Krithika Murali, and Eva Sapi. 2020. "Ex Vivo Murine Skin Model for B. burgdorferi Biofilm" Antibiotics 9, no. 9: 528. https://doi.org/10.3390/antibiotics9090528